Estimation of the absorption coefficients of two-layered media by a simple method using spatially and time-resolved reflectances

NASA Astrophysics Data System (ADS)

Shimada, M.; Sato, C.; Hoshi, Y.; Yamada, Y.

2009-08-01

Our newly developed method using spatially and time-resolved reflectances can easily estimate the absorption coefficients of each layer in a two-layered medium if the thickness of the upper layer and the reduced scattering coefficients of the two layers are known a priori. We experimentally validated this method using phantoms and examined its possibility of estimating the absorption coefficients of the tissues in human heads. In the case of a homogeneous plastic phantom (polyacetal block), the absorption coefficient estimated by our method agreed well with that obtained by a conventional method. Also, in the case of two-layered phantoms, our method successfully estimated the absorption coefficients of the two layers. Furthermore, the absorption coefficients of the extracerebral and cerebral tissue inside human foreheads were estimated under the assumption that the human heads were two-layered media. It was found that the absorption coefficients of the cerebral tissues were larger than those of the extracerebral tissues.

Mechanical properties of electrospun bilayer fibrous membranes as potential scaffolds for tissue engineering.

PubMed

Pu, Juan; Komvopoulos, Kyriakos

2014-06-01

Bilayer fibrous membranes of poly(l-lactic acid) (PLLA) were fabricated by electrospinning, using a parallel-disk mandrel configuration that resulted in the sequential deposition of a layer with fibers aligned across the two parallel disks and a layer with randomly oriented fibers, both layers deposited in a single process step. Membrane structure and fiber alignment were characterized by scanning electron microscopy and two-dimensional fast Fourier transform. Because of the intricacies of the generated electric field, bilayer membranes exhibited higher porosity than single-layer membranes consisting of randomly oriented fibers fabricated with a solid-drum collector. However, despite their higher porosity, bilayer membranes demonstrated generally higher elastic modulus, yield strength and toughness than single-layer membranes with random fibers. Bilayer membrane deformation at relatively high strain rates comprised multiple abrupt microfracture events characterized by discontinuous fiber breakage. Bilayer membrane elongation yielded excessive necking of the layer with random fibers and remarkable fiber stretching (on the order of 400%) in the layer with fibers aligned in the stress direction. In addition, fibers in both layers exhibited multiple localized necking, attributed to the nonuniform distribution of crystalline phases in the fibrillar structure. The high membrane porosity, good mechanical properties, and good biocompatibility and biodegradability of PLLA (demonstrated in previous studies) make the present bilayer membranes good scaffold candidates for a wide range of tissue engineering applications. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Voxel-based measurement sensitivity of spatially resolved near-infrared spectroscopy in layered tissues

NASA Astrophysics Data System (ADS)

Niwayama, Masatsugu

2018-03-01

We quantitatively investigated the measurement sensitivity of spatially resolved spectroscopy (SRS) across six tissue models: cerebral tissue, a small animal brain, the forehead of a fetus, an adult brain, forearm muscle, and thigh muscle. The optical path length in the voxel of the model was analyzed using Monte Carlo simulations. It was found that the measurement sensitivity can be represented as the product of the change in the absorption coefficient and the difference in optical path length in two states with different source-detector distances. The results clarified the sensitivity ratio between the surface layer and the deep layer at each source-detector distance for each model and identified changes in the deep measurement area when one of the detectors was close to the light source. A comparison was made with the results from continuous-wave spectroscopy. The study also identified measurement challenges that arise when the surface layer is inhomogeneous. Findings on the measurement sensitivity of SRS at each voxel and in each layer can support the correct interpretation of measured values when near-infrared oximetry or functional near-infrared spectroscopy is used to investigate different tissue structures.

Modular assembly of thick multifunctional cardiac patches

PubMed Central

Fleischer, Sharon; Shapira, Assaf; Feiner, Ron; Dvir, Tal

2017-01-01

In cardiac tissue engineering cells are seeded within porous biomaterial scaffolds to create functional cardiac patches. Here, we report on a bottom-up approach to assemble a modular tissue consisting of multiple layers with distinct structures and functions. Albumin electrospun fiber scaffolds were laser-patterned to create microgrooves for engineering aligned cardiac tissues exhibiting anisotropic electrical signal propagation. Microchannels were patterned within the scaffolds and seeded with endothelial cells to form closed lumens. Moreover, cage-like structures were patterned within the scaffolds and accommodated poly(lactic-co-glycolic acid) (PLGA) microparticulate systems that controlled the release of VEGF, which promotes vascularization, or dexamethasone, an anti-inflammatory agent. The structure, morphology, and function of each layer were characterized, and the tissue layers were grown separately in their optimal conditions. Before transplantation the tissue and microparticulate layers were integrated by an ECM-based biological glue to form thick 3D cardiac patches. Finally, the patches were transplanted in rats, and their vascularization was assessed. Because of the simple modularity of this approach, we believe that it could be used in the future to assemble other multicellular, thick, 3D, functional tissues. PMID:28167795

The peri-esophageal connective tissue layers and related compartments: visualization by histology and magnetic resonance imaging.

PubMed

Weijs, T J; Goense, L; van Rossum, P S N; Meijer, G J; van Lier, A L H M W; Wessels, F J; Braat, M N G; Lips, I M; Ruurda, J P; Cuesta, M A; van Hillegersberg, R; Bleys, R L A W

2017-02-01

An organized layer of connective tissue coursing from aorta to esophagus was recently discovered in the mediastinum. The relations with other peri-esophageal fascias have not been described and it is unclear whether this layer can be visualized by non-invasive imaging. This study aimed to provide a comprehensive description of the peri-esophageal fascias and determine whether the connective tissue layer between aorta and esophagus can be visualized by magnetic resonance imaging (MRI). First, T2-weighted MRI scanning of the thoracic region of a human cadaver was performed, followed by histological examination of transverse sections of the peri-esophageal tissue between the thyroid gland and the diaphragm. Secondly, pretreatment motion-triggered MRI scans were prospectively obtained from 34 patients with esophageal cancer and independently assessed by two radiologists for the presence and location of the connective tissue layer coursing from aorta to esophagus. A layer of connective tissue coursing from the anterior aspect of the descending aorta to the left lateral aspect of the esophagus, with a thin extension coursing to the right pleural reflection, was visualized ex vivo in the cadaver on MR images, macroscopic tissue sections, and after histologic staining, as well as on in vivo MR images. The layer connecting esophagus and aorta was named 'aorto-esophageal ligament' and the layer connecting aorta to the right pleural reflection 'aorto-pleural ligament'. These connective tissue layers divides the posterior mediastinum in an anterior compartment containing the esophagus, (carinal) lymph nodes and vagus nerve, and a posterior compartment, containing the azygos vein, thoracic duct and occasionally lymph nodes. The anterior compartment was named 'peri-esophageal compartment' and the posterior compartment 'para-aortic compartment'. The connective tissue layers superior to the aortic arch and at the diaphragm corresponded with the currently available anatomic descriptions. This study confirms the existence of the previously described connective tissue layer coursing from aorta to esophagus, challenging the long-standing paradigm that no such structure exists. A comprehensive, detailed description of the peri-esophageal fascias is provided and, furthermore, it is shown that the connective tissue layer coursing from aorta to esophagus can be visualized in vivo by MRI. © 2016 Anatomical Society.

Finite element modeling of mitral leaflet tissue using a layered shell approximation

PubMed Central

Ratcliffe, Mark B.; Guccione, Julius M.

2012-01-01

The current study presents a finite element model of mitral leaflet tissue, which incorporates the anisotropic material response and approximates the layered structure. First, continuum mechanics and the theory of layered composites are used to develop an analytical representation of membrane stress in the leaflet material. This is done with an existing anisotropic constitutive law from literature. Then, the concept is implemented in a finite element (FE) model by overlapping and merging two layers of transversely isotropic membrane elements in LS-DYNA, which homogenizes the response. The FE model is then used to simulate various biaxial extension tests and out-of-plane pressure loading. Both the analytical and FE model show good agreement with experimental biaxial extension data, and show good mutual agreement. This confirms that the layered composite approximation presented in the current study is able to capture the exponential stiffening seen in both the circumferential and radial directions of mitral leaflets. PMID:22971896

Specialized probes based on hydroxyapatite calcium for heart tissues research by atomic force microscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhukov, Mikhail, E-mail: cloudjyk@yandex.ru; Golubok, Alexander; Institute for Analytical Instrumentation, Russian Academy of Sciences

The new specialized AFM-probes with hydroxyapatite structures for atomic force microscopy of heart tissues calcification were created and studied. A process of probe fabrication is demonstrated. The adhesive forces between specialized hydroxyapatite probe and endothelium/subendothelial layers were investigated. It was found that the adhesion forces are significantly higher for the subendothelial layers. We consider that it is connected with the formation and localization of hydroxyapatite in the area of subendothelial layers of heart tissues. In addition, the roughness analysis and structure visualization of the endothelial surface of the heart tissue were carried out. The results show high efficiency of createdmore » specialized probes at study a calcinations process of the aortic heart tissues.« less

Ultrastructural studies on the boundary tissue of the seminiferous tubules of different mammals.

PubMed

Cieciura, L; Jaszczuk-Jarosz, B; Pietrzkowska, K

1988-01-01

The aims of our studies were to compare the ultrastructure of the boundary tissue of seminiferous tubules of various mammals (rat, mouse, hamster, guinea pig, rabbit, ram, bull and man). Visual analysis of electron micrographs revealed the similarity of structure of all layers at investigated animals. The boundary tissue consists of 4 layers: 1) amorphous inner lamina, 2) cellular inner lamina, 3) amorphous outer lamina, 4) cellular outer lamina. The outer lamina of boundary tissue of rat, mouse and hamster revealed in histochemical reactions meshes resembling honey-combs. The wall of seminiferous canalicules of bull and ram consists of more bigger and different structure than one at the other laboratory animals. The most different structure of boundary tissue in man was observed. The capillary vessels penetrate in the myofibroblastic layer, when comparted to that found in other mammals on the surface of the wall.

New Details of the Human Corneal Limbus Revealed With Second Harmonic Generation Imaging.

PubMed

Park, Choul Yong; Lee, Jimmy K; Zhang, Cheng; Chuck, Roy S

2015-09-01

To report novel findings of the human corneal limbus by using second harmonic generation (SHG) imaging. Corneal limbus was imaged by using an inverted two-photon excitation fluorescence microscope. Laser (Ti:Sapphire) was tuned at 850 nm for two-photon excitation. Backscatter signals of SHG and autofluorescence (AF) were collected through a 425/30-nm emission filter and a 525/45-emission filter, respectively. Multiple, consecutive, and overlapping image stacks (z-stack) were acquired for the corneal limbal area. Two novel collagen structures were revealed by SHG imaging at the limbus: an anterior limbal cribriform layer and presumed anchoring fibers. Anterior limbal cribriform layer is an intertwined reticular collagen architecture just beneath the limbal epithelial niche and is located between the peripheral cornea and Tenon's/scleral tissue. Autofluorescence imaging revealed high vascularity in this structure. Central to the anterior limbal cribriform layer, radial strands of collagen were found to connect the peripheral cornea to the limbus. These presumed anchoring fibers have both collagen and elastin and were found more extensively in the superficial layers than deep layer and were absent in very deep limbus near Schlemm's canal. By using SHG imaging, new details of the collagen architecture of human corneal limbal area were elucidated. High resolution images with volumetric analysis revealed two novel collagen structures.

Isolation of tissue layers in hermatypic corals by N-acetylcysteine: morphological and proteomic examinations

NASA Astrophysics Data System (ADS)

Peng, S.-E.; Luo, Y.-J.; Huang, H.-J.; Lee, I.-T.; Hou, L.-S.; Chen, W.-N. U.; Fang, L.-S.; Chen, C.-S.

2008-03-01

Corals are diploblastic in body pattern and include two tissue layers, the epidermis and gastrodermis, interconnected by an acellular matrix mesoglea. During development, cells in these tissue layers differentiate morphologically and functionally. In most hermatypic corals, the gastrodermis further develops an ability to associate with microalgae dinoflagellates. This endosymbiosis occurs inside specific host gastrodermal cells, and its mechanism still remains unclear notwithstanding decades of research. The delay in progress is partly due to the difficulty in separating the gastrodermis and its symbionts from the epidermis for detailed cellular and biochemical investigations. The present study reports a simple method to separate these two tissue layers in hermatypic corals using the reducing agent, N-acetylcysteine (NAC). Efficient tissue and proteomic isolations are demonstrated by microscopy and two-dimensional SDS polyacrylamide gel electrophoresis (2D SDS-PAGE). The NAC treatment was able to separate tissue layers without inducing protein degradation. Furthermore, the sensitivity of protein detection greatly increases in the isolated tissue layers. The application of the present technique provides future research on endosymbiosis and coral development with a tool for higher accuracy and sensitivity.

Photochemical bonding of epithelial cell-seeded collagen lattice to rat muscle layer for esophageal tissue engineering: a pilot study

NASA Astrophysics Data System (ADS)

Chan, Barbara P.; Sato, M.; Vacanti, Joseph P.; Kochevar, Irene E.; Redmond, Robert W.

2005-04-01

Bilayered tube structures consist of epithelial cell-seeded collagen lattice and muscle layer have been fabricated for esophageal tissue engineering. Good adhesion between layers in order to facilitate cell infiltration and neovascularization in the collagen lattice is required. Previous efforts include using other bioglues such as fibrin glue and silicone tube as the physical support. However, the former is subjected to chances of transmitting blood-born infectious disease and is time consuming while the latter requires a second surgical procedure. The current project aimed to bond the cell-seeded collagen lattice to muscle layer using photochemical bonding, which has previously been demonstrated a rapid and non-thermal procedure in bonding collagenous tissues. Rat esophageal epithelial cells were seeded on collagen lattice and together with the latissimus dorsi muscle layer, were exposed to a photosensitizer rose Bengal at the bonding surface. An argon laser was used to irradiate the approximated layers. Bonding strength was measured during the peeling test of the collagen layer from the muscle layer. Post-bonding cell viability was assessed using a modified NADH-diaphorase microassay. A pilot in vivo study was conducted by directly bonding the cell-seeded collagen layer onto the muscle flap in rats and the structures were characterized histologically. Photochemical bonding was found to significantly increase the adherence at the bonding interface without compromising the cell viability. This indicates the feasibility of using the technique to fabricate multi-layered structures in the presence of living cells. The pilot animal study demonstrated integration of the collagen lattice with the muscle layer at the bonding interface although the subsequent surgical manipulation disturbed the integration at some region. This means that an additional procedure removing the tube could be avoided if the approximation and thus the bonding are optimized. Cell infiltration and neovascularization were also evident demonstrating that direct bonding of engineered tissue structures in particular those with low processability such as collagen lattice to the host tissue is feasible.

Developing visible and near-infrared reflectance spectroscopy to detect changes of the dermal collagen concentration

NASA Astrophysics Data System (ADS)

Wang, Chiao-Yi; Liao, Andy Ying Chi; Sung, Kung Bin

2018-02-01

Collagen provides skin structure integrity and its concentration is related to the severity of scars. The objective of this study is to develop a hand-held and relatively inexpensive system to detect changes of the dermal collagen concentration in vivo. Diffuse reflectance spectroscopy and two-layer diffusion model have often been used to quantify the collagen concentration and other optical properties of the skin. However, the influences of fat and muscle, which are just below the dermis, have not been thoroughly investigated. We applied Monte Carlo simulations to find source-detector separations most sensitive to changes in collagen absorption and identify four wavelengths between 650 nm and 1000 nm suitable for separating influences of other chromophores including melanin, oxyhemoglobin and deoxyhemoglobin. Our tissue model consisted of at least three layers including the epidermis, dermis and subcutaneous fat with an optional forth layer representing the muscle. Results showed that the reflectance of the three-layered tissue model differed significantly from that of the two-layered tissue model, and the additional muscle layer might also influence the reflectance depending on the thickness of the fat layer. In addition, whether scattering coefficients of the epidermis and dermis were the same significantly affected the reflectance. Differences in reflectance due to changes in the collagen concentration were distinct from those due to changes in scattering coefficients and other chromophores. Further in-vivo experiments are ongoing to to validate the proposed approach.

Sodium alginate hydrogel-based bioprinting using a novel multinozzle bioprinting system.

PubMed

Song, Seung-Joon; Choi, Jaesoon; Park, Yong-Doo; Hong, Soyoung; Lee, Jung Joo; Ahn, Chi Bum; Choi, Hyuk; Sun, Kyung

2011-11-01

Bioprinting is a technology for constructing bioartificial tissue or organs of complex three-dimensional (3-D) structure with high-precision spatial shape forming ability in larger scale than conventional tissue engineering methods and simultaneous multiple components composition ability. It utilizes computer-controlled 3-D printer mechanism or solid free-form fabrication technologies. In this study, sodium alginate hydrogel that can be utilized for large-dimension tissue fabrication with its fast gelation property was studied regarding material-specific printing technique and printing parameters using a multinozzle bioprinting system developed by the authors. A sodium alginate solution was prepared with a concentration of 1% (wt/vol), and 1% CaCl(2) solution was used as cross-linker for the gelation. The two materials were loaded in each of two nozzles in the multinozzle bioprinting system that has a total of four nozzles of which the injection speed can be independently controlled. A 3-D alginate structure was fabricated through layer-by-layer printing. Each layer was formed through two phases of printing, the first phase with the sodium alginate solution and the second phase with the calcium chloride solution, in identical printing pattern and speed condition. The target patterns were lattice shaped with 2-mm spacing and two different line widths. The nozzle moving speed was 6.67 mm/s, and the injection head speed was 10 µm/s. For the two different line widths, two injection needles with inner diameters of 260 and 410 µm were used. The number of layers accumulated was five in this experiment. By varying the nozzle moving speed and the injection speed, various pattern widths could be achieved. The feasibility of sodium alginate hydrogel free-form formation by alternate printing of alginate solution and sodium chloride solution was confirmed in the developed multinozzle bioprinting system. © 2011, Copyright the Authors. Artificial Organs © 2011, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

Fabrication and preliminary study of a biomimetic tri-layer tubular graft based on fibers and fiber yarns for vascular tissue engineering.

PubMed

Wu, Tong; Zhang, Jialing; Wang, Yuanfei; Li, Dandan; Sun, Binbin; El-Hamshary, Hany; Yin, Meng; Mo, Xiumei

2018-01-01

Designing a biomimetic and functional tissue-engineered vascular graft has been urgently needed for repairing and regenerating defected vascular tissues. Utilizing a multi-layered vascular scaffold is commonly considered an effective way, because multi-layered scaffolds can easily simulate the structure and function of natural blood vessels. Herein, we developed a novel tri-layer tubular graft consisted of Poly(L-lactide-co-caprolactone)/collagen (PLCL/COL) fibers and Poly(lactide-co-glycolide)/silk fibroin (PLGA/SF) yarns via a three-step electrospinning method. The tri-layer vascular graft consisted of PLCL/COL aligned fibers in inner layer, PLGA/SF yarns in middle layer, and PLCL/COL random fibers in outer layer. Each layer possessed tensile mechanical strength and elongation, and the entire tubular structure provided tensile and compressive supports. Furthermore, the human umbilical vein endothelial cells (HUVECs) and smooth muscle cells (SMCs) proliferated well on the materials. Fluorescence staining images demonstrated that the axially aligned PLCL/COL fibers prearranged endothelium morphology in lumen and the circumferential oriented PLGA/SF yarns regulated SMCs organization along the single yarns. The outside PLCL/COL random fibers performed as the fixed layer to hold the entire tubular structure. The in vivo results showed that the tri-layer vascular graft supported cell infiltration, scaffold biodegradation and abundant collagen production after subcutaneous implantation for 10weeks, revealing the optimal biocompatibility and tissue regenerative capability of the tri-layer graft. Therefore, the specially designed tri-layer vascular graft will be beneficial to vascular reconstruction. Copyright © 2017. Published by Elsevier B.V.

Multilayer porous UHMWPE scaffolds for bone defects replacement.

PubMed

Maksimkin, A V; Senatov, F S; Anisimova, N Yu; Kiselevskiy, M V; Zalepugin, D Yu; Chernyshova, I V; Tilkunova, N A; Kaloshkin, S D

2017-04-01

Reconstruction of the structural integrity of the damaged bone tissue is an urgent problem. UHMWPE may be potentially used for the manufacture of porous implants simulating as closely as possible the porous cancellous bone tissue. But the extremely high molecular weight of the polymer does not allow using traditional methods of foaming. Porous and multilayer UHMWPE scaffolds with nonporous bulk layer and porous layer that mimics cancellous bone architecture were obtained by solid-state mixing, thermopressing and washing in subcritical water. Structural and mechanical properties of the samples were studied. Porous UHMWPE samples were also studied in vitro and in vivo. The pores of UHMWPE scaffold are open and interconnected. Volume porosity of the obtained samples was 79±2%; the pore size range was 80-700μm. Strong connection of the two layers in multilayer UHMWPE scaffolds was observed with decreased number of fusion defects. Functionality of implants based on multilayer UHMWPE scaffolds is provided by the fixation of scaffolds in the bone defect through ingrowths of the connective tissue into the pores, which ensures the maintenance of the animals' mobility. Copyright © 2016 Elsevier B.V. All rights reserved.

Influence of the Dermis Thickness on the Results of the Skin Treatment with Monopolar and Bipolar Radiofrequency Currents.

PubMed

Kruglikov, Ilja L

2016-01-01

Electrically layered tissue structure significantly modifies distribution of radiofrequency (RF) current in the dermis and in the subcutaneous adipose tissue comparing to that in a homogeneous medium. On the basis of the simple model of RF current distribution in a two-layer skin containing dermis and subcutis, we assess the influence of the dermal thickness on the current density in different skin layers. Under other equal conditions, current density in the dermis is higher for the skin having thinner dermis. This contradicts the main paradigm of the RF theory stating that treatment results are mainly dependent on the maximal temperature reached in a target tissue, since the best short- and long-term clinical results of RF application to the skin were reported in the areas having thicker dermis. To resolve this contradiction, it is proposed that the long-term effect of RF can be realized through a structural modification of the subcutaneous fat depot adjacent to the treated skin area. Stimulation of these cells located near the interface dermis/subcutis will demand the concentration of applied RF energy in this area and will require the optimal arrangement of RF electrodes on the skin surface.

Influence of the Dermis Thickness on the Results of the Skin Treatment with Monopolar and Bipolar Radiofrequency Currents

PubMed Central

2016-01-01

Electrically layered tissue structure significantly modifies distribution of radiofrequency (RF) current in the dermis and in the subcutaneous adipose tissue comparing to that in a homogeneous medium. On the basis of the simple model of RF current distribution in a two-layer skin containing dermis and subcutis, we assess the influence of the dermal thickness on the current density in different skin layers. Under other equal conditions, current density in the dermis is higher for the skin having thinner dermis. This contradicts the main paradigm of the RF theory stating that treatment results are mainly dependent on the maximal temperature reached in a target tissue, since the best short- and long-term clinical results of RF application to the skin were reported in the areas having thicker dermis. To resolve this contradiction, it is proposed that the long-term effect of RF can be realized through a structural modification of the subcutaneous fat depot adjacent to the treated skin area. Stimulation of these cells located near the interface dermis/subcutis will demand the concentration of applied RF energy in this area and will require the optimal arrangement of RF electrodes on the skin surface. PMID:27493952

3D structural patterns in scalable, elastomeric scaffolds guide engineered tissue architecture.

PubMed

Kolewe, Martin E; Park, Hyoungshin; Gray, Caprice; Ye, Xiaofeng; Langer, Robert; Freed, Lisa E

2013-08-27

Microfabricated elastomeric scaffolds with 3D structural patterns are created by semiautomated layer-by-layer assembly of planar polymer sheets with through-pores. The mesoscale interconnected pore architectures governed by the relative alignment of layers are shown to direct cell and muscle-like fiber orientation in both skeletal and cardiac muscle, enabling scale up of tissue constructs towards clinically relevant dimensions. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fabrication of 3D Reconstituted Organoid Arrays by DNA-programmed Assembly of Cells (DPAC)

PubMed Central

Todhunter, Michael E; Weber, Robert J; Farlow, Justin; Jee, Noel Y; Cerchiari, Alec E; Gartner, Zev J

2016-01-01

Tissues are the organizational units of function in metazoan organisms. Tissues comprise an assortment of cellular building blocks, soluble factors, and extracellular matrix (ECM) that are composed into specific three dimensional (3D) structures. The capacity to reconstitute tissues in vitro with the structural complexity observed in vivo is key to understanding processes such as morphogenesis, homeostasis, and disease. In this unit, we describe DNA-programmed Assembly of Cells (DPAC), a method to fabricate viable, functional arrays of organoid-like tissues within 3D ECM gels. In DPAC, dissociated cells are chemically functionalized with degradable oligonucleotide “velcro,” allowing rapid, specific, and reversible cell adhesion to a two-dimensional (2D) template patterned with complementary DNA. An iterative assembly process builds up organoids, layer-by-layer, from this initial 2D template and into the third dimension. Cleavage of the DNA releases the completed array of tissues that are captured and fully embedded in ECM gels for culture and observation. DPAC controls the size, shape, composition, and spatial heterogeneity of organoids, and permits positioning constituent cells with single-cell resolution even within cultures several centimeters long. PMID:27622567

Depth-resolved fluorescence of biological tissue

NASA Astrophysics Data System (ADS)

Wu, Yicong; Xi, Peng; Cheung, Tak-Hong; Yim, So Fan; Yu, Mei-Yung; Qu, Jianan Y.

2005-06-01

The depth-resolved autofluorescence ofrabbit oral tissue, normal and dysplastic human ectocervical tissue within l20μm depth were investigated utilizing a confocal fluorescence spectroscopy with the excitations at 355nm and 457nm. From the topmost keratinizing layer of oral and ectocervical tissue, strong keratin fluorescence with the spectral characteristics similar to collagen was observed. The fluorescence signal from epithelial tissue between the keratinizing layer and stroma can be well resolved. Furthermore, NADH and FADfluorescence measured from the underlying non-keratinizing epithelial layer were strongly correlated to the tissue pathology. This study demonstrates that the depth-resolved fluorescence spectroscopy can reveal fine structural information on epithelial tissue and potentially provide more accurate diagnostic information for determining tissue pathology.

Biofabricated Structures Reconstruct Functional Urinary Bladders in Radiation-injured Rat Bladders.

PubMed

Imamura, Tetsuya; Shimamura, Mitsuru; Ogawa, Teruyuki; Minagawa, Tomonori; Nagai, Takashi; Silwal Gautam, Sudha; Ishizuka, Osamu

2018-05-08

The ability to repair damaged urinary bladders through the application of bone marrow-derived cells is in the earliest stages of development. We investigated the application of bone marrow-derived cells to repair radiation-injured bladders. We used a three-dimensional (3D) bioprinting robot system to biofabricate bone marrow-derived cell structures. We then determined if the biofabricated structures could restore the tissues and functions of radiation-injured bladders. The bladders of female 10-week-old Sprague-Dawley (SD) rats were irradiated with 2-Gy once a week for 5 weeks. Adherent and proliferating bone marrow-derived cells harvested from the femurs of male 17-week-old green fluorescence protein-transfected Tg-SD rats were cultured in collagen-coated flasks. Bone marrow-derived cell spheroids were formed in 96-well plates. Three layers of spheroids were assembled by the bioprinter onto a 9x9 microneedle array. The assembled spheroids were perfusion cultured for 7 days, and then the microneedle array was removed. Two weeks after the last radiation treatment, the biofabricated structures were transplanted into an incision on the anterior wall of the bladders (n=10). Control rats received the same surgery but without the biofabricated structures (sham-structure, n=12). At 2 and 4 weeks after surgery, the sham-structure control bladder tissues exhibited disorganized smooth muscle layers, decreased nerve cells, and significant fibrosis with increased presence of fibrosis-marker P4HB-positive cells and hypoxia-marker HIF1α-positive cells. The transplanted structures survived within the recipient tissues, and blood vessels extended within them from the recipient tissues. The bone marrow-derived cells in the structures differentiated into smooth muscle cells and formed smooth muscle clusters. The recipient tissues near the transplanted structures had distinct smooth muscle layers and reconstructed nerve cells, and only minimal fibrosis with decreased presence of P4HB- and HIF1α-positive cells. At 4 weeks after surgery, the sham-structure control rats exhibited significant urinary frequency symptoms with irregular and short voiding intervals, and low micturition volumes. In contrast, the structure-transplanted rats had regular micturition with longer voiding intervals and higher micturition volumes compared to the control rats. Further, the residual volume of the structure-transplanted rats was lower than for the controls. Therefore, transplantation of biofabricated bone marrow-derived cell structures reconstructed functional bladders.

Raman spectroscopic analysis of human skin tissue sections ex-vivo: evaluation of the effects of tissue processing and dewaxing

NASA Astrophysics Data System (ADS)

Ali, Syed M.; Bonnier, Franck; Tfayli, Ali; Lambkin, Helen; Flynn, Kathleen; McDonagh, Vincent; Healy, Claragh; Clive Lee, T.; Lyng, Fiona M.; Byrne, Hugh J.

2013-06-01

Raman spectroscopy coupled with K-means clustering analysis (KMCA) is employed to elucidate the biochemical structure of human skin tissue sections and the effects of tissue processing. Both hand and thigh sections of human cadavers were analyzed in their unprocessed and formalin-fixed, paraffin-processed (FFPP), and subsequently dewaxed forms. In unprocessed sections, KMCA reveals clear differentiation of the stratum corneum (SC), intermediate underlying epithelium, and dermal layers for sections from both anatomical sites. The SC is seen to be relatively rich in lipidic content; the spectrum of the subjacent layers is strongly influenced by the presence of melanin, while that of the dermis is dominated by the characteristics of collagen. For a given anatomical site, little difference in layer structure and biochemistry is observed between samples from different cadavers. However, the hand and thigh sections are consistently differentiated for all cadavers, largely based on lipidic profiles. In dewaxed FFPP samples, while the SC, intermediate, and dermal layers are clearly differentiated by KMCA of Raman maps of tissue sections, the lipidic contributions to the spectra are significantly reduced, with the result that respective skin layers from different anatomical sites become indistinguishable. While efficient at removing the fixing wax, the tissue processing also efficiently removes the structurally similar lipidic components of the skin layers. In studies of dermatological processes in which lipids play an important role, such as wound healing, dewaxed samples are therefore not appropriate. Removal of the lipids does however accentuate the spectral features of the cellular and protein components, which may be more appropriate for retrospective analysis of disease progression and biochemical analysis using tissue banks.

Optical property measurement from layered biological media

NASA Astrophysics Data System (ADS)

Muller, Matthew R.

1998-12-01

Near infrared (NIR) photon reflectance spectroscopy is applied to measurement of blood concentration and its oxygen saturation within biological tissue. The measurement relies upon the changes in photon absorption of hemoglobin in the tissue as changes occur in the hemoglobin concentration and oxygen content. In the present study, NIR light is introduced at the skin surface and the optical properties (absorption and scattering) within the underlying tissue are determined from the resulting surface reflectance. Typically the tissue is modeled as a homogeneous mixture of bloodless tissue and blood, and the model incorporates the physical relationship between the surface reflectance and the optical properties of the tissue. The skin and underlying tissue, although heterogeneous, have a characteristic layered structure. These layers can be differentiated optically. The modeling and the inverse problem of measuring the optical properties in each of the tissue layers from the surface reflectance have been the subject of much attention by a number of investigators. Nonetheless, quantification of the relationship between surface reflectance and the optical properties of layered tissue has not been well understood nor well described. In the forward problem, tissue optical properties yield surface reflectance profiles (SRPs). Surface reflectance profiles, or SRPs, from diffusive media consisting of two layers are calculated using numerical solutions to the Boltzmann equation. Experimental SRPs are also measured in vitro from a test medium and in vivo from the calf of human subjects. This study provides a new approach to solving the inverse problem of determining optical properties from SRPs. To solve the inverse problem, an effective diffusion constant (Ke) is determined for the layered media. The Ke is the diffusion constant of an equivalent homogeneous medium which best fits the SRP of the layered medium. The departure from Ke of the SRP for a layered media is captured concisely, and Ke becomes a tool in describing the layered optical properties. This approach is applied clinically to measure changes in the blood concentration and oxygenation measured in vivo from normals and patients with peripheral vascular disease. A significant finding from the modeling was to identify the functional relationship of Ke to the top and lower layer diffusion constants, and the top layer thickness. When applied to in vitro measurements from media containing homogeneous layers with known optical properties, this functional relationship predicted Ke within the 95% confidence interval of the measured Ke. For the in vivo measurements, changes in K e with exercise are consistent with expected exercise physiology. With the incorporation of the known optical absorbance of hemoglobin in the presence of oxygen, the SRPs provide a means to measure the oxygen saturation of a deep tissue layer from the surface light reflectance.

Mueller matrix mapping of biological polycrystalline layers using reference wave

NASA Astrophysics Data System (ADS)

Dubolazov, A.; Ushenko, O. G.; Ushenko, Yu. O.; Pidkamin, L. Y.; Sidor, M. I.; Grytsyuk, M.; Prysyazhnyuk, P. V.

2018-01-01

The paper consists of two parts. The first part is devoted to the short theoretical basics of the method of differential Mueller-matrix description of properties of partially depolarizing layers. It was provided the experimentally measured maps of differential matrix of the 1st order of polycrystalline structure of the histological section of brain tissue. It was defined the statistical moments of the 1st-4th orders, which characterize the distribution of matrix elements. In the second part of the paper it was provided the data of statistic analysis of birefringence and dichroism of the histological sections of mice liver tissue (normal and with diabetes). It were defined the objective criteria of differential diagnostics of diabetes.

Anatomy and histology of the transverse humeral ligament.

PubMed

Snow, Brian J; Narvy, Steven J; Omid, Reza; Atkinson, Roscoe D; Vangsness, C Thomas

2013-10-01

The classic literature describes the transverse humeral ligament (THL) as a distinct anatomic structure with a role in biceps tendon stability; however, recent literature suggests that it is not a distinct anatomic structure. The purpose of this study was to evaluate the gross and microscopic anatomy of the THL, including a specific investigation of the histology of this ligament. Thirty frozen, embalmed cadaveric specimens were dissected to determine the gross anatomy of the THL. Seven specimens were evaluated histologically for the presence of mechanoreceptors and free nerve endings. Two tissue layers were identified in the area described as the THL. In the deep layer, fibers of the subscapularis tendon were found to span the bicipital groove with contributions from the coracohumeral ligament and the supraspinatus tendon. Superficial to this layer was a fibrous fascial covering consisting of distinct bands of tissue. Neurohistology staining revealed the presence of free nerve endings but no mechanoreceptors. This study's findings demonstrate that the THL is a distinct structure continuous with the rotator cuff tendons and the coracohumeral ligament. The finding of free nerve endings in the THL suggests a potential role as a shoulder pain generator. Copyright 2013, SLACK Incorporated.

Segmenting Brain Tissues from Chinese Visible Human Dataset by Deep-Learned Features with Stacked Autoencoder

PubMed Central

Zhao, Guangjun; Wang, Xuchu; Niu, Yanmin; Tan, Liwen; Zhang, Shao-Xiang

2016-01-01

Cryosection brain images in Chinese Visible Human (CVH) dataset contain rich anatomical structure information of tissues because of its high resolution (e.g., 0.167 mm per pixel). Fast and accurate segmentation of these images into white matter, gray matter, and cerebrospinal fluid plays a critical role in analyzing and measuring the anatomical structures of human brain. However, most existing automated segmentation methods are designed for computed tomography or magnetic resonance imaging data, and they may not be applicable for cryosection images due to the imaging difference. In this paper, we propose a supervised learning-based CVH brain tissues segmentation method that uses stacked autoencoder (SAE) to automatically learn the deep feature representations. Specifically, our model includes two successive parts where two three-layer SAEs take image patches as input to learn the complex anatomical feature representation, and then these features are sent to Softmax classifier for inferring the labels. Experimental results validated the effectiveness of our method and showed that it outperformed four other classical brain tissue detection strategies. Furthermore, we reconstructed three-dimensional surfaces of these tissues, which show their potential in exploring the high-resolution anatomical structures of human brain. PMID:27057543

Segmenting Brain Tissues from Chinese Visible Human Dataset by Deep-Learned Features with Stacked Autoencoder.

PubMed

Zhao, Guangjun; Wang, Xuchu; Niu, Yanmin; Tan, Liwen; Zhang, Shao-Xiang

2016-01-01

Cryosection brain images in Chinese Visible Human (CVH) dataset contain rich anatomical structure information of tissues because of its high resolution (e.g., 0.167 mm per pixel). Fast and accurate segmentation of these images into white matter, gray matter, and cerebrospinal fluid plays a critical role in analyzing and measuring the anatomical structures of human brain. However, most existing automated segmentation methods are designed for computed tomography or magnetic resonance imaging data, and they may not be applicable for cryosection images due to the imaging difference. In this paper, we propose a supervised learning-based CVH brain tissues segmentation method that uses stacked autoencoder (SAE) to automatically learn the deep feature representations. Specifically, our model includes two successive parts where two three-layer SAEs take image patches as input to learn the complex anatomical feature representation, and then these features are sent to Softmax classifier for inferring the labels. Experimental results validated the effectiveness of our method and showed that it outperformed four other classical brain tissue detection strategies. Furthermore, we reconstructed three-dimensional surfaces of these tissues, which show their potential in exploring the high-resolution anatomical structures of human brain.

Bessel light sheet structured illumination microscopy

NASA Astrophysics Data System (ADS)

Noshirvani Allahabadi, Golchehr

Biomedical study researchers using animals to model disease and treatment need fast, deep, noninvasive, and inexpensive multi-channel imaging methods. Traditional fluorescence microscopy meets those criteria to an extent. Specifically, two-photon and confocal microscopy, the two most commonly used methods, are limited in penetration depth, cost, resolution, and field of view. In addition, two-photon microscopy has limited ability in multi-channel imaging. Light sheet microscopy, a fast developing 3D fluorescence imaging method, offers attractive advantages over traditional two-photon and confocal microscopy. Light sheet microscopy is much more applicable for in vivo 3D time-lapsed imaging, owing to its selective illumination of tissue layer, superior speed, low light exposure, high penetration depth, and low levels of photobleaching. However, standard light sheet microscopy using Gaussian beam excitation has two main disadvantages: 1) the field of view (FOV) of light sheet microscopy is limited by the depth of focus of the Gaussian beam. 2) Light-sheet images can be degraded by scattering, which limits the penetration of the excitation beam and blurs emission images in deep tissue layers. While two-sided sheet illumination, which doubles the field of view by illuminating the sample from opposite sides, offers a potential solution, the technique adds complexity and cost to the imaging system. We investigate a new technique to address these limitations: Bessel light sheet microscopy in combination with incoherent nonlinear Structured Illumination Microscopy (SIM). Results demonstrate that, at visible wavelengths, Bessel excitation penetrates up to 250 microns deep in the scattering media with single-side illumination. Bessel light sheet microscope achieves confocal level resolution at a lateral resolution of 0.3 micron and an axial resolution of 1 micron. Incoherent nonlinear SIM further reduces the diffused background in Bessel light sheet images, resulting in confocal quality images in thick tissue. The technique was applied to live transgenic zebra fish tg(kdrl:GFP), and the sub-cellular structure of fish vasculature genetically labeled with GFP was captured in 3D. The superior speed of the microscope enables us to acquire signal from 200 layers of a thick sample in 4 minutes. The compact microscope uses exclusively off-the-shelf components and offers a low-cost imaging solution for studying small animal models or tissue samples.

Bioactive Nano-Fibrous Scaffolds for Bone and Cartilage Tissue Engineering

NASA Astrophysics Data System (ADS)

Feng, Kai

Scaffolds that can mimic the structural features of natural extracellular matrix and can deliver biomolecules in a controlled fashion may provide cells with a favorable microenvironment to facilitate tissue regeneration. Biodegradable nanofibrous scaffolds with interconnected pore network have previously been developed in our laboratory to mimic collagen matrix and advantageously support both bone and cartilage regeneration. This dissertation project aims to expand both the structural complexity and the biomolecule delivery capacity of such biomimetic scaffolds for tissue engineering. We first developed a nanofibrous scaffold that can release an antibiotic (doxycycline) with a tunable release rate and a tunable dosage, which was demonstrated to be able to inhibit bacterial growth over a prolonged time period. We then developed a nanofibrous tissue-engineciing scaffold that can release basic fibroblast growth factor (bFGF) in a spatially and temporally controlled fashion. In a mouse subcutaneous implantation model, the bFGF-releasing scaffold was shown to enhance cell penetration, tissue ingrowth and angiogenesis. It was also found that both the dose and the release rate of bFGF play roles in the biologic function of the scaffold. After that, we developed a nanofibrous PLLA scaffold that can release both bone morphogenetic protein 7 (BMP-7) and platelet-derived growth factor (PDGF) with distinct dosages and release kinetics. It was demonstrated that BMP-7 and PDGF could synergistically enhance bone regeneration using a mouse ectopic bone formation model and a rat periodontal fenestration defect regeneration model. The regeneration outcome was dependent on the dosage, the ratio and the release kinetics of the two growth factors. Last, we developed an anisotropic composite scaffold with an upper layer mimicking the superficial zone of cartilage and a lower layer mimicking the middle zone of cartilage. The thin superficial layer was fabricated using an electrospinning technique to support a more parallel ECM orientation to the cartilage surface. The lower layer was fabricated using a phase-separation technique to support a more isotropic ECM distribution. Human bone marrow-derived mesenchymal stem cells (hMSCs) were seeded on this complex scaffold and cultured under chondrogenic conditions. The results showed that the composite scaffold was indeed able to support anisotropic cartilage tissue structure formation.

Modal response of a computational vocal fold model with a substrate layer of adipose tissue.

PubMed

Jones, Cameron L; Achuthan, Ajit; Erath, Byron D

2015-02-01

This study demonstrates the effect of a substrate layer of adipose tissue on the modal response of the vocal folds, and hence, on the mechanics of voice production. Modal analysis is performed on the vocal fold structure with a lateral layer of adipose tissue. A finite element model is employed, and the first six mode shapes and modal frequencies are studied. The results show significant changes in modal frequencies and substantial variation in mode shapes depending on the strain rate of the adipose tissue. These findings highlight the importance of considering adipose tissue in computational vocal fold modeling.

The effect of abdominal wall morphology on ultrasonic pulse distortion. Part II. Simulations.

PubMed

Mast, T D; Hinkelman, L M; Orr, M J; Waag, R C

1998-12-01

Wavefront propagation through the abdominal wall was simulated using a finite-difference time-domain implementation of the linearized wave propagation equations for a lossless, inhomogeneous, two-dimensional fluid as well as a simplified straight-ray model for a two-dimensional absorbing medium. Scanned images of six human abdominal wall cross sections provided the data for the propagation media in the simulations. The images were mapped into regions of fat, muscle, and connective tissue, each of which was assigned uniform sound speed, density, and absorption values. Propagation was simulated through each whole specimen as well as through each fat layer and muscle layer individually. Wavefronts computed by the finite-difference method contained arrival time, energy level, and wave shape distortion similar to that in measurements. Straight-ray simulations produced arrival time fluctuations similar to measurements but produced much smaller energy level fluctuations. These simulations confirm that both fat and muscle produce significant wavefront distortion and that distortion produced by fat sections differs from that produced by muscle sections. Spatial correlation of distortion with tissue composition suggests that most major arrival time fluctuations are caused by propagation through large-scale inhomogeneities such as fatty regions within muscle layers, while most amplitude and waveform variations are the result of scattering from smaller inhomogeneities such as septa within the subcutaneous fat. Additional finite-difference simulations performed using uniform-layer models of the abdominal wall indicate that wavefront distortion is primarily caused by tissue structures and inhomogeneities rather than by refraction at layer interfaces or by variations in layer thicknesses.

Phase-resolved reflectance spectroscopy on layered turbid media

NASA Astrophysics Data System (ADS)

Hielscher, Andreas H.; Liu, Hanli; Chance, Britton; Tittel, Frank K.; Jacques, Steven L.

1995-05-01

In this study, we investigate the influence of layered tissue structures on the phase-resolved reflectance. As a particular example, we consider the affect of the skin, skull, and meninges on noninvasive blood oxygenation determination of the brain. In this case, it's important to know how accurate one can measure the absorption coefficient of the brain through the enclosing layers of different tissues. Experiments were performed on layered gelatin tissue phantoms and the results compared to diffusion theory. It is shown that when a high absorbing medium is placed on top of a low absorbing medium, the absorption coefficient of the lower layer is accessible. In the inverse case, where a low absorbing medium is placed on top of a high absorbing medium, the absorption coefficient of the underlying medium can only be determined if the differences in the absorption coefficient are small, or the top layer is very thin. Investigations on almost absorption and scattering free layers, like the cerebral fluid filled arachnoid, reveal that the determination of the absorption coefficient is barely affected by these kinds of structures.

Mechanisms of lamellar collagen formation in connective tissues.

PubMed

Ghazanfari, Samaneh; Khademhosseini, Ali; Smit, Theodoor H

2016-08-01

The objective of tissue engineering is to regenerate functional tissues. Engineering functional tissues requires an understanding of the mechanisms that guide the formation and evolution of structure in the extracellular matrix (ECM). In particular, the three-dimensional (3D) collagen fiber arrangement is important as it is the key structural determinant that provides mechanical integrity and biological function. In this review, we survey the current knowledge on collagen organization mechanisms that can be applied to create well-structured functional lamellar tissues and in particular intervertebral disc and cornea. Thus far, the mechanisms behind the formation of cross-aligned collagen fibers in the lamellar structures is not fully understood. We start with cell-induced collagen alignment and strain-stabilization behavior mechanisms which can explain a single anisotropically aligned collagen fiber layer. These mechanisms may explain why there is anisotropy in a single layer in the first place. However, they cannot explain why a consecutive collagen layer is laid down with an alternating alignment. Therefore, we explored another mechanism, called liquid crystal phasing. While dense concentrations of collagen show such behavior, there is little evidence that the conditions for liquid crystal phasing are actually met in vivo. Instead, lysyl aldehyde-derived collagen cross-links have been found essential for correct lamellar matrix deposition. Furthermore, we suggest that supra-cellular (tissue-level) shear stress may be instrumental in the alignment of collagen fibers. Understanding the potential mechanisms behind the lamellar collagen structure in connective tissues will lead to further improvement of the regeneration strategies of functional complex lamellar tissues. Copyright © 2016 Elsevier Ltd. All rights reserved.

New physiologically-relevant liver tissue model based on hierarchically cocultured primary rat hepatocytes with liver endothelial cells.

PubMed

Xiao, Wenjin; Perry, Guillaume; Komori, Kikuo; Sakai, Yasuyuki

2015-11-01

To develop an in vitro liver tissue equivalent, hepatocytes should be cocultured with liver non-parenchymal cells to mimic the in vivo physiological microenvironments. In this work, we describe a physiologically-relevant liver tissue model by hierarchically organizing layers of primary rat hepatocytes and human liver sinusoidal endothelial cells (TMNK-1) on an oxygen-permeable polydimethylsiloxane (PDMS) membrane, which facilitates direct oxygenation by diffusion through the membrane. This in vivo-mimicking hierarchical coculture was obtained by simply proceeding the overlay of TMNK-1 cells on the hepatocyte layer re-formed on the collagen immobilized PDMS membranes. The comparison of hepatic functionalities was achieved between coculture and sandwich culture with Matrigel, in the presence and absence of direct oxygenation. A complete double-layered structure of functional liver cells with vertical contact between hepatocytes and TMNK-1 was successfully constructed in the coculture with direct oxygen supply and was well-maintained for 14 days. The hepatocytes in this hierarchical culture exhibited improved survival, functional bile canaliculi formation, cellular level polarization and maintenance of metabolic activities including Cyp1A1/2 activity and albumin production. By contrast, the two cell populations formed discontinuous monolayers on the same surfaces in the non-oxygen-permeable cultures. These results demonstrate that (i) the direct oxygenation through the PDMS membranes enables very simple formation of a hierarchical structure consisting of a hepatocyte layer and a layer of TMNK-1 and (ii) we may include other non-parenchymal cells in this format easily, which can be widely applicable to other epithelial organs.

INTERLAYER MICROMECHANICS OF THE AORTIC HEART VALVE LEAFLET

PubMed Central

Buchanan, Rachel M.; Sacks, Michael S.

2014-01-01

While the mechanical behaviors of the fibrosa and ventricularis layers of the aortic valve (AV) leaflet are understood, little information exists on their mechanical interactions mediated by the GAG-rich central spongiosa layer. Parametric simulations of the interlayer interactions of the AV leaflets in flexure utilized a tri-layered finite element (FE) model of circumferentially oriented tissue sections to investigate inter-layer sliding hypothesized to occur. Simulation results indicated that the leaflet tissue functions as a tightly bonded structure when the spongiosa effective modulus was at least 25% that of the fibrosa and ventricularis layers. Novel studies that directly measured transmural strain in flexure of AV leaflet tissue specimens validated these findings. Interestingly, a smooth transmural strain distribution indicated that the layers of the leaflet indeed act as a bonded unit, consistent with our previous observations (Stella and Sacks, 2007) of a large number of transverse collagen fibers interconnecting the fibrosa and ventricularis layers. Additionally, when the tri-layered FE model was refined to match the transmural deformations, a layer-specific bimodular material model (resulting in four total moduli) accurately matched the transmural strain and moment-curvature relations simultaneously. Collectively, these results provide evidence, contrary to previous assumptions, that the valve layers function as a bonded structure in the low-strain flexure deformation mode. Most likely, this results directly from the transverse collagen fibers that bind the layers together to disable physical sliding and maintain layer residual stresses. Further, the spongiosa may function as a general dampening layer while the AV leaflets deforms as a homogenous structure despite its heterogeneous architecture. PMID:24292631

Three-Dimensional Computer-Assisted Two-Layer Elastic Models of the Face.

PubMed

Ueda, Koichi; Shigemura, Yuka; Otsuki, Yuki; Fuse, Asuka; Mitsuno, Daisuke

2017-11-01

To make three-dimensional computer-assisted elastic models for the face, we decided on five requirements: (1) an elastic texture like skin and subcutaneous tissue; (2) the ability to take pen marking for incisions; (3) the ability to be cut with a surgical knife; (4) the ability to keep stitches in place for a long time; and (5) a layered structure. After testing many elastic solvents, we have made realistic three-dimensional computer-assisted two-layer elastic models of the face and cleft lip from the computed tomographic and magnetic resonance imaging stereolithographic data. The surface layer is made of polyurethane and the inner layer is silicone. Using this elastic model, we taught residents and young doctors how to make several typical local flaps and to perform cheiloplasty. They could experience realistic simulated surgery and understand three-dimensional movement of the flaps.

Assessment of microcirculation dynamics during cutaneous wound healing phases in vivo using optical microangiography

PubMed Central

Yousefi, Siavash; Qin, Jia; Dziennis, Suzan; Wang, Ruikang K.

2014-01-01

Abstract. Cutaneous wound healing consists of multiple overlapping phases starting with blood coagulation following incision of blood vessels. We utilized label-free optical coherence tomography and optical microangiography (OMAG) to noninvasively monitor healing process and dynamics of microcirculation system in a mouse ear pinna wound model. Mouse ear pinna is composed of two layers of skin separated by a layer of cartilage and because its total thickness is around 500 μm, it can be utilized as an ideal model for optical imaging techniques. These skin layers are identical to human skin structure except for sweat ducts and glands. Microcirculatory system responds to the wound injury by recruiting collateral vessels to supply blood flow to hypoxic region. During the inflammatory phase, lymphatic vessels play an important role in the immune response of the tissue and clearing waste from interstitial fluid. In the final phase of wound healing, tissue maturation, and remodeling, the wound area is fully closed while blood vessels mature to support the tissue cells. We show that using OMAG technology allows noninvasive and label-free monitoring and imaging each phase of wound healing that can be used to replace invasive tissue sample histology and immunochemistry technologies. PMID:25036212

Convergence and Extrusion Are Required for Normal Fusion of the Mammalian Secondary Palate

PubMed Central

Kim, Seungil; Lewis, Ace E.; Singh, Vivek; Ma, Xuefei; Adelstein, Robert; Bush, Jeffrey O.

2015-01-01

The fusion of two distinct prominences into one continuous structure is common during development and typically requires integration of two epithelia and subsequent removal of that intervening epithelium. Using confocal live imaging, we directly observed the cellular processes underlying tissue fusion, using the secondary palatal shelves as a model. We find that convergence of a multi-layered epithelium into a single-layer epithelium is an essential early step, driven by cell intercalation, and is concurrent to orthogonal cell displacement and epithelial cell extrusion. Functional studies in mice indicate that this process requires an actomyosin contractility pathway involving Rho kinase (ROCK) and myosin light chain kinase (MLCK), culminating in the activation of non-muscle myosin IIA (NMIIA). Together, these data indicate that actomyosin contractility drives cell intercalation and cell extrusion during palate fusion and suggest a general mechanism for tissue fusion in development. PMID:25848986

Fabrication and characterization of two-layered nanofibrous membrane for guided bone and tissue regeneration application.

PubMed

Masoudi Rad, Maryam; Nouri Khorasani, Saied; Ghasemi-Mobarakeh, Laleh; Prabhakaran, Molamma P; Foroughi, Mohammad Reza; Kharaziha, Mahshid; Saadatkish, Niloufar; Ramakrishna, Seeram

2017-11-01

Membranes used in dentistry act as a barrier to prevent invasion of intruder cells to defected area and obtains spaces that are to be subsequently filled with new bone and provide required bone volume for implant therapy when there is insufficient volume of healthy bone at implant site. In this study a two-layered bioactive membrane were fabricated by electrospinning whereas one layer provides guided bone regeneration (GBR) and fabricated using poly glycerol sebacate (PGS)/polycaprolactone (PCL) and Beta tri-calcium phosphate (β-TCP) (5, 10 and 15%) and another one containing PCL/PGS and chitosan acts as guided tissue regeneration (GTR). The morphology, chemical, physical and mechanical characterizations of the membranes were studied using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), tensile testing, then biodegradability and bioactivity properties were evaluated. In vitro cell culture study was also carried out to investigate proliferation and mineralization of cells on different membranes. Transmission electron microscope (TEM) and SEM results indicated agglomeration of β-TCP nanoparticles in the structure of nanofibers containing 15% β-TCP. Moreover by addition of β-TCP from 5% to 15%, contact angle decreased due to hydrophilicity of nanoparticles and bioactivity was found to increase. Mechanical properties of the membrane increased by incorporation of 5% and 10% of β-TCP in the structure of nanofibers, while addition of 15% of β-TCP was found to deteriorate mechanical properties of nanofibers. Although the presence of 5% and 10% of nanoparticles in the nanofibers increased proliferation of cells on GBR layer, cell proliferation was observed to decrease by addition of 15% β-TCP in the structure of nanofibers which is likely due to agglomeration of nanoparticles in the nanofiber structure. Our overall results revealed PCL/PGS containing 10% β-TCP could be selected as the optimum GBR membrane in view point of physical and mechanical properties along with cell behavior. PCL/PGS nanofibers containing 10% β-TCP were electrospun on the GTR layer for fabrication of final membrane. Addition of chitosan in the structure of PCL/PGS nanofibers was found to decrease fiber diameter, contact angle and porosity which are favorable for GTR layer. Two-layered dental membrane fabricated in this study can serve as a suitable substrate for application in dentistry as it provides appropriate osteoconductivity and flexibility along with barrier properties. Copyright © 2017 Elsevier B.V. All rights reserved.

Synthetic, multi-layer, self-oscillating vocal fold model fabrication.

PubMed

Murray, Preston R; Thomson, Scott L

2011-12-02

Sound for the human voice is produced via flow-induced vocal fold vibration. The vocal folds consist of several layers of tissue, each with differing material properties. Normal voice production relies on healthy tissue and vocal folds, and occurs as a result of complex coupling between aerodynamic, structural dynamic, and acoustic physical phenomena. Voice disorders affect up to 7.5 million annually in the United States alone and often result in significant financial, social, and other quality-of-life difficulties. Understanding the physics of voice production has the potential to significantly benefit voice care, including clinical prevention, diagnosis, and treatment of voice disorders. Existing methods for studying voice production include in vivo experimentation using human and animal subjects, in vitro experimentation using excised larynges and synthetic models, and computational modeling. Owing to hazardous and difficult instrument access, in vivo experiments are severely limited in scope. Excised larynx experiments have the benefit of anatomical and some physiological realism, but parametric studies involving geometric and material property variables are limited. Further, they are typically only able to be vibrated for relatively short periods of time (typically on the order of minutes). Overcoming some of the limitations of excised larynx experiments, synthetic vocal fold models are emerging as a complementary tool for studying voice production. Synthetic models can be fabricated with systematic changes to geometry and material properties, allowing for the study of healthy and unhealthy human phonatory aerodynamics, structural dynamics, and acoustics. For example, they have been used to study left-right vocal fold asymmetry, clinical instrument development, laryngeal aerodynamics, vocal fold contact pressure, and subglottal acoustics (a more comprehensive list can be found in Kniesburges et al.) Existing synthetic vocal fold models, however, have either been homogenous (one-layer models) or have been fabricated using two materials of differing stiffness (two-layer models). This approach does not allow for representation of the actual multi-layer structure of the human vocal folds that plays a central role in governing vocal fold flow-induced vibratory response. Consequently, one- and two-layer synthetic vocal fold models have exhibited disadvantages such as higher onset pressures than what are typical for human phonation (onset pressure is the minimum lung pressure required to initiate vibration), unnaturally large inferior-superior motion, and lack of a "mucosal wave" (a vertically-traveling wave that is characteristic of healthy human vocal fold vibration). In this paper, fabrication of a model with multiple layers of differing material properties is described. The model layers simulate the multi-layer structure of the human vocal folds, including epithelium, superficial lamina propria (SLP), intermediate and deep lamina propria (i.e., ligament; a fiber is included for anterior-posterior stiffness), and muscle (i.e., body) layers. Results are included that show that the model exhibits improved vibratory characteristics over prior one- and two-layer synthetic models, including onset pressure closer to human onset pressure, reduced inferior-superior motion, and evidence of a mucosal wave.

Tissue bioengineering and artificial organs.

PubMed

Llames, Sara; García, Eva; Otero Hernández, Jesús; Meana, Alvaro

2012-01-01

The scarcity of organs and tissues for transplant and the need of immunosuppressive drugs to avoid rejection constitute two reasons that justify organ and tissue production in the laboratory. Tissue engineering based tissues (TE) could allow to regenerate the whole organ from a fragment or even to produce several organs from an organ donor for grafting purposes. TE is based in: (1) the ex vivo expansion of cells, (2) the seeding of these expanded cells in tridimensional structures that mimic physiological conditions and, (3) grafting the prototype. In order to graft big structures it is necessary that the organ or tissue produced "ex vivo" bears a vascular tree to ensure the nutrition of its deep layers. At present, no technology has been developed to provide this vascular tree to TE derived products. Thus, these tissues must be thin enough to acquire nutrients during the first days by diffusion from surrounding tissues. This fact constitutes nowadays the greatest limitation of technologies for organ development in the laboratory.In this chapter, all these problems and their possible solutions are commented. Also, the present status of TE techniques in the regeneration of different organ systems is reviewed.

In vitro reconstruction of hybrid vascular tissue. Hierarchic and oriented cell layers.

PubMed

Kanda, K; Matsuda, T; Oka, T

1993-01-01

Hybrid vascular tissue was hierarchically reconstructed in vitro. A hybrid medial layer composed of type I collagen gel, in which SMCs derived from a mongrel dog were embedded, was formed on the inner surface of a compliant porous polyurethane graft (internal diameter = 3 mm). Endothelial cells (ECs) from the same animal were seeded and cultured on the hybrid media to build an intimal layer. Subsequently, hierarchically structured grafts constructed in this manner were subjected to pulsatile flow (flow rate: 8.5 ml/min; frequency: 60 rpm; amplitude: 5% of graft outer diameter) of culture medium (Medium 199 supplemented with 20% fetal calf serum). After stress loading for as long as 10 days, tissues were morphologically investigated with a light microscope and a scanning electron microscope. Inner surfaces of the hybrid tissues were covered with EC monolayers that aligned along the direction of the flow (i.e., longitudinally). However, SMCs beneath the intima aligned in the circumferential direction. These cellular orientations resembled those in native muscular arteries. The pulsatile stress loaded hybrid tissue mimicked native muscular arteries with respect to hierarchic structure and cellular orientation. In vitro mechanical stress loading on a hybrid graft might provide a high degree of integrity in terms of tissue structure that promises high tolerance toward hydrodynamic stress and regulation of vasomotor tone upon implantation.

Triple-Layer Vascular Grafts Fabricated by Combined E-Jet 3D Printing and Electrospinning.

PubMed

Huang, Ruiying; Gao, Xiangkai; Wang, Jian; Chen, Haoxiang; Tong, Chunyi; Tan, Yongjun; Tan, Zhikai

2018-05-29

Small-diameter tissue-engineered vascular grafts are urgently needed for clinic arterial substitute. To simulate the structures and functions of natural blood vessels, we designed a novel triple-layer poly(ε-caprolactone) (PCL) fibrous vascular graft by combining E-jet 3D printing and electrospinning techniques. The resultant vascular graft consisted of an interior layer comprising 3D-printed highly aligned strong fibers, a middle layer made by electrospun densely fibers, and an exterior structure composed of mixed fibers fabricated by co-electrospraying. The biocompatible triple-layer graft was used for in vivo implantation, and results demonstrated that the longitudinally-aligned fibers within the lumen of the graft could enhance the proliferation and migration of endothelial cells, while maintained good mechanical properties. The exterior layer provided a pathway that encouraged cells to migrate into the scaffold after implantation. This experimental graft overcame the limitations of conventionally electrospun vascular grafts of inadequate porosity and lowly cell penetration. The unique structure of the triple-layer vascular graft promoted cell growth and infiltration in vivo, thus provided an encouraging substitute for in situ tissue engineering.

3D tissue formation by stacking detachable cell sheets formed on nanofiber mesh.

PubMed

Kim, Min Sung; Lee, Byungjun; Kim, Hong Nam; Bang, Seokyoung; Yang, Hee Seok; Kang, Seong Min; Suh, Kahp-Yang; Park, Suk-Hee; Jeon, Noo Li

2017-03-23

We present a novel approach for assembling 3D tissue by layer-by-layer stacking of cell sheets formed on aligned nanofiber mesh. A rigid frame was used to repeatedly collect aligned electrospun PCL (polycaprolactone) nanofiber to form a mesh structure with average distance between fibers 6.4 µm. When human umbilical vein endothelial cells (HUVECs), human foreskin dermal fibroblasts, and skeletal muscle cells (C2C12) were cultured on the nanofiber mesh, they formed confluent monolayers and could be handled as continuous cell sheets with areas 3 × 3 cm 2 or larger. Thicker 3D tissues have been formed by stacking multiple cell sheets collected on frames that can be nested (i.e. Matryoshka dolls) without any special tools. When cultured on the nanofiber mesh, skeletal muscle, C2C12 cells oriented along the direction of the nanofibers and differentiated into uniaxially aligned multinucleated myotube. Myotube cell sheets were stacked (upto 3 layers) in alternating or aligned directions to form thicker tissue with ∼50 µm thickness. Sandwiching HUVEC cell sheets with two dermal fibroblast cell sheets resulted in vascularized 3D tissue. HUVECs formed extensive networks and expressed CD31, a marker of endothelial cells. Cell sheets formed on nanofiber mesh have a number of advantages, including manipulation and stacking of multiple cell sheets for constructing 3D tissue and may find applications in a variety of tissue engineering applications.

Analytical modeling of polarization transformation of laser radiation of various spectral ranges by birefringent structures

NASA Astrophysics Data System (ADS)

Motrich, A. V.; Ushenko, O. G.

2018-01-01

The results of statistical dependence and correlation structures of two-dimensional Mueller matrix elements in various spectral regions of laser radiation by changes in the distribution of orientations of optical axes and birefringence of protein crystals. Namely, a two-wave ("red-blue") approach - layer of biological tissues irradiated by He-Ne laser (λ1 = 0,63μm ) and He-Cd laser (λ1 = 0,41μm )was used Conducted analysis of polarimetric sensitivity was made, a state of polarization points that contain volumetric structures of biological objects to spectral region of laser radiation was detected.

Differential 3D Mueller-matrix mapping of optically anisotropic depolarizing biological layers

NASA Astrophysics Data System (ADS)

Ushenko, O. G.; Grytsyuk, M.; Ushenko, V. O.; Bodnar, G. B.; Vanchulyak, O.; Meglinskiy, I.

2018-01-01

The paper consists of two parts. The first part is devoted to the short theoretical basics of the method of differential Mueller-matrix description of properties of partially depolarizing layers. It was provided the experimentally measured maps of differential matrix of the 2nd order of polycrystalline structure of the histological section of rectum wall tissue. It was defined the values of statistical moments of the1st-4th orders, which characterize the distribution of matrix elements. In the second part of the paper it was provided the data of statistic analysis of birefringence and dichroism of the histological sections of connecting component of vagina wall tissue (normal and with prolapse). It were defined the objective criteria of differential diagnostics of pathologies of vagina wall.

A multilayer scaffold design with spatial arrangement of cells to modulate esophageal tissue growth.

PubMed

Soliman, Sherif; Laurent, Julie; Kalenjian, Lena; Burnette, Kalia; Hedberg, Bert; La Francesca, Saverio

2018-05-02

Esophageal diseases may require resectioning of the damaged portion. The current standard of care requires the replacement of the esophagus with the stomach or the intestine. Such procedures have high rates of mortality and morbidity; therefore, the use of alternative conduits is needed. A tissue engineering approach that allows for the regeneration of esophageal tissues would have significant clinical application. A cell-seeded synthetic scaffold could replace the resected part of the esophagus and elicit tissue regrowth. In order to ideally recreate a functioning esophagus, its two crucial tissue layers should be induced: an epithelium on the luminal surface and a muscle layer on the exterior surface. To create a bioengineered esophagus with both tissue layers, a multilayer (ML) tubular scaffold design was considered. Luminal and exterior layers were electrospun with broad pore size to promote penetration and proliferation of mesenchymal stem cells on the lumen and smooth muscle cells on the external. These two layers would be separated by a thin layer with substantially narrower pore size intended to act as a barrier for the two cell types. This ML scaffold design was achieved via electrospinning by tuning the solution and the process parameters. Analysis of the scaffold demonstrated that this tuning enabled the production of three integrated layers with distinguishable microstructures and good mechanical integrity. In vitro validation was conducted on the separated unilayer components of the ML scaffold. The resultant proof-of-concept ML scaffold design could possibly support the spatial arrangement of cells needed to promote esophageal tissue regeneration. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2018. © 2018 Wiley Periodicals, Inc.

Optical measurements of absorption changes in two-layered diffusive media

NASA Astrophysics Data System (ADS)

Fabbri, Francesco; Sassaroli, Angelo; Henry, Michael E.; Fantini, Sergio

2004-04-01

We have used Monte Carlo simulations for a two-layered diffusive medium to investigate the effect of a superficial layer on the measurement of absorption variations from optical diffuse reflectance data processed by using: (a) a multidistance, frequency-domain method based on diffusion theory for a semi-infinite homogeneous medium; (b) a differential-pathlength-factor method based on a modified Lambert-Beer law for a homogeneous medium and (c) a two-distance, partial-pathlength method based on a modified Lambert-Beer law for a two-layered medium. Methods (a) and (b) lead to a single value for the absorption variation, whereas method (c) yields absorption variations for each layer. In the simulations, the optical coefficients of the medium were representative of those of biological tissue in the near-infrared. The thickness of the first layer was in the range 0.3-1.4 cm, and the source-detector distances were in the range 1-5 cm, which is typical of near-infrared diffuse reflectance measurements in tissue. The simulations have shown that (1) method (a) is mostly sensitive to absorption changes in the underlying layer, provided that the thickness of the superficial layer is ~0.6 cm or less; (2) method (b) is significantly affected by absorption changes in the superficial layer and (3) method (c) yields the absorption changes for both layers with a relatively good accuracy of ~4% for the superficial layer and ~10% for the underlying layer (provided that the absorption changes are less than 20-30% of the baseline value). We have applied all three methods of data analysis to near-infrared data collected on the forehead of a human subject during electroconvulsive therapy. Our results suggest that the multidistance method (a) and the two-distance partial-pathlength method (c) may better decouple the contributions to the optical signals that originate in deeper tissue (brain) from those that originate in more superficial tissue layers.

Characterization of vibrissa germinative cells: transition of cell types.

PubMed

Osada, A; Kobayashi, K

2001-12-01

Germinative cells, small cell masses attached to the stalks of dermal papillae that are able to differentiate into the hair shaft and inner root sheath, form follicular bulb-like structures when co-cultured with dermal papilla cells. We studied the growth characteristics of germinative cells to determine the cell types in the vibrissa germinative tissue. Germinative tissues, attaching to dermal papillae, were cultured on 3T3 feeder layers. The cultured keratinocytes were harvested and transferred, equally and for two passages, onto lined dermal papilla cells (LDPC) and/or 3T3 feeder layers. The resulting germinative cells were classified into three types in the present experimental condition. Type 1 cells grow very well on either feeder layer, whereas Type 3 cells scarcely grow on either feeder layer. Type 2 cells are very conspicuous and are reversible. They grow well on 3T3 but growth is suppressed on LDPC feeder layers. The Type 2 cells that grow well on 3T3 feeder layers, however, are suppressed when transferred onto LDPC and the Type 2 cells that are suppressed on LDPC begin to grow again on 3T3. The transition of one cell type to another in vitro and the cell types that these germinative cell types correspond to in vivo is discussed. It was concluded that stem cells or their close progenitors reside in the germinative tissues of the vibrissa bulb except at late anagen-early catagen.

S-layer fusion protein as a tool functionalizing emulsomes and CurcuEmulsomes for antibody binding and targeting

PubMed Central

Ucisik, Mehmet H.; Küpcü, Seta; Breitwieser, Andreas; Gelbmann, Nicola; Schuster, Bernhard; Sleytr, Uwe B.

2015-01-01

Selective targeting of tumor cells by nanoparticle-based drug delivery systems is highly desirable because it maximizes the drug concentration at the desired target while simultaneously protecting the surrounding healthy tissues. Here, we show a design for smart nanocarriers based on a biomimetic approach that utilizes the building principle of virus envelope structures. Emulsomes and CurcuEmulsomes comprising a tripalmitin solid core surrounded by phospholipid layers are modified by S-layer proteins that self-assemble into a two-dimensional array to form a surface layer. One significant advantage of this nanoformulation is that it increases the solubility of the lipophilic anti-cancer agent curcumin in the CurcuEmulsomes by a factor of 2700. In order to make the emulsomes specific for IgG, the S-layer protein is fused with two protein G domains. This S-layer fusion protein preserves its recrystallization characteristics, forming an ordered surface layer (square lattice with 13 nm unit-by-unit distance). The GG domains are presented in a predicted orientation and exhibit a selective binding affinity for IgG. PMID:25734967

Pericarditis

MedlinePlus

... properly. The sac is made of two thin layers of tissue that enclose your heart. Between the two layers is a small amount of fluid. This fluid keeps the layers from rubbing against each other and causing friction. ...

Effects of scaffold architecture on mechanical characteristics and osteoblast response to static and perfusion bioreactor cultures.

PubMed

Bartnikowski, Michal; Klein, Travis J; Melchels, Ferry P W; Woodruff, Maria A

2014-07-01

Tissue engineering focuses on the repair and regeneration of tissues through the use of biodegradable scaffold systems that structurally support regions of injury while recruiting and/or stimulating cell populations to rebuild the target tissue. Within bone tissue engineering, the effects of scaffold architecture on cellular response have not been conclusively characterized in a controlled-density environment. We present a theoretical and practical assessment of the effects of polycaprolactone (PCL) scaffold architectural modifications on mechanical and flow characteristics as well as MC3T3-E1 preosteoblast cellular response in an in vitro static plate and custom-designed perfusion bioreactor model. Four scaffold architectures were contrasted, which varied in inter-layer lay-down angle and offset between layers, while maintaining a structural porosity of 60 ± 5%. We established that as layer angle was decreased (90° vs. 60°) and offset was introduced (0 vs. 0.5 between layers), structural stiffness, yield stress, strength, pore size, and permeability decreased, while computational fluid dynamics-modeled wall shear stress was increased. Most significant effects were noted with layer offset. Seeding efficiencies in static culture were also dramatically increased due to offset (∼ 45% to ∼ 86%), with static culture exhibiting a much higher seeding efficiency than perfusion culture. Scaffold architecture had minimal effect on cell response in static culture. However, architecture influenced osteogenic differentiation in perfusion culture, likely by modifying the microfluidic environment. © 2014 Wiley Periodicals, Inc.

Label-free multiphoton microscopy reveals altered tissue architecture in hippocampal sclerosis.

PubMed

Uckermann, Ortrud; Galli, Roberta; Leupold, Susann; Coras, Roland; Meinhardt, Matthias; Hallmeyer-Elgner, Susanne; Mayer, Thomas; Storch, Alexander; Schackert, Gabriele; Koch, Edmund; Blümcke, Ingmar; Steiner, Gerald; Kirsch, Matthias

2017-01-01

The properties and structure of tissue can be visualized without labeling or preparation by multiphoton microscopy combining coherent anti-Stokes Raman scattering (CARS), addressing lipid content, second harmonic generation (SHG) showing collagen, and two-photon excited fluorescence (TPEF) of endogenous fluorophores. We compared samples of sclerotic and nonsclerotic human hippocampus to detect pathologic changes in the brain of patients with pharmacoresistant temporomesial epilepsy (n = 15). Multiphoton microscopy of cryosections and bulk tissue revealed hippocampal layering and micromorphologic details in accordance with reference histology: CARS displayed white and gray matter layering and allowed the assessment of axonal myelin. SHG visualized blood vessels based on adventitial collagen. In addition, corpora amylacea (CoA) were found to be SHG-active. Pyramidal cell bodies were characterized by intense cytoplasmic endogenous TPEF. Furthermore, diffuse TPEF around blood vessels was observed that co-localized with positive albumin immunohistochemistry and might indicate degeneration-associated vascular leakage. We present a label-free and fast optical approach that analyzes pathologic aspects of HS. Hippocampal layering, loss of pyramidal cells, and presence of CoA indicative of sclerosis are visualized. Label-free multiphoton microscopy has the potential to extend the histopathologic armamentarium for ex vivo assessment of changes of the hippocampal formation on fresh tissue and prospectively in vivo. Wiley Periodicals, Inc. © 2016 International League Against Epilepsy.

General theory of skin reinforcement.

PubMed

Kruglikov, Ilja L; Scherer, Philipp E

2017-01-01

Macroscopic mechanical properties of human skin in vivo cannot be considered independent of adjacent subcutaneous white adipose tissue (sWAT). The layered system skin/sWAT appears as the hierarchical structural composite in which single layers behave as fiber-reinforced structures. Effective macroscopic mechanical properties of such composites are mainly determined either by the properties of the skin or by those of the sWAT, dependent on the conditions of mechanical loading. Mechanical interactions between the skin and the adjacent sWAT associated with a mismatch in the mechanical moduli of these two layers can lead to production of the skin wrinkles. Reinforcement of the composite skin/sWAT can take place in different ways. It can be provided through reorientation of collagen fibers under applied loading, through production of new bonds between existing collagen fibers and through induction of additional collagen structures. Effectiveness of this type of reinforcement is strongly dependent on the type of mechanical loading. Different physical interventions induce the reinforcement of at least one of these two layers, thus increasing the effective macroscopic stiffness of the total composite. At the same time, the standalone reinforcement of the skin appears to be less effective to achieve a delay or a reduction of the apparent signs of skin aging relative to the reinforcement of the sWAT.

Optical coherence tomography monitoring of vocal fold femtosecond laser microsurgery

NASA Astrophysics Data System (ADS)

Wisweh, Henning; Merkel, Ulrich; Hüller, Ann-Kristin; Lüerßen, Kathrin; Lubatschowski, Holger

2007-07-01

Surgery of benign pathological alterations of the vocal folds results in permanent disphonia if the bounderies of the vocal fold layers are disregarded. Precise cutting with a femtosecond laser (fs-laser) combined with simultanous imaging of the layered structure enables accurate resections with respect to the layer boundaries. Earlier works demonstrated the capability of optical coherence tomography (OCT) for utilization on vocal folds. The layered structure can be imaged with a spatial resolution of 10-20μm up to a depth of 1.5mm. The performance of fs-laser cutting was analyzed on extracted porcine vocal folds with OCT monitoring. Histopathological sections of the same processed samples could be well correlated with the OCT images. With adequate laser parameters thermal effects induced only negligable damage to the processed tissue. The dimensions of the thermal necrosis were determined to be smaller than 1μm. OCT contolled fs-laser cutting of porcine vocal fold tissue in the μm range with minimal tissue damage is presented.

Hemodynamic monitoring in different cortical layers with a single fiber optical system

NASA Astrophysics Data System (ADS)

Yu, Linhui; Noor, M. Sohail; Kiss, Zelma H. T.; Murari, Kartikeya

2018-02-01

Functional monitoring of highly-localized deep brain structures is of great interest. However, due to light scattering, optical methods have limited depth penetration or can only measure from a large volume. In this research, we demonstrate continuous measurement of hemodynamics in different cortical layers in response to thalamic deep brain stimulation (DBS) using a single fiber optical system. A 200-μm-core-diameter multimode fiber is used to deliver and collect light from tissue. The fiber probe can be stereotaxically implanted into the brain region of interest at any depth to measure the di use reflectance spectra from a tissue volume of 0.02-0.03 mm3 near the fiber tip. Oxygenation is then extracted from the reflectance spectra using an algorithm based on Monte Carlo simulations. Measurements were performed on the surface (cortical layer I) and at 1.5 mm depth (cortical layer VI) of the motor cortex in anesthetized rats with thalamic DBS. Preliminary results revealed the oxygenation changes in response to DBS. Moreover, the baseline as well as the stimulus-evoked change in oxygenation were different at the two depths of cortex.

Applying macromolecular crowding to 3D bioprinting: fabrication of 3D hierarchical porous collagen-based hydrogel constructs.

PubMed

Ng, Wei Long; Goh, Min Hao; Yeong, Wai Yee; Naing, May Win

2018-02-27

Native tissues and/or organs possess complex hierarchical porous structures that confer highly-specific cellular functions. Despite advances in fabrication processes, it is still very challenging to emulate the hierarchical porous collagen architecture found in most native tissues. Hence, the ability to recreate such hierarchical porous structures would result in biomimetic tissue-engineered constructs. Here, a single-step drop-on-demand (DOD) bioprinting strategy is proposed to fabricate hierarchical porous collagen-based hydrogels. Printable macromolecule-based bio-inks (polyvinylpyrrolidone, PVP) have been developed and printed in a DOD manner to manipulate the porosity within the multi-layered collagen-based hydrogels by altering the collagen fibrillogenesis process. The experimental results have indicated that hierarchical porous collagen structures could be achieved by controlling the number of macromolecule-based bio-ink droplets printed on each printed collagen layer. This facile single-step bioprinting process could be useful for the structural design of collagen-based hydrogels for various tissue engineering applications.

Amorphous surface layer versus transient amorphous precursor phase in bone - A case study investigated by solid-state NMR spectroscopy.

PubMed

Von Euw, Stanislas; Ajili, Widad; Chan-Chang, Tsou-Hsi-Camille; Delices, Annette; Laurent, Guillaume; Babonneau, Florence; Nassif, Nadine; Azaïs, Thierry

2017-09-01

The presence of an amorphous surface layer that coats a crystalline core has been proposed for many biominerals, including bone mineral. In parallel, transient amorphous precursor phases have been proposed in various biomineralization processes, including bone biomineralization. Here we propose a methodology to investigate the origin of these amorphous environments taking the bone tissue as a key example. This study relies on the investigation of a bone tissue sample and its comparison with synthetic calcium phosphate samples, including a stoichiometric apatite, an amorphous calcium phosphate sample, and two different biomimetic apatites. To reveal if the amorphous environments in bone originate from an amorphous surface layer or a transient amorphous precursor phase, a combined solid-state nuclear magnetic resonance (NMR) experiment has been used. The latter consists of a double cross polarization 1 H→ 31 P→ 1 H pulse sequence followed by a 1 H magnetization exchange pulse sequence. The presence of an amorphous surface layer has been investigated through the study of the biomimetic apatites; while the presence of a transient amorphous precursor phase in the form of amorphous calcium phosphate particles has been mimicked with the help of a physical mixture of stoichiometric apatite and amorphous calcium phosphate. The NMR results show that the amorphous and the crystalline environments detected in our bone tissue sample belong to the same particle. The presence of an amorphous surface layer that coats the apatitic core of bone apatite particles has been unambiguously confirmed, and it is certain that this amorphous surface layer has strong implication on bone tissue biogenesis and regeneration. Questions still persist on the structural organization of bone and biomimetic apatites. The existing model proposes a core/shell structure, with an amorphous surface layer coating a crystalline bulk. The accuracy of this model is still debated because amorphous calcium phosphate (ACP) environments could also arise from a transient amorphous precursor phase of apatite. Here, we provide an NMR spectroscopy methodology to reveal the origin of these ACP environments in bone mineral or in biomimetic apatite. The 1 H magnetization exchange between protons arising from amorphous and crystalline domains shows unambiguously that an ACP layer coats the apatitic crystalline core of bone et biomimetic apatite platelets. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

In vivo soft tissue differentiation by diffuse reflectance spectroscopy: preliminary results

NASA Astrophysics Data System (ADS)

Zam, Azhar; Stelzle, Florian; Tangermann-Gerk, Katja; Adler, Werner; Nkenke, Emeka; Neukam, Friedrich Wilhelm; Schmidt, Michael; Douplik, Alexandre

Remote laser surgery does not provide haptic feedback to operate layer by layer and preserve vulnerable anatomical structures like nerve tissue or blood vessels. The aim of this study is identification of soft tissue in vivo by diffuse reflectance spectroscopy to set the base for a feedback control system to enhance nerve preservation in oral and maxillofacial laser surgery. Various soft tissues can be identified by diffuse reflectance spectroscopy in vivo. The results may set the base for a feedback system to prevent nerve damage during oral and maxillofacial laser surgery.

Human cartilage tissue fabrication using three-dimensional inkjet printing technology.

PubMed

Cui, Xiaofeng; Gao, Guifang; Yonezawa, Tomo; Dai, Guohao

2014-06-10

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and regenerative medicine. With digital control cells, scaffolds, and growth factors can be precisely deposited to the desired two-dimensional (2D) and three-dimensional (3D) locations rapidly. Therefore, this technology is an ideal approach to fabricate tissues mimicking their native anatomic structures. In order to engineer cartilage with native zonal organization, extracellular matrix composition (ECM), and mechanical properties, we developed a bioprinting platform using a commercial inkjet printer with simultaneous photopolymerization capable for 3D cartilage tissue engineering. Human chondrocytes suspended in poly(ethylene glycol) diacrylate (PEGDA) were printed for 3D neocartilage construction via layer-by-layer assembly. The printed cells were fixed at their original deposited positions, supported by the surrounding scaffold in simultaneous photopolymerization. The mechanical properties of the printed tissue were similar to the native cartilage. Compared to conventional tissue fabrication, which requires longer UV exposure, the viability of the printed cells with simultaneous photopolymerization was significantly higher. Printed neocartilage demonstrated excellent glycosaminoglycan (GAG) and collagen type II production, which was consistent with gene expression. Therefore, this platform is ideal for accurate cell distribution and arrangement for anatomic tissue engineering.

Basic Components of Vascular Connective Tissue and Extracellular Matrix.

PubMed

Halper, Jaroslava

2018-01-01

Though the composition of the three layers constituting the blood vessel wall varies among the different types of blood vessels, and some layers may even be missing in capillaries, certain basic components, and properties are shared by all blood vessels, though each histologically distinct layer contains a unique complement of extracellular components, growth factors and cytokines, and cell types as well. The structure and composition of vessel layers informs and is informed by the function of the particular blood vessel. The adaptation of the composition and the resulting function of the extracellular matrix (ECM) to changes in circulation/blood flow and a variety of other extravascular stimuli can be characterized as remodeling spearheaded by vascular cells. There is a surprising amount of cell traffic among the three layers. It starts with endothelial cell mediated transmigration of inflammatory cells from the bloodstream into the subendothelium, and then into tissue adjoining the blood vessel. Smooth muscle cells and a variety of adventitial cells reside in tunica media and tunica externa, respectively. The latter cells are a mixture of progenitor/stem cells, fibroblasts, myofibroblasts, pericytes, macrophages, and dendritic cells and respond to endothelial injury by transdifferentiation as they travel into the two inner layers, intima and media for corrective mission in the ECM composition. This chapter addresses the role of various vascular cell types and ECM components synthesized by them in maintenance of normal structure and in their contribution to major pathological processes, such as atherosclerosis, organ fibrosis, and diabetic retinopathy. © 2018 Elsevier Inc. All rights reserved.

Platform technology for scalable assembly of instantaneously functional mosaic tissues

PubMed Central

Zhang, Boyang; Montgomery, Miles; Davenport-Huyer, Locke; Korolj, Anastasia; Radisic, Milica

2015-01-01

Engineering mature tissues requires a guided assembly of cells into organized three-dimensional (3D) structures with multiple cell types. Guidance is usually achieved by microtopographical scaffold cues or by cell-gel compaction. The assembly of individual units into functional 3D tissues is often time-consuming, relying on cell ingrowth and matrix remodeling, whereas disassembly requires an invasive method that includes either matrix dissolution or mechanical cutting. We invented Tissue-Velcro, a bio-scaffold with a microfabricated hook and loop system. The assembly of Tissue-Velcro preserved the guided cell alignment realized by the topographical features in the 2D scaffold mesh and allowed for the instant establishment of coculture conditions by spatially defined stacking of cardiac cell layers or through endothelial cell coating. The assembled cardiac 3D tissue constructs were immediately functional as measured by their ability to contract in response to electrical field stimulation. Facile, on-demand tissue disassembly was demonstrated while preserving the structure, physical integrity, and beating function of individual layers. PMID:26601234

Dehomogenized Elastic Properties of Heterogeneous Layered Materials in AFM Indentation Experiments.

PubMed

Lee, Jia-Jye; Rao, Satish; Kaushik, Gaurav; Azeloglu, Evren U; Costa, Kevin D

2018-06-05

Atomic force microscopy (AFM) is used to study mechanical properties of biological materials at submicron length scales. However, such samples are often structurally heterogeneous even at the local level, with different regions having distinct mechanical properties. Physical or chemical disruption can isolate individual structural elements but may alter the properties being measured. Therefore, to determine the micromechanical properties of intact heterogeneous multilayered samples indented by AFM, we propose the Hybrid Eshelby Decomposition (HED) analysis, which combines a modified homogenization theory and finite element modeling to extract layer-specific elastic moduli of composite structures from single indentations, utilizing knowledge of the component distribution to achieve solution uniqueness. Using finite element model-simulated indentation of layered samples with micron-scale thickness dimensions, biologically relevant elastic properties for incompressible soft tissues, and layer-specific heterogeneity of an order of magnitude or less, HED analysis recovered the prescribed modulus values typically within 10% error. Experimental validation using bilayer spin-coated polydimethylsiloxane samples also yielded self-consistent layer-specific modulus values whether arranged as stiff layer on soft substrate or soft layer on stiff substrate. We further examined a biophysical application by characterizing layer-specific microelastic properties of full-thickness mouse aortic wall tissue, demonstrating that the HED-extracted modulus of the tunica media was more than fivefold stiffer than the intima and not significantly different from direct indentation of exposed media tissue. Our results show that the elastic properties of surface and subsurface layers of microscale synthetic and biological samples can be simultaneously extracted from the composite material response to AFM indentation. HED analysis offers a robust approach to studying regional micromechanics of heterogeneous multilayered samples without destructively separating individual components before testing. Copyright © 2018 Biophysical Society. Published by Elsevier Inc. All rights reserved.

The influence of elevated CO2 on non-structural carbohydrate distribution and fructan accumulation in wheat canopies

NASA Technical Reports Server (NTRS)

Smart, D. R.; Chatterton, N. J.; Bugbee, B.

1994-01-01

We grew 2.4 m2 wheat canopies in a large growth chamber under high photosynthetic photon flux (1000 micromoles m-2 s-1) and using two CO2 concentrations, 360 and 1200 micromoles mol-1. Photosynthetically active radiation (400-700 nm) was attenuated slightly faster through canopies grown in 360 micromoles mol-1 than through canopies grown in 1200 micromoles mol-1, even though high-CO2 canopies attained larger leaf area indices. Tissue fractions were sampled from each 5-cm layer of the canopies. Leaf tissue sampled from the tops of canopies grown in 1200 micromoles mol-1 accumulated significantly more total non-structural carbohydrate, starch, fructan, sucrose, and glucose (p < 0.05) than for canopies grown in 360 micromoles mol-1. Non-structural carbohydrate did not significantly increase in the lower canopy layers of the elevated CO2 treatment. Elevated CO2 induced fructan synthesis in all leaf tissue fractions, but fructan formation was greatest in the uppermost leaf area. A moderate temperature reduction of 10 degrees C over 5 d increased starch, fructan and glucose levels in canopies grown in 1200 micromoles mol-1, but concentrations of sucrose and fructose decreased slightly or remained unchanged. Those results may correspond with the use of fructosyl-residues and release of glucose when sucrose is consumed in fructan synthesis.

Two-layer tissue engineered urethra using oral epithelial and muscle derived cells.

PubMed

Mikami, Hiroshi; Kuwahara, Go; Nakamura, Nobuyuki; Yamato, Masayuki; Tanaka, Masatoshi; Kodama, Shohta

2012-05-01

We fabricated novel tissue engineered urethral grafts using autologously harvested oral cells. We report their viability in a canine model. Oral tissues were harvested by punch biopsy and divided into mucosal and muscle sections. Epithelial cells from mucosal sections were cultured as epithelial cell sheets. Simultaneously muscle derived cells were seeded on collagen mesh matrices to form muscle cell sheets. At 2 weeks the sheets were joined and tubularized to form 2-layer tissue engineered urethras, which were autologously grafted to surgically induced urethral defects in 10 dogs in the experimental group. Tissue engineered grafts were not applied to the induced urethral defect in control dogs. The dogs were followed 12 weeks postoperatively. Urethrogram and histological examination were done to evaluate the grafting outcome. We successfully fabricated 2-layer tissue engineered urethras in vitro and transplanted them in dogs in the experimental group. The 12-week complication-free rate was significantly higher in the experimental group than in controls. Urethrogram confirmed urethral patency without stricture in the complication-free group at 12 weeks. Histologically urethras in the transplant group showed a stratified epithelial layer overlying well differentiated submucosa. In contrast, urethras in controls showed severe fibrosis without epithelial layer formation. Two-layer tissue engineered urethras were engineered using cells harvested by minimally invasive oral punch biopsy. Results suggest that this technique can encourage regeneration of a functional urethra. Copyright © 2012 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

A multistage selective weighting method for improved microwave breast tomography.

PubMed

Shahzad, Atif; O'Halloran, Martin; Jones, Edward; Glavin, Martin

2016-12-01

Microwave tomography has shown potential to successfully reconstruct the dielectric properties of the human breast, thereby providing an alternative to other imaging modalities used in breast imaging applications. Considering the costly forward solution and complex iterative algorithms, computational complexity becomes a major bottleneck in practical applications of microwave tomography. In addition, the natural tendency of microwave inversion algorithms to reward high contrast breast tissue boundaries, such as the skin-adipose interface, usually leads to a very slow reconstruction of the internal tissue structure of human breast. This paper presents a multistage selective weighting method to improve the reconstruction quality of breast dielectric properties and minimize the computational cost of microwave breast tomography. In the proposed two stage approach, the skin layer is approximated using scaled microwave measurements in the first pass of the inversion algorithm; a numerical skin model is then constructed based on the estimated skin layer and the assumed dielectric properties of the skin tissue. In the second stage of the algorithm, the skin model is used as a priori information to reconstruct the internal tissue structure of the breast using a set of temporal scaling functions. The proposed method is evaluated on anatomically accurate MRI-derived breast phantoms and a comparison with the standard single-stage technique is presented. Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Multimodal, 3D pathology-mimicking bladder phantom for evaluation of cystoscopic technologies (Conference Presentation)

NASA Astrophysics Data System (ADS)

Smith, Gennifer T.; Lurie, Kristen L.; Zlatev, Dimitar V.; Liao, Joseph C.; Ellerbee, Audrey K.

2016-02-01

Optical coherence tomography (OCT) and blue light cystoscopy (BLC) have shown significant potential as complementary technologies to traditional white light cystoscopy (WLC) for early bladder cancer detection. Three-dimensional (3D) organ-mimicking phantoms provide realistic imaging environments for testing new technology designs, the diagnostic potential of systems, and novel image processing algorithms prior to validation in real tissue. Importantly, the phantom should mimic features of healthy and diseased tissue as they appear under WLC, BLC, and OCT, which are sensitive to tissue color and structure, fluorescent contrast, and optical scattering of subsurface layers, respectively. We present a phantom posing the hollow shape of the bladder and fabricated using a combination of 3D-printing and spray-coating with Dragon Skin (DS) (Smooth-On Inc.), a highly elastic polymer to mimic the layered structure of the bladder. Optical scattering of DS was tuned by addition of titanium dioxide, resulting in scattering coefficients sufficient to cover the human bladder range (0.49 to 2.0 mm^-1). Mucosal vasculature and tissue coloration were mimicked with elastic cord and red dye, respectively. Urethral access was provided through a small hole excised from the base of the phantom. Inserted features of bladder pathology included altered tissue color (WLC), fluorescence emission (BLC), and variations in layered structure (OCT). The phantom surface and underlying material were assessed on the basis of elasticity, optical scattering, layer thicknesses, and qualitative image appearance. WLC, BLC, and OCT images of normal and cancerous features in the phantom qualitatively matched corresponding images from human bladders.

Reference Models for Multi-Layer Tissue Structures

DTIC Science & Technology

2016-09-01

simulation,  finite   element  analysis 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF PAGES 19a. NAME OF RESPONSIBLE PERSON USAMRMC...Physiologically realistic, fully specimen-specific, nonlinear reference models. Tasks. Finite element analysis of non-linear mechanics of cadaver...models. Tasks. Finite element analysis of non-linear mechanics of multi-layer tissue regions of human subjects. Deliverables. Partially subject- and

Microfabricated 3D Scaffolds for Tissue Engineering Applications

DTIC Science & Technology

2005-01-01

coated layers as sacrificial material for subsequent SU-8 layers (Figure 5b). (a) (b) Figure 5. Four-level SU-8 structure realized with a single...connective tissue progenitor cells on micro-textured polydimethylsiloxane surfaces. Journal of Biomedical Materials Research 2002; 62:499-506. 7. Ratner BD...Applications DISTRIBUTION: Approved for public release, distribution unlimited This paper is part of the following report: TITLE: Materials Research Society

Knitting for heart valve tissue engineering

PubMed Central

Ayad, Nadia; Wojciechowska, Dorota; Zielińska, Dorota; Struszczyk, Marcin H.; Latif, Najma; Yacoub, Magdi

Knitting is a versatile technology which offers a large portfolio of products and solutions of interest in heart valve (HV) tissue engineering (TE). One of the main advantages of knitting is its ability to construct complex shapes and structures by precisely assembling the yarns in the desired position. With this in mind, knitting could be employed to construct a HV scaffold that closely resembles the authentic valve. This has the potential to reproduce the anisotropic structure that is characteristic of the heart valve with the yarns, in particular the 3-layered architecture of the leaflets. These yarns can provide oriented growth of cells lengthwise and consequently enable the deposition of extracellular matrix (ECM) proteins in an oriented manner. This technique, therefore, has a potential to provide a functional knitted scaffold, but to achieve that textile engineers need to gain a basic understanding of structural and mechanical aspects of the heart valve and in addition, tissue engineers must acquire the knowledge of tools and capacities that are essential in knitting technology. The aim of this review is to provide a platform to consolidate these two fields as well as to enable an efficient communication and cooperation among these two research areas. PMID:29043276

Interwoven Aligned Conductive Nanofiber Yarn/Hydrogel Composite Scaffolds for Engineered 3D Cardiac Anisotropy.

PubMed

Wu, Yaobin; Wang, Ling; Guo, Baolin; Ma, Peter X

2017-06-27

Mimicking the anisotropic cardiac structure and guiding 3D cellular orientation play a critical role in designing scaffolds for cardiac tissue regeneration. Significant advances have been achieved to control cellular alignment and elongation, but it remains an ongoing challenge for engineering 3D cardiac anisotropy using these approaches. Here, we present a 3D hybrid scaffold based on aligned conductive nanofiber yarns network (NFYs-NET, composition: polycaprolactone, silk fibroin, and carbon nanotubes) within a hydrogel shell for mimicking the native cardiac tissue structure, and further demonstrate their great potential for engineering 3D cardiac anisotropy for cardiac tissue engineering. The NFYs-NET structures are shown to control cellular orientation and enhance cardiomyocytes (CMs) maturation. 3D hybrid scaffolds were then fabricated by encapsulating NFYs-NET layers within hydrogel shell, and these 3D scaffolds performed the ability to promote aligned and elongated CMs maturation on each layer and individually control cellular orientation on different layers in a 3D environment. Furthermore, endothelialized myocardium was constructed by using this hybrid strategy via the coculture of CMs on NFYs-NET layer and endothelial cells within hydrogel shell. Therefore, these 3D hybrid scaffolds, containing NFYs-NET layer inducing cellular orientation, maturation, and anisotropy and hydrogel shell providing a suitable 3D environment for endothelialization, has great potential in engineering 3D cardiac anisotropy.

A review of rapid prototyping techniques for tissue engineering purposes.

PubMed

Peltola, Sanna M; Melchels, Ferry P W; Grijpma, Dirk W; Kellomäki, Minna

2008-01-01

Rapid prototyping (RP) is a common name for several techniques, which read in data from computer-aided design (CAD) drawings and manufacture automatically three-dimensional objects layer-by-layer according to the virtual design. The utilization of RP in tissue engineering enables the production of three-dimensional scaffolds with complex geometries and very fine structures. Adding micro- and nanometer details into the scaffolds improves the mechanical properties of the scaffold and ensures better cell adhesion to the scaffold surface. Thus, tissue engineering constructs can be customized according to the data acquired from the medical scans to match the each patient's individual needs. In addition RP enables the control of the scaffold porosity making it possible to fabricate applications with desired structural integrity. Unfortunately, every RP process has its own unique disadvantages in building tissue engineering scaffolds. Hence, the future research should be focused on the development of RP machines designed specifically for fabrication of tissue engineering scaffolds, although RP methods already can serve as a link between tissue and engineering.

Contribution of collagen and elastin fibers to the mechanical behavior of an abdominal connective tissue.

PubMed

Levillain, A; Orhant, M; Turquier, F; Hoc, T

2016-08-01

The linea alba is a complex structure commonly involved in hernia formation. Knowledge of its mechanical behavior is essential to design suitable meshes and reduce the risk of recurrence. The aim of this study was to investigate the relationships between the mechanical properties of the linea alba and the organization of collagen and elastin fibers. For that purpose, longitudinal and transversal samples were removed from four porcine and three human linea alba, to perform tensile tests under a biphotonic confocal microscope, in each direction. Microscopic observation revealed a tissue composed of two layers, made of transversal collagen fibers in the dorsal side and oblique collagen fibers in the ventral side. This particular architecture led to an anisotropic mechanical behavior, with higher stress in the transversal direction. During loading, oblique fibers of the ventral layer reoriented toward the tensile axis in both directions, while fibers of the dorsal layer remained in the transversal direction. This rotation of oblique fibers progressively increased the stiffness of the tissue and induced a non-linear stress-stretch relation. Elastin fibers formed a layer covering the collagen fibers and followed their movement, suggesting that they ensure their elastic recoil. All of these results demonstrated the strong relationships between the microstructure and the mechanical behavior of the linea alba. Copyright © 2016 Elsevier Ltd. All rights reserved.

The clinical significance of the atrial subendocardial smooth muscle layer and cardiac myofibroblasts in human atrial tissue with valvular atrial fibrillation.

PubMed

Park, Jae Hyung; Pak, Hui-Nam; Lee, Sak; Park, Han Ki; Seo, Jeong-Wook; Chang, Byung-Chul

2013-01-01

The existence of myofibroblasts (MFBs) and the role of subendocardial smooth muscle (SSM) layer of human atrial tissue in atrial fibrillation (AF) have not yet been elucidated. We hypothesized that the SSM layer and MFB play some roles in atrial structural remodeling and maintenance of valvular AF in patients who undergo cardiac surgery. We analyzed immunohistochemical staining of left atrial (LA) appendage tissues taken from 17 patients with AF and 15 patients remaining in sinus rhythm (SR) who underwent cardiac surgery (male 50.0%, 54.1 ± 14.2 years old, valve surgery 87.5%). SSM was quantified by α-smooth muscle actin (α-SMA) stain excluding vascular structure. MFB was defined as α-SMA+ cells with disorganized Connexin 43-positive gap junctions in Sirius red-positive fibrotic area. The SSM layer of atrium was significantly thicker in patients with AF than in those with SR (P=.0091). Patients with SSM layer ≥ 14 μm had a larger LA size (P=.0006) and greater fibrotic area (P=.0094) than those patients whose SSM layer <14 μm. MFBs were found in 7 of 17 (41.2%) patients with AF and 2 of 15 (13.3%) in SR group (P=.0456) in SSM area, colocalized with Periodic Acid-Schiff (PAS) stain-positive glycogen storage cells (95.5%). SSM layer was closely related to the existence of AF, degrees of atrial remodeling, and fibrosis in patients who underwent open heart surgery. We found that MFB does exist in SSM layer of human atrial tissue co-localized with PAS-positive cells. Crown Copyright © 2013. Published by Elsevier Inc. All rights reserved.

Autofluorescence spectroscopy for multimodal tissues characterization in colitis-associated cancer murine model

NASA Astrophysics Data System (ADS)

Dorez, Hugo; Sablong, Raphaël.; Canaple, Laurence; Saint-Jalmes, Hervé; Gaillard, Sophie; Moussata, Driffa; Beuf, Olivier

2015-07-01

The purpose of this research project is to assess mice colon wall, using three optical modalities (conventional endoscopy, confocal endomicroscopy and optical spectroscopy) and endoluminal MRI. The study is done in the context of inflammatory bowel disease and colorectal cancer that represent 13% of new cases of cancer, every year in western countries. An optical spectroscopic bench (autofluorescence and reflectance) was developed with a flexible fiber probe. This latter has been combined with a mini multi-purpose rigid endoscope and a confocal endomicroscope. The optical modalities were first used in vivo on SWISS mice. Then, a specific optical a phantom (containing two layers of distinct fluorophores) was developed in order to evaluate our two-channel spectroscopic probe as a basic depth-sensitive measurement tool. The preliminary results show the feasibility to combine such modalities in the same in vivo protocol. Conventional endoscopy is useful to depict inflammation along colon wall. Confocal endomicroscopy provides high-contrasted images of microvascularization. Measured optical spectra both depend on biochemical tissue content and layered structure of the medium. The light collected from one channel is not similar to the other, in terms of intensity and spectroscopic profile as the interaction with the medium observed volume is different. A comparative analysis of the spectra based on our in vitro model exhibits a strong correlation between simple index extracted from spectral data and two main phantom characteristics (fluorophore concentrations and superficial layer thickness). This work suggests that this technique could contribute to assess tissues alterations through autofluorescence spectroscopic measurement under endoscopy.

Multiaxial mechanical response and constitutive modeling of esophageal tissues: Impact on esophageal tissue engineering.

PubMed

Sommer, Gerhard; Schriefl, Andreas; Zeindlinger, Georg; Katzensteiner, Andreas; Ainödhofer, Herwig; Saxena, Amulya; Holzapfel, Gerhard A

2013-12-01

Congenital defects of the esophagus are relatively frequent, with 1 out of 2500 babies suffering from such a defect. A new method of treatment by implanting tissue engineered esophagi into newborns is currently being developed and tested using ovine esophagi. For the reconstruction of the biological function of native tissues with engineered esophagi, their cellular structure as well as their mechanical properties must be considered. Since very limited mechanical and structural data for the esophagus are available, the aim of this study was to investigate the multiaxial mechanical behavior of the ovine esophagus and the underlying microstructure. Therefore, uniaxial tensile, biaxial tensile and extension-inflation tests on esophagi were performed. The underlying microstructure was examined in stained histological sections through standard optical microscopy techniques. Moreover, the uniaxial ultimate tensile strength and residual deformations of the tissue were determined. Both the mucosa-submucosa and the muscle layers showed nonlinear and anisotropic mechanical behavior during uniaxial, biaxial and inflation testing. Cyclical inflation of the intact esophageal tube caused marked softening of the passive esophagi in the circumferential direction. The rupture strength of the mucosa-submucosa layer was much higher than that of the muscle layer. Overall, the ovine esophagus showed a heterogeneous and anisotropic behavior with different mechanical properties for the individual layers. The intact and layer-specific multiaxial properties were characterized using a well-known three-dimensional microstructurally based strain-energy function. This novel and complete set of data serves the basis for a better understanding of tissue remodeling in diseased esophagi and can be used to perform computer simulations of surgical interventions or medical-device applications. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Shock Wave Propagation in Functionally Graded Mineralized Tissue

NASA Astrophysics Data System (ADS)

Nelms, Matthew; Hodo, Wayne; Livi, Ken; Browning, Alyssa; Crawford, Bryan; Rajendran, A. M.

2017-06-01

In this investigation, the effects of shock wave propagation in bone-like biomineralized tissue was investigated. The Alligator gar (Atractosteus spatula) exoskeleton is comprised of many disparate scales that provide a biological analog for potential design of flexible protective material systems. The gar scale is identified as a two-phase, (1) hydroxyapatite mineral and (2) collagen protein, biological composite with two distinct layers where a stiff, ceramic-like ganoine overlays a soft, highly ductile ganoid bone. Previous experimentations has shown significant softening under compressive loading and an asymmetrical stress-strain response for analogous mineralized tissues. The structural features, porosity, and elastic modulus were determined from high-resolution scanning electron microscopy, 3D micro-tomography, and dynamic nanoindentation experiments to develop an idealized computational model for FE simulations. The numerical analysis employed Gurson's yield criterion to determine the influence of porosity and pressure on material strength. Functional gradation of elastic moduli and certain structural features, such as the sawtooth interface, are explicitly modeled to study the plate impact shock profile for a full 3-D analysis using ABAQUS finite element software.

Polyimide-based intracortical neural implant with improved structural stiffness

NASA Astrophysics Data System (ADS)

Lee, Kee-Keun; He, Jiping; Singh, Amarjit; Massia, Stephen; Ehteshami, Gholamreza; Kim, Bruce; Raupp, Gregory

2004-01-01

A novel structure for chronically implantable cortical electrodes using polyimide bio-polymer was devised, which provides both flexibility for micro-motion compliance between brain tissues and the skull and at the brain/implant interface and stiffness for better surgical handling. A 5-10 µm thick silicon backbone layer was attached to the tip of the electrode to enhance the structural stiffness. This stiff segment was then followed by a 1 mm flexible segment without a silicon backbone layer. The fabricated implants have tri-shanks with five recording sites (20 µm × 20 µm) and two vias of 40 µm × 40 µm on each shank. In vitro cytotoxicity tests of prototype implants revealed no adverse toxic effects on cells. Bench test impedance values were assessed, resulting in an average impedance value of ~2 MOmega at 1 KHz. For a 5 µm thick silicon backbone electrode, the stiffness of polyimide-based electrodes was increased ten times over that of electrodes without the silicon backbone layer. Furthermore, polyimide-based electrodes with 5 µm and 10 µm thick silicon backbone layer penetrated pia of rat brain without buckling that has been observed in implants without silicon reinforcement.

Measurements of surface layer of the articular cartilage using microscopic techniques

NASA Astrophysics Data System (ADS)

Ryniewicz, A. M.; Ryniewicz, A.; Ryniewicz, W.; Gaska, A.

2010-07-01

The articular cartilage is the structure that directly cooperates tribologically in biobearing. It belongs to the connective tissues and in the joints it assumes two basic forms: hyaline cartilage that builds joint surfaces and fibrocartilage which may create joint surfaces. From this fibrocartilage are built semilunar cartilage and joint disc are built as well. The research of articular cartilage have been done in macro, micro and nano scale. In all these measurement areas characteristic features occur which can identify biobearing tribology. The aim of the research was the identification of surface layer of articular cartilage by means of scanning electron microscopy (SEM) and atom force microscopy (AFM) and the analysis of topography of these layers. The material used in the research of surface layer was the animal articular cartilage: hyaline cartilage and fibrocartilage.

Development of Multiple-Frequency Ultrasonic Imaging System Using Multiple Resonance Piezoelectric Transducer

NASA Astrophysics Data System (ADS)

Akiyama, Iwaki; Yoshizumi, Natsuki; Saito, Shigemi; Wada, Yuji; Koyama, Daisuke; Nakamura, Kentaro

2012-07-01

The authors have developed a multiple frequency imaging system using a multiple resonance transducer (MRT) consisting of 1-3 composite materials with a low mechanical quality factor Q bonded together. The MRT has a structure consisting of thin and thick piezoelectric plates, two matching layers, and a backing layer. This makes it possible to obtain B-mode images of satisfactory resolution using ultrasonic pulses owing to their short duration. In this paper, the vibration property of the MRT derived through equivalent-circuit analysis is first shown. By utilizing the result, an MRT capable of transmitting ultrasonic pulses for generation of the images of biological tissues with satisfactory resolution is designed and prototyped. Setting the prototype transducer in the mechanical sector probe of commercial ultrasonic diagnosis equipment, the speckle reduction effect is demonstrated using images of various phantoms to mimic biological tissues and a human thyroid.

Raman microscopy of freeze-dried mouse eyeball-slice in conjunction with the "in vivo cryotechnique".

PubMed

Terada, Nobuo; Ohno, Nobuhiko; Saitoh, Sei; Fujii, Yasuhisa; Ohguro, Hiroshi; Ohno, Shinichi

2007-07-01

The wavelength of Raman-scattered light depends on the molecular composition of the substance. This is the first attempt to acquire Raman spectra of a mouse eyeball removed from a living mouse, in which the eyeball was preserved using the "in vivo cryotechnique" followed by freeze-drying. Eyeballs were cryofixed using a rapid freezing cryotechnique, and then sliced in the cryostat machine. The slices were sandwiched between glass slides, freeze-dried, and analyzed with confocal Raman microscopy. Important areas including various eyeball tissue layers were selected using bright-field microscopy, and then the Raman spectra were obtained at 240 locations. Four typical patterns of Raman spectra were electronically mapped on the specimen images obtained by the bright-field microscopy. Tissue organization was confirmed by embedding the same eyeball slice used for Raman spectra into epoxy resin and the thick sections were prepared with the inverted capsule method. Each Raman spectral pattern represents a different histological layer in the eyeball which was mapped by comparing the images of toluidine blue staining and Raman mapping with different colors. In the choroid and pigment cell layer, the Raman spectrum had two peaks, corresponding to melanin. Some of the peaks of the Raman spectra obtained from the blood vessels in sclera and the photoreceptor layer were similar to those obtained from the purified hemoglobin and rhodopsin proteins, respectively. Our experimental protocol can distinguish different tissue components with Raman microscopy; therefore, this method can be very useful for examining the distribution of a biological structures and/or chemical components in rapidly frozen freeze-dried tissue.

Three-dimensional fuse deposition modeling of tissue-simulating phantom for biomedical optical imaging

NASA Astrophysics Data System (ADS)

Dong, Erbao; Zhao, Zuhua; Wang, Minjie; Xie, Yanjun; Li, Shidi; Shao, Pengfei; Cheng, Liuquan; Xu, Ronald X.

2015-12-01

Biomedical optical devices are widely used for clinical detection of various tissue anomalies. However, optical measurements have limited accuracy and traceability, partially owing to the lack of effective calibration methods that simulate the actual tissue conditions. To facilitate standardized calibration and performance evaluation of medical optical devices, we develop a three-dimensional fuse deposition modeling (FDM) technique for freeform fabrication of tissue-simulating phantoms. The FDM system uses transparent gel wax as the base material, titanium dioxide (TiO2) powder as the scattering ingredient, and graphite powder as the absorption ingredient. The ingredients are preheated, mixed, and deposited at the designated ratios layer-by-layer to simulate tissue structural and optical heterogeneities. By printing the sections of human brain model based on magnetic resonance images, we demonstrate the capability for simulating tissue structural heterogeneities. By measuring optical properties of multilayered phantoms and comparing with numerical simulation, we demonstrate the feasibility for simulating tissue optical properties. By creating a rat head phantom with embedded vasculature, we demonstrate the potential for mimicking physiologic processes of a living system.

Laser autofluorescence polarimetry of optically anisotropic structures of biological tissues in cancer diagnostics

NASA Astrophysics Data System (ADS)

Ushenko, Yu. A.

2015-06-01

The results of a new physical study of polarization manifestations of laser autofluorescence of optically anisotropic structures in human female reproductive tissues are presented. A Mueller-matrix model of describing the complex anisotropy (linear and circular birefringence, linear and circular dichroism) of such biological layers is proposed. Interrelations between mechanisms of optical anisotropy and polarization manifestations of laser autofluorescence of histological layers of the uterine cervix tissue in different spectral regions are determined. Magnitudes and variation ranges of statistical moments from the first to the fourth order describing the distributions of azimuthally stable elements of Mueller matrices of autofluorescence in human female reproductive tissues in different physiological states are found. The informative value of the proposed method is determined and the differentiation of histological biopsy sections of benign (dysplasia) and malignant (adenocarcinoma) uterine cervix tumors is implemented for the first time.

Modeling and Characterization of Capacitive Elements With Tissue as Dielectric Material for Wireless Powering of Neural Implants.

PubMed

Erfani, Reza; Marefat, Fatemeh; Sodagar, Amir M; Mohseni, Pedram

2018-05-01

This paper reports on the modeling and characterization of capacitive elements with tissue as the dielectric material, representing the core building block of a capacitive link for wireless power transfer to neural implants. Each capacitive element consists of two parallel plates that are aligned around the tissue layer and incorporate a grounded, guarded, capacitive pad to mitigate the adverse effect of stray capacitances and shield the plates from external interfering electric fields. The plates are also coated with a biocompatible, insulating, coating layer on the inner side of each plate in contact with the tissue. A comprehensive circuit model is presented that accounts for the effect of the coating layers and is validated by measurements of the equivalent capacitance as well as impedance magnitude/phase of the parallel plates over a wide frequency range of 1 kHz-10 MHz. Using insulating coating layers of Parylene-C at a thickness of and Parylene-N at a thickness of deposited on two sets of parallel plates with different sizes and shapes of the guarded pad, our modeling and characterization results accurately capture the effect of the thickness and electrical properties of the coating layers on the behavior of the capacitive elements over frequency and with different tissues.

Reinforcement of mono- and bi-layer poly(ethylene glycol) hydrogels with a fibrous collagen scaffold

PubMed Central

Kinneberg, K. R. C.; Nelson, A.; Stender, M.; Aziz, A. H.; Mozdzen, L. C.; Harley, B. A. C.; Bryant, S. J.; Ferguson, V. L.

2015-01-01

Biomaterial-based tissue engineering strategies hold great promise for osteochondral tissue repair. Yet significant challenges remain in joining highly dissimilar materials to achieve a biomimetic, mechanically robust design for repairing interfaces between soft tissue and bone. This study sought to improve interfacial properties and function in a bilayer, multi-phase hydrogel interpenetrated with a fibrous collagen scaffold. ‘Soft’ 10% (w/w) and ‘stiff’ 30% (w/w) PEGDM was formed into mono- or bilayer hydrogels possessing a sharp diffusional interface. Hydrogels were evaluated as single- (hydrogel only) or multi-phase (hydrogel+fibrous scaffold penetrating throughout the stiff layer and extending >500μm into the soft layer). Including a fibrous scaffold into both soft and stiff single-phase hydrogels significantly increased tangent modulus and toughness and decreased lateral expansion under compressive loading. In multi-phase hydrogels, finite element simulations predict substantially reduced stress and strain gradients across the soft—stiff hydrogel interface. When combining two low moduli constituent material, composites theory poorly predicts the observed, large modulus increases. These results suggest material structure associated with the fibrous scaffold penetrating within the PEG hydrogel as the major contributor to improved properties and function – the hydrogel bore compressive loads and the 3D fibrous scaffold was loaded in tension thus resisting lateral expansion. PMID:26001970

Altered vocal fold kinematics in synthetic self-oscillating models that employ adipose tissue as a lateral boundary condition.

NASA Astrophysics Data System (ADS)

Saidi, Hiba; Erath, Byron D.

2015-11-01

The vocal folds play a major role in human communication by initiating voiced sound production. During voiced speech, the vocal folds are set into sustained vibrations. Synthetic self-oscillating vocal fold models are regularly employed to gain insight into flow-structure interactions governing the phonation process. Commonly, a fixed boundary condition is applied to the lateral, anterior, and posterior sides of the synthetic vocal fold models. However, physiological observations reveal the presence of adipose tissue on the lateral surface between the thyroid cartilage and the vocal folds. The goal of this study is to investigate the influence of including this substrate layer of adipose tissue on the dynamics of phonation. For a more realistic representation of the human vocal folds, synthetic multi-layer vocal fold models have been fabricated and tested while including a soft lateral layer representative of adipose tissue. Phonation parameters have been collected and are compared to those of the standard vocal fold models. Results show that vocal fold kinematics are affected by adding the adipose tissue layer as a new boundary condition.

Raman Monte Carlo simulation for light propagation for tissue with embedded objects

NASA Astrophysics Data System (ADS)

Periyasamy, Vijitha; Jaafar, Humaira Bte; Pramanik, Manojit

2018-02-01

Monte Carlo (MC) stimulation is one of the prominent simulation technique and is rapidly becoming the model of choice to study light-tissue interaction. Monte Carlo simulation for light transport in multi-layered tissue (MCML) is adapted and modelled with different geometry by integrating embedded objects of various shapes (i.e., sphere, cylinder, cuboid and ellipsoid) into the multi-layered structure. These geometries would be useful in providing a realistic tissue structure such as modelling for lymph nodes, tumors, blood vessels, head and other simulation medium. MC simulations were performed on various geometric medium. Simulation of MCML with embedded object (MCML-EO) was improvised for propagation of the photon in the defined medium with Raman scattering. The location of Raman photon generation is recorded. Simulations were experimented on a modelled breast tissue with tumor (spherical and ellipsoidal) and blood vessels (cylindrical). Results were presented in both A-line and B-line scans for embedded objects to determine spatial location where Raman photons were generated. Studies were done for different Raman probabilities.

Enhanced nutrient transport improves the depth-dependent properties of tri-layered engineered cartilage constructs with zonal co-culture of chondrocytes and MSCs.

PubMed

Kim, Minwook; Farrell, Megan J; Steinberg, David R; Burdick, Jason A; Mauck, Robert L

2017-08-01

Biomimetic design in cartilage tissue engineering is a challenge given the complexity of the native tissue. While numerous studies have generated constructs with near-native bulk properties, recapitulating the depth-dependent features of native tissue remains a challenge. Furthermore, limitations in nutrient transport and matrix accumulation in engineered constructs hinders maturation within the central core of large constructs. To overcome these limitations, we fabricated tri-layered constructs that recapitulate the depth-dependent cellular organization and functional properties of native tissue using zonally derived chondrocytes co-cultured with MSCs. We also introduced porous hollow fibers (HFs) and HFs/cotton threads to enhance nutrient transport. Our results showed that tri-layered constructs with depth-dependent organization and properties could be fabricated. The addition of HFs or HFs/threads improved matrix accumulation in the central core region. With HF/threads, the local modulus in the deep region of tri-layered constructs nearly matched that of native tissue, though the properties in the central regions remained lower. These constructs reproduced the zonal organization and depth-dependent properties of native tissue, and demonstrate that a layer-by-layer fabrication scheme holds promise for the biomimetic repair of focal cartilage defects. Articular cartilage is a highly organized tissue driven by zonal heterogeneity of cells, extracellular matrix proteins and fibril orientations, resulting in depth-dependent mechanical properties. Therefore, the recapitulation of the functional properties of native cartilage in a tissue engineered construct requires such a biomimetic design of the morphological organization, and this has remained a challenge in cartilage tissue engineering. This study demonstrates that a layer-by-layer fabrication scheme, including co-cultures of zone-specific articular CHs and MSCs, can reproduce the depth-dependent characteristics and mechanical properties of native cartilage while minimizing the need for large numbers of chondrocytes. In addition, introduction of a porous hollow fiber (combined with a cotton thread) enhanced nutrient transport and depth-dependent properties of the tri-layered construct. Such a tri-layered construct may provide critical advantages for focal cartilage repair. These constructs hold promise for restoring native tissue structure and function, and may be beneficial in terms of zone-to-zone integration with adjacent host tissue and providing more appropriate strain transfer after implantation. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Surgical anatomy of the posterior liver surface: the retrohepatic lamina as the basis for mobilisation of the right liver.

PubMed

Macchi, Veronica; Porzionato, Andrea; Bardini, Romeo; Picardi, Edgardo Enrico Edoardo; De Caro, Raffaele

2013-10-01

During right hepatectomies, dissection of the bare area is performed to obtain mobilisation of the liver. Fifty computed tomography scans of the upper abdomen of patients were examined. Specimens of supramesocolic compartment were sampled from 10 un-embalmed cadavers. Macrosections were cut for histotopographic study. In four cadavers, in situ dissection of the posterior liver surface was performed. The hepatophrenic tissue showed a stratigraphic organisation resulting from the juxtaposition of thin layer of dense connective tissue corresponding to the inferior diaphragmatic fascia (mean thickness is 30 ± 4 μm); variable amount of fibroadipose tissue corresponding to retroperitoneal fibroadipose tissue (mean thickness is 34 ± 8 μm); two connective layers with nets of flat cells forming a fusion fascia, the retrohepatic lamina (mean thickness 24 ± 6 μm); and layer of connective tissue corresponding to the hepatic capsule. The juxta-caval portion of the retrohepatic lamina, connecting the right and left sides of the caval groove, forms the inferior vena cava ligament. During dissection, fluid injection developed a preferential plane between the two layers of the retrohepatic lamina, close to the hepatic surface, and no major or minor vessels were ever found along this plane. During right hepatectomy, to reduce the risk of dissemination of tumour cells, the dissection plane should be performed between the two layers of the retrohepatic lamina.

Ultrastructural and molecular confirmation of the development of Sarcocystis neurona tissue cysts in the central nervous system of southern sea otters (Enhydra lutris nereis).

PubMed

Miller, M A; Barr, B C; Nordhausen, R; James, E R; Magargal, S L; Murray, M; Conrad, P A; Toy-Choutka, S; Jessup, D A; Grigg, M E

2009-10-01

In 2004, three wild sea otters were diagnosed with putative Sarcocystis neurona-associated meningoencephalitis by histopathology and immunohistochemistry. Schizonts, free merozoites and tissue cysts were observed in the brains of all three infected animals. Tissue cysts walls from sea otter 1 (SO1) stained positively using anti-S. neurona polyclonal antiserum. However, positive staining does not preclude infection by closely related or cross-reactive tissue cyst-forming coccidian parasites. Two immature tissue cysts in the brain of SO1 were examined using transmission electron microscopy. Ultrastructural features included cyst walls with thin villous projections up to 1 microm long with tapered ends and a distinctive, electron-dense outer lining layer composed of linearly-arranged, semi-circular structures with a "hobnailed" surface contour. Small numbers of microtubules extended down through the villi into the underlying granular layer. Metrocytes were short and plump with an anterior apical complex, 22 sub-pellicular microtubules, numerous free ribosomes and no rhoptries. Some metrocytes appeared to be dividing, with two adjacent nuclear profiles. Collectively these ultrastructural features were compatible with developing protozoal cysts and were similar to prior descriptions of S. neurona tissue cysts. Panspecific 18S rDNA primers were utilized to identify protozoa infecting the brains of these otters and DNA amplification and additional sequencing at the ITS1 locus confirmed that all three otters were infected with S. neurona. No other Sarcocystis spp. were detected in the brains or skeletal muscles of these animals by immunohistochemistry or PCR. We believe this is the first ultrastructural and molecular confirmation of the development of S. neurona tissue cysts in the CNS of any animal.

Ultrastructural and molecular confirmation of the development of Sarcocystis neurona tissue cysts in the central nervous system of southern sea otters (Enhydra lutris nereis)

PubMed Central

Miller, M.A.; Barr, B.C.; Nordhausen, R.; James, E.R.; Magargal, S.L.; Murray, M.; Conrad, P.A.; Toy-Choutka, S.; Jessup, D.A.; Grigg, M.E.

2009-01-01

In 2004, three wild sea otters were diagnosed with putative Sarcocystis neurona-associated meningoencephalitis by histopathology and immunohistochemistry. Schizonts, free merozoites and tissue cysts were observed in the brains of all three infected animals. Tissue cysts from sea otter 1 (SO1) stained positively using anti-S. neurona polyclonal antiserum. However, positive staining does not preclude infection by closely related or cross-reactive tissue cyst-forming coccidian parasites. Two immature tissue cysts in the brain of SO1 were examined using transmission electron microscopy. Ultrastructural features included cyst walls with thin villous projections up to 1 μm long with tapered ends and a distinctive, electron-dense outer lining layer composed of linearly-arranged, semi-circular structures with a “hobnailed” surface contour. Small numbers of microtubules extended down through the villi into the underlying granular layer. Metrocytes were short and plump with an anterior apical complex, 22 subpellicular microtubules, numerous free ribosomes and no rhoptries. Some metrocytes appeared to be dividing, with two adjacent nuclear profiles. Collectively these ultrastructural features were compatible with developing protozoal cysts and were similar to prior descriptions of S. neurona tissue cysts. Panspecific 18S rDNA primers were utilized to identify protozoa infecting the brains of these otters and DNA amplification and additional sequencing at the ITS1 locus confirmed that all three otters were infected with S. neurona. No other Sarcocystis spp. were detected in the brains or skeletal muscles of these animals by immunohistochemistry or PCR. We believe this is the first ultrastructural and molecular confirmation of the development of S. neurona tissue cysts in the CNS of any animal. PMID:19527725

Two-photon excited fluorescence microscopy application for ex vivo investigation of ocular fundus samples

NASA Astrophysics Data System (ADS)

Peters, Sven; Hammer, Martin; Schweitzer, Dietrich

2011-07-01

Two-photon excited fluorescence (TPEF) imaging of ocular tissue has recently become a promising tool in ophthalmology for diagnostic and research purposes. The feasibility and the advantages of TPEF imaging, namely deeper tissue penetration and improved high-resolution imaging of microstructures, have been demonstrated lately using human ocular samples. The autofluorescence properties of endogenous fluorophores in ocular fundus tissue are well known from spectrophotometric analysis. But fluorophores, especially when it comes to fluorescence lifetime, typically display a dependence of their fluorescence properties on local environmental parameters. Hence, a more detailed investigation of ocular fundus autofluorescence ideally in vivo is of utmost interest. The aim of this study is to determine space-resolved the stationary and time-resolved fluorescence properties of endogenous fluorophores in ex vivo porcine ocular fundus samples by means of two-photon excited fluorescence spectrum and lifetime imaging microscopy (FSIM/FLIM). By our first results, we characterized the autofluorescence of individual anatomical structures of porcine retina samples excited at 760 nm. The fluorescence properties of almost all investigated retinal layers are relatively homogenous. But as previously unknown, ganglion cell bodies show a significantly shorter fluorescence lifetime compared to the adjacent mueller cells. Since all retinal layers exhibit bi-exponential autofluorescence decays, we were able to achieve a more precise characterization of fluorescence properties of endogenous fluorophores compared to a present in vivo FLIM approach by confocal scanning laser ophthalmoscope (cSLO).

Nanometer-sized extracellular matrix coating on polymer-based scaffold for tissue engineering applications.

PubMed

Uchida, Noriyuki; Sivaraman, Srikanth; Amoroso, Nicholas J; Wagner, William R; Nishiguchi, Akihiro; Matsusaki, Michiya; Akashi, Mitsuru; Nagatomi, Jiro

2016-01-01

Surface modification can play a crucial role in enhancing cell adhesion to synthetic polymer-based scaffolds in tissue engineering applications. Here, we report a novel approach for layer-by-layer (LbL) fabrication of nanometer-size fibronectin and gelatin (FN-G) layers on electrospun fibrous poly(carbonate urethane)urea (PCUU) scaffolds. Alternate immersions into the solutions of fibronectin and gelatin provided thickness-controlled FN-G nano-layers (PCUU(FN-G) ) which maintained the scaffold's 3D structure and width of fibrous bundle of PCUU as evidenced by scanning electron miscroscopy. The PCUU(FN-G) scaffold improved cell adhesion and proliferation of bladder smooth muscles (BSMCs) when compared to uncoated PCUU. The high affinity of PCUU(FN-G) for cells was further demonstrated by migration of adherent BSMCs from culture plates to the scaffold. Moreover, the culture of UROtsa cells, human urothelium-derived cell line, on PCUU(FN-G) resulted in an 11-15 μm thick multilayered cell structure with cell-to-cell contacts although many UROtsa cells died without forming cell connections on PCUU. Together these results indicate that this approach will aid in advancing the technology for engineering bladder tissues in vitro. Because FN-G nano-layers formation is based on nonspecific physical adsorption of fibronectin onto polymer and its subsequent interactions with gelatin, this technique may be applicable to other polymer-based scaffold systems for various tissue engineering/regenerative medicine applications. © 2015 Wiley Periodicals, Inc.

Analytical model of diffuse reflectance spectrum of skin tissue

NASA Astrophysics Data System (ADS)

Lisenko, S. A.; Kugeiko, M. M.; Firago, V. A.; Sobchuk, A. N.

2014-01-01

We have derived simple analytical expressions that enable highly accurate calculation of diffusely reflected light signals of skin in the spectral range from 450 to 800 nm at a distance from the region of delivery of exciting radiation. The expressions, taking into account the dependence of the detected signals on the refractive index, transport scattering coefficient, absorption coefficient and anisotropy factor of the medium, have been obtained in the approximation of a two-layer medium model (epidermis and dermis) for the same parameters of light scattering but different absorption coefficients of layers. Numerical experiments on the retrieval of the skin biophysical parameters from the diffuse reflectance spectra simulated by the Monte Carlo method show that commercially available fibre-optic spectrophotometers with a fixed distance between the radiation source and detector can reliably determine the concentration of bilirubin, oxy- and deoxyhaemoglobin in the dermis tissues and the tissue structure parameter characterising the size of its effective scatterers. We present the examples of quantitative analysis of the experimental data, confirming the correctness of estimates of biophysical parameters of skin using the obtained analytical expressions.

Depth-resolved fluorescence of human ectocervical tissue

NASA Astrophysics Data System (ADS)

Wu, Yicong; Xi, Peng; Cheung, Tak-Hong; Yim, So Fan; Yu, Mei-Yung; Qu, Jianan Y.

2005-04-01

The depth-resolved autofluorescence of normal and dysplastic human ectocervical tissue within 120um depth were investigated utilizing a portable confocal fluorescence spectroscopy with the excitations at 355nm and 457nm. From the topmost keratinizing layer of all ectocervical tissue samples, strong keratin fluorescence with the spectral characteristics similar to collagen was observed, which created serious interference in seeking the correlation between tissue fluorescence and tissue pathology. While from the underlying non-keratinizing epithelial layer, the measured NADH fluorescence induced by 355nm excitation and FAD fluorescence induced by 457nm excitation were strongly correlated to the tissue pathology. The ratios between NADH over FAD fluorescence increased statistically in the CIN epithelial relative to the normal and HPV epithelia, which indicated increased metabolic activity in precancerous tissue. This study demonstrates that the depth-resolved fluorescence spectroscopy can reveal fine structural information on epithelial tissue and potentially provide more accurate diagnostic information for determining tissue pathology.

Evaluation of the tissue reaction to a new bilayered collagen matrix in vivo and its translation to the clinic.

PubMed

Ghanaati, Shahram; Schlee, Markus; Webber, Matthew J; Willershausen, Ines; Barbeck, Mike; Balic, Ela; Görlach, Christoph; Stupp, Samuel I; Sader, Robert A; Kirkpatrick, C James

2011-02-01

This study evaluates a new collagen matrix that is designed with a bilayered structure in order to promote guided tissue regeneration and integration within the host tissue. This material induced a mild tissue reaction when assessed in a murine model and was well integrated within the host tissue, persisting in the implantation bed throughout the in vivo study. A more porous layer was rapidly infiltrated by host mesenchymal cells, while a layer designed to be a barrier allowed cell attachment and host tissue integration, but at the same time remained impermeable to invading cells for the first 30 days of the study. The tissue reaction was favorable, and unlike a typical foreign body response, did not include the presence of multinucleated giant cells, lymphocytes, or granulation tissue. In the context of translation, we show preliminary results from the clinical use of this biomaterial applied to soft tissue regeneration in the treatment of gingival tissue recession and exposed roots of human teeth. Such a condition would greatly benefit from guided tissue regeneration strategies. Our findings demonstrate that this material successfully promoted the ingrowth of gingival tissue and reversed gingival tissue recession. Of particular importance is the fact that the histological evidence from these human studies corroborates our findings in the murine model, with the barrier layer preventing unspecific tissue ingrowth, as the scaffold becomes infiltrated by mesenchymal cells from adjacent tissue into the porous layer. Also in the clinical situation no multinucleated giant cells, no granulation tissue and no evidence of a marked inflammatory response were observed. In conclusion, this bilayered matrix elicits a favorable tissue reaction, demonstrates potential as a barrier for preferential tissue ingrowth, and achieves a desirable therapeutic result when applied in humans for soft tissue regeneration.

Physical analysis on laser-induced cerebral damage

NASA Astrophysics Data System (ADS)

Luo, Xiaosen; Liu, Jiangang; Tao, Chunkan; Lan, Xiufeng; Cao, Lingyan; Pan, Weimin; Shen, Zhonghua; Lu, Jian; Ni, Xiaowu

2005-01-01

Experimental investigation on cerebral damage of adult SD rats induced by 532nm CW laser was performed. Tissue heat conductive equation was set up based on two-layered structure model. Finite difference algorithm was utilized to numerically simulate the temperature distribution in the brain tissue. Allowing for tissue response to temperature variation, free boundary model was used to discuss tissue thermal coagulation formation in brain. Experimental observations show that thermal coagulation and necrosis can be caused due to laser light absorption. The result of the calculation shows that the process of the thermal coagulation of the given mode comprises two stages: fast and slow. At the first stage, necrosis domain grows fast. Then necrosis domain growth becomes slower because of the competition between the heat diffusion into the surrounding undamaged tissue and the heat dissipation caused by blood perfusion. At the center of coagulation area no neuron was observed and at the transitional zone few nervous cells were seen by microscope. The research can provide reference data for developing clinical therapy of some kind of encephalic diseases by using 532nm laser, and for making cerebral infarction models in animal experiment.

Two-layer optical model of skin for early, non-invasive detection of wound development on the diabetic foot

NASA Astrophysics Data System (ADS)

Yudovsky, Dmitry; Nouvong, Aksone; Schomacker, Kevin; Pilon, Laurent

2010-02-01

Foot ulceration is a debilitating comorbidity of diabetes that may result in loss of mobility and amputation. Optical detection of cutaneous tissue changes due to inflammation and necrosis at the preulcer site could constitute a preventative strategy. A commercial hyperspectral oximetry system was used to measure tissue oxygenation on the feet of diabetic patients. A previously developed predictive index was used to differentiate preulcer tissue from surrounding healthy tissue with a sensitivity of 92% and specificity of 80%. To improve prediction accuracy, an optical skin model was developed treating skin as a two-layer medium and explicitly accounting for (i) melanin content and thickness of the epidermis, (ii) blood content and hemoglobin saturation of the dermis, and (iii) tissue scattering in both layers. Using this forward model, an iterative inverse method was used to determine the skin properties from hyperspectral images of preulcerative areas. The use of this information in lowering the false positive rate was discussed.

Blood flow mechanics and oxygen transport and delivery in the retinal microcirculation: multiscale mathematical modeling and numerical simulation.

PubMed

Causin, Paola; Guidoboni, Giovanna; Malgaroli, Francesca; Sacco, Riccardo; Harris, Alon

2016-06-01

The scientific community continues to accrue evidence that blood flow alterations and ischemic conditions in the retina play an important role in the pathogenesis of ocular diseases. Many factors influence retinal hemodynamics and tissue oxygenation, including blood pressure, blood rheology, oxygen arterial permeability and tissue metabolic demand. Since the influence of these factors on the retinal circulation is difficult to isolate in vivo, we propose here a novel mathematical and computational model describing the coupling between blood flow mechanics and oxygen ([Formula: see text]) transport in the retina. Albeit in a simplified manner, the model accounts for the three-dimensional anatomical structure of the retina, consisting in a layered tissue nourished by an arteriolar/venular network laying on the surface proximal to the vitreous. Capillary plexi, originating from terminal arterioles and converging into smaller venules, are embedded in two distinct tissue layers. Arteriolar and venular networks are represented by fractal trees, whereas capillary plexi are represented using a simplified lumped description. In the model, [Formula: see text] is transported along the vasculature and delivered to the tissue at a rate that depends on the metabolic demand of the various tissue layers. First, the model is validated against available experimental results to identify baseline conditions. Then, a sensitivity analysis is performed to quantify the influence of blood pressure, blood rheology, oxygen arterial permeability and tissue oxygen demand on the [Formula: see text] distribution within the blood vessels and in the tissue. This analysis shows that: (1) systemic arterial blood pressure has a strong influence on the [Formula: see text] profiles in both blood and tissue; (2) plasma viscosity and metabolic consumption rates have a strong influence on the [Formula: see text] tension at the level of the retinal ganglion cells; and (3) arterial [Formula: see text] permeability has a strong influence on the [Formula: see text] saturation in the retinal arterioles.

Engineering cartilaginous grafts using chondrocyte-laden hydrogels supported by a superficial layer of stem cells.

PubMed

Mesallati, Tariq; Buckley, Conor T; Kelly, Daniel J

2017-05-01

During postnatal joint development, progenitor cells that reside in the superficial region of articular cartilage first drive the rapid growth of the tissue and later help direct the formation of mature hyaline cartilage. These developmental processes may provide directions for the optimal structuring of co-cultured chondrocytes (CCs) and multipotent stromal/stem cells (MSCs) required for engineering cartilaginous tissues. The objective of this study was to engineer cartilage grafts by recapitulating aspects of joint development where a population of superficial progenitor cells drives the development of the tissue. To this end, MSCs were either self-assembled on top of CC-laden agarose gels (structured co-culture) or were mixed with CCs before being embedded in an agarose hydrogel (mixed co-culture). Porcine infrapatellar fat pad-derived stem cells (FPSCs) and bone marrow-derived MSCs (BMSCs) were used as sources of progenitor cells. The DNA, sGAG and collagen content of a mixed co-culture of FPSCs and CCs was found to be lower than the combined content of two control hydrogels seeded with CCs and FPSCs only. In contrast, a mixed co-culture of BMSCs and CCs led to increased proliferation and sGAG and collagen accumulation. Of note was the finding that a structured co-culture, at the appropriate cell density, led to greater sGAG accumulation than a mixed co-culture for both MSC sources. In conclusion, assembling MSCs onto CC-laden hydrogels dramatically enhances the development of the engineered tissue, with the superficial layer of progenitor cells driving CC proliferation and cartilage ECM production, mimicking certain aspects of developing cartilage. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

Construction and histological analysis of a 3D human arterial wall model containing vasa vasorum using a layer-by-layer technique.

PubMed

Shima, Fumiaki; Narita, Hirokazu; Hiura, Ayami; Shimoda, Hiroshi; Akashi, Mitsuru

2017-03-01

There is considerable global demand for three-dimensional (3D) functional tissues which mimic our native organs and tissues for use as in vitro drug screening systems and in regenerative medicine. In particular, there has been an increasing number of patients who suffer from arterial diseases such as arteriosclerosis. As such, in vitro 3D arterial wall models that can evaluate the effects of novel medicines and a novel artificial graft for the treatment are required. In our previous study, we reported the rapid construction of 3D tissues by employing a layer-by-layer (LbL) technique and revealed their potential applications in the pharmaceutical fields and tissue engineering. In this study, we successfully constructed a 3D arterial wall model containing vasa vasorum by employing a LbL technique for the first time. The cells were coated with extracellular matrix nanofilms and seeded into a culture insert using a cell accumulation method. This model had a three-layered hierarchical structure: a fibroblast layer, a smooth muscle layer, and an endothelial layer, which resembled the native arterial wall. Our method could introduce vasa vasorum into a fibroblast layer in vitro and the 3D arterial wall model showed barrier function which was evaluated by immunostaining and transendothelial electrical resistance measurement. Furthermore, electron microscopy observations revealed that the vasa vasorum was composed of single-layered endothelial cells, and the endothelial tubes were surrounded by the basal lamina, which are known to promote maturation and stabilization in native blood capillaries. These models should be useful for tissue engineering, regenerative medicine, and pharmaceutical applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 814-823, 2017. © 2016 Wiley Periodicals, Inc.

Retinal Structure of Birds of Prey Revealed by Ultra-High Resolution Spectral-Domain Optical Coherence Tomography

PubMed Central

Ruggeri, Marco; Major, James C.; McKeown, Craig; Knighton, Robert W.; Puliafito, Carmen A.

2010-01-01

Purpose. To reveal three-dimensional (3-D) information about the retinal structures of birds of prey in vivo. Methods. An ultra-high resolution spectral-domain optical coherence tomography (SD-OCT) system was built for in vivo imaging of retinas of birds of prey. The calibrated imaging depth and axial resolution of the system were 3.1 mm and 2.8 μm (in tissue), respectively. 3-D segmentation was performed for calculation of the retinal nerve fiber layer (RNFL) map. Results. High-resolution OCT images were obtained of the retinas of four species of birds of prey: two diurnal hawks (Buteo platypterus and Buteo brachyurus) and two nocturnal owls (Bubo virginianus and Strix varia). These images showed the detailed retinal anatomy, including the retinal layers and the structure of the deep and shallow foveae. The calculated thickness map showed the RNFL distribution. Traumatic injury to one bird's retina was also successfully imaged. Conclusions. Ultra-high resolution SD-OCT provides unprecedented high-quality 2-D and 3-D in vivo visualization of the retinal structures of birds of prey. SD-OCT is a powerful imaging tool for vision research in birds of prey. PMID:20554605

Study of the structure of 3-D composites based on carbon nanotubes in bovine serum albumin matrix by X-ray microtomography

NASA Astrophysics Data System (ADS)

Ignatov, D.; Zhurbina, N.; Gerasimenko, A.

2017-01-01

3-D composites are widely used in tissue engineering. A comprehensive analysis by X-ray microtomography was conducted to study the structure of the 3-D composites. Comprehensive analysis of the structure of the 3-D composites consisted of scanning, image reconstruction of shadow projections, two-dimensional and three-dimensional visualization of the reconstructed images and quantitative analysis of the samples. Experimental samples of composites were formed by laser vaporization of the aqueous dispersion BSA and single-walled (SWCNTs) and multi-layer (MWCNTs) carbon nanotubes. The samples have a homogeneous structure over the entire volume, the percentage of porosity of 3-D composites based on SWCNTs and MWCNTs - 16.44%, 28.31%, respectively. An average pore diameter of 3-D composites based on SWCNTs and MWCNTs - 45 μm 93 μm. 3-D composites based on carbon nanotubes in bovine serum albumin matrix can be used in tissue engineering of bone and cartilage, providing cell proliferation and blood vessel sprouting.

3D Printing and 3D Bioprinting in Pediatrics.

PubMed

Vijayavenkataraman, Sanjairaj; Fuh, Jerry Y H; Lu, Wen Feng

2017-07-13

Additive manufacturing, commonly referred to as 3D printing, is a technology that builds three-dimensional structures and components layer by layer. Bioprinting is the use of 3D printing technology to fabricate tissue constructs for regenerative medicine from cell-laden bio-inks. 3D printing and bioprinting have huge potential in revolutionizing the field of tissue engineering and regenerative medicine. This paper reviews the application of 3D printing and bioprinting in the field of pediatrics.

Investigation of changes in fractal dimension from layered retinal structures of healthy and diabetic eyes with optical coherence tomography

NASA Astrophysics Data System (ADS)

Gao, Wei; Zakharov, Valery P.; Myakinin, Oleg O.; Bratchenko, Ivan A.; Artemyev, Dmitry N.; Kornilin, Dmitry V.

2015-07-01

Optical coherence tomography (OCT) is usually employed for the measurement of retinal thickness characterizing the structural changes of tissue. However, fractal dimension (FD) could also character the structural changes of tissue. Therefore, fractal dimension changes may provide further information regarding cellular layers and early damage in ocular diseases. We investigated the possibility of OCT in detecting changes in fractal dimension from layered retinal structures. OCT images were obtained from diabetic patients without retinopathy (DM, n = 38 eyes) or mild diabetic retinopathy (MDR, n = 43 eyes) and normal healthy subjects (Controls, n = 74 eyes). Fractal dimension was calculated using the differentiate box counting methodology. We evaluated the usefulness of quantifying fractal dimension of layered structures in the detection of retinal damage. Generalized estimating equations considering within-subject intereye relations were used to test for differences between the groups. A modified p value of <0.001 was considered statistically significant. Receiver operating characteristic (ROC) curves were constructed to describe the ability of fractal dimension to discriminate between the eyes of DM, MDR and healthy eyes. Significant decreases of fractal dimension were observed in all layers in the MDR eyes compared with controls except in the inner nuclear layer (INL). Significant decreases of fractal dimension were also observed in all layers in the MDR eyes compared with DM eyes. The highest area under receiver operating characteristic curve (AUROC) values estimated for fractal dimension were observed for the outer plexiform layer (OPL) and outer segment photoreceptors (OS) when comparing MDR eyes with controls. The highest AUROC value estimated for fractal dimension were also observed for the retinal nerve fiber layer (RNFL) and OS when comparing MDR eyes with DM eyes. Our results suggest that fractal dimension of the intraretinal layers may provide useful information to differentiate pathological from healthy eyes. Further research is warranted to determine how this approach may be used to improve diagnosis of early retinal neurodegeneration.

Effects of printing-induced interfaces on localized strain within 3D printed hydrogel structures.

PubMed

Christensen, Kyle; Davis, Brian; Jin, Yifei; Huang, Yong

2018-08-01

Additive manufacturing, or 3D printing, is a promising approach for the fabrication of biological structures for regenerative medicine applications using tissue-like materials such as hydrogels. Herein, inkjet printing is implemented as a model droplet-based 3D printing technology for which interfaces have been shown to form between printed lines within printed layers of hydrogel structures. Experimental samples with interfaces in two orientations are fabricated by inkjet printing and control samples with and without interfaces are fabricated by extrusion printing and casting, respectively. The formation of partial and full interfaces is modeled in terms of printing conditions and gelation parameters, and an approach to predicting the ratio of interfacial area to the total contact area between two adjacent lines is presented. Digital image correlation is used to determine strain distributions and identify regions of increased localized deformation for samples under uniaxial tension. Despite the presence of interfaces in inkjet-printed samples, strain distributions are found to be homogeneous regardless of interface orientation, which may be attributed to the multi-layer nature of samples. Conversely, single-layer extrusion-printed samples exhibit localized regions of increased deformation between printed lines, indicating delamination along interfaces. The effective stiffness, failure strength, and failure strain of inkjet-printed samples are found to be dependent on the orientation of interfaces within layers. Specifically, inkjet-printed samples in which tensile forces pull apart interfaces exhibit significantly decreased mechanical properties compared to cast samples. Copyright © 2018 Elsevier B.V. All rights reserved.

Losartan Attenuates Degradation of Aorta and Lung Tissue Micromechanics in a Mouse Model of Severe Marfan Syndrome

PubMed Central

Lee, Jia-Jye; Galatioto, Josephine; Rao, Satish; Ramirez, Francesco; Costa, Kevin D.

2018-01-01

Marfan syndrome (MFS) is an autosomal dominant disease of the connective tissue due to mutations in the fibrillin-1 gene (FBN1). This study aimed at characterizing microelastic properties of the ascending aorta wall and lung parenchyma tissues from wild type (WT) and age-matched Fbn1 hypomorphic mice (Fbn1mgR/mgR mice) to identify tissue-specific biomechanical effects of aging and disease in MFS. Atomic force microscopy (AFM) was used to indent lung parenchyma and aortic wall tissues, using Hybrid Eshelby Decomposition analysis to extract layer-specific properties of the intima and media. The intima stiffened with age and was not different between WT and Fbn1mgR/mgR tissues, whereas the media layer of mutant aortas showed progressive structural and mechanical degradation with a modulus that was 50% softer than WT by 3.5 months of age. Similarly, mutant mice displayed progressive structural and mechanical deterioration of lung tissue, which was over 85% softer than WT by 3.5 months of age. Chronic treatment with the angiotensin type I receptor antagonist, losartan, attenuated the aorta and lung tissue degradation, resulting in structural and mechanical properties not significantly different from age-matched WT controls. By revealing micromechanical softening of elastin-rich aorta and lung tissues with disease progression in fibrillin-1 deficient mice, our findings support the use of losartan as a prophylactic treatment that may abrogate the life-threatening symptoms of MFS. PMID:27090893

Losartan Attenuates Degradation of Aorta and Lung Tissue Micromechanics in a Mouse Model of Severe Marfan Syndrome.

PubMed

Lee, Jia-Jye; Galatioto, Josephine; Rao, Satish; Ramirez, Francesco; Costa, Kevin D

2016-10-01

Marfan syndrome (MFS) is an autosomal dominant disease of the connective tissue due to mutations in the fibrillin-1 gene (FBN1). This study aimed at characterizing microelastic properties of the ascending aortic wall and lung parenchyma tissues from wild type (WT) and age-matched Fbn1 hypomorphic mice (Fbn1(mgR/mgR) mice) to identify tissue-specific biomechanical effects of aging and disease in MFS. Atomic force microscopy was used to indent lung parenchyma and aortic wall tissues, using Hybrid Eshelby Decomposition analysis to extract layer-specific properties of the intima and media. The intima stiffened with age and was not different between WT and Fbn1(mgR/mgR) tissues, whereas the media layer of MFS aortas showed progressive structural and mechanical degradation with a modulus that was 50% softer than WT by 3.5 months of age. Similarly, MFS mice displayed progressive structural and mechanical deterioration of lung tissue, which was over 85% softer than WT by 3.5 months of age. Chronic treatment with the angiotensin type I receptor antagonist, losartan, attenuated the aorta and lung tissue degradation, resulting in structural and mechanical properties not significantly different from age-matched WT controls. By revealing micromechanical softening of elastin-rich aorta and lung tissues with disease progression in fibrillin-1 deficient mice, our findings support the use of losartan as a prophylactic treatment that may abrogate the life-threatening symptoms of MFS.

Bone marrow mesenchymal stem cells accelerate the hyperglycemic refractory wound healing by inhibiting an excessive inflammatory response.

PubMed

Nan, Wenbin; Xu, Zhihao; Chen, Zhibin; Yuan, Xin; Lin, Juntang; Feng, Huigen; Lian, Jie; Chen, Hongli

2017-05-01

The aim of the present study was to evaluate the healing effect of bone marrow-derived mesenchymal stem cells administered to hyperglycemia model mice with skin wounds, and to explore the underlying mechanism contributing to their effects in promoting refractory wound healing. A full‑thickness skin wound mouse model was established, and refers to a wound of the skin and subcutaneous tissue. The mice were randomly divided into three groups: Blank control group, hyperglycemic group and a hyperglycemic group treated with stem cells. Wound healing was monitored and the wound‑healing rate was determined at 3, 6, 9, and 12 days following trauma. The structure of the organization of new skin tissue was observed by hematoxylin and eosin staining, and expression levels of the inflammatory cytokines interleukin (IL)‑6 and tumor necrosis factor (TNF)‑α were determined from 1 to 6 days following trauma. The wound healing of the hyperglycemic group was slower than that of the blank group, and the hyperglycemic mice treated with stem cells presented faster healing than the hyperglycemia group. The horny layer and granular layer of the skin were thinner and incomplete in the new skin tissue of the hyperglycemic group, whereas the new skin wound tissue basal layer was flat and demonstrated better fusion with the wound edge in the other two groups. The expression of inflammatory cytokines (IL‑6 and TNF‑α) was significantly increased in all three groups, with continuously higher expression in the hyperglycemic group and decreased expression in the other two groups over time. Hyperglycemia refractory wounds are likely related to the excessive expression of inflammatory cytokines surrounding the wound area. Stem cells may be able to alleviate the excessive inflammatory reaction in the wound tissue of hyperglycemic mice, so as to promote wound healing.

Multiscale image analysis reveals structural heterogeneity of the cell microenvironment in homotypic spheroids.

PubMed

Schmitz, Alexander; Fischer, Sabine C; Mattheyer, Christian; Pampaloni, Francesco; Stelzer, Ernst H K

2017-03-03

Three-dimensional multicellular aggregates such as spheroids provide reliable in vitro substitutes for tissues. Quantitative characterization of spheroids at the cellular level is fundamental. We present the first pipeline that provides three-dimensional, high-quality images of intact spheroids at cellular resolution and a comprehensive image analysis that completes traditional image segmentation by algorithms from other fields. The pipeline combines light sheet-based fluorescence microscopy of optically cleared spheroids with automated nuclei segmentation (F score: 0.88) and concepts from graph analysis and computational topology. Incorporating cell graphs and alpha shapes provided more than 30 features of individual nuclei, the cellular neighborhood and the spheroid morphology. The application of our pipeline to a set of breast carcinoma spheroids revealed two concentric layers of different cell density for more than 30,000 cells. The thickness of the outer cell layer depends on a spheroid's size and varies between 50% and 75% of its radius. In differently-sized spheroids, we detected patches of different cell densities ranging from 5 × 10 5 to 1 × 10 6  cells/mm 3 . Since cell density affects cell behavior in tissues, structural heterogeneities need to be incorporated into existing models. Our image analysis pipeline provides a multiscale approach to obtain the relevant data for a system-level understanding of tissue architecture.

Flexible bipolar nanofibrous membranes for improving gradient microstructure in tendon-to-bone healing.

PubMed

Li, Xiaoxi; Cheng, Ruoyu; Sun, Zhiyong; Su, Wei; Pan, Guoqing; Zhao, Song; Zhao, Jinzhong; Cui, Wenguo

2017-10-01

Enthesis is a specialized tissue interface between the tendon and bone. Enthesis structure is very complex because of gradient changes in its composition and structure. There is currently no strategy to create a suitable environment and to regenerate the gradual-changing enthesis because of the modular complexities between two tissue types. Herein, a dual-layer organic/inorganic flexible bipolar fibrous membrane (BFM) was successfully fabricated by electrospinning to generate biomimetic non-mineralized fibrocartilage and mineralized fibrocartilage in tendon-to-bone integration of enthesis. The growth of the in situ apatite nanoparticle layer was induced on the nano hydroxyapatite-poly-l-lactic acid (nHA-PLLA) fibrous layer in simulated body solution, and the poly-l-lactic acid (PLLA) fibrous layer retained its original properties to induce tendon regeneration. The in vivo results showed that BFM significantly increased the area of glycosaminoglycan staining at the tendon-bone interface and improved collagen organization when compared to the simplex fibrous membrane (SFM) of PLLA. Implanting the bipolar membrane also induced bone formation and fibrillogenesis as assessed by micro-CT and histological analysis. Biomechanical testing showed that the BFM group had a greater ultimate load-to-failure and stiffness than the SFM group at 12weeks after surgery. Therefore, this flexible bipolar nanofibrous membrane improves the healing and regeneration process of the enthesis in rotator cuff repair. In this study, we generated a biomimetic dual-layer organic/inorganic flexible bipolar fibrous membrane by sequential electrospinning and in situ biomineralization, producing integrated bipolar fibrous membranes of PLLA fibrous membrane as the upper layer and nHA-PLLA fibrous membrane as the lower layer to mimic non-mineralized fibrocartilage and mineralized fibrocartilage in tendon-to-bone integration of enthesis. Flexible bipolar nanofibrous membranes could be easily fabricated with gradient microstructure for enthesis regeneration in rotator cuff tears. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

On the genetic control of planar growth during tissue morphogenesis in plants.

PubMed

Enugutti, Balaji; Kirchhelle, Charlotte; Schneitz, Kay

2013-06-01

Tissue morphogenesis requires extensive intercellular communication. Plant organs are composites of distinct radial cell layers. A typical layer, such as the epidermis, is propagated by stereotypic anticlinal cell divisions. It is presently unclear what mechanisms coordinate cell divisions relative to the plane of a layer, resulting in planar growth and maintenance of the layer structure. Failure in the regulation of coordinated growth across a tissue may result in spatially restricted abnormal growth and the formation of a tumor-like protrusion. Therefore, one way to approach planar growth control is to look for genetic mutants that exhibit localized tumor-like outgrowths. Interestingly, plants appear to have evolved quite robust genetic mechanisms that govern these aspects of tissue morphogenesis. Here we provide a short summary of the current knowledge about the genetics of tumor formation in plants and relate it to the known control of coordinated cell behavior within a tissue layer. We further portray the integuments of Arabidopsis thaliana as an excellent model system to study the regulation of planar growth. The value of examining this process in integuments was established by the recent identification of the Arabidopsis AGC VIII kinase UNICORN as a novel growth suppressor involved in the regulation of planar growth and the inhibition of localized ectopic growth in integuments and other floral organs. An emerging insight is that misregulation of central determinants of adaxial-abaxial tissue polarity can lead to the formation of spatially restricted multicellular outgrowths in several tissues. Thus, there may exist a link between the mechanisms regulating adaxial-abaxial tissue polarity and planar growth in plants.

Microscopic functional anatomy: Integumentary system: Chapter 17

USGS Publications Warehouse

Elliott, Diane G.; Ostrander, Gary K.

2000-01-01

Many of the features of the fish integument can only be observed microscopically. Because there are over 20,000 living fishes, mostly higher bony fishes (teleosts), a great diversity exists in the microscopic anatomy of the integument. This chapter presents several examples from varied taxonomic groups to illustrate the variation in morphological features. As in all vertebrate epidermis, the fundamental structural unit is the epithelial cell. This is the only constant feature, as a great diversity of cell types exists in the various fish taxa. Some of these include apocrine mucous cells and a variety of other secretory cells, ionocytes, sensory cells, and wandering cells such as leukocytes. The dermis consists essentially of two sets of collagen fibers arranged in opposing geodesic spirals around the body. The dermis of most fishes is divided into two major layers. The upper (outer) layer, the stratum spongiosum or stratum laxum, is a loose network of connective tissue, whereas the lower layer, the stratum compactum, is a dense layer consisting primarily of orthogonal collagen bands. There are also specialized dermal elements such as chromatophores scales, and fin rays.

Computer-Aided Process Planning for the Layered Fabrication of Porous Scaffold Matrices

NASA Astrophysics Data System (ADS)

Starly, Binil

Rapid Prototyping (RP) technology promises to have a tremendous impact on the design and fabrication of porous tissue replacement structures for applications in tissue engineering and regenerative medicine. The layer-by-layer fabrication technology enables the design of patient-specific medical implants and complex structures for diseased tissue replacement strategies. Combined with advancements in imaging modalities and bio-modeling software, physicians can engage themselves in advanced solutions for craniofacial and mandibular reconstruction. For example, prior to the advancement of RP technologies, solid titanium parts used as implants for mandibular reconstruction were fashioned out of molding or CNC-based machining processes (Fig. 3.1). Titanium implants built using this process are often heavy, leading to increased patient discomfort. In addition, the Young's modulus of titanium is almost five times that of healthy cortical bone resulting in stress shielding effects [1,2]. With the advent of CAD/CAM-based tools, the virtual reconstruction of the implants has resulted in significant design improvements. The new generation of implants can be porous, enabling the in-growth of healthy bone tissue for additional implant fixation and stabilization. Newer implants would conform to the external shape of the defect site that is intended to be filled in. More importantly, the effective elastic modulus of the implant can be designed to match that of surrounding tissue. Ideally, the weight of the implant can be designed to equal the weight of the tissue that is being replaced resulting in increased patient comfort. Currently, such porous structures for reconstruction can only be fabricated using RP-based metal fabrication technologies such as Electron Beam Melting (EBM), Selective Laser Sintering (SLS®), and 3D™ Printing processes.

Cultivation and phenotypic characterization of rabbit epithelial cells expanded ex vivo from fresh and cryopreserved limbal and oral mucosal explants.

PubMed

Promprasit, Daranee; Bumroongkit, Kanokkan; Tocharus, Chainarong; Mevatee, Umnat; Tananuvat, Napaporn

2015-03-01

To compare the morphology of cultured rabbit epithelial sheets and the expression of stem cells with differentiated cell markers of cultivated epithelial cells from fresh and cryopreserved limbal and oral mucosal biopsies. Six New Zealand white rabbits were divided into two groups of three, from which limbal and oral mucosal biopsies were taken. Harvested tissues from each rabbit were brought to immediate cultivation, while another set of tissues was cryopreserved. Cultivation was performed by the explant culture technique using human amniotic membrane as a culture substrate, co-culturing with 3T3 fibroblasts and using the air-lifting method. Cells were cultured for three weeks; then cultured epithelial sheets were stained with hematoxylin-eosin and examined for expression patterns of p63, keratin 3 (K3) and connexin 43 (Cx43). Cryopreservation was carried out using the vitrification method. Tissues were preserved in liquid nitrogen using 25% dimethyl sulfoxide combined with 25% propylene glycol in Dulbecco's Modified Eagle's Medium containing 20% fetal bovine serum. After two months, the tissues were warmed, cultured and stained using the same processes as for fresh tissue cultures. Cultivation of fresh limbal and fresh oral mucosal tissues showed epithelial stratification, with two to five cell layers. Immunohistochemical staining showed p63-positive cells in basal and intermediate cell layers. K3 staining was observed in cells in the suprabasal layer, while expression of Cx43 was scattered throughout all layers of the epithelia. All culture sheets expressed p63, K3 and Cx43 with the exception of one sheet from the oral mucosal culture that was p63-negative. Cultured epithelial sheets from cryopreserved tissues showed results similar to those from fresh tissue culture. This study found that cells in cultivated fresh limbal and oral mucosal tissues had similar morphology to cells in cultivated cryopreserved limbal and oral mucosal tissues, both containing a heterogeneous population of cells including stem cells and differentiated cells.

Wound diagnostics with microwaves.

PubMed

Schertlen, Ralph; Pivit, Florian; Wiesbeck, Werner

2002-01-01

The reflection of electromagnetic waves on material surfaces is very depending on the electric and magnetic properties of these materials, on their structure and on the surface texture. Therefore the different layers and dielectric properties of healthy and unsound body tissue also show different reflection behavior towards incidentating electromagnetic waves. By analyzing the reflected signals of incident electromagnetic waves, it is possible to get information about the inner structure of the reflecting body tissue. This effect could then be used for a contactless analysis of body tissue e.g. to gain crucial medical information about healing processes. In this paper the results of several full wave simulations of various tissue structures are presented and the significance and usability of this method is shown.

3D Printing and 3D Bioprinting in Pediatrics

PubMed Central

Vijayavenkataraman, Sanjairaj; Fuh, Jerry Y H; Lu, Wen Feng

2017-01-01

Additive manufacturing, commonly referred to as 3D printing, is a technology that builds three-dimensional structures and components layer by layer. Bioprinting is the use of 3D printing technology to fabricate tissue constructs for regenerative medicine from cell-laden bio-inks. 3D printing and bioprinting have huge potential in revolutionizing the field of tissue engineering and regenerative medicine. This paper reviews the application of 3D printing and bioprinting in the field of pediatrics. PMID:28952542

The layer concept: utilization in determining the pain generators, pathology and how structure determines treatment.

PubMed

Draovitch, Peter; Edelstein, Jaime; Kelly, Bryan T

2012-03-01

The level of understanding of pain in the non-arthritic hip has made significant strides in the last couple of decades beginning with the discoveries of Reinhold Ganz, MD. However, even with the detection of subtle bony abnormalities, including femoroacetabular impingement, a clinician's ability to differentiate pain generators in the hip has been ambiguous. Deciphering the etiology of the pathology versus the pain generator is essential in prescribing the proper treatment. The Layer Concept developed by Dr. Bryan Kelly, is a systematic means of determining which structures about the hip are the source of the pathology, which are the pain generators and how to then best implement treatment. Four layers will be discussed in this article. Layer I, the osseous layer, Layer II, the inert tissue layer, Layer III, the contractile layer and Layer IV, the neuromechanical layer.

Characterization of southern yellow pine bark layers by Attenuated Total Reflectance (ATR) and Fourier Transform Infrared (FT-IR) Spectroscopy

Treesearch

Thomas L. Eberhardt

2009-01-01

The outer bark (rhytidome) of the southern yellow pines is a complex structure comprised of alternating layers of obliterated phloem and periderm tissues, with the latter comprised of three layers, those being phellem, phellogen, and phelloderm. An attenuated total reflectance (ATR) sampling accessory, coupled with a Fourier transform infrared (FTIR) spectrometer,...

3D visualization of middle ear structures

NASA Astrophysics Data System (ADS)

Vogel, Uwe; Schmitt, Thomas

1998-06-01

The achievement of volume geometry data from middle ear structures and surrounding components performs a necessary supposition for the finite element simulation of the vibrational and transfer characteristics of the ossicular chain. So far those models base on generalized figures and size data from anatomy textbooks or particular manual and one- or two-dimensional distance measurements of single ossicles, mostly obtained by light microscopy, respectively. Therefore the goal of this study is to create a procedure for complete three-dimensional imaging of real middle ear structures (tympanic membrane, ossicles, ligaments) in vitro or even in vivo. The main problems are their microscopic size with relevant structures from 10 micrometer to 5 mm, representing various tissue properties (bone, soft tissue). Additionally, these structures are surrounded by the temporal bone, the most solid bone of the human body. Generally there exist several established diagnostic tools for medical imaging that could be used for geometry data acquisition, e.g., X-ray computed tomography and magnetic resonance imaging. Basically they image different tissue parameters, either bony structures (ossicles), or soft tissue (tympanic membrane, ligaments). But considering this application those standard techniques allow low spatial resolution only, usually in the 0.5 - 1mm range, at least in one spatial direction. Thus particular structures of the middle ear region could even be missed completely because of their spatial location. In vitro there is a way out by collecting three complete data sets, each distinguished by 90 degree rotation of a cube-shaped temporal bone specimen. That allows high-resolution imaging in three orthogonal planes, which essentially supports the three-dimensional interpolation of the unknown elements, starting from the regularly set elements of the cubic grid with an edge extension given by the original two-dimensional matrix. A different approach represents the application of a micro- tomographic imaging device. Therefore an X-ray beam focused down to few microns passes the object in a tomographic arrangement. Subsequently the slices become reconstructed. Generally spatial resolution down to 10 micrometer may be obtained by using this procedure. But there exist few devices only, it is not available as standard equipment. The best results concerning spatial resolution should be achieved by applying conventional histologic sectioning techniques. Of course the target will become destroyed during the procedure. It is cut into sections (e.g., 10 micrometer thick), every layer is stained, and the image acquired and stored by a digital still-camera with appropriate resolution (e.g., 2024 X 3036). Three-dimensional reconstruction is done with the computer. The staining allows visual selection of bones and soft tissues, resolutions down to 10 micrometer are possible without target segmentation. But there arise some practical problems. Mainly the geometric context of the layers is affected by the cutting procedure, especially if cutting bone. Another problem performs the adjustment of the -- possibly distorted -- slices to each other. Artificial markers are necessary, which could allow automatic adjustment too. But the introduction and imaging of the markers is difficult inside the temporal bone specimen, that is interspersed by several cavities. Of course the internal target structures must not be destroyed by the marker introduction. Furthermore the embedding compound could disturb the image acquisition, e.g., by optical scattering of paraffin. A related alternative is given by layered ablation/grinding and imaging of the top layer. This saves the geometric consistency, but requires very tricky and time-consuming embedding procedures. Both approaches require considerable expenditures. The possible approaches are evaluated in detail and first results are compared. So far none of the above-mentioned procedures has been established as a standard tool for three-dimensional geometry data acquisition of the middle ear. Otherwise the establishment of a high-resolution imaging technique for those structures, even in vivo, would be of high interest in diagnostics, anatomy and middle ear modeling and research at all.

The influence of tissue layering on microwave thermographic measurements.

PubMed

Hawley, M S; Conway, J; Anderson, A P; Cudd, P A

1988-01-01

Non-invasive thermal imaging and temperature measurement by microwave radiometry has been investigated for medical diagnostic applications and monitoring hyperthermia treatment of cancer, in the context of heterogeneous body structure. The temperature measured by a radiometer is a function of the emission and propagation of microwaves in tissue and the receiving characteristics of the radiometric probe. Propagation of microwaves in lossy media was analysed by a spectral diffraction approach. Extension of this technique via a cascade transmission line model provides an efficient algorithm for predicting the field patterns of aperture antennas contacting multi-layered tissue. A coherent radiative transfer analysis was used to relate the field pattern of a radiating antenna to its receiving characteristics when used as a radiometer probe, leading to a method for simulating radiometric data. Measurements and simulations were used to assess the effect of overlying fat layers upon radiometer response to temperature hot spots in muscle-type media. Results suggest that dielectric layering in tissue greatly influences measured temperatures and should be accounted for in the interpretation of radiometric data.

The structure and possible functions of the milkfish Chanos chanos adipose eyelid.

PubMed

Chang, C-H; Chiao, C-C; Yan, H Y

2009-07-01

Basic histological sections (with different staining methods) and scanning electron microscopy (SEM) examinations showed that there were three distinctive layers in the adipose eyelid of milkfish Chanos chanos, which is found in the cephalie region and covers the entire eye. The outer and inner layers were epithelial tissues and the middle layer was composed of connective tissue formed by type I collagen fibrils. No adipose tissue was found in any of the three layers of the so-called adipose eyelid. Examination by transmission spectrophotometer showed that the adipose tissue could filter out ambient light with a wavelength shorter than 305 nm. A photoretinoscope was used to investigate whether the adipose eyelid influenced the mechanism of eye focusing. Eye diopter values did not differ before or after eyelid removal, which indicated that the adipose eyelid did not play a role in eye focusing. In light of these findings, it is suggested that the adipose eyelid serves to block exposure of harmful ultraviolet light into eyes and may also to offer some protection against impact to the eye in the aquatic environment.

SAM-based Cell Transfer to Photopatterned Hydrogels for Microengineering Vascular-Like Structures

PubMed Central

Sadr, Nasser; Zhu, Mojun; Osaki, Tatsuya; Kakegawa, Takahiro; Yang, Yunzhi; Moretti, Matteo; Fukuda, Junji; Khademhosseini, Ali

2011-01-01

A major challenge in tissue engineering is to reproduce the native 3D microvascular architecture fundamental for in vivo functions. Current approaches still lack a network of perfusable vessels with native 3D structural organization. Here we present a new method combining self-assembled monolayer (SAM)-based cell transfer and gelatin methacrylate hydrogel photopatterning techniques for microengineering vascular structures. Human umbilical vein cell (HUVEC) transfer from oligopeptide SAM-coated surfaces to the hydrogel revealed two SAM desorption mechanisms: photoinduced and electrochemically triggered. The former, occurs concomitantly to hydrogel photocrosslinking, and resulted in efficient (>97%) monolayer transfer. The latter, prompted by additional potential application, preserved cell morphology and maintained high transfer efficiency of VE-cadherin positive monolayers over longer culture periods. This approach was also applied to transfer HUVECs to 3D geometrically defined vascular-like structures in hydrogels, which were then maintained in perfusion culture for 15 days. As a step toward more complex constructs, a cell-laden hydrogel layer was photopatterned around the endothelialized channel to mimic the vascular smooth muscle structure of distal arterioles. This study shows that the coupling of the SAM-based cell transfer and hydrogel photocrosslinking could potentially open up new avenues in engineering more complex, vascularized tissue constructs for regenerative medicine and tissue engineering applications. PMID:21802723

Tough Adhesives for Diverse Wet Surfaces

PubMed Central

Li, J.; Celiz, A. D.; Yang, J.; Yang, Q.; Wamala, I.; Whyte, W.; Seo, B. R.; Vasilyev, N. V.; Vlassak, J. J.; Suo, Z.; Mooney, D. J.

2018-01-01

Adhesion to wet and dynamic surfaces, including biological tissues, is important in many fields, but has proven extremely challenging. Existing adhesives are either cytotoxic, adhere weakly to tissues, or cannot be utilized in wet environments. We report a bio-inspired design for adhesives consisting of two layers: an adhesive surface and a dissipative matrix. The former adheres to the substrate by electrostatic interactions, covalent bonds, and physical interpenetration. The latter amplifies energy dissipation through hysteresis. The two layers synergistically lead to higher adhesion energy on wet surfaces than existing adhesives. Adhesion occurs within minutes, independent of blood exposure, and compatible with in vivo dynamic movements. This family of adhesives may be useful in many areas of application, including tissue adhesives, wound dressings and tissue repair. PMID:28751604

Multi-responsive hydrogels for drug delivery and tissue engineering applications

PubMed Central

Knipe, Jennifer M.; Peppas, Nicholas A.

2014-01-01

Multi-responsive hydrogels, or ‘intelligent’ hydrogels that respond to more than one environmental stimulus, have demonstrated great utility as a regenerative biomaterial in recent years. They are structured biocompatible materials that provide specific and distinct responses to varied physiological or externally applied stimuli. As evidenced by a burgeoning number of investigators, multi-responsive hydrogels are endowed with tunable, controllable and even biomimetic behavior well-suited for drug delivery and tissue engineering or regenerative growth applications. This article encompasses recent developments and challenges regarding supramolecular, layer-by-layer assembled and covalently cross-linked multi-responsive hydrogel networks and their application to drug delivery and tissue engineering. PMID:26816625

Coherent Timescales and Mechanical Structure of Multicellular Aggregates.

PubMed

Yu, Miao; Mahtabfar, Aria; Beelen, Paul; Demiryurek, Yasir; Shreiber, David I; Zahn, Jeffrey D; Foty, Ramsey A; Liu, Liping; Lin, Hao

2018-06-05

Multicellular aggregates are an excellent model system to explore the role of tissue biomechanics in specifying multicellular reorganization during embryonic developments and malignant invasion. Tissue-like spheroids, when subjected to a compressive force, are known to exhibit liquid-like behaviors at long timescales (hours), largely because of cell rearrangements that serve to effectively dissipate the applied stress. At short timescales (seconds to minutes), before cell rearrangement, the mechanical behavior is strikingly different. The current work uses shape relaxation to investigate the structural characteristics of aggregates and discovers two coherent timescales: one on the order of seconds, the other tens of seconds. These timescales are universal, conserved across a variety of tested species, and persist despite great differences in other properties such as tissue surface tension and adhesion. A precise mathematical theory is used to correlate the timescales with mechanical properties and reveals that aggregates have a relatively strong envelope and an unusually "soft" interior (weak bulk elastic modulus). This characteristic is peculiar, considering that both layers consist of identical units (cells), but is consistent with the fact that this structure can engender both structural integrity and the flexibility required for remodeling. In addition, tissue surface tension, elastic modulus, and viscosity are proportional to each other. Considering that these tissue-level properties intrinsically derive from cellular-level properties, the proportionalities imply precise coregulation of the latter and in particular of the tension on the cell-medium and cell-cell interfaces. Copyright © 2018 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Atomic layer deposition to prevent metal transfer from implants: An X-ray fluorescence study

NASA Astrophysics Data System (ADS)

Bilo, Fabjola; Borgese, Laura; Prost, Josef; Rauwolf, Mirjam; Turyanskaya, Anna; Wobrauschek, Peter; Kregsamer, Peter; Streli, Christina; Pazzaglia, Ugo; Depero, Laura E.

2015-12-01

We show that Atomic Layer Deposition is a suitable coating technique to prevent metal diffusion from medical implants. The metal distribution in animal bone tissue with inserted bare and coated Co-Cr alloys was evaluated by means of micro X-ray fluorescence mapping. In the uncoated implant, the migration of Co and Cr particles from the bare alloy in the biological tissues is observed just after one month and the number of particles significantly increases after two months. In contrast, no metal diffusion was detected in the implant coated with TiO2. Instead, a gradient distribution of the metals was found, from the alloy surface going into the tissue. No significant change was detected after two months of aging. As expected, the thicker is the TiO2 layer, the lower is the metal migration.

Rapid engineering of endothelial cell-lined vascular-like structures in in situ crosslinkable hydrogels.

PubMed

Kageyama, Tatsuto; Kakegawa, Takahiro; Osaki, Tatsuya; Enomoto, Junko; Ito, Taichi; Nittami, Tadashi; Fukuda, Junji

2014-06-01

Fabrication of perfusable vascular networks in vitro is one of the most critical challenges in the advancement of tissue engineering. Because cells consume oxygen and nutrients during the fabrication process, a rapid fabrication approach is necessary to construct cell-dense vital tissues and organs, such as the liver. In this study, we propose a rapid molding process using an in situ crosslinkable hydrogel and electrochemical cell transfer for the fabrication of perfusable vascular structures. The in situ crosslinkable hydrogel was composed of hydrazide-modified gelatin (gelatin-ADH) and aldehyde-modified hyaluronic acid (HA-CHO). By simply mixing these two solutions, the gelation occurred in less than 20 s through the formation of a stable hydrazone bond. To rapidly transfer cells from a culture surface to the hydrogel, we utilized a zwitterionic oligopeptide, which forms a self-assembled molecular layer on a gold surface. Human umbilical vein endothelial cells adhering on a gold surface via the oligopeptide layer were transferred to the hydrogel within 5 min, along with electrochemical desorption of the oligopeptides. This approach was applicable to cylindrical needles 200-700 µm in diameter, resulting in the formation of perfusable microchannels where the internal surface was fully enveloped with the transferred endothelial cells. The entire fabrication process was completed within 10 min, including 20 s for the hydrogel crosslinking and 5 min for the electrochemical cell transfer. This rapid fabrication approach may provide a promising strategy to construct perfusable vasculatures in cell-dense tissue constructs and subsequently allow cells to organize complicated and fully vascularized tissues while preventing hypoxic cell injury.

The primate fovea: Structure, function and development.

PubMed

Bringmann, Andreas; Syrbe, Steffen; Görner, Katja; Kacza, Johannes; Francke, Mike; Wiedemann, Peter; Reichenbach, Andreas

2018-03-30

A fovea is a pitted invagination in the inner retinal tissue (fovea interna) that overlies an area of photoreceptors specialized for high acuity vision (fovea externa). Although the shape of the vertebrate fovea varies considerably among the species, there are two basic types. The retina of many predatory fish, reptilians, and birds possess one (or two) convexiclivate fovea(s), while the retina of higher primates contains a concaviclivate fovea. By refraction of the incoming light, the convexiclivate fovea may function as image enlarger, focus indicator, and movement detector. By centrifugal displacement of the inner retinal layers, which increases the transparency of the central foveal tissue (the foveola), the primate fovea interna improves the quality of the image received by the central photoreceptors. In this review, we summarize ‒ with the focus on Müller cells of the human and macaque fovea ‒ data regarding the structure of the primate fovea, discuss various aspects of the optical function of the fovea, and propose a model of foveal development. The "Müller cell cone" of the foveola comprises specialized Müller cells which do not support neuronal activity but may serve optical and structural functions. In addition to the "Müller cell cone", structural stabilization of the foveal morphology may be provided by the 'z-shaped' Müller cells of the fovea walls, via exerting tractional forces onto Henle fibers. The spatial distribution of glial fibrillary acidic protein may suggest that the foveola and the Henle fiber layer are subjects to mechanical stress. During development, the foveal pit is proposed to be formed by a vertical contraction of the centralmost Müller cells. After widening of the foveal pit likely mediated by retracting astrocytes, Henle fibers are formed by horizontal contraction of Müller cell processes in the outer plexiform layer and the centripetal displacement of photoreceptors. A better understanding of the molecular, cellular, and mechanical factors involved in the developmental morphogenesis and the structural stabilization of the fovea may help to explain the (patho-) genesis of foveal hypoplasia and macular holes. Copyright © 2018 Elsevier Ltd. All rights reserved.

Human umbilical cord mesenchymal stromal cells in a sandwich approach for osteochondral tissue engineering

PubMed Central

Wang, Limin; Zhao, Liang; Detamore, Michael S.

2013-01-01

Cell sources and tissue integration between cartilage and bone regions are critical to successful osteochondral regeneration. In this study, human umbilical cord mesenchymal stromal cells (hUCMSCs), derived from Wharton’s jelly, were introduced to the field of osteochondral tissue engineering and a new strategy for osteochondral integration was developed by sandwiching a layer of cells between chondrogenic and osteogenic constructs before suturing them together. Specifically, hUCMSCs were cultured in biodegradable poly-l-lactic acid scaffolds for 3 weeks in either chondrogenic or osteogenic medium to differentiate cells toward cartilage or bone lineages, respectively. A highly concentrated cell solution containing undifferentiated hUCMSCs was pasted onto the surface of the bone layer at week 3 and the two layers were then sutured together to form an osteochondral composite for another 3 week culture period. Chondrogenic and osteogenic differentiation was initiated during the first 3 weeks, as evidenced by the expression of type II collagen and runt-related transcription factor 2 genes, respectively, and continued with the increase of extracellular matrix during the last 3 weeks. Histological and immunohistochemical staining, such as for glycosaminoglycans, type I collagen and calcium, revealed better integration and transition of these matrices between two layers in the composite group containing sandwiched cells compared to other control composites. These results suggest that hUCMSCs may be a suitable cell source for osteochondral regeneration, and the strategy of sandwiching cells between two layers may facilitate scaffold and tissue integration. PMID:21953869

Interaction of Vascular Smooth Muscle Cells Under Low Shear Stress

NASA Technical Reports Server (NTRS)

Seidel, Charles L.

1998-01-01

The blood vessel wall consists of three cellular layers, an outer adventitial, a middle medial and an inner intimal layer. When the blood vessel forms in the embryo it begins as a tube composed of a single cell type called endothelial cells. Over time, other cells are recruited from the surrounding tissue to form additional layers on the outer surface of the endothelial tube. The cells that are recruited are called mesenchymal cells. Mesenchymal cells are responsible for the production of connective tissue that holds the blood vessel together and for developing into vascular smooth muscle cells that are responsible for regulating the diameter of the vessel (1) and therefore, blood flow. In a fully developed blood vessel, the endothelial cells make- up the majority of cells in the intimal layer while the mesenchymal cells make-up the majority of cells in the medial and adventitial layers. Within the medial layer of a mature vessel, cells are organized into multiple circular layers of alternating bands of connective tissue and cells. The cell layer is composed of a mixture of mesenchymal cells that have not developed into smooth muscle cells and fully developed smooth muscle cells (2). The assembly and organization of complex tissues is directed in part by a signaling system composed of proteins on the cell surface called adhesion molecules. Adhesion molecules enable cells to recognize each other as well as the composition of the connective tissue in which they reside (3). It was hypothesized that the different cell types that compose the vascular wall possess different adhesion molecules that enable them to recognize each other and through this recognition system, form the complex layered organization of the vascular wall. In other words, the layered organization is an intrinsic property of the cells. If this hypothesis is correct then the different cells that make up the vessel wall, when mixed together, should organize themselves into a layered structure resembling an intact blood vessel. Experiments described below were designed to test this hypothesis.

Monte Carlo and discrete-ordinate simulations of spectral radiances in a coupled air-tissue system.

PubMed

Hestenes, Kjersti; Nielsen, Kristian P; Zhao, Lu; Stamnes, Jakob J; Stamnes, Knut

2007-04-20

We perform a detailed comparison study of Monte Carlo (MC) simulations and discrete-ordinate radiative-transfer (DISORT) calculations of spectral radiances in a 1D coupled air-tissue (CAT) system consisting of horizontal plane-parallel layers. The MC and DISORT models have the same physical basis, including coupling between the air and the tissue, and we use the same air and tissue input parameters for both codes. We find excellent agreement between radiances obtained with the two codes, both above and in the tissue. Our tests cover typical optical properties of skin tissue at the 280, 540, and 650 nm wavelengths. The normalized volume scattering function for internal structures in the skin is represented by the one-parameter Henyey-Greenstein function for large particles and the Rayleigh scattering function for small particles. The CAT-DISORT code is found to be approximately 1000 times faster than the CAT-MC code. We also show that the spectral radiance field is strongly dependent on the inherent optical properties of the skin tissue.

Application of photo-magnetic therapy for treatment of skin radiation damage in rats.

PubMed

Simonova-Pushkar, L I; Gertman, V Z; Bilogurova, L V

2014-09-01

To improve methods of prevention and treatment of local radiation injury to the skin using the photomagnetic therapy. Materials and methods. Study was conducted on 60 male Wistar rats with 180-200 g bodyweight. The femoral area right hind limb of rats was locally irradiated by X-ray unit at a dose of 80.0 Gy. Exposed animals were divided into 2 groups: control and experimental. The rats of the experimental group received 2 courses of photo-magnetic therapy on the irradiated skin. The observations were carried out for 60 days. Methods - clinical, histological and statistical. Results. Local irradiation of rat skin causes the development of radiation ulcers in 60-70 % of the animals with the destruction of the structure in all layers of the skin. Spontaneous healing of radiation ulcer lasts at least two months with no complete skin recovery. Photo-magnetic therapy applied immediately after irradiation resulted in two-folddecrease of frequency of radiation ulcer incidence, accelerated the complete healing for 3 weeks and to ameliorated their progress. Histological examination showed that the photo-magnetic therapy reduced the extent of damage to all layers of the skin with restoration of epidermis and dermis structure and reduced the degree of inflammatory and destructive processes in the dermis. Conclusions. Photo-magnetic therapy produces a significant positive treatment effect by significantly reducing the inflammatory and destructive processes in all layers of the skin, stimulates the blood flow recovery in damaged tissue both with fibroblast proliferation and synthesis activation of native collagen fibers and other components of connective tissue, so almost a month accelerates ulcer healing radiation. L. I. Simonova-Pushkar, V. Z. Gertman, L. V. Bilogurova.

Tunable hydrogel composite with two-step processing in combination with innovative hardware upgrade for cell-based three-dimensional bioprinting.

PubMed

Wüst, Silke; Godla, Marie E; Müller, Ralph; Hofmann, Sandra

2014-02-01

Three-dimensional (3-D) bioprinting is the layer-by-layer deposition of biological material with the aim of achieving stable 3-D constructs for application in tissue engineering. It is a powerful tool for the spatially directed placement of multiple materials and/or cells within the 3-D sample. Encapsulated cells are protected by the bioink during the printing process. Very few materials are available that fulfill requirements for bioprinting as well as provide adequate properties for cell encapsulation during and after the printing process. A hydrogel composite including alginate and gelatin precursors was tuned with different concentrations of hydroxyapatite (HA) and characterized in terms of rheology, swelling behavior and mechanical properties to assess the versatility of the system. Instantaneous as well as long-term structural integrity of the printed hydrogel was achieved with a two-step mechanism combining the thermosensitive properties of gelatin with chemical crosslinking of alginate. Novel syringe tip heaters were developed for improved temperature control of the bioink to avoid clogging. Human mesenchymal stem cells mixed into the hydrogel precursor survived the printing process and showed high cell viability of 85% living cells after 3 days of subsequent in vitro culture. HA enabled the visualization of the printed structures with micro-computed tomography. The inclusion of HA also favors the use of the bioink for bone tissue engineering applications. By adding factors other than HA, the composite could be used as a bioink for applications in drug delivery, microsphere deposition or soft tissue engineering. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Scanning electron microscopy of echinoid podia.

PubMed

Florey, E; Cahill, M A

1982-01-01

Tube feet of the sea urchin Strongylocentrotus franciscanus were studied with the scanning electron microscope (SEM). By use of fractured preparations it was possible to obtain views of all components of the layered tube-foot wall. The outer epithelium was found to bear tufts of cilia possibly belonging to sensory cells. The nerve plexus was clearly revealed as being composed of bundles of varicose axons. The basal lamina, which covers the outer and inner surfaces of the connective tissue layer, was found to be a mechanically resistant and elastic membrane. The connective tissue appears as dense bundles of (collagen) fibers. The luminal epithelium (coelothelium) is a single layer of flagellated collar cells. There is no indication that the muscle fibers, which insert on the inner basal lamina of the connective tissue layer are innervated by axons from the basi-epithelial nerve plexus. The results agree with previous conclusions concerning tube-foot structure based on transmission electron microscopy, and provide additional information, particularly with regard to the outer and inner epithelia.

Characterization of mechanical properties of lamellar structure of the aortic wall: Effect of aging.

PubMed

Taghizadeh, Hadi; Tafazzoli-Shadpour, Mohammad

2017-01-01

Arterial wall tissues are sensitive to their mechanical surroundings and remodel their structure and mechanical properties when subjected to mechanical stimuli such as increased arterial pressure. Such remodeling is evident in hypertension and aging. Aging is characterized by stiffening of the artery wall which is assigned to disturbed elastin function and increased collagen content. To better understand and provide new insight on microstructural changes induced by aging, the lamellar model of the aortic media was utilized to characterize and compare wall structure and mechanical behavior of the young and old human thoracic aortic samples. Such model regards arterial media as two sets of alternating concentric layers, namely sheets of elastin and interlamellar layers. Histological and biaxial tests were performed and microstructural features and stress-strain curves of media were evaluated in young and old age groups. Then using optimization algorithms and hyperelastic constitutive equations the stress-strain curves of layers were evaluated for both age groups. Results indicated slight elevation in the volume fraction of interlamellar layer among old subjects most probably due to age related collagen deposition. Aging indicated substantial stiffening of interlamellar layers accompanied by noticeable softening of elastic lamellae. The general significant stiffening of old samples were attributed to both increase of volume fraction of interlamellar layers and earlier recruitment of collagen fibers during load bearing due to functional loss of elastin within wall lamellae. Mechanical characterization of lamellar structure of wall media is beneficial in study of arterial remodeling in response to alternated mechanical environment in aging and clinical conditions through coupling of wall microstructure and mechanical behavior. Copyright © 2016 Elsevier Ltd. All rights reserved.

Cell reintegration: Stray epithelial cells make their way home.

PubMed

Wilson, Tyler J; Bergstralh, Dan T

2017-06-01

Ongoing work shows that misplaced epithelial cells have the capacity to reintegrate back into tissue layers. This movement appears to underlie tissue stability and may also control aspects of tissue structure. A recent study reveals that cell reintegration in at least one tissue, the Drosophila follicular epithelium, is based on adhesion molecules that line lateral cell surfaces. In this article we will review these observations, discuss their implications for epithelial tissue development and maintenance, and identify future directions for study. © 2017 WILEY Periodicals, Inc.

The narwhal (Monodon monoceros) cementum-dentin junction: a functionally graded biointerphase.

PubMed

Grandfield, Kathryn; Chattah, Netta Lev-Tov; Djomehri, Sabra; Eidelmann, Naomi; Eichmiller, Frederick C; Webb, Samuel; Schuck, P James; Nweeia, Martin; Ho, Sunita P

2014-08-01

In nature, an interface between dissimilar tissues is often bridged by a graded zone, and provides functional properties at a whole organ level. A perfect example is a "biological interphase" between stratified cementum and dentin of a narwhal tooth. This study highlights the graded structural, mechanical, and chemical natural characteristics of a biological interphase known as the cementum-dentin junction layer and their effect in resisting mechanical loads. From a structural perspective, light and electron microscopy techniques illustrated the layer as a wide 1000-2000 μm graded zone consisting of higher density continuous collagen fiber bundles from the surface of cementum to dentin, that parallels hygroscopic 50-100 μm wide collagenous region in human teeth. The role of collagen fibers was evident under compression testing during which the layer deformed more compared to cementum and dentin. This behavior is reflected through site-specific nanoindentation indicating a lower elastic modulus of 2.2 ± 0.5 GPa for collagen fiber bundle compared to 3 ± 0.4 GPa for mineralized regions in the layer. Similarly, microindentation technique illustrated lower hardness values of 0.36 ± 0.05 GPa, 0.33 ± 0.03 GPa, and 0.3 ± 0.07 GPa for cementum, dentin, and cementum-dentin layer, respectively. Biochemical analyses including Raman spectroscopy and synchrotron-source microprobe X-ray fluorescence demonstrated a graded composition across the interface, including a decrease in mineral-to-matrix and phosphate-to-carbonate ratios, as well as the presence of tidemark-like bands with Zn. Understanding the structure-function relationships of wider tissue interfaces can provide insights into natural tissue and organ function. © IMechE 2014.

Ligament flow during drop-on-demand inkjet printing of bioink containing living cells

NASA Astrophysics Data System (ADS)

Zhang, Mengyun; Krishnamoorthy, Srikumar; Song, Hongtao; Zhang, Zhengyi; Xu, Changxue

2017-03-01

Organ printing utilizes tissue spheroids or filaments as building blocks to fabricate three-dimensional (3D) functional tissues and organs based on a layer-by-layer manufacturing mechanism. These fabricated tissues and organs are envisioned as alternatives to replace the damaged human tissues and organs, which is emerging as a promising solution to solve the organ donor shortage problem being faced all over the world. Inkjetting, one of the key technologies in organ printing, has been widely developed because of its moderate fabrication cost, good process controllability, and scale-up potentials. There are several key steps towards inkjet-based organ printing: generation of droplets from bioink, fabrication of 3D cellular structures, and post-printing tissue fusion and maturation. The droplet formation process is the first step, affecting the overall feasibility of the envisioned organ printing technology. This paper focuses on the ligament flow of the droplet formation process during inkjet printing of bioink containing living cells and its corresponding effect on post-printing cell viability and cell distribution. It is found that (1) two types of ligament flow are observed: at 30 V (Type I), the ligament flow has two different directions at the locations near the nozzle orifice and the forming droplet; at 60 V (Type II), the ligament flow directions are the same at both locations; (2) compared to Type II, fewer cells are ejected into the primary droplets in Type I, because some cells move back into the nozzle driven by the ligament flow in the positive z direction; and (3) cell viability in both Type I and Type II is around 90% without a significant difference. The resulting knowledge will benefit precise control of printing dynamics during inkjet printing of viscoelastic bioink for 3D biofabrication applications.

Osteoblasts generate harder, stiffer, and more delamination-resistant mineralized tissue on titanium than on polystyrene, associated with distinct tissue micro- and ultrastructure.

PubMed

Saruwatari, Lei; Aita, Hideki; Butz, Frank; Nakamura, Hiromi K; Ouyang, Jianyong; Yang, Yang; Chiou, Wen-An; Ogawa, Takahiro

2005-11-01

This study revealed that osteoblasts generate harder, stiffer, and more delamination-resistant mineralized tissue on titanium than on the tissue culture polystyrene, associated with modulated gene expression, uniform mineralization, well-crystallized interfacial calcium-phosphate layer, and intensive collagen deposition. Knowledge of this titanium-induced alteration of osteogenic potential leading to enhanced intrinsic biomechanical properties of mineralized tissue provides novel opportunities and implications for understanding and improving bone-titanium integration and engineering physiomechanically tolerant bone. Bone-titanium integration is a biological phenomenon characterized by continuous generation and preservation of peri-implant bone and serves as endosseous anchors against endogenous and exogenous loading, of which mechanisms are poorly understood. This study determines the intrinsic biomechanical properties and interfacial strength of cultured mineralized tissue on titanium and characterizes the tissue structure as possible contributing factors in biomechanical modulation. Rat bone marrow-derived osteoblastic cells were cultured either on a tissue culture-grade polystyrene dish or titanium-coated polystyrene dish having comparable surface topography. Nano-indentation and nano-scratch tests were undertaken on mineralized tissues cultured for 28 days to evaluate its hardness, elastic modulus, and critical load (force required to delaminate tissue). Gene expression was analyzed using RT-PCR. The tissue structural properties were examined by scanning electron microscopy (SEM), collagen colorimetry and localization with Sirius red stain, mineral quantification, and localization with von Kossa stain and transmission electron microscopy (TEM). Hardness and elastic modulus of mineralized tissue on titanium were three and two times greater, respectively, than those on the polystyrene. Three times greater force was required to delaminate the tissue on titanium than that on the polystyrene. SEM of the polystyrene culture displayed a porous structure consisting of fibrous and globular components, whereas the titanium tissue culture appeared to be uniformly solid. Cell proliferation was remarkably reduced on titanium. Microscopic observations revealed that the mineralized tissue on titanium was composed of uniform collagen-supported mineralization from the titanium interface to the outer surface, with intensive collagen deposition at tissue-titanium interface. In contrast, tissue on the polystyrene was characterized by collagen-deficient mineralization at the polystyrene interface and calcium-free collagenous matrix formation in the outer tissue area. Such characteristic microstructure of titanium-associated tissue was corresponded with upregulated gene expression of collagen I and III, osteopontin, and osteocalcin mRNA. Cross-sectional TEM revealed the apposition of a high-contrast and well-crystallized calcium phosphate layer at the titanium interface but not at the polystyrene interface. Culturing osteoblasts on titanium, compared with polystyrene, enhances the hardness, elastic modulus, and interfacial strength of mineralized tissue to a higher degree. Titanium per se possesses an ability to alter cellular phenotypes and tissue micro- and ultrastructure that result in enhanced intrinsic biomechanical properties of mineralized tissue.

Bi-layered collagen nano-structured membrane prototype collagen matrix 10826® for soft tissue regeneration in rabbits: an in vivo ultra-structural study of the early healing phase.

PubMed

De Santis, D; Menchini Fabris, G B; Lotti, J; Palumbo, C; Ferretti, M; Castellani, R; Lotti, T; Zanotti, G; Gelpi, F; Covani, C; Nocini, P F

Collagen Matrix (CM) 10826 is a nanostructured bi-layered collagen membrane obtained from type I and III porcine collagen, which in vitro has shown to have the potential to be a substitute and/or stimulant for soft oral tissue regeneration. The objective of this study was to evaluate the in vivo potential and safety of this membrane for soft tissue regeneration in the early stage of wound healing. Two soft tissue wounds (test and control) were created on the back skin of 5 rabbits (female New Zealand White Rabbits specific pathogen free). All wounds were protected by a special poly-tetra-fluoro-ethylene (PTFE) healing camera. On each rabbit on the test side CM-10826 was used, while on the control side conventional treatment (an autologous pedicle graft) was performed. The healing process was observed clinically after 2 and 6 days, and Magnetic Resonance Imaging (MRI) was performed after this period. After 7 days, animals were sacrificed and specimens were analyzed with light optic microscopy (LM), Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM). These in vivo trials on rabbits confirmed that CM-10826 is well tolerated, without signs of histological inflammatory reaction and proved to be able to accelerate the spontaneous repair of the skin defect taken as the control. The light-optic and ultra-microscopy of serial biopsies showed that the new matrix is biocompatible and is able to function as a scaffold inducing soft tissue regeneration. In conclusion this study demonstrates that CM-10826 promote early soft tissue regeneration and suggests it is a potential constituent for human autologous keratinocytes seeded derma bioequivalent. It protects the wound from injuries and bacterial contamination accelerating healing process. As a clinical relevance, we consider that the quality of life of patients will be improved avoiding the use of major autologous grafts, reducing the hospitalization time and morbidity.

The structure of the conus arteriosus of the sturgeon (Acipenser naccarii) heart: II. The myocardium, the subepicardium, and the conus-aorta transition.

PubMed

Icardo, José M; Colvee, Elvira; Cerra, Maria C; Tota, Bruno

2002-12-01

Sturgeons constitute a family of living "fossil" fish whose heart is related to that of other ancient fish and the elasmobranches. We have undertaken a systematic study of the structure of the sturgeon heart aimed at unraveling the relationship between the heart structure and the adaptive evolutionary changes. In a related paper, data were presented on the conus valves and the subendocardium. Here, the structure of the conus myocardium, the subepicardial tissue, and the conus-aorta transition were studied by conventional light, transmission, and scanning electron microscopy. In addition, actin localization by fluorescent phalloidin was used. The conus myocardium is organized into bundles whose spatial organization changes along the conus length. The variable orientation of the myocardial cell bundles may be effective in emptying the conus lumen during contraction and in preventing reflux of blood. Myocardial cell bundles are separated by loose connective tissue that contains collagen and elastin fibers, vessels, and extremely flat cells separating the cell bundles and enclosing blood vessels and collagen fibers. The ultrastructure of the myocardial cells was found to be very similar to that of other fish groups, suggesting that it is largely conservative. The subepicardium is characterized by the presence of nodular structures that contain lympho-hemopoietic (thymus-like) tissue in the young sturgeons and a large number of lymphocytes after the sturgeons reach sexual maturity. This tissue is likely implicated in the establishment and maintenance of the immune responses. The intrapericardial ventral aorta shows a middle layer of circumferentially oriented cells and internal and external layers with cells oriented longitudinally. Elastin fibers completely surround each smooth muscle cell, and the spaces between the different layers are occupied by randomly arranged collagen bundles. The intrapericardial segment of the ventral aorta is a true transitional segment whose structural characteristics are different from those of both the conus subendocardium and the rest of the ventral aorta. Copyright 2002 Wiley-Liss, Inc.

Bioprinting toward organ fabrication: challenges and future trends.

PubMed

Ozbolat, Ibrahim T; Yu, Yin

2013-03-01

Tissue engineering has been a promising field of research, offering hope for bridging the gap between organ shortage and transplantation needs. However, building three-dimensional (3-D) vascularized organs remains the main technological barrier to be overcome. Organ printing, which is defined as computer-aided additive biofabrication of 3-D cellular tissue constructs, has shed light on advancing this field into a new era. Organ printing takes advantage of rapid prototyping (RP) technology to print cells, biomaterials, and cell-laden biomaterials individually or in tandem, layer by layer, directly creating 3-D tissue-like structures. Here, we overview RP-based bioprinting approaches and discuss the current challenges and trends toward fabricating living organs for transplant in the near future.

Detection of abnormalities in the superficial zone of cartilage repaired using a tissue engineered construct derived from synovial stem cells.

PubMed

Ando, Wataru; Fujie, Hiromichi; Moriguchi, Yu; Nansai, Ryosuke; Shimomura, Kazunori; Hart, David A; Yoshikawa, Hideki; Nakamura, Norimasa

2012-09-28

The present study investigated the surface structure and mechanical properties of repair cartilage generated from a tissue engineered construct (TEC) derived from synovial mesenchymal stem cells at six months post-implantation compared to those of uninjured cartilage. TEC-mediated repair tissue was cartilaginous with Safranin O staining, and had comparable macro-scale compressive properties with uninjured cartilage. However, morphological assessments revealed that the superficial zone of TEC-mediated tissue was more fibrocartilage-like, in contrast to the middle or deep zones that were more hyaline cartilage-like with Safranin O staining. Histological scoring of the TEC-mediated tissue was significantly lower in the superficial zone than in the middle and deep zones. Scanning electron microscopy showed a thick tangential bundle of collagen fibres at the most superficial layer of uninjured cartilage, while no corresponding structure was detected at the surface of TEC-mediated tissue. Immunohistochemical analysis revealed that PRG4 was localised in the superficial area of uninjured cartilage, as well as the TEC-mediated tissue. Friction testing showed that the lubrication properties of the two tissues was similar, however, micro-indentation analysis revealed that the surface stiffness of the TEC-repair tissue was significantly lower than that of uninjured cartilage. Permeability testing indicated that the TEC-mediated tissue exhibited lower water retaining capacity than did uninjured cartilage, specifically at the superficial zone. Thus, TEC-mediated tissue exhibited compromised mechanical properties at the superficial zone, properties which need improvement in the future for maintenance of long term repair cartilage integrity.

Optical Coherence Tomography

NASA Astrophysics Data System (ADS)

Huang, David

Optical coherence tomography (OCT) is a new method for noninvasive cross-sectional imaging in biological systems. In OCT, the longitudinal locations of tissue structures are determined by measuring the time-of-flight delays of light backscattered from these structures. The optical delays are measured by low coherence interferometry. Information on lateral position is provided by transverse scanning of the probe beam. The two dimensional map of optical scattering from internal tissue microstructures is then represented in a false-color or grayscale image. OCT is the optical analog of ultrasonic pulse-echo imaging, but with greatly improved spatial resolutions (a few microns). This thesis describes the development of this new high resolution tomographic imaging technology and the demonstration of its use in a variety of tissues under both in vitro and in vivo conditions. In vitro OCT ranging and imaging studies were performed using human ocular and arterial tissues, two clinically relevant examples of transparent and turbid media, respectively. In the anterior eye, precise measurements of cornea and anterior chamber dimensions were made. In the arterial specimens, the differentiation between fatty -calcified and fibromuscular tissues was demonstrated. In vivo OCT imaging in the retina and optic nerve head in human subjects was also performed. The delineation of retinal layers, which has not been possible with other noninvasive imaging techniques, is demonstrated in these OCT images. OCT has high spatial resolution but limited penetration into turbid tissue. It has potential for diagnostic applications where high resolution is needed and optical access is available, such as in the eye, skin, surgically exposed tissues, and surfaces that can be reached by various catheters and endoscopic probes. In particular, the measurement of fine retinal structures promises improvements in the diagnosis and management of glaucoma, macular edema and other vitreo-retinal diseases. (Copies available exclusively from MIT Libraries, Rm. 14-0551, Cambridge, MA 02139-4307. Ph. 617-253-5668; Fax 617-253 -1690.).

Simple preparation of plant epidermal tissue for laser microdissection and downstream quantitative proteome and carbohydrate analysis

PubMed Central

Falter, Christian; Ellinger, Dorothea; von Hülsen, Behrend; Heim, René; Voigt, Christian A.

2015-01-01

The outwardly directed cell wall and associated plasma membrane of epidermal cells represent the first layers of plant defense against intruding pathogens. Cell wall modifications and the formation of defense structures at sites of attempted pathogen penetration are decisive for plant defense. A precise isolation of these stress-induced structures would allow a specific analysis of regulatory mechanism and cell wall adaption. However, methods for large-scale epidermal tissue preparation from the model plant Arabidopsis thaliana, which would allow proteome and cell wall analysis of complete, laser-microdissected epidermal defense structures, have not been provided. We developed the adhesive tape – liquid cover glass technique (ACT) for simple leaf epidermis preparation from A. thaliana, which is also applicable on grass leaves. This method is compatible with subsequent staining techniques to visualize stress-related cell wall structures, which were precisely isolated from the epidermal tissue layer by laser microdissection (LM) coupled to laser pressure catapulting. We successfully demonstrated that these specific epidermal tissue samples could be used for quantitative downstream proteome and cell wall analysis. The development of the ACT for simple leaf epidermis preparation and the compatibility to LM and downstream quantitative analysis opens new possibilities in the precise examination of stress- and pathogen-related cell wall structures in epidermal cells. Because the developed tissue processing is also applicable on A. thaliana, well-established, model pathosystems that include the interaction with powdery mildews can be studied to determine principal regulatory mechanisms in plant–microbe interaction with their potential outreach into crop breeding. PMID:25870605

Simple preparation of plant epidermal tissue for laser microdissection and downstream quantitative proteome and carbohydrate analysis.

PubMed

Falter, Christian; Ellinger, Dorothea; von Hülsen, Behrend; Heim, René; Voigt, Christian A

2015-01-01

The outwardly directed cell wall and associated plasma membrane of epidermal cells represent the first layers of plant defense against intruding pathogens. Cell wall modifications and the formation of defense structures at sites of attempted pathogen penetration are decisive for plant defense. A precise isolation of these stress-induced structures would allow a specific analysis of regulatory mechanism and cell wall adaption. However, methods for large-scale epidermal tissue preparation from the model plant Arabidopsis thaliana, which would allow proteome and cell wall analysis of complete, laser-microdissected epidermal defense structures, have not been provided. We developed the adhesive tape - liquid cover glass technique (ACT) for simple leaf epidermis preparation from A. thaliana, which is also applicable on grass leaves. This method is compatible with subsequent staining techniques to visualize stress-related cell wall structures, which were precisely isolated from the epidermal tissue layer by laser microdissection (LM) coupled to laser pressure catapulting. We successfully demonstrated that these specific epidermal tissue samples could be used for quantitative downstream proteome and cell wall analysis. The development of the ACT for simple leaf epidermis preparation and the compatibility to LM and downstream quantitative analysis opens new possibilities in the precise examination of stress- and pathogen-related cell wall structures in epidermal cells. Because the developed tissue processing is also applicable on A. thaliana, well-established, model pathosystems that include the interaction with powdery mildews can be studied to determine principal regulatory mechanisms in plant-microbe interaction with their potential outreach into crop breeding.

Validity of the semi-infinite tumor model in diffuse reflectance spectroscopy for epithelial cancer diagnosis: a Monte Carlo study

NASA Astrophysics Data System (ADS)

Zhu, Caigang; Liu, Quan

2011-08-01

The accurate understanding of optical properties of human tissues plays an important role in the optical diagnosis of early epithelial cancer. Many inverse models used to determine the optical properties of a tumor have assumed that the tumor was semi-infinite, which infers infinite width and length but finite thickness. However, this simplified assumption could lead to large errors for small tumor, especially at the early stages. We used a modified Monte Carlo code, which is able to simulate light transport in a layered tissue model with buried tumor-like targets, to investigate the validity of the semi-infinite tumor assumption in two common epithelial tissue models: a squamous cell carcinoma (SCC) tissue model and a basal cell carcinoma (BCC) tissue model. The SCC tissue model consisted of three layers, i.e. the top epithelium, the middle tumor and the bottom stroma. The BCC tissue model also consisted of three layers, i.e. the top epidermis, the middle tumor and the bottom dermis. Diffuse reflectance was simulated for two common fiber-optic probes. In one probe, both source and detector fibers were perpendicular to the tissue surface; while in the other, both fibers were tilted at 45 degrees relative to the normal axis of the tissue surface. It was demonstrated that the validity of the semi-infinite tumor model depends on both the fiber-optic probe configuration and the tumor dimensions. Two look-up tables, which relate the validity of the semi-infinite tumor model to the tumor width in terms of the source-detector separation, were derived to guide the selection of appropriate tumor models and fiber optic probe configuration for the optical diagnosis of early epithelial cancers.

Mesh-based Monte Carlo code for fluorescence modeling in complex tissues with irregular boundaries

NASA Astrophysics Data System (ADS)

Wilson, Robert H.; Chen, Leng-Chun; Lloyd, William; Kuo, Shiuhyang; Marcelo, Cynthia; Feinberg, Stephen E.; Mycek, Mary-Ann

2011-07-01

There is a growing need for the development of computational models that can account for complex tissue morphology in simulations of photon propagation. We describe the development and validation of a user-friendly, MATLAB-based Monte Carlo code that uses analytically-defined surface meshes to model heterogeneous tissue geometry. The code can use information from non-linear optical microscopy images to discriminate the fluorescence photons (from endogenous or exogenous fluorophores) detected from different layers of complex turbid media. We present a specific application of modeling a layered human tissue-engineered construct (Ex Vivo Produced Oral Mucosa Equivalent, EVPOME) designed for use in repair of oral tissue following surgery. Second-harmonic generation microscopic imaging of an EVPOME construct (oral keratinocytes atop a scaffold coated with human type IV collagen) was employed to determine an approximate analytical expression for the complex shape of the interface between the two layers. This expression can then be inserted into the code to correct the simulated fluorescence for the effect of the irregular tissue geometry.

A sparsity-based simplification method for segmentation of spectral-domain optical coherence tomography images

NASA Astrophysics Data System (ADS)

Meiniel, William; Gan, Yu; Olivo-Marin, Jean-Christophe; Angelini, Elsa

2017-08-01

Optical coherence tomography (OCT) has emerged as a promising image modality to characterize biological tissues. With axio-lateral resolutions at the micron-level, OCT images provide detailed morphological information and enable applications such as optical biopsy and virtual histology for clinical needs. Image enhancement is typically required for morphological segmentation, to improve boundary localization, rather than enrich detailed tissue information. We propose to formulate image enhancement as an image simplification task such that tissue layers are smoothed while contours are enhanced. For this purpose, we exploit a Total Variation sparsity-based image reconstruction, inspired by the Compressed Sensing (CS) theory, but specialized for images with structures arranged in layers. We demonstrate the potential of our approach on OCT human heart and retinal images for layers segmentation. We also compare our image enhancement capabilities to the state-of-the-art denoising techniques.

Laminated rare earth structure and method of making

DOEpatents

Senor, David J [West Richland, WA; Johnson, Roger N [Richland, WA; Reid, Bruce D [Pasco, WA; Larson, Sandra [Richland, WA

2002-07-30

A laminated structure having two or more layers, wherein at least one layer is a metal substrate and at least one other layer is a coating comprising at least one rare earth element. For structures having more than two layers, the coating and metal substrate layers alternate. In one embodiment of the invention, the structure is a two-layer laminate having a rare earth coating electrospark deposited onto a metal substrate. In another embodiment of the invention, the structure is a three-layer laminate having the rare earth coating electrospark deposited onto a first metal substrate and the coating subsequently abonded to a second metal substrate. The bonding of the coating to the second metal substrate may be accomplished by hot pressing, hot rolling, high deformation rate processing, or combinations thereof. The laminated structure may be used in nuclear components where reactivity control or neutron absorption is desired and in non-nuclear applications such as magnetic and superconducting films.

Evidence for a Structural Role for Acid-Fast Lipids in Oocyst Walls of Cryptosporidium, Toxoplasma, and Eimeria

PubMed Central

Bushkin, G. Guy; Motari, Edwin; Carpentieri, Andrea; Dubey, Jitender P.; Costello, Catherine E.; Robbins, Phillips W.; Samuelson, John

2013-01-01

ABSTRACT Coccidia are protozoan parasites that cause significant human disease and are of major agricultural importance. Cryptosporidium spp. cause diarrhea in humans and animals, while Toxoplasma causes disseminated infections in fetuses and untreated AIDS patients. Eimeria is a major pathogen of commercial chickens. Oocysts, which are the infectious form of Cryptosporidium and Eimeria and one of two infectious forms of Toxoplasma (the other is tissue cysts in undercooked meat), have a multilayered wall. Recently we showed that the inner layer of the oocyst walls of Toxoplasma and Eimeria is a porous scaffold of fibers of β-1,3-glucan, which are also present in fungal walls but are absent from Cryptosporidium oocyst walls. Here we present evidence for a structural role for lipids in the oocyst walls of Cryptosporidium, Toxoplasma, and Eimeria. Briefly, oocyst walls of each organism label with acid-fast stains that bind to lipids in the walls of mycobacteria. Polyketide synthases similar to those that make mycobacterial wall lipids are abundant in oocysts of Toxoplasma and Eimeria and are predicted in Cryptosporidium. The outer layer of oocyst wall of Eimeria and the entire oocyst wall of Cryptosporidium are dissolved by organic solvents. Oocyst wall lipids are complex mixtures of triglycerides, some of which contain polyhydroxy fatty acyl chains like those present in plant cutin or elongated fatty acyl chains like mycolic acids. We propose a two-layered model of the oocyst wall (glucan and acid-fast lipids) that resembles the two-layered walls of mycobacteria (peptidoglycan and acid-fast lipids) and plants (cellulose and cutin). PMID:24003177

Differentiation of the intracellular structure of slow- versus fast-twitch muscle fibers through evaluation of the dielectric properties of tissue

PubMed Central

Sanchez, B; Li, J; Bragos, R; Rutkove, S B

2014-01-01

Slow-twitch (type 1) skeletal muscle fibers have markedly greater mitochondrial content than fast-twitch (type 2) fibers. Accordingly, we sought to determine whether the dielectric properties of these two fiber types differed, consistent with their distinct intracellular morphologies. The longitudinal and transverse dielectric spectrum of the ex vivo rat soleus (a predominantly type 1 muscle) and the superficial layers of rat gastrocnemius (predominantly type 2) (n = 15) were measured in the 1 kHz–10 MHz frequency range and modeled to a resistivity Cole–Cole function. Major differences were especially apparent in the dielectric spectrum in the 1 to 10 MHz range. Specifically, the gastrocnemius demonstrated a well-defined, higher center frequency than the soleus muscle, whereas the soleus muscle showed a greater difference in the modeled zero and infinite resistivities than the gastrocnemius. These findings are consistent with the fact that soleus tissue has larger and more numerous mitochondria than gastrocnemius. Evaluation of tissue at high frequency could provide a novel approach for assessing intracellular structure in health and disease. PMID:24743385

Differentiation of the intracellular structure of slow- versus fast-twitch muscle fibers through evaluation of the dielectric properties of tissue

NASA Astrophysics Data System (ADS)

Sanchez, B.; Li, J.; Bragos, R.; Rutkove, S. B.

2014-05-01

Slow-twitch (type 1) skeletal muscle fibers have markedly greater mitochondrial content than fast-twitch (type 2) fibers. Accordingly, we sought to determine whether the dielectric properties of these two fiber types differed, consistent with their distinct intracellular morphologies. The longitudinal and transverse dielectric spectrum of the ex vivo rat soleus (a predominantly type 1 muscle) and the superficial layers of rat gastrocnemius (predominantly type 2) (n = 15) were measured in the 1 kHz-10 MHz frequency range and modeled to a resistivity Cole-Cole function. Major differences were especially apparent in the dielectric spectrum in the 1 to 10 MHz range. Specifically, the gastrocnemius demonstrated a well-defined, higher center frequency than the soleus muscle, whereas the soleus muscle showed a greater difference in the modeled zero and infinite resistivities than the gastrocnemius. These findings are consistent with the fact that soleus tissue has larger and more numerous mitochondria than gastrocnemius. Evaluation of tissue at high frequency could provide a novel approach for assessing intracellular structure in health and disease.

Melanoma thickness measurement in two-layer tissue phantoms using pulsed photothermal radiometry (PPTR)

NASA Astrophysics Data System (ADS)

Wang, Tianyi; Qiu, Jinze; Paranjape, Amit; Milner, Thomas E.

2009-02-01

Melanoma is a malignant tumor of melanocytes which are found predominantly in skin. Melanoma is one of the rarer types of skin cancer but causes the majority of skin cancer related deaths. The staging of malignant melanoma using Breslow thickness is important because of the relationship to survival rate after five years. Pulsed photothermal radiometry (PPTR) is based on the time-resolved acquisition of infrared (IR) emission from a sample after pulsed laser exposure. PPTR can be used to investigate the relationship between melanoma thickness and detected radiometric temperature using two-layer tissue phantoms. We used a Monte Carlo simulation to mimic light transport in melanoma and employed a three-dimensional heat transfer model to obtain simulated radiometric temperature increase and, in comparison, we also conducted PPTR experiments to confirm our simulation results. Simulation and experimental results show similar trends: thicker absorbing layers corresponding to deeper lesions produce slower radiometric temperature decays. A quantitative relationship exists between PPTR radiometric temperature decay time and thickness of the absorbing layer in tissue phantoms.

Uniform Deposition of Protein Incorporated Mineral Layer on Three-Dimensional Porous Polymer Scaffolds

PubMed Central

Segvich, Sharon; Smith, Hayes C.; Luong, Linh N.; Kohn, David H.

2009-01-01

Inorganic–organic hybrid materials designed to facilitate bone tissue regeneration use a calcium phosphate mineral layer to encourage cell adhesion, proliferation, and osteogenic differentiation. Mineral formed on porous materials is often discontinuous through the thickness of the scaffold. This study aimed to uniformly coat the pores of three-dimensional (3D) porous, polymer scaffolds with a bone-like mineral layer in addition to uniformly incorporating a model protein within this mineral layer. A filtration system designed to induce simulated body fluid flow through the interstices of 3D polylactic-co-glycolic acid scaffolds (10-mm diameter × 2-mm thickness) illustrated that a uniform, continuous mineral layer can be precipitated on the pore surfaces of a 3D porous structure within 5 days. MicroCT analysis showed increased mineral volume percent (MV%) (7.86 ± 3.25 MV%, p = 0.029) and continuous mineralization of filtered scaffolds compared with two static control groups (floating, 0.16 ± 0.26 MV% and submerged, 0.20 ± 0.01 MV%). Furthermore, the system was effective in coprecipitating a model protein, bone sialoprotein (BSA), within the mineral layer. A 10-fold increase in BSA incorporation was seen when coprecipitated filtered scaffolds (1308 ± 464 μg) were compared to a submerged static control group (139 ± 45 μg), p < 0.001. Confocal microscopy visually confirmed uniform coprecipitation of BSA throughout the thickness of the filtration scaffolds. The designed system enables 3D mineralization through the thickness of porous materials, and provides the option of including coprecipitated biomolecular cues within the mineral layer. This approach of providing a 3D conductive and osteoinductive environment could be conducive to bone tissue regeneration. PMID:17618505

[Tissue engineering of urinary bladder using acellular matrix].

PubMed

Glybochko, P V; Olefir, Yu V; Alyaev, Yu G; Butnaru, D V; Bezrukov, E A; Chaplenko, A A; Zharikova, T M

2017-04-01

Tissue engineering has become a new promising strategy for repairing damaged organs of the urinary system, including the bladder. The basic idea of tissue engineering is to integrate cellular technology and advanced bio-compatible materials to replace or repair tissues and organs. of the study is the objective reflection of the current trends and advances in tissue engineering of the bladder using acellular matrix through a systematic search of preclinical and clinical studies of interest. Relevant studies, including those on methods of tissue engineering of urinary bladder, was retrieved from multiple databases, including Scopus, Web of Science, PubMed, Embase. The reference lists of the retrieved review articles were analyzed for the presence of the missing relevant publications. In addition, a manual search for registered clinical trials was conducted in clinicaltrials.gov. Following the above search strategy, a total of 77 eligible studies were selected for further analysis. Studies differed in the types of animal models, supporting structures, cells and growth factors. Among those, studies using cell-free matrix were selected for a more detailed analysis. Partial restoration of urothelium layer was observed in most studies where acellular grafts were used for cystoplasty, but no the growth of the muscle layer was observed. This is the main reason why cellular structures are more commonly used in clinical practice.

Improved Visualization of Glaucomatous Retinal Damage Using High-speed Ultrahigh-Resolution Optical Coherence Tomography

PubMed Central

Mumcuoglu, Tarkan; Wollstein, Gadi; Wojtkowski, Maciej; Kagemann, Larry; Ishikawa, Hiroshi; Gabriele, Michelle L.; Srinivasan, Vivek; Fujimoto, James G.; Duker, Jay S.; Schuman, Joel S.

2009-01-01

Purpose To test if improving optical coherence tomography (OCT) resolution and scanning speed improves the visualization of glaucomatous structural changes as compared with conventional OCT. Design Prospective observational case series. Participants Healthy and glaucomatous subjects in various stages of disease. Methods Subjects were scanned at a single visit with commercially available OCT (StratusOCT) and high-speed ultrahigh-resolution (hsUHR) OCT. The prototype hsUHR OCT had an axial resolution of 3.4 μm (3 times higher than StratusOCT), with an A-scan rate of 24 000 hertz (60 times faster than StratusOCT). The fast scanning rate allowed the acquisition of novel scanning patterns such as raster scanning, which provided dense coverage of the retina and optic nerve head. Main Outcome Measures Discrimination of retinal tissue layers and detailed visualization of retinal structures. Results High-speed UHR OCT provided a marked improvement in tissue visualization as compared with StratusOCT. This allowed the identification of numerous retinal layers, including the ganglion cell layer, which is specifically prone to glaucomatous damage. Fast scanning and the enhanced A-scan registration properties of hsUHR OCT provided maps of the macula and optic nerve head with unprecedented detail, including en face OCT fundus images and retinal nerve fiber layer thickness maps. Conclusion High-speed UHR OCT improves visualization of the tissues relevant to the detection and management of glaucoma. PMID:17884170

The Architecture of the Connective Tissue in the Musculoskeletal System—An Often Overlooked Functional Parameter as to Proprioception in the Locomotor Apparatus

PubMed Central

van der Wal, Jaap

2009-01-01

The architecture of the connective tissue, including structures such as fasciae, sheaths, and membranes, is more important for understanding functional meaning than is more traditional anatomy, whose anatomical dissection method neglects and denies the continuity of the connective tissue as integrating matrix of the body. The connective tissue anatomy and architecture exhibits two functional tendencies that are present in all areas of the body in different ways and relationships. In body cavities, the “disconnecting” quality of shaping space enables mobility; between organs and body parts, the “connecting” dimension enables functional mechanical interactions. In the musculoskeletal system, those two features of the connective tissue are also present. They cannot be found by the usual analytic dissection procedures. An architectural description is necessary. This article uses such a methodologic approach and gives such a description for the lateral elbow region. The result is an alternative architectural view of the anatomic substrate involved in the transmission and conveyance of forces over synovial joints. An architectural description of the muscular and connective tissue organized in series with each other to enable the transmission of forces over these dynamic entities is more appropriate than is the classical concept of “passive” force-guiding structures such as ligaments organized in parallel to actively force-transmitting structures such as muscles with tendons. The discrimination between so-called joint receptors and muscle receptors is an artificial distinction when function is considered. Mechanoreceptors, also the so-called muscle receptors, are arranged in the context of force circumstances—that is, of the architecture of muscle and connective tissue rather than of the classical anatomic structures such as muscle, capsules, and ligaments. In the lateral cubital region of the rat, a spectrum of mechanosensitive substrate occurs at the transitional areas between regular dense connective tissue layers and the muscle fascicles organized in series with them. This substrate exhibits features of type and location of the mechanosensitive nerve terminals that usually are considered characteristic for “joint receptors” as well as for “muscle receptors.” The receptors for proprioception are concentrated in those areas where tensile stresses are conveyed over the elbow joint. Structures cannot be divided into either joint receptors or muscle receptors when muscular and collagenous connective tissue structures function in series to maintain joint integrity and stability. In vivo, those connective tissue structures are strained during movements of the skeletal parts, those movements in turn being induced and led by tension in muscular tissue. In principle, because of the architecture, receptors can also be stimulated by changes in muscle tension without skeletal movement, or by skeletal movement without change in muscle tension. A mutual relationship exists between structure (and function) of the mechanoreceptors and the architecture of the muscular and regular dense connective tissue. Both are instrumental in the coding of proprioceptive information to the central nervous system. PMID:21589740

SAM-based cell transfer to photopatterned hydrogels for microengineering vascular-like structures.

PubMed

Sadr, Nasser; Zhu, Mojun; Osaki, Tatsuya; Kakegawa, Takahiro; Yang, Yunzhi; Moretti, Matteo; Fukuda, Junji; Khademhosseini, Ali

2011-10-01

A major challenge in tissue engineering is to reproduce the native 3D microvascular architecture fundamental for in vivo functions. Current approaches still lack a network of perfusable vessels with native 3D structural organization. Here we present a new method combining self-assembled monolayer (SAM)-based cell transfer and gelatin methacrylate hydrogel photopatterning techniques for microengineering vascular structures. Human umbilical vein cell (HUVEC) transfer from oligopeptide SAM-coated surfaces to the hydrogel revealed two SAM desorption mechanisms: photoinduced and electrochemically triggered. The former, occurs concomitantly to hydrogel photocrosslinking, and resulted in efficient (>97%) monolayer transfer. The latter, prompted by additional potential application, preserved cell morphology and maintained high transfer efficiency of VE-cadherin positive monolayers over longer culture periods. This approach was also applied to transfer HUVECs to 3D geometrically defined vascular-like structures in hydrogels, which were then maintained in perfusion culture for 15 days. As a step toward more complex constructs, a cell-laden hydrogel layer was photopatterned around the endothelialized channel to mimic the vascular smooth muscle structure of distal arterioles. This study shows that the coupling of the SAM-based cell transfer and hydrogel photocrosslinking could potentially open up new avenues in engineering more complex, vascularized tissue constructs for regenerative medicine and tissue engineering applications. Copyright © 2011 Elsevier Ltd. All rights reserved.

Uric Acid Spherulites in the Reflector Layer of Firefly Light Organ

PubMed Central

Goh, King-Siang; Sheu, Hwo-Shuenn; Hua, Tzu-En; Kang, Mei-Hua; Li, Chia-Wei

2013-01-01

Background In firefly light organs, reflector layer is a specialized tissue which is believed to play a key role for increasing the bioluminescence intensity through reflection. However, the nature of this unique tissue remains elusive. In this report, we investigated the role, fine structure and nature of the reflector layer in the light organ of adult Luciola cerata. Principal Findings Our results indicated that the reflector layer is capable of reflecting bioluminescence, and contains abundant uric acid. Electron microscopy (EM) demonstrated that the cytosol of the reflector layer's cells is filled with densely packed spherical granules, which should be the uric acid granules. These granules are highly regular in size (∼700 nm in diameter), and exhibit a radial internal structure. X-ray diffraction (XRD) analyses revealed that an intense single peak pattern with a d-spacing value of 0.320 nm is specifically detected in the light organ, and is highly similar to the diffraction peak pattern and d-spacing value of needle-formed crystals of monosodium urate monohydrate. However, the molar ratio evaluation of uric acid to various cations (K+, Na+, Ca2+ and Mg2+) in the light organ deduced that only a few uric acid molecules were in the form of urate salts. Thus, non-salt uric acid should be the source of the diffraction signal detected in the light organ. Conclusions In the light organ, the intense single peak diffraction signal might come from a unique needle-like uric acid form, which is different from other known structures of non-salt uric acid form. The finding of a radial structure in the granules of reflector layer implies that the spherical uric acid granules might be formed by the radial arrangement of needle-formed packing matter. PMID:23441187

Induction and differentiation of human induced pluripotent stem cells into functional cardiomyocytes on a compartmented monolayer of gelatin nanofibers

NASA Astrophysics Data System (ADS)

Tang, Yadong; Liu, Li; Li, Junjun; Yu, Leqian; Wang, Li; Shi, Jian; Chen, Yong

2016-07-01

Extensive efforts have been devoted to develop new substrates for culture and differentiation of human induced pluripotent stem cells (hiPSCs) toward cardiac cell-based assays. A more exciting prospect is the construction of cardiac tissue for robust drug screening and cardiac tissue repairing. Here, we developed a patch method by electrospinning and crosslinking of monolayer gelatin nanofibers on a honeycomb frame made of poly(ethylene glycol) diacrylate (PEGDA). The monolayer of the nanofibrous structure can support cells with minimal exogenous contact and a maximal efficiency of cell-medium exchange whereas a single hiPSC colony can be uniformly formed in each of the honeycomb compartments. By modulating the treatment time of the ROCK inhibitor Y-27632, the shape of the hiPSC colony could be controlled from a flat layer to a hemisphere. Afterwards, the induction and differentiation of hiPSCs were achieved on the same patch, leading to a uniform cardiac layer with homogeneous contraction. This cardiac layer could then be used for extracellular recording with a commercial multi-electrode array, showing representative field potential waveforms of matured cardiac tissues with appropriate drug responses.Extensive efforts have been devoted to develop new substrates for culture and differentiation of human induced pluripotent stem cells (hiPSCs) toward cardiac cell-based assays. A more exciting prospect is the construction of cardiac tissue for robust drug screening and cardiac tissue repairing. Here, we developed a patch method by electrospinning and crosslinking of monolayer gelatin nanofibers on a honeycomb frame made of poly(ethylene glycol) diacrylate (PEGDA). The monolayer of the nanofibrous structure can support cells with minimal exogenous contact and a maximal efficiency of cell-medium exchange whereas a single hiPSC colony can be uniformly formed in each of the honeycomb compartments. By modulating the treatment time of the ROCK inhibitor Y-27632, the shape of the hiPSC colony could be controlled from a flat layer to a hemisphere. Afterwards, the induction and differentiation of hiPSCs were achieved on the same patch, leading to a uniform cardiac layer with homogeneous contraction. This cardiac layer could then be used for extracellular recording with a commercial multi-electrode array, showing representative field potential waveforms of matured cardiac tissues with appropriate drug responses. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr04545f

Novel Methods of Automated Quantification of Gap Junction Distribution and Interstitial Collagen Quantity from Animal and Human Atrial Tissue Sections

PubMed Central

Yan, Jiajie; Thomson, Justin K.; Wu, Xiaomin; Zhao, Weiwei; Pollard, Andrew E.; Ai, Xun

2014-01-01

Background Gap junctions (GJs) are the principal membrane structures that conduct electrical impulses between cardiac myocytes while interstitial collagen (IC) can physically separate adjacent myocytes and limit cell-cell communication. Emerging evidence suggests that both GJ and interstitial structural remodeling are linked to cardiac arrhythmia development. However, automated quantitative identification of GJ distribution and IC deposition from microscopic histological images has proven to be challenging. Such quantification is required to improve the understanding of functional consequences of GJ and structural remodeling in cardiac electrophysiology studies. Methods and Results Separate approaches were employed for GJ and IC identification in images from histologically stained tissue sections obtained from rabbit and human atria. For GJ identification, we recognized N-Cadherin (N-Cad) as part of the gap junction connexin 43 (Cx43) molecular complex. Because N-Cad anchors Cx43 on intercalated discs (ID) to form functional GJ channels on cell membranes, we computationally dilated N-Cad pixels to create N-Cad units that covered all ID-associated Cx43 pixels on Cx43/N-Cad double immunostained confocal images. This approach allowed segmentation between ID-associated and non-ID-associated Cx43. Additionally, use of N-Cad as a unique internal reference with Z-stack layer-by-layer confocal images potentially limits sample processing related artifacts in Cx43 quantification. For IC quantification, color map thresholding of Masson's Trichrome blue stained sections allowed straightforward and automated segmentation of collagen from non-collagen pixels. Our results strongly demonstrate that the two novel image-processing approaches can minimize potential overestimation or underestimation of gap junction and structural remodeling in healthy and pathological hearts. The results of using the two novel methods will significantly improve our understanding of the molecular and structural remodeling associated functional changes in cardiac arrhythmia development in aged and diseased hearts. PMID:25105669

Morphologic differentiation of colon carcinoma cell lines HT-29 and HT-29KM in rotating-wall vessels

NASA Technical Reports Server (NTRS)

Goodwin, T. J.; Jessup, J. M.; Wolf, D. A.

1992-01-01

A new low shear stress microcarrier culture system has been developed at NASA's Johnson Space Center that permits three-dimensional tissue culture. Two established human colon adenocarcinoma cell lines, HT-29, an undifferentiated, and HT-29KM, a stable, moderately differentiated subline of HT-29, were grown in new tissue culture bioreactors called Rotating-Wall Vessels (RWVs). RWVs are used in conjunction with multicellular cocultivation to develop a unique in vitro tissue modeling system. Cells were cultivated on Cytodex-3 microcarrier beads, with and without mixed normal human colonic fibroblasts, which served as the mesenchymal layer. Culture of the tumor lines in the absence of fibroblasts produced spheroidlike growth and minimal differentiation. In contrast, when tumor lines were co-cultivated with normal colonic fibroblasts, initial growth was confined to the fibroblast population until the microcarriers were covered. The tumor cells then commenced proliferation at an accelerated rate, organizing themselves into three-dimensional tissue masses that achieved 1.0- to 1.5-cm diameters. The masses displayed glandular structures, apical and internal glandular microvilli, tight intercellular junctions, desmosomes, cellular polarity, sinusoid development, internalized mucin, and structural organization akin to normal colon crypt development. Differentiated samples were subjected to transmission and scanning electron microscopy and histologic analysis, revealing embryoniclike mesenchymal cells lining the areas around the growth matrices. Necrosis was minimal throughout the tissue masses. These data suggest that the RWV affords a new model for investigation and isolation of growth, regulatory, and structural processes within neoplastic and normal tissue.

Medical applications of model-based dynamic thermography

NASA Astrophysics Data System (ADS)

Nowakowski, Antoni; Kaczmarek, Mariusz; Ruminski, Jacek; Hryciuk, Marcin; Renkielska, Alicja; Grudzinski, Jacek; Siebert, Janusz; Jagielak, Dariusz; Rogowski, Jan; Roszak, Krzysztof; Stojek, Wojciech

2001-03-01

The proposal to use active thermography in medical diagnostics is promising in some applications concerning investigation of directly accessible parts of the human body. The combination of dynamic thermograms with thermal models of investigated structures gives attractive possibility to make internal structure reconstruction basing on different thermal properties of biological tissues. Measurements of temperature distribution synchronized with external light excitation allow registration of dynamic changes of local temperature dependent on heat exchange conditions. Preliminary results of active thermography applications in medicine are discussed. For skin and under- skin tissues an equivalent thermal model may be determined. For the assumed model its effective parameters may be reconstructed basing on the results of transient thermal processes. For known thermal diffusivity and conductivity of specific tissues the local thickness of a two or three layer structure may be calculated. Results of some medical cases as well as reference data of in vivo study on animals are presented. The method was also applied to evaluate the state of the human heart during the open chest cardio-surgical interventions. Reference studies of evoked heart infarct in pigs are referred, too. We see the proposed new in medical applications technique as a promising diagnostic tool. It is a fully non-invasive, clean, handy, fast and affordable method giving not only qualitative view of investigated surfaces but also an objective quantitative measurement result, accurate enough for many applications including fast screening of affected tissues.

A tissue-like culture system using microstructures: influence of extracellular matrix material on cell adhesion and aggregation.

PubMed

Knedlitschek, G; Schneider, F; Gottwald, E; Schaller, T; Eschbach, E; Weibezahn, K F

1999-02-01

Special microenvironmental conditions are required to induce and/or maintain specific qualities of differentiated cells. An important parameter is the three-dimensional tissue architecture that cannot be reproduced in conventional monolayer systems. Advanced tissue culture systems will meet many of these demands, but may reach their limits, especially when gradients of specific substances over distinct tissue layers must be established for long-term culture. These limitations may be overcome by incorporating microstructures into tissue-like culture systems. The microstructured cell support presented consists of a flat array of 625 cubic microcontainers with porous bottoms, in which cells can be supplied with specific media from both sides of the tissue layer. Permanent cell lines and primary rat hepatocytes have been used to test the culture system. In order to define reproducible conditions for tissue formation and for cell adherence to the structure, several ECM (extracellular matrix) components were tested for coating of microstructured substrata. The described tissue culture system offers great flexibility in adapting the cell support to specific needs.

Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes.

PubMed

Nanduri, Vibudha; Tattikota, Surendra Mohan; T, Avinash Raj; Sriramagiri, Vijaya Rama Rao; Kantipudi, Suma; Pande, Gopal

2014-06-01

Articular cartilage (AC) injuries and malformations are commonly noticed because of trauma or age-related degeneration. Many methods have been adopted for replacing or repairing the damaged tissue. Currently available AC repair methods, in several cases, fail to yield good-quality long-lasting results, perhaps because the reconstructed tissue lacks the cellular and matrix properties seen in hyaline cartilage (HC). To reconstruct HC tissue from 2-dimensional (2D) and 3-dimensional (3D) cultures of AC-derived human chondrocytes that would specifically exhibit the cellular and biochemical properties of the deep layer of HC. Descriptive laboratory study. Two-dimensional cultures of human AC-derived chondrocytes were established in classical medium (CM) and newly defined medium (NDM) and maintained for a period of 6 weeks. These cells were suspended in 2 mm-thick collagen I gels, placed in 24-well culture inserts, and further cultured up to 30 days. Properties of chondrocytes, grown in 2D cultures and the reconstructed 3D cartilage tissue, were studied by optical and scanning electron microscopic techniques, immunohistochemistry, and cartilage-specific gene expression profiling by reverse transcription polymerase chain reaction and were compared with those of the deep layer of native human AC. Two-dimensional chondrocyte cultures grown in NDM, in comparison with those grown in CM, showed more chondrocyte-specific gene activity and matrix properties. The NDM-grown chondrocytes in 3D cultures also showed better reproduction of deep layer properties of HC, as confirmed by microscopic and gene expression analysis. The method used in this study can yield cartilage tissue up to approximately 1.6 cm in diameter and 2 mm in thickness that satisfies the very low cell density and matrix composition properties present in the deep layer of normal HC. This study presents a novel and reproducible method for long-term culture of AC-derived chondrocytes and reconstruction of cartilage tissue with properties similar to the deep layer of HC in vitro. The HC tissue obtained by the method described can be used to develop an implantable product for the replacement of damaged or malformed AC, especially in younger patients where the lesions are caused by trauma or mechanical stress.

Enamel tissue engineering using subcultured enamel organ epithelial cells in combination with dental pulp cells.

PubMed

Honda, Masaki J; Shinmura, Yuka; Shinohara, Yoshinori

2009-01-01

We describe a strategy for the in vitro engineering of enamel tissue using a novel technique for culturing enamel organ epithelial (EOE) cells isolated from the enamel organ using 3T3-J2 cells as a feeder layer. These subcultured EOE cells retain the capacity to produce enamel structures over a period of extended culture. In brief, enamel organs from 6-month-old porcine third molars were dissociated into single cells and subcultured on 3T3-J2 feeder cell layers. These subcultured EOE cells were then seeded onto a collagen sponge in combination with primary dental pulp cells isolated at an early stage of crown formation, and these constructs were transplanted into athymic rats. After 4 weeks, complex enamel-dentin structures were detected in the implants. These results show that our culture technique maintained ameloblast lineage cells that were able to produce enamel in vivo. This novel subculture technique provides an important tool for tooth tissue engineering. Copyright 2008 S. Karger AG, Basel.

Recent Advances in Bioink Design for 3D Bioprinting of Tissues and Organs.

PubMed

Ji, Shen; Guvendiren, Murat

2017-01-01

There is a growing demand for alternative fabrication approaches to develop tissues and organs as conventional techniques are not capable of fabricating constructs with required structural, mechanical, and biological complexity. 3D bioprinting offers great potential to fabricate highly complex constructs with precise control of structure, mechanics, and biological matter [i.e., cells and extracellular matrix (ECM) components]. 3D bioprinting is an additive manufacturing approach that utilizes a "bioink" to fabricate devices and scaffolds in a layer-by-layer manner. 3D bioprinting allows printing of a cell suspension into a tissue construct with or without a scaffold support. The most common bioinks are cell-laden hydrogels, decellulerized ECM-based solutions, and cell suspensions. In this mini review, a brief description and comparison of the bioprinting methods, including extrusion-based, droplet-based, and laser-based bioprinting, with particular focus on bioink design requirements are presented. We also present the current state of the art in bioink design including the challenges and future directions.

Recent Advances in Bioink Design for 3D Bioprinting of Tissues and Organs

PubMed Central

Ji, Shen; Guvendiren, Murat

2017-01-01

There is a growing demand for alternative fabrication approaches to develop tissues and organs as conventional techniques are not capable of fabricating constructs with required structural, mechanical, and biological complexity. 3D bioprinting offers great potential to fabricate highly complex constructs with precise control of structure, mechanics, and biological matter [i.e., cells and extracellular matrix (ECM) components]. 3D bioprinting is an additive manufacturing approach that utilizes a “bioink” to fabricate devices and scaffolds in a layer-by-layer manner. 3D bioprinting allows printing of a cell suspension into a tissue construct with or without a scaffold support. The most common bioinks are cell-laden hydrogels, decellulerized ECM-based solutions, and cell suspensions. In this mini review, a brief description and comparison of the bioprinting methods, including extrusion-based, droplet-based, and laser-based bioprinting, with particular focus on bioink design requirements are presented. We also present the current state of the art in bioink design including the challenges and future directions. PMID:28424770

Polyethylenimine/silk fibroin multilayers deposited nanofibrics for cell culture.

PubMed

Ye, Xinguo; Li, Sheng; Chen, Xuanxuan; Zhan, Yingfei; Li, Xiaonan

2017-01-01

Scaffold with good three-dimensional (3D) structure and appropriate surface modification is essential to tissue regeneration in the treatment of tissue or organ failure. Silk fibroin (SF) is a promising scaffolding material with high biocompatibility, cytocompatibility, biodegradability and flexibility. In this study, positively charged polyethylenimine (PEI) and negatively charged SF assembled alternately onto cellulose nanofibrous substrates hydrolyzed from electrospun cellulose acetate nanofibrous mats. The obtained nanofibrous membranes modified with multiple layers of PEI/SF were characterized by field emission scanning electron microscopy, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy and thermogravimetric analysis. L929 cells were applied to examine the cytocompatibility of PEI/SF coated membranes. The results demonstrated that the nanofibrous membranes after modification with multiple layers of PEI/SF maintained 3D nanofibrous structure, and cells cultured on them showed good adherence and spreading on them as well, which indicated that PEI/SF coated membranes had potential application in tissue engineering. Copyright © 2016 Elsevier B.V. All rights reserved.

Silicon Carbide Nanoparticles as an Effective Bioadhesive to Bond Collagen Containing Composite Gel Layers for Tissue Engineering Applications.

PubMed

Attalla, Rana; Ling, Celine S N; Selvaganapathy, Ponnambalam Ravi

2018-03-01

Additive manufacturing via layer-by-layer adhesive bonding holds much promise for scalable manufacturing of tissue-like constructs, specifically scaffolds with integrated vascular networks for tissue engineering applications. However, there remains a lack of effective adhesives capable of composite layer fusion without affecting the integrity of patterned features. Here, the use of silicon carbide is introduced as an effective adhesive to achieve strong bonding (0.39 ± 0.03 kPa) between hybrid hydrogel films composed of alginate and collagen. The techniques have allowed us to fabricate multilayered, heterogeneous constructs with embedded high-resolution microchannels (150 µm-1 mm) that are precisely interspaced (500-600 µm). Hydrogel layers are effectively bonded with silicon carbide nanoparticles without blocking the hollow microchannels and high cell viability (90.61 ± 3.28%) is maintained within the scaffold. Nanosilica is also tested and found to cause clogging of smaller microchannels when used for interlayer bonding, but is successfully used to attach synthetic polymers (e.g., Tygon) to the hydrogels (32.5 ± 2.12 mN bond strength). This allows us to form inlet and outlet interconnections to the gel constructs. This ability to integrate hollow channel networks into bulk soft material structures for perfusion can be useful in 3D tissue engineering applications. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Additive manufacturing: From implants to organs.

PubMed

Douglas, Tania S

2014-05-12

Additive manufacturing (AM) constructs 3D objects layer by layer under computer control from 3D models. 3D printing is one example of this kind of technology. AM offers geometric flexibility in its products and therefore allows customisation to suit individual needs. Clinical success has been shown with models for surgical planning, implants, assistive devices and scaffold-based tissue engineering. The use of AM to print tissues and organs that mimic nature in structure and function remains an elusive goal, but has the potential to transform personalised medicine, drug development and scientific understanding of the mechanisms of disease. 

Model based recovery of histological parameters starting from reflectance spectra of the colon

NASA Astrophysics Data System (ADS)

Hidovic-Rowe, Dzena; Claridge, Ela

2005-06-01

Colon cancer alters the tissue macro-architecture. Changes include increase in blood content and distortion of the collagen matrix, which affect the reflectance spectra of the colon and its colouration. We have developed a physics-based model for predicting colon tissue spectra. The colon structure is represented by three layers: mucosa, submucosa and smooth muscle. Each layer is represented by parameters defining its optical properties: molar concentration and absorption coefficients of haemoglobins, describing absorption of light; size and density of collagen fibres; refractive index of the medium and collagen fibres, describing light scattering; and layer thicknesses. Spectra were calculated using the Monte Carlo method. The output of the model was compared to experimental data comprising 50 spectra acquired in vivo from normal tissue. The extracted histological parameters showed good agreement with known values. An experiment was carried out to study the differences between normal and abnormal tissue. These were characterised by increased blood content and decreased collagen density, which is consistent with known differences between normal and abnormal tissue. This suggests that histological quantities of the colon could be computed from its reflectance spectra. The method is likely to have diagnostic value in the early detection of colon cancer.

Spiral-wave dynamics in a mathematical model of human ventricular tissue with myocytes and Purkinje fibers.

PubMed

Nayak, Alok Ranjan; Panfilov, A V; Pandit, Rahul

2017-02-01

We present systematic numerical studies of the possible effects of the coupling of human endocardial and Purkinje cells at cellular and two-dimensional tissue levels. We find that the autorhythmic-activity frequency of the Purkinje cell in a composite decreases with an increase in the coupling strength; this can even eliminate the autorhythmicity. We observe a delay between the beginning of the action potentials of endocardial and Purkinje cells in a composite; such a delay increases as we decrease the diffusive coupling, and eventually a failure of transmission occurs. An increase in the diffusive coupling decreases the slope of the action-potential-duration-restitution curve of an endocardial cell in a composite. By using a minimal model for the Purkinje network, in which we have a two-dimensional, bilayer tissue, with a layer of Purkinje cells on top of a layer of endocardial cells, we can stabilize spiral-wave turbulence; however, for a sparse distribution of Purkinje-ventricular junctions, at which these two layers are coupled, we can also obtain additional focal activity and many complex transient regimes. We also present additional effects resulting from the coupling of Purkinje and endocardial layers and discuss the relation of our results to the studies performed in anatomically accurate models of the Purkinje network.

Spiral-wave dynamics in a mathematical model of human ventricular tissue with myocytes and Purkinje fibers

NASA Astrophysics Data System (ADS)

Nayak, Alok Ranjan; Panfilov, A. V.; Pandit, Rahul

2017-02-01

We present systematic numerical studies of the possible effects of the coupling of human endocardial and Purkinje cells at cellular and two-dimensional tissue levels. We find that the autorhythmic-activity frequency of the Purkinje cell in a composite decreases with an increase in the coupling strength; this can even eliminate the autorhythmicity. We observe a delay between the beginning of the action potentials of endocardial and Purkinje cells in a composite; such a delay increases as we decrease the diffusive coupling, and eventually a failure of transmission occurs. An increase in the diffusive coupling decreases the slope of the action-potential-duration-restitution curve of an endocardial cell in a composite. By using a minimal model for the Purkinje network, in which we have a two-dimensional, bilayer tissue, with a layer of Purkinje cells on top of a layer of endocardial cells, we can stabilize spiral-wave turbulence; however, for a sparse distribution of Purkinje-ventricular junctions, at which these two layers are coupled, we can also obtain additional focal activity and many complex transient regimes. We also present additional effects resulting from the coupling of Purkinje and endocardial layers and discuss the relation of our results to the studies performed in anatomically accurate models of the Purkinje network.

Emerging Techniques in Stratified Designs and Continuous Gradients for Tissue Engineering of Interfaces

PubMed Central

Dormer, Nathan H.; Berkland, Cory J.; Detamore, Michael S.

2013-01-01

Interfacial tissue engineering is an emerging branch of regenerative medicine, where engineers are faced with developing methods for the repair of one or many functional tissue systems simultaneously. Early and recent solutions for complex tissue formation have utilized stratified designs, where scaffold formulations are segregated into two or more layers, with discrete changes in physical or chemical properties, mimicking a corresponding number of interfacing tissue types. This method has brought forth promising results, along with a myriad of regenerative techniques. The latest designs, however, are employing “continuous gradients” in properties, where there is no discrete segregation between scaffold layers. This review compares the methods and applications of recent stratified approaches to emerging continuously graded methods. PMID:20411333

Synthesis, crystal structure and optical properties of two new layered cadmium iodates: Cd(IO{sub 3})X (X=Cl, OH)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yang, Bing-Ping, E-mail: ybp@fjirsm.ac.cn; Mao, Jiang-Gao

Systematic explorations of new compounds in the cadmium iodate system by hydrothermal reactions led to two layered iodates, namely, Cd(IO{sub 3})X (X=Cl, OH). Cd(IO{sub 3})Cl crystallizes in the orthorhombic space group Cmca (No. 64) whereas Cd(IO{sub 3})(OH) crystallizes in the orthorhombic space group Pnma (No. 62). Cd(IO{sub 3})Cl displays a unique double layered structure composed of {sup 1}{sub ∞}[Cd−O{sub 3}Cl]{sub n} chains. Cadmium octahedrons form a 1D chain along the a-axis through edge sharing, and such chains are further interconnected via IO{sub 3} groups to form a special double layer on (020) plane. Cd(IO{sub 3})(OH) also exhibits a layered structuremore » that is composed of cadmium cations, IO{sub 3} groups and hydroxyl ions. Within a layer, chains of CdO{sub 6} edge-shared octahedra are observed along the b-axis. And these chains are connected by IO{sub 3} groups into a layer parallel to the bc plane. Spectroscopic characterizations, elemental analysis, and thermogravimetric analysis for the reported two compounds are also presented. - Graphical abstract: Two new layered cadmium iodates Cd(IO{sub 3})X (X=Cl, OH) are reported. Cd(IO{sub 3})Cl features a unique double layered structure whereas Cd(IO{sub 3})(OH) displays an ordinary layered structure. - Highlights: • Two new layered cadmium iodates Cd(IO{sub 3})X (X=Cl, OH) are reported. • Cd(IO{sub 3})Cl features a unique double layered structure. • Cd(IO{sub 3})(OH) displays an ordinary layered structure. • The spectroscopic and thermal properties have been studied in detail.« less

Fabrication of Orientation-Controlled 3D Tissues Using a Layer-by-Layer Technique and 3D Printed a Thermoresponsive Gel Frame.

PubMed

Tsukamoto, Yoshinari; Akagi, Takami; Shima, Fumiaki; Akashi, Mitsuru

2017-06-01

Herein, we report the fabrication of orientation-controlled tissues similar to heart and nerve tissues using a cell accumulation and three-dimensional (3D) printing technique. We first evaluated the 3D shaping ability of hydroxybutyl chitosan (HBC), a thermoresponsive polymer, by using a robotic dispensing 3D printer. HBC polymer could be laminated to a height of 1124 ± 14 μm. Based on this result, we fabricated 3D gel frames of various shapes, such as square, triangular, rectangular, and circular, for shape control of 3D tissue and then normal human cardiac fibroblasts (NHCFs) coated with extracellular matrix nanofilms were seeded in the frames. Observation of shape-controlled tissues after 1 day of cultivation showed that the orientation of fibroblasts was in one direction when a short-sided, thin, rectangular-shaped frame was used. Next, we tried to fabricate orientation-controlled tissue with a vascular network by coculturing NHCF and normal human cardiac microvascular endothelial cells. As a consequence of cultivation for 4 days, observation of cocultured tissue confirmed aligned cells and blood capillaries in orientation-controlled tissue. Our results clearly demonstrated that it would be possible to control the cell orientation by controlling the shape of the tissues by combining a cell accumulation technique and a 3D printing system. The results of this study suggest promising strategies for the fabrication of oriented 3D tissues in vitro. These tissues, mimicking native organ structures, such as muscle and nerve tissue with a cell alignment structure, would be useful for tissue engineering, regenerative medicine, and pharmaceutical applications.

Improvement of depth resolution on photoacoustic imaging using multiphoton absorption

NASA Astrophysics Data System (ADS)

Yamaoka, Yoshihisa; Fujiwara, Katsuji; Takamatsu, Tetsuro

2007-07-01

Commercial imaging systems, such as computed tomography and magnetic resonance imaging, are frequently used powerful tools for observing structures deep within the human body. However, they cannot precisely visualized several-tens micrometer-sized structures for lack of spatial resolution. In this presentation, we propose photoacoustic imaging using multiphoton absorption technique to generate ultrasonic waves as a means of improving depth resolution. Since the multiphoton absorption occurs at only the focus point and the employed infrared pulses deeply penetrate living tissues, it enables us to extract characteristic features of structures embedded in the living tissue. When nanosecond pulses from a 1064-nm Nd:YAG laser were focused on Rhodamine B/chloroform solution (absorption peak: 540 nm), the peak intensity of the generated photoacoustic signal was proportional to the square of the input pulse energy. This result shows that the photoacoustic signals can be induced by the two-photon absorption of infrared nanosecond pulse laser and also can be detected by a commercial low-frequency MHz transducer. Furthermore, in order to evaluate the depth resolution of multiphoton-photoacoustic imaging, we investigated the dependence of photoacoustic signal on depth position using a 1-mm-thick phantom in a water bath. We found that the depth resolution of two-photon photoacoustic imaging (1064 nm) is greater than that of one-photon photoacoustic imaging (532 nm). We conclude that evolving multiphoton-photoacoustic imaging technology renders feasible the investigation of biomedical phenomena at the deep layer in living tissue.

In vivo imaging of coral tissue and skeleton with optical coherence tomography

PubMed Central

Wentzel, Camilla; Jacques, Steven L.; Wagner, Michael

2017-01-01

Application of optical coherence tomography (OCT) for in vivo imaging of tissue and skeleton structure of intact living corals enabled the non-invasive visualization of coral tissue layers (endoderm versus ectoderm), skeletal cavities and special structures such as mesenterial filaments and mucus release from intact living corals. Coral host chromatophores containing green fluorescent protein-like pigment granules appeared hyper-reflective to near-infrared radiation allowing for excellent optical contrast in OCT and a rapid characterization of chromatophore size, distribution and abundance. In vivo tissue plasticity could be quantified by the linear contraction velocity of coral tissues upon illumination resulting in dynamic changes in the live coral tissue surface area, which varied by a factor of 2 between the contracted and expanded state of a coral. Our study provides a novel view on the in vivo organization of coral tissue and skeleton and highlights the importance of microstructural dynamics for coral ecophysiology. PMID:28250104

High-throughput bone and cartilage micropellet manufacture, followed by assembly of micropellets into biphasic osteochondral tissue.

PubMed

Babur, Betul Kul; Futrega, Kathryn; Lott, William B; Klein, Travis Jacob; Cooper-White, Justin; Doran, Michael Robert

2015-09-01

Engineered biphasic osteochondral tissues may have utility in cartilage defect repair. As bone-marrow-derived mesenchymal stem/stromal cells (MSC) have the capacity to make both bone-like and cartilage-like tissues, they are an ideal cell population for use in the manufacture of osteochondral tissues. Effective differentiation of MSC to bone-like and cartilage-like tissues requires two unique medium formulations and this presents a challenge both in achieving initial MSC differentiation and in maintaining tissue stability when the unified osteochondral tissue is subsequently cultured in a single medium formulation. In this proof-of-principle study, we used an in-house fabricated microwell platform to manufacture thousands of micropellets formed from 166 MSC each. We then characterized the development of bone-like and cartilage-like tissue formation in the micropellets maintained for 8-14 days in sequential combinations of osteogenic or chondrogenic induction medium. When bone-like or cartilage-like micropellets were induced for only 8 days, they displayed significant phenotypic changes when the osteogenic or chondrogenic induction medium, respectively, was swapped. Based on these data, we developed an extended 14-day protocol for the pre-culture of bone-like and cartilage-like micropellets in their respective induction medium. Unified osteochondral tissues were formed by layering 12,000 osteogenic micropellets and 12,000 chondrogenic micropellets into a biphasic structure and then further culture in chondrogenic induction medium. The assembled tissue was cultured for a further 8 days and characterized via histology. The micropellets had amalgamated into a continuous structure with distinctive bone-like and cartilage-like regions. This proof-of-concept study demonstrates the feasibility of micropellet assembly for the formation of osteochondral-like tissues for possible use in osteochondral defect repair.

Layer by Layer Three-dimensional Tissue Epitaxy by Cell-Laden Hydrogel Droplets

PubMed Central

Moon, SangJun; Hasan, Syed K.; Song, Young S.; Xu, Feng; Keles, Hasan Onur; Manzur, Fahim; Mikkilineni, Sohan; Hong, Jong Wook; Nagatomi, Jiro; Haeggstrom, Edward; Khademhosseini, Ali

2010-01-01

The ability to bioengineer three-dimensional (3D) tissues is a potentially powerful approach to treat diverse diseases such as cancer, loss of tissue function, or organ failure. Traditional tissue engineering methods, however, face challenges in fabricating 3D tissue constructs that resemble the native tissue microvasculature and microarchitectures. We have developed a bioprinter that can be used to print 3D patches of smooth muscle cells (5 mm × 5 mm × 81 μm) encapsulated within collagen. Current inkjet printing systems suffer from loss of cell viability and clogging. To overcome these limitations, we developed a system that uses mechanical valves to print high viscosity hydrogel precursors containing cells. The bioprinting platform that we developed enables (i) printing of multilayered 3D cell-laden hydrogel structures (16.2 μm thick per layer) with controlled spatial resolution (proximal axis: 18.0 ± 7.0 μm and distal axis: 0.5 ± 4.9 μm), (ii) high-throughput droplet generation (1 s per layer, 160 droplets/s), (iii) cell seeding uniformity (26 ± 2 cells/mm2 at 1 million cells/mL, 122 ± 20 cells/mm2 at 5 million cells/mL, and 216 ± 38 cells/mm2 at 10 million cells/mL), and (iv) long-term viability in culture (>90%, 14 days). This platform to print 3D tissue constructs may be beneficial for regenerative medicine applications by enabling the fabrication of printed replacement tissues. PMID:19586367

Multilayer solar cell waveguide structures containing metamaterials

NASA Astrophysics Data System (ADS)

Hamouche, Houria.; Shabat, Mohammed. M.; Schaadt, Daniel M.

2017-01-01

Multilayer antireflection coating structures made from silicon and metamaterials are designed and investigated using the Transfer Matrix Method (TMM). The Transfer Matrix Method is a very useful algorithm for the analysis of periodic structures. We investigate in this paper two anti-reflection coating structures for silicon solar cells with a metamaterial film layer. In the first structure, the metamaterial film layer is sandwiched between a semi-infinite glass cover layer and a semi-infinite silicon substrate layer. The second structure consists of a four layers, a pair of metamaterial-dielectric layer with opposite real part of refractive indices, is placed between the two semi-infinite cover and substrate. We have simulated the absorptivity property of the structures for adjustable thicknesses by using MAPLE software. The absorptivity of the structures achieves greater than 80% for incident electromagnetic wave of transverse magnetic (TM) polarization.

Inverse Monte Carlo method in a multilayered tissue model for diffuse reflectance spectroscopy

NASA Astrophysics Data System (ADS)

Fredriksson, Ingemar; Larsson, Marcus; Strömberg, Tomas

2012-04-01

Model based data analysis of diffuse reflectance spectroscopy data enables the estimation of optical and structural tissue parameters. The aim of this study was to present an inverse Monte Carlo method based on spectra from two source-detector distances (0.4 and 1.2 mm), using a multilayered tissue model. The tissue model variables include geometrical properties, light scattering properties, tissue chromophores such as melanin and hemoglobin, oxygen saturation and average vessel diameter. The method utilizes a small set of presimulated Monte Carlo data for combinations of different levels of epidermal thickness and tissue scattering. The path length distributions in the different layers are stored and the effect of the other parameters is added in the post-processing. The accuracy of the method was evaluated using Monte Carlo simulations of tissue-like models containing discrete blood vessels, evaluating blood tissue fraction and oxygenation. It was also compared to a homogeneous model. The multilayer model performed better than the homogeneous model and all tissue parameters significantly improved spectral fitting. Recorded in vivo spectra were fitted well at both distances, which we previously found was not possible with a homogeneous model. No absolute intensity calibration is needed and the algorithm is fast enough for real-time processing.

Radiometric Thermometry for Wearable Deep Tissue Monitoring

NASA Astrophysics Data System (ADS)

Momenroodaki, Parisa

Microwave thermometry is an attractive non-invasive method for measuring internal body temperature. This approach has the potential of enabling a wearable device that can continuously monitor core body temperature. There are a number of health-related applications in both diagnostics and therapy, which can benefit from the knowledge of core body temperature. However,there are a limited number of device solutions, which are usually not wearable or cannot continuously monitor internal body temperature non-invasively. In this thesis, a possible path toward implementing such a thermometer is presented. The device operates in the "quiet" frequency band of 1.4 GHz which is chosen as a compromise between sensing depth and radio frequency interference (RFI). A major challenge in microwave thermometry is detecting small temperature variations of deep tissue layers from surface (skin) measurements. The type and thickness of tissue materials significantly affect the design of the probe, which has the function of receiving black-body radiation from tissues beneath it and coupling the power to a sensitive radiometric receiver. High dielectric constant contrast between skin, fat (/bone), and muscle layers suggests structures with dominant tangential component of the electric field, such as a patch or slot. Adding a layer of low-loss,low-dielectric constant superstrate can further reduce the contribution of superficial tissue layers in the received thermal noise. Several probe types are designed using a full-wave electromagnetic simulator, with a goal of maximizing the power reception from deep tissue layers. The designs are validated with a second software tool and various measurements. A stable, narrow-band, and highly sensitive radiometer is developed, enabling the device to operate in a non-shielded RF environment.To use the microwave thermometer in a RF congested environment, not only narrow-band probe and radiometers are used but an additional probe is introduced for observing the environmental interference. By applying an adaptive filter, the effect of RFI is mitigated in long-term measurements. Several solid and liquid tissue phantoms, required for accurate modeling of the probe and human body interaction, are also developed. The concept of human body microwave thermometry is validated through several measurements on the single-layer and multiple-layer tissue phantoms as well as on the surface of the human body, specifically on the cheek where the internal temperature can easily be changed and independently measured with a thermocouple. Measurement results prove the capability of the device in tracking the temperature of buried tissue layer phantoms to within 0.2K, as well as monitoring internal human body temperature.

Wet-chemical synthesis and applications of non-layer structured two-dimensional nanomaterials

PubMed Central

Tan, Chaoliang; Zhang, Hua

2015-01-01

Non-layer structured nanomaterials with single- or few-layer thickness have two-dimensional sheet-like structures and possess intriguing properties. Recent years have seen major advances in development of a host of non-layer structured ultrathin two-dimensional nanomaterials such as noble metals, metal oxides and metal chalcogenides. The wet-chemical synthesis has emerged as the most promising route towards high-yield and mass production of such nanomaterials. These nanomaterials are now finding increasing applications in a wide range of areas including catalysis, energy production and storage, sensor and nanotherapy, to name but a few. PMID:26303763

Two layer structure for reinforcing pothole repair

NASA Astrophysics Data System (ADS)

Yuan, Wei; Yuan, Kuo-Yao; Zou, Linhua; Yang, Jenn-Ming; Ju, Jiann-Wen; Kao, Wei; Carlson, Larry

2013-04-01

We have applied dicyclopentadiene (DCPD) resin for reinforcing pothole patch materials due to its unique properties - low cost, low viscosity at beginning and ultra-toughness after curing, chemical compatibility with tar, tunable curing profile through catalyst design. In this paper, we have designed a two layer structure - well compacted base layer and DCPD reinforced 1-1.5" top layer - for pothole repair. By choosing two graded asphalt mixes, a porous top layer and fully compacted base layer was prepared after compaction and ready for DCPD resin infiltration. The DCPD curing and infiltration profile within this porous top layer was measured with thermocouples. The rutting resistance was tested with home-made wheel rutter. The cage effect due to the p-DCPD wrapping was characterized with wheel penetration test. The results showed that this two layer structure pothole repair has greatly improved properties and can be used for pothole repair to increase the service life.

Determination of absorption changes from moments of distributions of times of flight of photons: optimization of measurement conditions for a two-layered tissue model.

PubMed

Liebert, Adam; Wabnitz, Heidrun; Elster, Clemens

2012-05-01

Time-resolved near-infrared spectroscopy allows for depth-selective determination of absorption changes in the adult human head that facilitates separation between cerebral and extra-cerebral responses to brain activation. The aim of the present work is to analyze which combinations of moments of measured distributions of times of flight (DTOF) of photons and source-detector separations are optimal for the reconstruction of absorption changes in a two-layered tissue model corresponding to extra- and intra-cerebral compartments. To this end we calculated the standard deviations of the derived absorption changes in both layers by considering photon noise and a linear relation between the absorption changes and the DTOF moments. The results show that the standard deviation of the absorption change in the deeper (superficial) layer increases (decreases) with the thickness of the superficial layer. It is confirmed that for the deeper layer the use of higher moments, in particular the variance of the DTOF, leads to an improvement. For example, when measurements at four different source-detector separations between 8 and 35 mm are available and a realistic thickness of the upper layer of 12 mm is assumed, the inclusion of the change in mean time of flight, in addition to the change in attenuation, leads to a reduction of the standard deviation of the absorption change in the deeper tissue layer by a factor of 2.5. A reduction by another 4% can be achieved by additionally including the change in variance.

OCT/PS-OCT imaging of brachial plexus neurovascular structures

NASA Astrophysics Data System (ADS)

Raphael, David T.; Zhang, Jun; Zhang, Yaoping; Chen, Zhongping; Miller, Carol; Zhou, Li

2004-07-01

Introduction: Optical coherence tomography (OCT) allows high-resolution imaging (less than 10 microns) of tissue structures. A pilot study with OCT and polarization-sensitive OCT (PS-OCT) was undertaken to image ex-vivo neurovascular structures (vessels, nerves) of the canine brachial plexus. Methods: OCT is an interferometry-based optical analog of B-mode ultrasound, which can image through non-transparent biological tissues. With approval of the USC Animal Care and Use Committee, segments of the supra- and infraclavicular brachial plexus were excised from euthanized adult dogs, and the ex-vivo specimens were placed in cold pH-buffered physiologic solution. An OCT beam, in micrometer translational steps, scanned the fixed-position bisected specimens in transverse and longitudinal views. Two-dimensional images were obtained from identified arteries and nerves, with specific sections of interest stained with hematoxylin-eosin for later imaging through a surgical microscope. Results: with the beam scan direction transverse to arteries, the resulting OCT images showed an identifiable arterial lumen and arterial wall tissue layers. By comparison, transverse beam OCT images of nerves revealed a multitude of smaller nerve bundles contained within larger circular-shaped fascicles. PS-OCT imaging was helpful in showing the characteristic birefringence exhibited by arrayed neural structures. Discussion: High-resolution OCT imaging may be useful in the optical identification of neurovascular structures during attempted regional nerve blockade. If incorporated into a needle-shaped catheter endoscope, such a technology could prevent intraneural and intravascular injections immediately prior to local anesthetic injection. The major limitation of OCT is that it can form a coherent image of tissue structures only to a depth of 1.5 - 2 mm.

Influence of head size on the development of metallic wear and on the characteristics of carbon layers in metal-on-metal hip joints

PubMed Central

Sprecher, Christoph M; Wimmer, Markus A; Milz, Stefan; Taeger, Georg

2009-01-01

Background and purpose Particles originating from the articulating surfaces of hip endoprostheses often induce an inflammatory response, which can be related to implant failure. We therefore analyzed the metal content in capsular tissue from 44 McKee-Farrar metal-on-metal hip prostheses (with 3 different head sizes) and we also analyzed the morphological structure of layers located on articulating surfaces. Methods Atomic absorption spectrometry (AAS) was used to analyze the metal content in capsular tissue. Visually detectable carbon layers located on the articulating surfaces were evaluated using scanning electron microscopy (SEM), energy-dispersive Xray spectroscopy (EDX), and X-ray photoelectron spectroscopy (XPS). Results Metallic debris was detected in all capsular tissue samples but no statistically significant differences in metal content were found in relation to implant head size. The morphological characteristics of the different layer zones allowed an exact analysis of contact and non-contact areas. Furthermore, surface layers appear to have a protective function because they can prevent sharp-edged particles from damaging the prostheses surface. Interpretation The implant head size does not appear to influence the amount of metallic debris. The layers obviously act like a lubricating agent because the protection function does not occur in regions without layers where the metal surface often shows numerous scratches. As layers are not generated immediately after the implantation of hip prostheses, these findings may at least partially explain the high amount of wear early after implantation. PMID:19421914

Cytologic appearance of retinal cells included in a fine-needle aspirate of a meningioma around the optic nerve of a dog.

PubMed

Tvedten, Harold; Hillström, Anna

2013-06-01

A 6-year-old Wirehair Dachshund had a meningioma around the optic nerve that caused exophthalmos. A benign mesenchymal tumor was suspected based on the cytologic pattern of a fine-needle aspirate, and a meningioma was diagnosed by histopathologic examination. In addition to the meningioma cells, the cytologic smears included groups of cells from apparently 4 layers of normal retina. In particular, uniform rod-shaped structures in the cytologic sample could suggest rod-shaped bacteria, but these structures were identified as cylindrical outer segments of photoreceptor rod cells. Other retinal structures recognized included pigmented epithelial layer cells with their uniquely formed pigment granules, the characteristic bi-lobed, cleaved nuclei from the outer nuclear layer, and nerve tissue likely from the outer plexiform layer of the retina. © 2013 American Society for Veterinary Clinical Pathology.

Failure mechanisms of additively manufactured porous biomaterials: Effects of porosity and type of unit cell.

PubMed

Kadkhodapour, J; Montazerian, H; Darabi, A Ch; Anaraki, A P; Ahmadi, S M; Zadpoor, A A; Schmauder, S

2015-10-01

Since the advent of additive manufacturing techniques, regular porous biomaterials have emerged as promising candidates for tissue engineering scaffolds owing to their controllable pore architecture and feasibility in producing scaffolds from a variety of biomaterials. The architecture of scaffolds could be designed to achieve similar mechanical properties as in the host bone tissue, thereby avoiding issues such as stress shielding in bone replacement procedure. In this paper, the deformation and failure mechanisms of porous titanium (Ti6Al4V) biomaterials manufactured by selective laser melting from two different types of repeating unit cells, namely cubic and diamond lattice structures, with four different porosities are studied. The mechanical behavior of the above-mentioned porous biomaterials was studied using finite element models. The computational results were compared with the experimental findings from a previous study of ours. The Johnson-Cook plasticity and damage model was implemented in the finite element models to simulate the failure of the additively manufactured scaffolds under compression. The computationally predicted stress-strain curves were compared with the experimental ones. The computational models incorporating the Johnson-Cook damage model could predict the plateau stress and maximum stress at the first peak with less than 18% error. Moreover, the computationally predicted deformation modes were in good agreement with the results of scaling law analysis. A layer-by-layer failure mechanism was found for the stretch-dominated structures, i.e. structures made from the cubic unit cell, while the failure of the bending-dominated structures, i.e. structures made from the diamond unit cells, was accompanied by the shearing bands of 45°. Copyright © 2015 Elsevier Ltd. All rights reserved.

Thermal Inkjet Printing in Tissue Engineering and Regenerative Medicine

PubMed Central

Cui, Xiaofeng; Boland, Thomas; D’Lima, Darryl D.; Lotz, Martin K.

2013-01-01

With the advantages of high throughput, digital control, and highly accurate placement of cells and biomaterial scaffold to the desired 2D and 3D locations, bioprinting has great potential to develop promising approaches in translational medicine and organ replacement. The most recent advances in organ and tissue bioprinting based on the thermal inkjet printing technology are described in this review. Bioprinting has no or little side effect to the printed mammalian cells and it can conveniently combine with gene transfection or drug delivery to the ejected living systems during the precise placement for tissue construction. With layer-by-layer assembly, 3D tissues with complex structures can be printed using scanned CT or MRI images. Vascular or nerve systems can be enabled simultaneously during the organ construction with digital control. Therefore, bioprinting is the only solution to solve this critical issue in thick and complex tissues fabrication with vascular system. Collectively, bioprinting based on thermal inkjet has great potential and broad applications in tissue engineering and regenerative medicine. This review article introduces some important patents related to bioprinting living systems and the bioprinting in tissue engineering field. PMID:22436025

Endomembrane proteomics reveals putative enzymes involved in cell wall metabolism in wheat grain outer layers

PubMed Central

Chateigner-Boutin, Anne-Laure; Suliman, Muhtadi; Bouchet, Brigitte; Alvarado, Camille; Lollier, Virginie; Rogniaux, Hélène; Guillon, Fabienne; Larré, Colette

2015-01-01

Cereal grain outer layers fulfil essential functions for the developing seed such as supplying energy and providing protection. In the food industry, the grain outer layers called ‘the bran’ is valuable since it is rich in dietary fibre and other beneficial nutriments. The outer layers comprise several tissues with a high content in cell wall material. The cell wall composition of the grain peripheral tissues was investigated with specific probes at a stage of active cell wall synthesis. Considerable wall diversity between cell types was revealed. To identify the cellular machinery involved in cell wall synthesis, a subcellular proteomic approach was used targeting the Golgi apparatus where most cell wall polysaccharides are synthesized. The tissues were dissected into outer pericarp and intermediate layers where 822 and 1304 proteins were identified respectively. Many carbohydrate-active enzymes were revealed: some in the two peripheral grain fractions, others only in one tissue. Several protein families specific to one fraction and with characterized homologs in other species might be related to the specific detection of a polysaccharide in a particular cell layer. This report provides new information on grain cell walls and its biosynthesis in the valuable outer tissues, which are poorly studied so far. A better understanding of the mechanisms controlling cell wall composition could help to improve several quality traits of cereal products (e.g. dietary fibre content, biomass conversion to biofuel). PMID:25769308

Polarized light imaging specifies the anisotropy of light scattering in the superficial layer of a tissue

PubMed Central

Jacques, Steven L.; Roussel, Stéphane; Samatham, Ravikant

2016-01-01

Abstract. This report describes how optical images acquired using linearly polarized light can specify the anisotropy of scattering (g) and the ratio of reduced scattering [μs′=μs(1−g)] to absorption (μa), i.e., N′=μs′/μa. A camera acquired copolarized (HH) and crosspolarized (HV) reflectance images of a tissue (skin), which yielded images based on the intensity (I=HH+HV) and difference (Q=HH−HV) of reflectance images. Monte Carlo simulations generated an analysis grid (or lookup table), which mapped Q and I into a grid of g versus N′, i.e., g(Q,I) and N′(Q,I). The anisotropy g is interesting because it is sensitive to the submicrometer structure of biological tissues. Hence, polarized light imaging can monitor shifts in the submicrometer (50 to 1000 nm) structure of tissues. The Q values for forearm skin on two subjects (one Caucasian, one pigmented) were in the range of 0.046±0.007 (24), which is the mean±SD for 24 measurements on 8 skin sites×3 visible wavelengths, 470, 524, and 625 nm, which indicated g values of 0.67±0.07 (24). PMID:27165546

Polyurethane foam/nano hydroxyapatite composite as a suitable scaffold for bone tissue regeneration.

PubMed

Meskinfam, M; Bertoldi, S; Albanese, N; Cerri, A; Tanzi, M C; Imani, R; Baheiraei, N; Farokhi, M; Farè, S

2018-01-01

In bone tissue regeneration, the use of biomineralized scaffolds to create the 3D porous structure needed for well-fitting with defect size and appropriate cell interactions, is a promising alternative to autologous and heterologous bone grafts. Biomineralized polyurethane (PU) foams are here investigated as scaffold for bone tissue regeneration. Biomineralization of the foams was carried out by activation of PU surface by a two steps procedure performed for different times (1 to 4 weeks). Scaffolds were investigated for morphological, chemico-physical and mechanical properties, as well as for in vitro interaction with rat Bone Marrow Mesenchymal Stem Cells (BMSCs). Untreated and biomineralized PU samples showed a homogenous morphology and regular pore size (average Ø=407μm). Phase and structure of formed calcium phosphates (CaPs) layer onto the PU foam were analyzed by Fourier Transform Infrared spectroscopy and X-ray diffraction, proving the formation of bone-like nano hydroxyapatite. Biomineralization caused a significant increase of mechanical properties of treated foams compared to untreated ones. Biomineralization also affected the PU scaffold cytocompatibility providing a more appropriate surface for cell attachment and proliferation. Considering the obtained results, the proposed scaffold can be considered suitable for bone tissue regeneration. Copyright © 2017 Elsevier B.V. All rights reserved.

3D bioprinting of biomimetic aortic vascular constructs with self-supporting cells.

PubMed

Kucukgul, Can; Ozler, S Burce; Inci, Ilyas; Karakas, Ezgi; Irmak, Ster; Gozuacik, Devrim; Taralp, Alpay; Koc, Bahattin

2015-04-01

Cardiovascular diseases are the leading cause of deaths throughout the world. Vascular diseases are mostly treated with autografts and blood vessel transplantations. However, traditional grafting methods have several problems including lack of suitable harvest sites, additional surgical costs for harvesting procedure, pain, infection, lack of donors, and even no substitutes at all. Recently, tissue engineering and regenerative medicine approaches are used to regenerate damaged or diseased tissues. Most of the tissue engineering investigations have been based on the cell seeding into scaffolds by providing a suitable environment for cell attachment, proliferation, and differentiation. Because of the challenges such as difficulties in seeding cells spatially, rejection, and inflammation of biomaterials used, the recent tissue engineering studies focus on scaffold-free techniques. In this paper, the development of novel computer aided algorithms and methods are developed for 3D bioprinting of scaffold-free biomimetic macrovascular structures. Computer model mimicking a real human aorta is generated using imaging techniques and the proposed computational algorithms. An optimized three-dimensional bioprinting path planning are developed with the proposed self-supported model. Mouse embryonic fibroblast (MEF) cell aggregates and support structures (hydrogels) are 3D bioprinted layer-by-layer according to the proposed self-supported method to form an aortic tissue construct. © 2014 Wiley Periodicals, Inc.

Structure and function of the digestive system of solen grandis dunker

NASA Astrophysics Data System (ADS)

Sheng, Xiuzhen; Zhan, Wenbin; Ren, Sulian

2003-10-01

Structure and function of the digestive system of a bivalve mollusc, Solen grandis, were studied using light microscopy and histochemical methods. The wall of digestive tube consists of four layers: the mucosal epithelium, connective tissue, muscular and fibrosa or serosa (only in the portion of rectum) from the inner to the outer. The ciliated columnar epithelial cells, dispersed by cup-shaped mucous cells, rest on a thin base membrane. There are abundant blood spaces in connective tissue layer. The digestive diverticula are composed of multi-branched duct and digestive tubules. The digestive tubules are lined with digestive and basophilic secretory cells, and surrounded by a layer of smooth muscle fibers and connective tissues. Activities of acid and alkaline phosphatases, esterase and lipase are detected in the digestive cells, and the epithelia of stomach and intestine, suggesting that these cells are capable of intracellular digesting of food materials and absorbing. Besides, acid phosphatase and esterase activities are present in the posterior portion of esophagus. Phagocytes are abundant in blood spaces and the lumens of stomach and intestine, containing brown granules derived from the engulfed food materials. The present work indicates that phagocytes play important roles in ingestion and digestion of food materials, which is supported as well by the activities of acid phosphatase, esterase and lipase detected in blood spaces.

Deep learning classifier with optical coherence tomography images for early dental caries detection

NASA Astrophysics Data System (ADS)

Karimian, Nima; Salehi, Hassan S.; Mahdian, Mina; Alnajjar, Hisham; Tadinada, Aditya

2018-02-01

Dental caries is a microbial disease that results in localized dissolution of the mineral content of dental tissue. Despite considerable decline in the incidence of dental caries, it remains a major health problem in many societies. Early detection of incipient lesions at initial stages of demineralization can result in the implementation of non-surgical preventive approaches to reverse the demineralization process. In this paper, we present a novel approach combining deep convolutional neural networks (CNN) and optical coherence tomography (OCT) imaging modality for classification of human oral tissues to detect early dental caries. OCT images of oral tissues with various densities were input to a CNN classifier to determine variations in tissue densities resembling the demineralization process. The CNN automatically learns a hierarchy of increasingly complex features and a related classifier directly from training data sets. The initial CNN layer parameters were randomly selected. The training set is split into minibatches, with 10 OCT images per batch. Given a batch of training patches, the CNN employs two convolutional and pooling layers to extract features and then classify each patch based on the probabilities from the SoftMax classification layer (output-layer). Afterward, the CNN calculates the error between the classification result and the reference label, and then utilizes the backpropagation process to fine-tune all the layer parameters to minimize this error using batch gradient descent algorithm. We validated our proposed technique on ex-vivo OCT images of human oral tissues (enamel, cortical-bone, trabecular-bone, muscular-tissue, and fatty-tissue), which attested to effectiveness of our proposed method.

Phantom with pulsatile arteries to investigate the influence of blood vessel depth on pulse oximeter signal strength.

PubMed

Stuban, Norbert; Niwayama, Masatsugu; Santha, Hunor

2012-01-01

This paper describes a three-layer head phantom with artificial pulsating arteries at five different depths (1.2 mm, 3.7 mm, 6.8 mm, 9.6 mm and 11.8 mm). The structure enables formation of spatially and temporally varying tissue properties similar to those of living tissues. In our experiment, pressure pulses were generated in the arteries by an electronically controlled pump. The physical and optical parameters of the layers and the liquid in the artificial arteries were similar to those of real tissues and blood. The amplitude of the pulsating component of the light returning from the phantom tissues was measured at each artery depth mentioned above. The build-up of the in-house-developed pulse oximeter used for performing the measurements and the physical layout of the measuring head are described. The radiant flux generated by the LED on the measuring head was measured to be 1.8 mW at 910 nm. The backscattered radiant flux was measured, and found to be 0.46 nW (0.26 ppm), 0.55 nW (0.31 ppm), and 0.18 nW (0.10 ppm) for the 1.2 mm, 3.7 mm and 6.8 mm arteries, respectively. In the case of the 9.6 mm and 11.8 mm arteries, useful measurement data were not obtained owing to weak signals. We simulated the phantom with the arteries at the above-mentioned five depths and at two additional ones (2.5 mm and 5.3 mm in depth) using the Monte Carlo method. The measurement results were verified by the simulation results. We concluded that in case of 11 mm source-detector separation the arteries at a depth of about 2.5 mm generate the strongest pulse oximeter signal level in a tissue system comprising three layers of thicknesses: 1.5 mm (skin), 5.0 mm (skull), and >50 mm (brain).

Phantom with Pulsatile Arteries to Investigate the Influence of Blood Vessel Depth on Pulse Oximeter Signal Strength

PubMed Central

Stuban, Norbert; Niwayama, Masatsugu; Santha, Hunor

2012-01-01

This paper describes a three-layer head phantom with artificial pulsating arteries at five different depths (1.2 mm, 3.7 mm, 6.8 mm, 9.6 mm and 11.8 mm). The structure enables formation of spatially and temporally varying tissue properties similar to those of living tissues. In our experiment, pressure pulses were generated in the arteries by an electronically controlled pump. The physical and optical parameters of the layers and the liquid in the artificial arteries were similar to those of real tissues and blood. The amplitude of the pulsating component of the light returning from the phantom tissues was measured at each artery depth mentioned above. The build-up of the in-house-developed pulse oximeter used for performing the measurements and the physical layout of the measuring head are described. The radiant flux generated by the LED on the measuring head was measured to be 1.8 mW at 910 nm. The backscattered radiant flux was measured, and found to be 0.46 nW (0.26 ppm), 0.55 nW (0.31 ppm), and 0.18 nW (0.10 ppm) for the 1.2 mm, 3.7 mm and 6.8 mm arteries, respectively. In the case of the 9.6 mm and 11.8 mm arteries, useful measurement data were not obtained owing to weak signals. We simulated the phantom with the arteries at the above-mentioned five depths and at two additional ones (2.5 mm and 5.3 mm in depth) using the Monte Carlo method. The measurement results were verified by the simulation results. We concluded that in case of 11 mm source-detector separation the arteries at a depth of about 2.5 mm generate the strongest pulse oximeter signal level in a tissue system comprising three layers of thicknesses: 1.5 mm (skin), 5.0 mm (skull), and >50 mm (brain). PMID:22368501

Imaging of oral pathological tissue using optical coherence tomography

NASA Astrophysics Data System (ADS)

Canjau, Silvana; Todea, Carmen; Sinescu, Cosmin; Duma, Virgil-Florin; Topala, Florin I.; Podoleanu, Adrian G.

2014-01-01

Oral squamous cell carcinoma (OSCC) constitutes 90% of oral cancer. Early detection is a cornerstone to improve survival. Interaction of light with tissues may highlight changes in tissue structure and metabolism. We propose optical coherence tomography (OCT), as a non-invasive diagnosis method, being a new high-resolution optical technique that permits tri-dimensional (3-D), real-time imaging of near surface abnormalities in complex tissues. In this study half of the excisional biopsy was directed to the pathologist and the other half was assigned for OCT investigation. Histopathology validated the results. Areas of OSCC of the buccal mucosa were identified in the OCT images. The elements obserced included extensive epithelial down-growth, the disruption of the basement membrane, with areas of erosion, an epithelial layer that was highly variable in thickness and invasion into the sub-epithelial layers. Therefore, OCT appears to be a highly promising imaging modality.

In vivo measurement of skin surface strain and sub-surface layer deformation induced by natural tissue stretching.

PubMed

Maiti, Raman; Gerhardt, Lutz-Christian; Lee, Zing S; Byers, Robert A; Woods, Daniel; Sanz-Herrera, José A; Franklin, Steve E; Lewis, Roger; Matcher, Stephen J; Carré, Matthew J

2016-09-01

Stratum corneum and epidermal layers change in terms of thickness and roughness with gender, age and anatomical site. Knowledge of the mechanical and tribological properties of skin associated with these structural changes are needed to aid in the design of exoskeletons, prostheses, orthotics, body mounted sensors used for kinematics measurements and in optimum use of wearable on-body devices. In this case study, optical coherence tomography (OCT) and digital image correlation (DIC) were combined to determine skin surface strain and sub-surface deformation behaviour of the volar forearm due to natural tissue stretching. The thickness of the epidermis together with geometry changes of the dermal-epidermal junction boundary were calculated during change in the arm angle, from flexion (90°) to full extension (180°). This posture change caused an increase in skin surface Lagrange strain, typically by 25% which induced considerable morphological changes in the upper skin layers evidenced by reduction of epidermal layer thickness (20%), flattening of the dermal-epidermal junction undulation (45-50% reduction of flatness being expressed as Ra and Rz roughness profile height change) and reduction of skin surface roughness Ra and Rz (40-50%). The newly developed method, DIC combined with OCT imaging, is a powerful, fast and non-invasive methodology to study structural skin changes in real time and the tissue response provoked by mechanical loading or stretching. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

SEM and EDS investigation of a pyrolytic carbon covered C/C composite maxillofacial implant retrieved from the human body after 8 years.

PubMed

Sebők, Béla; Kiss, Gábor; Szabó, Péter J; Rigler, Dániel; Molnár, Milán L; Dobos, Gábor; Réti, Ferenc; Szőcs, Hajnal; Joób, Arpád F; Bogdán, Sándor; Szabó, György

2013-03-01

The long term effect of the human body on a pyrolytic carbon covered C/C composite maxillofacial implant (CarBulat(Tm)) was investigated by comparing the structure, the surface morphology and composition of an implant retrieved after 8 years to a sterilized, but not implanted one. Although the thickness of the carbon fibres constituting the implants did not change during the 8 year period, the surface of the implant retrieved was covered with a thin surface layer not present on the unimplanted implant. The composition of this layer is identical to the composition of the underlying carbon fibres. Calcium can only be detected on the surface as a trace element implying that the new layer is not formed by bone tissue. Residual soft tissue penetrating the bulk material between the carbon fibre bunches was found on the retrieved implant indicating the importance of the surface morphology in tissue growth and adhering to implants.

Optical assessment of tissue anisotropy in ex vivo distended rat bladders

NASA Astrophysics Data System (ADS)

Alali, Sanaz; Aitken, Karen J.; Shröder, Annette; Bagli, Darius J.; Alex Vitkin, I.

2012-08-01

Microstructural remodelling in epithelial layers of various hollow organs, including changes in tissue anisotropy, are known to occur under mechanical distension and during disease processes. In this paper, we analyze how bladder distension alters wall anisotropy using polarized light imaging (followed by Mueller matrix decomposition). Optical retardance values of different regions of normal rat bladders under different distension pressures are derived. Then optical coherence tomography is used to measure local bladder wall thicknesses, enabling the calculation of the tissue birefringence maps as a measure of the tissue anisotropy. Selected two-photon microscopy is also performed to better understand the compositional origins of the obtained anisotropy results. The dome region of the bladder shows maximum birefringence when the bladder is distended to high pressures, whereas the ventral remains roughly isotropic during distension. In addition, the average anisotropy direction is longitudinal, along the urethra to dome. The derived wall anisotropy trends are based on birefringence as an intrinsic property of the tissue organization independent of its thickness, to aid in understanding the structure-functions relation in healthy bladders. These new insights into the wall microstructure of ex vivo distending bladders may help improve the functionality of the artificially engineered bladder tissues.

3D-printed membrane for guided tissue regeneration.

PubMed

Tayebi, Lobat; Rasoulianboroujeni, Morteza; Moharamzadeh, Keyvan; Almela, Thafar K D; Cui, Zhanfeng; Ye, Hua

2018-03-01

Three-dimensional (3D) printing is currently being intensely studied for a diverse set of applications, including the development of bioengineered tissues, as well as the production of functional biomedical materials and devices for dental and orthopedic applications. The aim of this study was to develop and characterize a 3D-printed hybrid construct that can be potentially suitable for guided tissue regeneration (GTR). For this purpose, the rheology analyses have been performed on different bioinks and a specific solution comprising 8% gelatin, 2% elastin and 0.5% sodium hyaluronate has been selected as the most suitable composition for printing a structured membrane for GTR application. Each membrane is composed of 6 layers with strand angles from the first layer to the last layer of 45, 135, 0, 90, 0 and 90°. Confirmed by 3D Laser Measuring imaging, the membrane has small pores on one side and large pores on the other to be able to accommodate different cells like osteoblasts, fibroblasts and keratinocytes on different sides. The ultimate cross-linked product is a 150μm thick flexible and bendable membrane with easy surgical handling. Static and dynamic mechanical testing revealed static tensile modules of 1.95±0.55MPa and a dynamic tensile storage modulus of 314±50kPa. Through seeding the membranes with fibroblast and keratinocyte cells, the results of in vitro tests, including histological analysis, tissue viability examinations and DAPI staining, indicated that the membrane has desirable in vitro biocompatibility. The membrane has demonstrated the barrier function of a GTR membrane by thorough separation of the oral epithelial layer from the underlying tissues. In conclusion, we have characterized a biocompatible and bio-resorbable 3D-printed structured gelatin/elastin/sodium hyaluronate membrane with optimal biostability, mechanical strength and surgical handling characteristics in terms of suturability for potential application in GTR procedures. Copyright © 2017 Elsevier B.V. All rights reserved.

The fine structure of human germ layers in vivo: clues to the early differentiation of embryonic stem cells in vitro.

PubMed

Sathananthan, Henry; Selvaraj, Kamala; Clark, Joan

2011-08-01

The fine structure of the three germ layers in human ectopic embryos (stage 7) have been documented by digital light and electron microscopy. The formation of ectoderm, endoderm and mesoderm and notochordal cells, and also the extraembryonic membranes, amnion and yolk sac, are imaged. The germ layers give rise to all the cells and tissues of the human body. Possible clues to the early differentiation of embryonic stem cells (ESC) in vitro were obtained, since these events are more or less mimicked in cultures of ESC derived from the inner cell mass of human blastocysts. The findings are discussed with reference to previous studies on the fine structure of ESC using the same technique. Copyright © 2011. Published by Elsevier Ltd.

High-frequency viscoelastic shear properties of vocal fold tissues: implications for vocal fold tissue engineering.

PubMed

Teller, Sean S; Farran, Alexandra J E; Xiao, Longxi; Jiao, Tong; Duncan, Randall L; Clifton, Rodney J; Jia, Xinqiao

2012-10-01

The biomechanical function of the vocal folds (VFs) depends on their viscoelastic properties. Many conditions can lead to VF scarring that compromises voice function and quality. To identify candidate replacement materials, the structure, composition, and mechanical properties of native tissues need to be understood at phonation frequencies. Previously, the authors developed the torsional wave experiment (TWE), a stress-wave-based experiment to determine the linear viscoelastic shear properties of small, soft samples. Here, the viscoelastic properties of porcine and human VFs were measured over a frequency range of 10-200 Hz. The TWE utilizes resonance phenomena to determine viscoelastic properties; therefore, the specimen test frequency is determined by the sample size and material properties. Viscoelastic moduli are reported at resonance frequencies. Structure and composition of the tissues were determined by histology and immunochemistry. Porcine data from the TWE are separated into two groups: a young group, consisting of fetal and newborn pigs, and an adult group, consisting of 6-9-month olds and 2+-year olds. Adult tissues had an average storage modulus of 2309±1394 Pa and a loss tangent of 0.38±0.10 at frequencies of 36-200 Hz. The VFs of young pigs were significantly more compliant, with a storage modulus of 394±142 Pa and a loss tangent of 0.40±0.14 between 14 and 30 Hz. No gender dependence was observed. Histological staining showed that adult porcine tissues had a more organized, layered structure than the fetal tissues, with a thicker epithelium and a more structured lamina propria. Elastin fibers in fetal VF tissues were immature compared to those in adult tissues. Together, these structural changes in the tissues most likely contributed to the change in viscoelastic properties. Adult human VF tissues, recovered postmortem from adult patients with a history of smoking or disease, had an average storage modulus of 756±439 Pa and a loss tangent of 0.42±0.10. Contrary to the results of some other investigators, no significant frequency dependence was observed. This lack of observable frequency dependence may be due to the modest frequency range of the experiments and the wide range of stiffnesses observed within nominally similar sample types.

High-Frequency Viscoelastic Shear Properties of Vocal Fold Tissues: Implications for Vocal Fold Tissue Engineering

PubMed Central

Teller, Sean S.; Farran, Alexandra J.E.; Xiao, Longxi; Jiao, Tong; Duncan, Randall L.

2012-01-01

The biomechanical function of the vocal folds (VFs) depends on their viscoelastic properties. Many conditions can lead to VF scarring that compromises voice function and quality. To identify candidate replacement materials, the structure, composition, and mechanical properties of native tissues need to be understood at phonation frequencies. Previously, the authors developed the torsional wave experiment (TWE), a stress-wave-based experiment to determine the linear viscoelastic shear properties of small, soft samples. Here, the viscoelastic properties of porcine and human VFs were measured over a frequency range of 10–200 Hz. The TWE utilizes resonance phenomena to determine viscoelastic properties; therefore, the specimen test frequency is determined by the sample size and material properties. Viscoelastic moduli are reported at resonance frequencies. Structure and composition of the tissues were determined by histology and immunochemistry. Porcine data from the TWE are separated into two groups: a young group, consisting of fetal and newborn pigs, and an adult group, consisting of 6–9-month olds and 2+-year olds. Adult tissues had an average storage modulus of 2309±1394 Pa and a loss tangent of 0.38±0.10 at frequencies of 36–200 Hz. The VFs of young pigs were significantly more compliant, with a storage modulus of 394±142 Pa and a loss tangent of 0.40±0.14 between 14 and 30 Hz. No gender dependence was observed. Histological staining showed that adult porcine tissues had a more organized, layered structure than the fetal tissues, with a thicker epithelium and a more structured lamina propria. Elastin fibers in fetal VF tissues were immature compared to those in adult tissues. Together, these structural changes in the tissues most likely contributed to the change in viscoelastic properties. Adult human VF tissues, recovered postmortem from adult patients with a history of smoking or disease, had an average storage modulus of 756±439 Pa and a loss tangent of 0.42±0.10. Contrary to the results of some other investigators, no significant frequency dependence was observed. This lack of observable frequency dependence may be due to the modest frequency range of the experiments and the wide range of stiffnesses observed within nominally similar sample types. PMID:22741523

Nondestructive measurement of esophageal biaxial mechanical properties utilizing sonometry

NASA Astrophysics Data System (ADS)

Aho, Johnathon M.; Qiang, Bo; Wigle, Dennis A.; Tschumperlin, Daniel J.; Urban, Matthew W.

2016-07-01

Malignant esophageal pathology typically requires resection of the esophagus and reconstruction to restore foregut continuity. Reconstruction options are limited and morbid. The esophagus represents a useful target for tissue engineering strategies based on relative simplicity in comparison to other organs. The ideal tissue engineered conduit would have sufficient and ideally matched mechanical tolerances to native esophageal tissue. Current methods for mechanical testing of esophageal tissues both in vivo and ex vivo are typically destructive, alter tissue conformation, ignore anisotropy, or are not able to be performed in fluid media. The aim of this study was to investigate biomechanical properties of swine esophageal tissues through nondestructive testing utilizing sonometry ex vivo. This method allows for biomechanical determination of tissue properties, particularly longitudinal and circumferential moduli and strain energy functions. The relative contribution of mucosal-submucosal layers and muscular layers are compared to composite esophagi. Swine thoracic esophageal tissues (n  =  15) were tested by pressure loading using a continuous pressure pump system to generate stress. Preconditioning of tissue was performed by pressure loading with the pump system and pre-straining the tissue to in vivo length before data was recorded. Sonometry using piezocrystals was utilized to determine longitudinal and circumferential strain on five composite esophagi. Similarly, five mucosa-submucosal and five muscular layers from thoracic esophagi were tested independently. This work on esophageal tissues is consistent with reported uniaxial and biaxial mechanical testing and reported results using strain energy theory and also provides high resolution displacements, preserves native architectural structure and allows assessment of biomechanical properties in fluid media. This method may be of use to characterize mechanical properties of tissue engineered esophageal constructs.

Bioorthogonal layer-by-layer encapsulation of pancreatic islets via hyperbranched polymers

PubMed Central

Gattás-Asfura, Kerim M.; Stabler, Cherie L.

2013-01-01

The encapsulation of viable tissues via layer-by-layer polymer assembly provides a versatile platform for cell surface engineering, with nanoscale control over capsule properties. Herein, we report the development of a hyperbranched polymer-based, ultrathin capsule architecture expressing bioorthogonal functionality and tailored physiochemical properties. Random carbodiimide-based condensation of 3,5-dicarboxyphenyl glycineamide on alginate yielded a highly branched polysaccharide with multiple, spatially restricted, and readily functionalizable terminal carboxylate moieties. Poly(ethylene glycol) (PEG) was utilized to link azido end groups to the structured alginate. Together with phosphine functionalized poly(amido amine) (PAMAM) dendrimer, nanoscale layer-by-layer coatings, covalently stabilized via Staudinger ligation, were assembled onto solid surfaces and pancreatic islets. The effects of electrostatic and/or bioorthogonal covalent interlayer interactions on the resulting coating efficiency and stability, as well as pancreatic islet viability and function, were studied. These hyperbranched polymers provide a flexible platform for the formation of covalently stabilized ultrathin coatings on viable cells and tissues. In addition, the hyperbranched nature of the polymers presents a highly functionalized surface capable of bioorthogonal conjugation of additional bioactive or labeling motifs. PMID:24063764

Comparison of different histological protocols for the preservation and quantification of the intestinal mucus layer in pigs.

PubMed

Röhe, Ilen; Hüttner, Friedrich Joseph; Plendl, Johanna; Drewes, Barbara; Zentek, Jürgen

2018-02-05

The histological characterization of the intestinal mucus layer is important for many scientific experiments investigating the interaction between intestinal microbiota, mucosal immune response and intestinal mucus production. The aim of this study was to examine and compare different fixation protocols for displaying and quantifying the intestinal mucus layer in piglets and to test which histomorphological parameters may correlate with the determined mucus layer thickness. Jejunal and colonal tissue samples of weaned piglets (n=10) were either frozen in liquid nitrogen or chemically fixed using methacarn solution. The frozen tissue samples were cryosectioned and subsequently postfixed using three different postfixatives: paraformaldehyde vapor, neutrally buffered formalin solution and ethanol solution. After dehydration, methacarn fixed tissues were embedded in paraffin wax. Both sections of cryopreserved and methacarn fixed tissue samples were stained with Alcian blue (AB)-PAS followed by the microscopically determination of the mucus layer thickness. Different pH values of the Alcian Blue staining solution and two mucus layer thickness measuring methods were compared. In addition, various histomorphological parameters of methacarn fixed tissue samples were evaluated including the number of goblet cells and the mucin staining area. Cryopreservation in combination with chemical postfixation led to mucus preservation in the colon of piglets allowing mucus thickness measurements. Mucus could be only partly preserved in cryosections of the jejunum impeding any quantitative description of the mucus layer thickness. The application of different postfixations, varying pH values of the AB solution and different mucus layer measuring methods led to comparable results regarding the mucus layer thickness. Methacarn fixation proved to be unsuitable for mucus depiction as only mucus patches were found in the jejunum or a detachment of the mucus layer from the epithelium was observed in the colon. Correlation analyses revealed that the proportion of the mucin staining area per crypt area (relative mucin staining) measured in methacarn fixed tissue samples corresponded to the colonal mucus layer thickness determined in cryopreserved tissue samples. In conclusion, the results showed that cryopreservation using liquid nitrogen followed by chemical postfixation and AB-PAS staining led to a reliable mucus preservation allowing a mucus thickness determination in the colon of pigs. Moreover, the detected relative mucin staining area may serve as a suitable histomorphological parameter for the assessment of the intestinal mucus layer thickness. The findings obtained in this study can be used for the implementation of an improved standard for the histological description of the mucus layer in the colon of pigs.

Aligned poly(L-lactic-co-e-caprolactone) electrospun microfibers and knitted structure: a novel composite scaffold for ligament tissue engineering.

PubMed

Vaquette, Cédryck; Kahn, Cyril; Frochot, Céline; Nouvel, Cécile; Six, Jean-Luc; De Isla, Natalia; Luo, Li-Hua; Cooper-White, Justin; Rahouadj, Rachid; Wang, Xiong

2010-09-15

We developed a novel technique involving knitting and electrospinning to fabricate a composite scaffold for ligament tissue engineering. Knitted structures were coated with poly(L-lactic-co-e-caprolactone) (PLCL) and then placed onto a rotating cylinder and a PLCL solution was electrospun onto the structure. Highly aligned 2-microm-diameter microfibers covered the space between the stitches and adhered to the knitted scaffolds. The stress-strain tensile curves exhibited an initial toe region similar to the tensile behavior of ligaments. Composite scaffolds had an elastic modulus (150 +/- 14 MPa) similar to the modulus of human ligaments. Biological evaluation showed that cells proliferated on the composite scaffolds and they spontaneously orientated along the direction of microfiber alignment. The microfiber architecture also induced a high level of extracellular matrix secretion, which was characterized by immunostaining. We found that cells produced collagen type I and type III, two main components found in ligaments. After 14 days of culture, collagen type III started to form a fibrous network. We fabricated a composite scaffold having the mechanical properties of the knitted structure and the morphological properties of the aligned microfibers. It is difficult to seed a highly macroporous structure with cells, however the technique we developed enabled an easy cell seeding due to presence of the microfiber layer. Therefore, these scaffolds presented attractive properties for a future use in bioreactors for ligament tissue engineering. (c) 2010 Wiley Periodicals, Inc.

Alignment hierarchies: engineering architecture from the nanometre to the micrometre scale.

PubMed

Kureshi, Alvena; Cheema, Umber; Alekseeva, Tijna; Cambrey, Alison; Brown, Robert

2010-12-06

Natural tissues are built of metabolites, soluble proteins and solid extracellular matrix components (largely fibrils) together with cells. These are configured in highly organized hierarchies of structure across length scales from nanometre to millimetre, with alignments that are dominated by anisotropies in their fibrillar matrix. If we are to successfully engineer tissues, these hierarchies need to be mimicked with an understanding of the interaction between them. In particular, the movement of different elements of the tissue (e.g. molecules, cells and bulk fluids) is controlled by matrix structures at distinct scales. We present three novel systems to introduce alignment of collagen fibrils, cells and growth factor gradients within a three-dimensional collagen scaffold using fluid flow, embossing and layering of construct. Importantly, these can be seen as different parts of the same hierarchy of three-dimensional structure, as they are all formed into dense collagen gels. Fluid flow aligns collagen fibrils at the nanoscale, embossed topographical features provide alignment cues at the microscale and introducing layered configuration to three-dimensional collagen scaffolds provides microscale- and mesoscale-aligned pathways for protein factor delivery as well as barriers to confine protein diffusion to specific spatial directions. These seemingly separate methods can be employed to increase complexity of simple extracellular matrix scaffolds, providing insight into new approaches to directly fabricate complex physical and chemical cues at different hierarchical scales, similar to those in natural tissues.

The functional response of bioactive titania-modified three-dimensional Ti-6Al-4V mesh structure toward providing a favorable pathway for intercellular communication and osteoincorporation.

PubMed

Nune, K C; Misra, R D K; Li, S J; Hao, Y L; Zhang, W

2016-10-01

The objective of the study is to fundamentally elucidate the biological response of 3D printed mesh structures subjected to plasma electrolytic oxidation process through the study of osteoblast functions. The cellular activity of plasma electrolytic-oxidized mesh structure was explored in terms of cell-to-cell communication involving proliferation, synthesis of extracellular and intracellular proteins, and mineralization. Upon plasma electrolytic oxidation of the mesh structure, a thin layer of bioactive titania with pore size 1-3 µm was nucleated on the surface. The combination of microporous bioactive titania and interconnected porous architecture provided the desired pathway for supply of nutrients and oxygen to cells and tissue and a favorable osteogenic microenvironment for tissue on-growth and in-growth, in relation to the unmodified mesh structure. The formation of a confluent layer as envisaged via electron microscopy and quantitative assessment of the expression level of proteins (actin, vinculin, and fibronectin) point toward the determining role of surface-modified mesh structure in modulating osteoblasts functions. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2488-2501, 2016. © 2016 Wiley Periodicals, Inc.

Cell Sheet-Based Tissue Engineering for Organizing Anisotropic Tissue Constructs Produced Using Microfabricated Thermoresponsive Substrates.

PubMed

Takahashi, Hironobu; Okano, Teruo

2015-11-18

In some native tissues, appropriate microstructures, including orientation of the cell/extracellular matrix, provide specific mechanical and biological functions. For example, skeletal muscle is made of oriented myofibers that is responsible for the mechanical function. Native artery and myocardial tissues are organized three-dimensionally by stacking sheet-like tissues of aligned cells. Therefore, to construct any kind of complex tissue, the microstructures of cells such as myotubes, smooth muscle cells, and cardiomyocytes also need to be organized three-dimensionally just as in the native tissues of the body. Cell sheet-based tissue engineering allows the production of scaffold-free engineered tissues through a layer-by-layer construction technique. Recently, using microfabricated thermoresponsive substrates, aligned cells are being harvested as single continuous cell sheets. The cell sheets act as anisotropic tissue units to build three-dimensional tissue constructs with the appropriate anisotropy. This cell sheet-based technology is straightforward and has the potential to engineer a wide variety of complex tissues. In addition, due to the scaffold-free cell-dense environment, the physical and biological cell-cell interactions of these cell sheet constructs exhibit unique cell behaviors. These advantages will provide important clues to enable the production of well-organized tissues that closely mimic the structure and function of native tissues, required for the future of tissue engineering. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Electronic structure of boron based single and multi-layer two dimensional materials

NASA Astrophysics Data System (ADS)

Miyazato, Itsuki; Takahashi, Keisuke

2017-09-01

Two dimensional nanosheets based on boron and Group VA elements are designed and characterized using first principles calculations. B-N, B-P, B-As, B-Sb, and B-Bi are found to possess honeycomb structures where formation energies indicate exothermic reactions. Contrary to B-N, the cases of B-P, B-As, B-Sb, and B-Bi nanosheets are calculated to possess narrow band gaps. In addition, calculations reveal that the electronegativity difference between B and Group VA elements in the designed materials is a good indicator to predict the charge transfer and band gap of the two dimensional materials. Hydrogen adsorption over defect-free B-Sb and B-Bi results in exothermic reactions, while defect-free B-N, B-P, and B-As result in endothermic reactions. The layerability of the designed two dimensional materials is also investigated where the electronic structure of two-layered two dimensional materials is strongly coupled with how the two dimensional materials are layered. Thus, one can consider that the properties of two dimensional materials can be controlled by the composition of two dimensional materials and the structure of layers.

Investigation of Migration and Differentiation of Human Mesenchymal Stem Cells on Five-Layered Collagenous Electrospun Scaffold Mimicking Native Cartilage Structure.

PubMed

Reboredo, Jenny W; Weigel, Tobias; Steinert, Andre; Rackwitz, Lars; Rudert, Maximilian; Walles, Heike

2016-09-01

Cartilage degeneration is the major cause of chronic pain, lost mobility, and reduced quality of life for over estimated 150 million osteoarthritis sufferers worldwide. Despite intensive research, none of the available therapies can restore the hyaline cartilage surface beyond just fibrous repair. To overcome these limitations, numerous cell-based approaches for cartilage repair are being explored that aim to provide an appropriate microenvironment for chondrocyte maintenance and differentiation of multipotent mesenchymal stem cells (MSCs) toward the chondrogenic lineage. Articular cartilage is composed of highly organized collagen network that entails the tissue into four distinct zones and each zone into three different regions based on differences in matrix morphology and biochemistry. Current cartilage implants cannot establish the hierarchical tissue organization that seems critical for normal cartilage function. Therefore, in this study, a structured, multilayered collagen scaffold designed for the replacement of damaged cartilage is presented that allows repopulation by host cells and synthesis of a new natural matrix. By using the electrospinning method, the potential to engineer a scaffold consisting of two different collagen types is obtained. With the developed collagen scaffold, a five-layered biomaterial is created that has the potency to induce the differentiation of human bone marrow derived MSCs toward the chondrogenic lineage. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Light source distribution and scattering phase function influence light transport in diffuse multi-layered media

NASA Astrophysics Data System (ADS)

Vaudelle, Fabrice; L'Huillier, Jean-Pierre; Askoura, Mohamed Lamine

2017-06-01

Red and near-Infrared light is often used as a useful diagnostic and imaging probe for highly scattering media such as biological tissues, fruits and vegetables. Part of diffusively reflected light gives interesting information related to the tissue subsurface, whereas light recorded at further distances may probe deeper into the interrogated turbid tissues. However, modelling diffusive events occurring at short source-detector distances requires to consider both the distribution of the light sources and the scattering phase functions. In this report, a modified Monte Carlo model is used to compute light transport in curved and multi-layered tissue samples which are covered with a thin and highly diffusing tissue layer. Different light source distributions (ballistic, diffuse or Lambertian) are tested with specific scattering phase functions (modified or not modified Henyey-Greenstein, Gegenbauer and Mie) to compute the amount of backscattered and transmitted light in apple and human skin structures. Comparisons between simulation results and experiments carried out with a multispectral imaging setup confirm the soundness of the theoretical strategy and may explain the role of the skin on light transport in whole and half-cut apples. Other computational results show that a Lambertian source distribution combined with a Henyey-Greenstein phase function provides a higher photon density in the stratum corneum than in the upper dermis layer. Furthermore, it is also shown that the scattering phase function may affect the shape and the magnitude of the Bidirectional Reflectance Distribution (BRDF) exhibited at the skin surface.

Model-based recovery of histological parameters from multispectral images of the colon

NASA Astrophysics Data System (ADS)

Hidovic-Rowe, Dzena; Claridge, Ela

2005-04-01

Colon cancer alters the macroarchitecture of the colon tissue. Common changes include angiogenesis and the distortion of the tissue collagen matrix. Such changes affect the colon colouration. This paper presents the principles of a novel optical imaging method capable of extracting parameters depicting histological quantities of the colon. The method is based on a computational, physics-based model of light interaction with tissue. The colon structure is represented by three layers: mucosa, submucosa and muscle layer. Optical properties of the layers are defined by molar concentration and absorption coefficients of haemoglobins; the size and density of collagen fibres; the thickness of the layer and the refractive indexes of collagen and the medium. Using the entire histologically plausible ranges for these parameters, a cross-reference is created computationally between the histological quantities and the associated spectra. The output of the model was compared to experimental data acquired in vivo from 57 histologically confirmed normal and abnormal tissue samples and histological parameters were extracted. The model produced spectra which match well the measured data, with the corresponding spectral parameters being well within histologically plausible ranges. Parameters extracted for the abnormal spectra showed the increase in blood volume fraction and changes in collagen pattern characteristic of the colon cancer. The spectra extracted from multi-spectral images of ex-vivo colon including adenocarcinoma show the characteristic features associated with normal and abnormal colon tissue. These findings suggest that it should be possible to compute histological quantities for the colon from the multi-spectral images.

MicroCT-Based Skeletal Models for Use in Tomographic Voxel Phantoms for Radiological Protection

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bolch, Wesley

The University of Florida (UF) proposes to develop two high-resolution image-based skeletal dosimetry models for direct use by ICRP Committee 2’s Task Group on Dose Calculation in their forthcoming Reference Voxel Male (RVM) and Reference Voxel Female (RVF) whole-body dosimetry phantoms. These two phantoms are CT-based, and thus do not have the image resolution to delineate and perform radiation transport modeling of the individual marrow cavities and bone trabeculae throughout their skeletal structures. Furthermore, new and innovative 3D microimaging techniques will now be required for the skeletal tissues following Committee 2’s revision of the target tissues of relevance for radiogenicmore » bone cancer induction. This target tissue had been defined in ICRP Publication 30 as a 10-μm cell layer on all bone surfaces of trabecular and cortical bone. The revised target tissue is now a 50-μm layer within the marrow cavities of trabecular bone only and is exclusive of the marrow adipocytes. Clearly, this new definition requires the use of 3D microimages of the trabecular architecture not available from past 2D optical studies of the adult skeleton. With our recent acquisition of two relatively young cadavers (males of age 18-years and 40-years), we will develop a series of reference skeletal models that can be directly applied to (1) the new ICRP reference voxel man and female phantoms developed for the ICRP, and (2) pediatric phantoms developed to target the ICRP reference children. Dosimetry data to be developed will include absorbed fractions for internal beta and alpha-particle sources, as well as photon and neutron fluence-to-dose response functions for direct use in external dosimetry studies of the ICRP reference workers and members of the general public« less

Genetics Home Reference: lattice corneal dystrophy type I

MedlinePlus

... have recurrent corneal erosions, which are caused by separation of particular layers of the cornea from one ... intricate network that forms in the spaces between cells and provides structural support to tissues. The protein ...

Analysis of the in vitro degradation and the in vivo tissue response to bi-layered 3D-printed scaffolds combining PLA and biphasic PLA/bioglass components - Guidance of the inflammatory response as basis for osteochondral regeneration.

PubMed

Barbeck, Mike; Serra, Tiziano; Booms, Patrick; Stojanovic, Sanja; Najman, Stevo; Engel, Elisabeth; Sader, Robert; Kirkpatrick, Charles James; Navarro, Melba; Ghanaati, Shahram

2017-12-01

The aim of the present study was the in vitro and in vivo analysis of a bi-layered 3D-printed scaffold combining a PLA layer and a biphasic PLA/bioglass G5 layer for regeneration of osteochondral defects in vivo Focus of the in vitro analysis was on the (molecular) weight loss and the morphological and mechanical variations after immersion in SBF. The in vivo study focused on analysis of the tissue reactions and differences in the implant bed vascularization using an established subcutaneous implantation model in CD-1 mice and established histological and histomorphometrical methods. Both scaffold parts kept their structural integrity, while changes in morphology were observed, especially for the PLA/G5 scaffold. Mechanical properties decreased with progressive degradation, while the PLA/G5 scaffolds presented higher compressive modulus than PLA scaffolds. The tissue reaction to PLA included low numbers of BMGCs and minimal vascularization of its implant beds, while the addition of G5 lead to higher numbers of BMGCs and a higher implant bed vascularization. Analysis revealed that the use of a bi-layered scaffold shows the ability to observe distinct in vivo response despite the physical proximity of PLA and PLA/G5 layers. Altogether, the results showed that the addition of G5 enables to reduce scaffold weight loss and to increase mechanical strength. Furthermore, the addition of G5 lead to a higher vascularization of the implant bed required as basis for bone tissue regeneration mediated by higher numbers of BMGCs, while within the PLA parts a significantly lower vascularization was found optimally for chondral regeneration. Thus, this data show that the analyzed bi-layered scaffold may serve as an ideal basis for the regeneration of osteochondral tissue defects. Additionally, the results show that it might be able to reduce the number of experimental animals required as it may be possible to analyze the tissue response to more than one implant in one experimental animal.

Raman spectroscopic evidence of tissue restructuring in heat-induced tissue fusion.

PubMed

Su, Lei; Cloyd, Kristy L; Arya, Shobhit; Hedegaard, Martin A B; Steele, Joseph A M; Elson, Daniel S; Stevens, Molly M; Hanna, George B

2014-09-01

Heat-induced tissue fusion via radio-frequency (RF) energy has gained wide acceptance clinically and here we present the first optical-Raman-spectroscopy study on tissue fusion samples in vitro. This study provides direct insights into tissue constituent and structural changes on the molecular level, exposing spectroscopic evidence for the loss of distinct collagen fibre rich tissue layers as well as the denaturing and restructuring of collagen crosslinks post RF fusion. These findings open the door for more advanced optical feedback-control methods and characterization during heat-induced tissue fusion, which will lead to new clinical applications of this promising technology. Copyright © 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.

GENE EXPRESSION CHANGES AFTER SEIZURE PRECONDITIONING IN THE THREE MAJOR HIPPOCAMPAL CELL LAYERS

PubMed Central

Borges, Karin; Shaw, Renee; Dingledine, Raymond

2008-01-01

Rodents experience hippocampal damage after status epilepticus (SE) mainly in pyramidal cells while sparing the dentate granule cell layer (DGCL). Hippocampal damage was prevented in rats that had been preconditioned by brief seizures on two consecutive days before SE. To identify neuroprotective genes and biochemical pathways changed after preconditioning we compared the effect of preconditioning on gene expression in the CA1 and CA3 pyramidal and DGCLs, harvested by laser capture microscopy. In the DGCL the expression of 632 genes was altered, compared to only 151 and 58 genes in CA1 and CA3 pyramidal cell layers. Most of the differentially expressed genes regulate tissue structure and intra- and extracellular signaling, including neurotransmission. A selective upregulation of energy metabolism transcripts occurred in CA1 pyramidal cells relative to the DGCL. These results reveal a broad transcriptional response of the DGCL to preconditioning, and suggest several mechanisms underlying the neuroprotective effect of preconditioning seizures. PMID:17239605

Structural changes in loaded equine tendons can be monitored by a novel spectroscopic technique

PubMed Central

Kostyuk, Oksana; Birch, Helen L; Mudera, Vivek; Brown, Robert A

2004-01-01

This study aimed to investigate the preferential collagen fibril alignment in unloaded and loaded tendons using elastic scattering spectroscopy. The device consisted of an optical probe, a pulsed light source (320–860 nm), a spectrometer and a PC. Two probes with either 2.75 mm or 300 μm source-detector separations were used to monitor deep and superficial layers, respectively. Equine superficial digital flexor tendons were subjected to ex vivo progressive tensional loading. Seven times more backscattered light was detected parallel rather than perpendicular to the tendon axis with the 2.75 mm separation probe in unloaded tendons. In contrast, using the 300 μm separation probe the plane of maximum backscatter (3-fold greater) was perpendicular to the tendon axis. There was no optical anisotropy in the cross-sectional plane of the tendon (i.e. the transversely cut tendon surface), with no structural anisotropy. During mechanical loading (9–14% strain) backscatter anisotropy increased 8.5- to 18.5-fold along the principal strain axis for 2.75 mm probe separation, but almost disappeared in the perpendicular plane (measured using the 300 μm probe separation). Optical (anisotropy) and mechanical (strain) measurements were highly correlated. We conclude that spatial anisotropy of backscattered light can be used for quantitative monitoring of collagen fibril alignment and tissue reorganization during loading, with the potential for minimally invasive real-time structural monitoring of fibrous tissues in normal, pathological or repairing tissues and in tissue engineering. PMID:14578479

Organic electrochemical transistors for cell-based impedance sensing

NASA Astrophysics Data System (ADS)

Rivnay, Jonathan; Ramuz, Marc; Leleux, Pierre; Hama, Adel; Huerta, Miriam; Owens, Roisin M.

2015-01-01

Electrical impedance sensing of biological systems, especially cultured epithelial cell layers, is now a common technique to monitor cell motion, morphology, and cell layer/tissue integrity for high throughput toxicology screening. Existing methods to measure electrical impedance most often rely on a two electrode configuration, where low frequency signals are challenging to obtain for small devices and for tissues with high resistance, due to low current. Organic electrochemical transistors (OECTs) are conducting polymer-based devices, which have been shown to efficiently transduce and amplify low-level ionic fluxes in biological systems into electronic output signals. In this work, we combine OECT-based drain current measurements with simultaneous measurement of more traditional impedance sensing using the gate current to produce complex impedance traces, which show low error at both low and high frequencies. We apply this technique in vitro to a model epithelial tissue layer and show that the data can be fit to an equivalent circuit model yielding trans-epithelial resistance and cell layer capacitance values in agreement with literature. Importantly, the combined measurement allows for low biases across the cell layer, while still maintaining good broadband signal.

Cooperation in group-structured populations with two layers of interactions

PubMed Central

Zhang, Yanling; Fu, Feng; Chen, Xiaojie; Xie, Guangming; Wang, Long

2015-01-01

Recently there has been a growing interest in studying multiplex networks where individuals are structured in multiple network layers. Previous agent-based simulations of games on multiplex networks reveal rich dynamics arising from interdependency of interactions along each network layer, yet there is little known about analytical conditions for cooperation to evolve thereof. Here we aim to tackle this issue by calculating the evolutionary dynamics of cooperation in group-structured populations with two layers of interactions. In our model, an individual is engaged in two layers of group interactions simultaneously and uses unrelated strategies across layers. Evolutionary competition of individuals is determined by the total payoffs accrued from two layers of interactions. We also consider migration which allows individuals to move to a new group within each layer. An approach combining the coalescence theory with the theory of random walks is established to overcome the analytical difficulty upon local migration. We obtain the exact results for all “isotropic” migration patterns, particularly for migration tuned with varying ranges. When the two layers use one game, the optimal migration ranges are proved identical across layers and become smaller as the migration probability grows. PMID:26632251

Electromechanical Coupling Factor of Breast Tissue as a Biomarker for Breast Cancer.

PubMed

Park, Kihan; Chen, Wenjin; Chekmareva, Marina A; Foran, David J; Desai, Jaydev P

2018-01-01

This research aims to validate a new biomarker of breast cancer by introducing electromechanical coupling factor of breast tissue samples as a possible additional indicator of breast cancer. Since collagen fibril exhibits a structural organization that gives rise to a piezoelectric effect, the difference in collagen density between normal and cancerous tissue can be captured by identifying the corresponding electromechanical coupling factor. The design of a portable diagnostic tool and a microelectromechanical systems (MEMS)-based biochip, which is integrated with a piezoresistive sensing layer for measuring the reaction force as well as a microheater for temperature control, is introduced. To verify that electromechanical coupling factor can be used as a biomarker for breast cancer, the piezoelectric model for breast tissue is described with preliminary experimental results on five sets of normal and invasive ductal carcinoma (IDC) samples in the 25-45 temperature range. While the stiffness of breast tissues can be captured as a representative mechanical signature which allows one to discriminate among tissue types especially in the higher strain region, the electromechanical coupling factor shows more distinct differences between the normal and IDC groups over the entire strain region than the mechanical signature. From the two-sample -test, the electromechanical coupling factor under compression shows statistically significant differences ( 0.0039) between the two groups. The increase in collagen density in breast tissue is an objective and reproducible characteristic of breast cancer. Although characterization of mechanical tissue property has been shown to be useful for differentiating cancerous tissue from normal tissue, using a single parameter may not be sufficient for practical usage due to inherent variation among biological samples. The portable breast cancer diagnostic tool reported in this manuscript shows the feasibility of measuring multiple parameters of breast tissue allowing for practical application.

Coupling of Gaussian electromagnetic pulse into a muscle-bone model of biological structure.

PubMed

Lin, J C; Lam, C K

1976-03-01

The effect of angle of incidence on the transmission electromagnetic pulse with Gaussion character in biological material is studied. The model assumed is a layer of soft tissue over a semi-infinite medium of boney structure governed by alpha dispersion. The numerical results demonstrate that the transmitted pulse strength is the greatest when the pulse is incident normally on the air-tissue interface. The coupling efficiency for a one microsecond pulse is three times as big as that for a ten microsecond pulse.

The Selective Centrifugation Ensures a Better In Vitro Isolation of ASCs and Restores a Soft Tissue Regeneration In Vivo.

PubMed

De Francesco, Francesco; Guastafierro, Antonio; Nicoletti, Gianfranco; Razzano, Sergio; Riccio, Michele; Ferraro, Giuseppe A

2017-05-12

Autologous fat grafting procedures in plastic surgery have been extensively used to reinforce soft tissue in congenital or acquired tissue impairments. With this background, the aim of this study is firstly to examine the impact of a selective centrifugation on existing adipose stem cells (ASCs) in terms of stemness profile maintenance and, secondly, to investigate the effect of restoring volume in reconstruction on patients affected by soft tissue damage. After centrifugation, the fat graft products were separated into two layers and subsequently examined in vitro for the expression of CD34, CD90, CD117, CD105, CD29, CD31, CD44, CD73, CD133, CD14 and CD45 markers by flow cytometry and gene expression analyses were performed for Sox2, WNT3A, END, CD44, FUT4, COLL1, CTNNB1, hbEGF, KRTLG, MMP2 and VIM genes. The results showed that in the middle-high density (MHD) layer there was a peak concentration of ASCs, compared to another layer obtained after centrifugation. Research carried out on patients under treatment for soft tissue regeneration using cells obtained from MHD layer selection will be fundamental in comparative analysis. These studies will lead to an adequate standardization of outcomes, provided that treatment is performed through cell selection. Therefore, a unique procedure in tissue reconstruction and regeneration through fat grafting is presented here.

Effect of hybrid layer on stress distribution in a premolar tooth restored with composite or ceramic inlay: an FEM study.

PubMed

Belli, Sema; Eskitaşcioglu, Gürcan; Eraslan, Oguz; Senawongse, Pisol; Tagami, Junji

2005-08-01

The aim of this finite elemental stress analysis study was to evaluate the effect of hybrid layer on distribution and amount of stress formed under occlusal loading in a premolar tooth restored with composite or ceramic inlay. The mandibular premolar tooth was selected as the model based on the anatomical measurements suggested by Wheeler. The analysis is performed by using a Pentium II IBM compatible computer with the SAP 2000 structural analysis program. Four different mathematical models including the following structures were evaluated: 1) composite inlay, adhesive resin, and tooth structure; 2) composite inlay, adhesive resin, hybrid layer, and tooth structure; 3) ceramic inlay, adhesive resin, and tooth structure; 4) ceramic inlay, adhesive resin, hybrid layer, and tooth structure. Loading was applied from the occlusal surface of the restoration, and shear stresses under loading were evaluated. The findings were drawn by the Saplot program, and the results were analyzed by graphical comparison method. The output indicated that the hybrid layer acts as a stress absorber in models 2 and 4. The hybrid layer has also changed mathematical values of stress on cavity floors in both restoration types. Ceramic inlay collected the stress inside the body of the material, but the composite inlay directly transferred the stress through dental tissues. As a result, it was concluded that the hybrid layer has an effect on stress distribution under loading in a premolar tooth model restored with composite or ceramic inlay. Copyright 2005 Wiley Periodicals, Inc.

Why we should care about soft tissue interfaces when applying ultrasonic diathermy: an experimental and computer simulation study.

PubMed

Omena, Thaís Pionório; Fontes-Pereira, Aldo José; Costa, Rejane Medeiros; Simões, Ricardo Jorge; von Krüger, Marco Antônio; Pereira, Wagner Coelho de Albuquerque

2017-01-01

One goal of therapeutic ultrasound is enabling heat generation in tissue. Ultrasound application protocols typically neglect these processes of absorption and backscatter/reflection at the skin/fat, fat/muscle, and muscle/bone interfaces. The aim of this study was to investigate the heating process at interfaces close to the transducer and the bone with the aid of computer simulation and tissue-mimicking materials (phantoms). The experimental setup consists of physiotherapeutic ultrasound equipment for irradiation, two layers of soft tissue-mimicking material, and one with and one without an additional layer of bone-mimicking material. Thermocouple monitoring is used in both cases. A computational model is used with the experimental parameters in a COMSOL® software platform. The experimental results show significant temperature rise (42 °C) at 10 mm depth, regardless of bone layer presence, diverging 3 °C from the simulated values. The probable causes are thermocouple and transducer heating and interface reverberations. There was no statistical difference in the experimental results with and without the cortical bone for the central thermocouple of the first interface [ t (38) = -1.52; 95% CI = -0.85, 0.12; p  = 14]. Temperature rise (>6 °C) close to the bone layer was lower than predicted (>21 °C), possibly because without the bone layer, thermocouples at 30 mm make contact with the water bath and convection intensifies heat loss; this factor was omitted in the simulation model. This work suggests that more attention should be given to soft tissue layer interfaces in ultrasound therapeutic procedures even in the absence of a close bone layer.

Functional and esthetic outcome enhancement of periodontal surgery by application of plastic surgery principles.

PubMed

Hürzeler, M B; Weng, D

1999-02-01

The closure of surgical wounds in a layer-by-layer fashion, a common principle of plastic surgery, is applied in this article to the field of periodontal surgery with the introduction of a new flap design. The suggested technique is indicated with all periodontal procedures that aim for hard and soft tissue augmentation (guided bone regeneration, mucogingival surgery, or plastic periodontal surgery) where passive, tension-free wound closure is fundamental for wound healing and a successful functional and esthetic outcome. By means of a series of incisions, buccal and lingual flaps are split several times; this results in a double-partial thickness flap and a coronally positioned palatal sliding flap, respectively. Thus, several tissue layers are obtained and the passive advancement of flaps becomes possible for the coverage of augmented areas. Wound closure with microsurgical suture material is accomplished in a multilayer approach, which ensures adaptation and closure of the outer tissue layers without any tension. Two case reports demonstrate the new plastic periodontal approach.

Ex vivo photometric and polarimetric multilayer characterization of human healthy colon by multispectral Mueller imaging.

PubMed

Pierangelo, Angelo; Manhas, Sandeep; Benali, Abdelali; Fallet, Clément; Antonelli, Maria-Rosaria; Novikova, Tatiana; Gayet, Brice; Validire, Pierre; De Martino, Antonello

2012-06-01

Healthy human colon samples were analyzed ex vivo with a multispectral imaging Mueller polarimeter operating from 500 to 700 nm in a backscattering configuration with diffuse light illumination impinging on the innermost tissue layer, the mucosa. The intensity and polarimetric responses were taken on whole tissues first and after progressive exfoliation of the outer layers afterwards. Moreover, these measurements were carried out with two different substrates (one bright and the other dark) successively placed beneath each sample, allowing a reasonably accurate evaluation of the contributions to the overall backscattered light by the various layers. For the shorter investigated wavelengths (500 to 550 nm) the major contribution comes from mucosa and submucosa, while for the longer wavelengths (650 to 700 nm) muscular tissue and fat also contribute significantly. The depolarization has also been studied and is found to be stronger in the red part of the spectrum, mainly due to the highly depolarizing power of the muscular and fat layers.

Toward soft-tissue elastography using digital holography to monitor surface acoustic waves

NASA Astrophysics Data System (ADS)

Li, Shiguang; Mohan, Karan D.; Sanders, William W.; Oldenburg, Amy L.

2011-11-01

Measuring the elasticity distribution inside the human body is of great interest because elastic abnormalities can serve as indicators of several diseases. We present a method for mapping elasticity inside soft tissues by imaging surface acoustic waves (SAWs) with digital holographic interferometry. With this method, we show that SAWs are consistent with Rayleigh waves, with velocities proportional to the square root of the elastic modulus greater than 2-40 kPa in homogeneous tissue phantoms. In two-layer phantoms, the SAW velocity transitions approximately from that of the lower layer to that of the upper layer as frequency is increased in agreement with the theoretical relationship between SAW dispersion and the depth-dependent stiffness profile. We also observed deformation in the propagation direction of SAWs above a stiff inclusion placed 8 mm below the surface. These findings demonstrate the potential for quantitative digital holography-based elastography of soft tissues as a noninvasive method for disease detection.

Methods and apparatus for microwave tissue welding for wound closure

NASA Technical Reports Server (NTRS)

Ngo, Phong H. (Inventor); Dusl, John R. (Inventor); Arndt, G. Dickey (Inventor); Phan, Chau T. (Inventor); Byerly, Diane L. (Inventor); Sognier, Marguerite A. (Inventor); Carl, James R. (Inventor)

2013-01-01

Methods and apparatus for joining biological tissue together are provided. In at least one specific embodiment, a method for joining biological tissue together can include applying a biological solder on a wound. A barrier layer can be disposed on the biological solder. An antenna can be located in proximate spatial relationship to the barrier layer. An impedance of the antenna can be matched to an impedance of the wound. Microwaves from a signal generator can be transmitted through the antenna to weld two or more biological tissue pieces of the wound together. A power of the microwaves can be adjusted by a control circuit disposed between the antenna and the signal generator. The heating profile within the tissue may be adjusted and controlled by the placement of metallic microspheres in or around the wound.

The collagenic architecture of human dura mater.

PubMed

Protasoni, Marina; Sangiorgi, Simone; Cividini, Andrea; Culuvaris, Gloria Tiffany; Tomei, Giustino; Dell'Orbo, Carlo; Raspanti, Mario; Balbi, Sergio; Reguzzoni, Marcella

2011-06-01

Human dura mater is the most external meningeal sheet surrounding the CNS. It provides an efficient protection to intracranial structures and represents the most important site for CSF turnover. Its intrinsic architecture is made up of fibrous tissue including collagenic and elastic fibers that guarantee the maintenance of its biophysical features. The recent technical advances in the repair of dural defects have allowed for the creation of many synthetic and biological grafts. However, no detailed studies on the 3D microscopic disposition of collagenic fibers in dura mater are available. The authors report on the collagenic 3D architecture of normal dura mater highlighting the orientation, disposition in 3 dimensions, and shape of the collagen fibers with respect to the observed layer. Thirty-two dura mater specimens were collected during cranial decompressive surgical procedures, fixed in 2.5% Karnovsky solution, and digested in 1 N NaOH solution. After a routine procedure, the specimens were observed using a scanning electron microscope. The authors distinguished the following 5 layers in the fibrous dura mater of varying thicknesses, orientation, and structures: bone surface, external median, vascular, internal median, and arachnoid layers. The description of the ultrastructural 3D organization of the different layers of dura mater will give us more information for the creation of synthetic grafts that are as similar as possible to normal dura mater. This description will be also related to the study of the neoplastic invasion.

Histopathologic and immunohistochemical features of capsular tissue around failed Ahmed glaucoma valves.

PubMed

Mahale, Alka; Fikri, Fatma; Al Hati, Khitam; Al Shahwan, Sami; Al Jadaan, Ibrahim; Al Katan, Hind; Khandekar, Rajiv; Maktabi, Azza; Edward, Deepak P

2017-01-01

Impervious encapsulation around Ahmed glaucoma valve (AGV) results in surgical failure raising intraocular pressure (IOP). Dysregulation of extracellular matrix (ECM) molecules and cellular factors might contribute to increased hydraulic resistance to aqueous drainage. Therefore, we examined these molecules in failed AGV capsular tissue. Immunostaining for ECM molecules (collagen I, collagen III, decorin, lumican, chondroitin sulfate, aggrecan and keratan sulfate) and cellular factors (αSMA and TGFβ) was performed on excised capsules from failed AGVs and control tenon's tissue. Staining intensity of ECM molecules was assessed using Image J. Cellular factors were assessed based on positive cell counts. Histopathologically two distinct layers were visible in capsules. The inner layer (proximal to the AGV) showed significant decrease in most ECM molecules compared to outer layer. Furthermore, collagen III (p = 0.004), decorin (p = 0.02), lumican (p = 0.01) and chondroitin sulfate (p = 0.02) was significantly less in inner layer compared to tenon's tissue. Outer layer labelling however was similar to control tenon's for most ECM molecules. Significantly increased cellular expression of αSMA (p = 0.02) and TGFβ (p = 0.008) was detected within capsular tissue compared to controls. Our results suggest profibrotic activity indicated by increased αSMA and TGFβ expression and decreased expression of proteoglycan (decorin and lumican) and glycosaminoglycans (chondroitin sulfate). Additionally, we observed decreased collagen III which might reflect increased myofibroblast contractility when coupled with increased TGFβ and αSMA expression. Together these events lead to tissue dysfunction potentially resulting in hydraulic resistance that may affect aqueous flow through the capsular wall.

Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty.

PubMed

Tu, Duong D; Chung, Yeun Goo; Gil, Eun Seok; Seth, Abhishek; Franck, Debra; Cristofaro, Vivian; Sullivan, Maryrose P; Di Vizio, Dolores; Gomez, Pablo; Adam, Rosalyn M; Kaplan, David L; Estrada, Carlos R; Mauney, Joshua R

2013-11-01

Acellular scaffolds derived from Bombyx mori silk fibroin were investigated for their ability to support functional tissue regeneration in a porcine model of augmentation cystoplasty. Two bi-layer matrix configurations were fabricated by solvent-casting/salt leaching either alone (Group 1) or in combination with silk film casting (Group 2) to yield porous foams buttressed by heterogeneous surface pore occlusions or homogenous silk films, respectively. Bladder augmentation was performed with each scaffold group (6 × 6 cm(2)) in juvenile Yorkshire swine for 3 m of implantation. Augmented animals exhibited high rates of survival (Group 1: 5/6, 83%; Group 2: 4/4, 100%) and voluntary voiding over the course of the study period. Urodynamic evaluations demonstrated mean increases in bladder capacity over pre-operative levels (Group 1: 277%; Group 2: 153%) which exceeded nonsurgical control gains (144%) encountered due to animal growth.In addition, animals augmented with both matrix configurations displayed increases in bladder compliance over pre-operative levels(Group 1: 357%; Group 2: 338%) similar to growth-related elevations observed in non-surgical controls (354%) [corrected]. Gross tissue evaluations revealed that both matrix configurations supported extensive de novo tissue formation throughout the entire original implantation site which exhibited ultimate tensile strength similar to nonsurgical counterparts. Histological and immunohistochemical analyses showed that both implant groups promoted comparable extents of smooth muscle regeneration and contractile protein (α-smooth muscle actin and SM22α) expression within defect sites similar to controls. Parallel evaluations demonstrated the formation of a transitional, multi-layered urothelium with prominent cytokeratin, uroplakin, and p63 protein expression in both matrix groups. De novo innervation and vascularization processes were evident in all regenerated tissues indicated by synaptophysin-positive neuronal cells and vessels lined with CD31 expressing endothelial cells. Ex vivo organ bath studies demonstrated that regenerated tissues supported by both silk matrices displayed contractile responses to carbachol, α,β-methylene-ATP, KCl, and electrical field stimulation similar to controls. Our data detail the ability of acellular silk scaffolds to support regeneration of innervated, vascularized smooth muscle and urothelial tissues within 3 m with structural, mechanical, and functional properties comparable to native tissue in a porcine model of bladder repair. © 2013 Elsevier Ltd. All rights reserved.

[Observation on the Histologic Structure of Multiceps multiceps in Artificially Infected Dogs].

PubMed

Shang, Qing-yan; Fan, Xi-ping; Zhang, Xiao-yu; Han, Jin-huan; Zhang, Qian; Sun, Xiao-ling

2015-06-01

To observe the microstructure and ultrastructure of Multiceps multiceps from the artificially infected dogs. METHEDS: Two male dogs were infected with the coenurus of M. multiceps from naturally-infected sheep (about 80-100 per dog). The adult worms of M. multiceps were recovered from the intestine, and fixed by the conventional method. The scolex, neck, immature proglottid, mature proglottid, and gravid proglottid were prepared for paraffin section and ultrathin sections with HE staining and uranyl acetate staining, and observed under light microscope and electron transmission microscope, respectively. Under light microscope, each proglottid consisted of cortical layer and parenchymal layer. The cortical layer was composed of microvilli, syncytium, and substrate layer. The parenchymal layer mainly consisted of muscle tissue, excretory system, and reproductive system. The microvilli layer of scolex was thinner than that of neck and mature proglottid, and the longest microvilli were mainly distributed in the binding site between the proglottids. The scolex was extremely muscular. The nervous system and excretory system were repeated in each proglottid. Mature proglottid had both male and female reproductive systems. Gravid proglottid had uterus and egg, and atrophic male reproductive organs. The special microstructure of Multiceps multiceps are that most microvilli in the cortex is cylindrical; the microvilli length in the binding sites between mature proglottids is longer than that of other parts.

Online quantitative analysis of multispectral images of human body tissues

NASA Astrophysics Data System (ADS)

Lisenko, S. A.

2013-08-01

A method is developed for online monitoring of structural and morphological parameters of biological tissues (haemoglobin concentration, degree of blood oxygenation, average diameter of capillaries and the parameter characterising the average size of tissue scatterers), which involves multispectral tissue imaging, image normalisation to one of its spectral layers and determination of unknown parameters based on their stable regression relation with the spectral characteristics of the normalised image. Regression is obtained by simulating numerically the diffuse reflectance spectrum of the tissue by the Monte Carlo method at a wide variation of model parameters. The correctness of the model calculations is confirmed by the good agreement with the experimental data. The error of the method is estimated under conditions of general variability of structural and morphological parameters of the tissue. The method developed is compared with the traditional methods of interpretation of multispectral images of biological tissues, based on the solution of the inverse problem for each pixel of the image in the approximation of different analytical models.

In vivo imaging of coral tissue and skeleton with optical coherence tomography.

PubMed

Wangpraseurt, Daniel; Wentzel, Camilla; Jacques, Steven L; Wagner, Michael; Kühl, Michael

2017-03-01

Application of optical coherence tomography (OCT) for in vivo imaging of tissue and skeleton structure of intact living corals enabled the non-invasive visualization of coral tissue layers (endoderm versus ectoderm), skeletal cavities and special structures such as mesenterial filaments and mucus release from intact living corals. Coral host chromatophores containing green fluorescent protein-like pigment granules appeared hyper-reflective to near-infrared radiation allowing for excellent optical contrast in OCT and a rapid characterization of chromatophore size, distribution and abundance. In vivo tissue plasticity could be quantified by the linear contraction velocity of coral tissues upon illumination resulting in dynamic changes in the live coral tissue surface area, which varied by a factor of 2 between the contracted and expanded state of a coral. Our study provides a novel view on the in vivo organization of coral tissue and skeleton and highlights the importance of microstructural dynamics for coral ecophysiology. © 2017 The Author(s).

Chitosan-collagen scaffolds with nano/microfibrous architecture for skin tissue engineering.

PubMed

Sarkar, Soumi Dey; Farrugia, Brooke L; Dargaville, Tim R; Dhara, Santanu

2013-12-01

In this study, a hierarchical nano/microfibrous chitosan/collagen scaffold that approximates structural and functional attributes of native extracellular matrix has been developed for applicability in skin tissue engineering. Scaffolds were produced by electrospinning of chitosan followed by imbibing of collagen solution, freeze-drying, and subsequent cross-linking of two polymers. Scanning electron microscopy showed formation of layered scaffolds with nano/microfibrous architechture. Physicochemical properties of scaffolds including tensile strength, swelling behavior, and biodegradability were found satisfactory for intended application. 3T3 fibroblasts and HaCaT keratinocytes showed good in vitro cellular response on scaffolds thereby indicating the matrices, cytocompatible nature. Scaffolds tested in an ex vivo human skin equivalent wound model, as a preliminary alternative to animal testing, showed keratinocyte migration and wound re-epithelization-a prerequisite for healing and regeneration. Taken together, the herein proposed chitosan/collagen scaffold, shows good potential for skin tissue engineering. Copyright © 2013 Wiley Periodicals, Inc., a Wiley Company.

Peripheral mineralization of a 3D biodegradable tubular construct as a way to enhance guidance stabilization in spinal cord injury regeneration.

PubMed

Oliveira, A L; Sousa, E C; Silva, N A; Sousa, N; Salgado, A J; Reis, R L

2012-11-01

Spinal cord injuries (SCI) present a major challenge to therapeutic development due to its complexity. Combinatorial approaches using biodegradable polymers that can simultaneously provide a tissue scaffold, a cell vehicle, and a reservoir for sustained drug delivery have shown very promising results. In our previous studies we have developed a novel hybrid system consisting of starch/poly-e-caprolactone (SPCL) semi-rigid tubular porous structure, based on a rapid prototyping technology, filled by a gellan gum hydrogel concentric core for the regeneration within spinal-cord injury sites. In the present work we intend to promote enhanced osteointegration on these systems by pre-mineralizing specifically the external surfaces of the SPCL tubular structures, though a biomimetic strategy, using a sodium silicate gel as nucleating agent. The idea is to create two different cell environments to promote axonal regeneration in the interior of the constructs while inducing osteogenic activity on its external surface. By using a Teflon cylinder to isolate the interior of the scaffold, it was possible to observe the formation of a bone-like poorly crystalline carbonated apatite layer continuously formed only in the external side of the tubular structure. This biomimetic layer was able to support the adhesion of Bone Marrow Mesenchymal Stem Cells, which have gone under cytoskeleton reorganization in the first hours of culture when compared to cells cultured on uncoated scaffolds. This strategy can be a useful route for locally stimulate bone tissue regeneration and facilitating early bone ingrowth.

Layer-by-Layer Bioprinting of Stem Cells for Retinal Tissue Regeneration

DTIC Science & Technology

2015-10-01

AWARD NUMBER: W81XWH-14-1-0522 TITLE: Layer-by-Layer Bioprinting of Stem Cells for Retinal Tissue Regeneration PRINCIPAL INVESTIGATOR...TITLE AND SUBTITLE Layer-by-Layer Bioprinting of Stem Cells for Retinal Tissue Regeneration 5a. CONTRACT NUMBER 5b. GRANT NUMBER W81XWH-14-1-0522 5c...bioprinting process using stem cells for retinal tissue regeneration. The LBL nature of the bioprinting process matches nicely with the native

Part I. Mechanisms of injury associated with extracorporeal shock wave lithotripsy; Part II. Exsolution of volatiles

NASA Astrophysics Data System (ADS)

Howard, Danny Dwayne

Part I - Shock waves are focused in extracorporeal shock wave lithotripsy (ESWL) machines to strengths sufficient to fracture kidney stones. Substantial side effects-most of them acute-have resulted from this procedure, including injury to soft tissue. The focusing of shock waves through various layers of tissue is a complex process which stimulates many bio-mechano-chemical responses.This thesis presents results of an in vitro study of the initial mechanical stimulus. Planar nitrocellulose membranes of order 10 um thick were used as models of thin tissue structures. Two modes of failure were recorded: Failure due to cavitation collapsing on or near the membranes, and failure induced by altering the structure of shock waves. Tests were done in water at and around F2 to characterize the extent of cavitation damage, and was found to be confined within the focal region, 1.2 cm along the axis of focus.Scattering media were used to simulate the effects of acoustic nonuniformity of tissue and to alter the structure of focusing shock waves. 40 um diameter (average) hollow glass spheres were added to ethylene glycol, glycerine and castor oil to vary the properties of the scattering media. Multiple layer samples of various types of phantom tissue were tested in degassed castor oil to gauge the validity of the scattering media. The scattering media and tissue samples increased the rise time decreased strain rate in a similar fashion. Membranes were damaged by the decreased strain rate and accumulated effects of the altered structure: After about 20 or so shocks immersed in the scattering media and after about 100 shocks behind the tissue samples. The mode of failure was tearing with multiple tears in some cases from about .1 cm to about 3 cm depending of the number of shocks and membrane thickness.Part II - This work examines the exsolution of volatiles-carbon dioxide from water-in a cylindrical test cell under different pressure conditions. Water was supersaturated with carbon dioxide under various pressures (620 to 1062 kPa), and depressurized rapidly to investigate how carbon dioxide is undissolved, exsolution, and its effects on the surrounding environment. Cavities grow as a result of convective diffusion: They move before depleting carbon dioxide in a given region. The radius of a cavity in this environment grows at a faster rate [...] than that of a cavity at rest [...]. Bubble growth rates were inferred by measuring the bulk liquid using high speed motion pictures. Water in the test-cell is accelerated as a result of buoyancy induced by cavity growth. Cavities are elliptical in shape and grow until mutual interaction causes them to fragment. Accelerations range from 10 to 100 g were measured with velocities ranging from 7 to 13 m/s.

Thermal inkjet printing in tissue engineering and regenerative medicine.

PubMed

Cui, Xiaofeng; Boland, Thomas; D'Lima, Darryl D; Lotz, Martin K

2012-08-01

With the advantages of high throughput, digital control, and highly accurate placement of cells and biomaterial scaffold to the desired 2D and 3D locations, bioprinting has great potential to develop promising approaches in translational medicine and organ replacement. The most recent advances in organ and tissue bioprinting based on the thermal inkjet printing technology are described in this review. Bioprinting has no or little side effect to the printed mammalian cells and it can conveniently combine with gene transfection or drug delivery to the ejected living systems during the precise placement for tissue construction. With layer-by-layer assembly, 3D tissues with complex structures can be printed using scanned CT or MRI images. Vascular or nerve systems can be enabled simultaneously during the organ construction with digital control. Therefore, bioprinting is the only solution to solve this critical issue in thick and complex tissues fabrication with vascular system. Collectively, bioprinting based on thermal inkjet has great potential and broad applications in tissue engineering and regenerative medicine. This review article introduces some important patents related to bioprinting of living systems and the applications of bioprinting in tissue engineering field.

Filtering properties of Thue-Morse nano-photonic crystals containing high-temperature superconductor

NASA Astrophysics Data System (ADS)

Talebzadeh, Robabeh; Bavaghar, Mehrdad

2018-05-01

In this paper, we introduced new design of quasi-periodic layered structures by choosing order two of ternary Thue-Morse structure. We considered Superconductor-dielectric photonic crystal with mirror symmetric as (ABSSAB)N(BASSBA)N composed of two kinds of nano-scale dielectric layers (A and B) and high-temperature superconductor layers where N is the number of period. This structure is assumed to be the free space. By using the transfer matrix method and the two fluid model, we theoretically study the transmission spectrum of ternary Thue-Morse superconducting photonic crystals with mirror symmetry and introduce this structure as a narrow optical filter. We showed that transmission peak so-called defect mode appears itself inside the transmission spectrum of suggested structure as same as defective layered structure. Also, we analyzed the influence of various related parameters such as the operating temperature of superconductor layer on position of defect mode. The redshift of defect mode with increasing the operating temperature was observed.

Platelet lysate-based pro-angiogenic nanocoatings.

PubMed

Oliveira, Sara M; Pirraco, Rogério P; Marques, Alexandra P; Santo, Vítor E; Gomes, Manuela E; Reis, Rui L; Mano, João F

2016-03-01

Human platelet lysate (PL) is a cost-effective and human source of autologous multiple and potent pro-angiogenic factors, such as vascular endothelial growth factor A (VEGF A), fibroblast growth factor b (FGF b) and angiopoietin-1. Nanocoatings previously characterized were prepared by layer-by-layer assembling incorporating PL with marine-origin polysaccharides and were shown to activate human umbilical vein endothelial cells (HUVECs). Within 20 h of incubation, the more sulfated coatings induced the HUVECS to the form tube-like structures accompanied by an increased expression of angiogenic-associated genes, such as angiopoietin-1 and VEGF A. This may be a cost-effective approach to modify 2D/3D constructs to instruct angiogenic cells towards the formation of neo-vascularization, driven by multiple and synergistic stimulations from the PL combined with sulfated polysaccharides. The presence, or fast induction, of a stable and mature vasculature inside 3D constructs is crucial for new tissue formation and its viability. This has been one of the major tissue engineering challenges, limiting the dimensions of efficient tissue constructs. Many approaches based on cells, growth factors, 3D bioprinting and channel incorporation have been proposed. Herein, we explored a versatile technique, layer-by-layer assembling in combination with platelet lysate (PL), that is a cost-effective source of many potent pro-angiogenic proteins and growth factors. Results suggest that the combination of PL with sulfated polyelectrolytes might be used to introduce interfaces onto 2D/3D constructs with potential to induce the formation of cell-based tubular structures. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Three-dimensional bioprinting in tissue engineering and regenerative medicine.

PubMed

Gao, Guifang; Cui, Xiaofeng

2016-02-01

With the advances of stem cell research, development of intelligent biomaterials and three-dimensional biofabrication strategies, highly mimicked tissue or organs can be engineered. Among all the biofabrication approaches, bioprinting based on inkjet printing technology has the promises to deliver and create biomimicked tissue with high throughput, digital control, and the capacity of single cell manipulation. Therefore, this enabling technology has great potential in regenerative medicine and translational applications. The most current advances in organ and tissue bioprinting based on the thermal inkjet printing technology are described in this review, including vasculature, muscle, cartilage, and bone. In addition, the benign side effect of bioprinting to the printed mammalian cells can be utilized for gene or drug delivery, which can be achieved conveniently during precise cell placement for tissue construction. With layer-by-layer assembly, three-dimensional tissues with complex structures can be printed using converted medical images. Therefore, bioprinting based on thermal inkjet is so far the most optimal solution to engineer vascular system to the thick and complex tissues. Collectively, bioprinting has great potential and broad applications in tissue engineering and regenerative medicine. The future advances of bioprinting include the integration of different printing mechanisms to engineer biphasic or triphasic tissues with optimized scaffolds and further understanding of stem cell biology.

A Robust Method to Generate Mechanically Anisotropic Vascular Smooth Muscle Cell Sheets for Vascular Tissue Engineering.

PubMed

Backman, Daniel E; LeSavage, Bauer L; Shah, Shivem B; Wong, Joyce Y

2017-06-01

In arterial tissue engineering, mimicking native structure and mechanical properties is essential because compliance mismatch can lead to graft failure and further disease. With bottom-up tissue engineering approaches, designing tissue components with proper microscale mechanical properties is crucial to achieve the necessary macroscale properties in the final implant. This study develops a thermoresponsive cell culture platform for growing aligned vascular smooth muscle cell (VSMC) sheets by photografting N-isopropylacrylamide (NIPAAm) onto micropatterned poly(dimethysiloxane) (PDMS). The grafting process is experimentally and computationally optimized to produce PNIPAAm-PDMS substrates optimal for VSMC attachment. To allow long-term VSMC sheet culture and increase the rate of VSMC sheet formation, PNIPAAm-PDMS surfaces were further modified with 3-aminopropyltriethoxysilane yielding a robust, thermoresponsive cell culture platform for culturing VSMC sheets. VSMC cell sheets cultured on patterned thermoresponsive substrates exhibit cellular and collagen alignment in the direction of the micropattern. Mechanical characterization of patterned, single-layer VSMC sheets reveals increased stiffness in the aligned direction compared to the perpendicular direction whereas nonpatterned cell sheets exhibit no directional dependence. Structural and mechanical anisotropy of aligned, single-layer VSMC sheets makes this platform an attractive microstructural building block for engineering a vascular graft to match the in vivo mechanical properties of native arterial tissue. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A Monte Carlo study of fluorescence generation probability in a two-layered tissue model

NASA Astrophysics Data System (ADS)

Milej, Daniel; Gerega, Anna; Wabnitz, Heidrun; Liebert, Adam

2014-03-01

It was recently reported that the time-resolved measurement of diffuse reflectance and/or fluorescence during injection of an optical contrast agent may constitute a basis for a technique to assess cerebral perfusion. In this paper, we present results of Monte Carlo simulations of the propagation of excitation photons and tracking of fluorescence photons in a two-layered tissue model mimicking intra- and extracerebral tissue compartments. Spatial 3D distributions of the probability that the photons were converted from excitation to emission wavelength in a defined voxel of the medium (generation probability) during their travel between source and detector were obtained for different optical properties in intra- and extracerebral tissue compartments. It was noted that the spatial distribution of the generation probability depends on the distribution of the fluorophore in the medium and is influenced by the absorption of the medium and of the fluorophore at excitation and emission wavelengths. Simulations were also carried out for realistic time courses of the dye concentration in both layers. The results of the study show that the knowledge of the absorption properties of the medium at excitation and emission wavelengths is essential for the interpretation of the time-resolved fluorescence signals measured on the surface of the head.

Automated 3D bioassembly of micro-tissues for biofabrication of hybrid tissue engineered constructs.

PubMed

Mekhileri, N V; Lim, K S; Brown, G C J; Mutreja, I; Schon, B S; Hooper, G J; Woodfield, T B F

2018-01-12

Bottom-up biofabrication approaches combining micro-tissue fabrication techniques with extrusion-based 3D printing of thermoplastic polymer scaffolds are emerging strategies in tissue engineering. These biofabrication strategies support native self-assembly mechanisms observed in developmental stages of tissue or organoid growth as well as promoting cell-cell interactions and cell differentiation capacity. Few technologies have been developed to automate the precise assembly of micro-tissues or tissue modules into structural scaffolds. We describe an automated 3D bioassembly platform capable of fabricating simple hybrid constructs via a two-step bottom-up bioassembly strategy, as well as complex hybrid hierarchical constructs via a multistep bottom-up bioassembly strategy. The bioassembly system consisted of a fluidic-based singularisation and injection module incorporated into a commercial 3D bioprinter. The singularisation module delivers individual micro-tissues to an injection module, for insertion into precise locations within a 3D plotted scaffold. To demonstrate applicability for cartilage tissue engineering, human chondrocytes were isolated and micro-tissues of 1 mm diameter were generated utilising a high throughput 96-well plate format. Micro-tissues were singularised with an efficiency of 96.0 ± 5.1%. There was no significant difference in size, shape or viability of micro-tissues before and after automated singularisation and injection. A layer-by-layer approach or aforementioned bottom-up bioassembly strategy was employed to fabricate a bilayered construct by alternatively 3D plotting a thermoplastic (PEGT/PBT) polymer scaffold and inserting pre-differentiated chondrogenic micro-tissues or cell-laden gelatin-based (GelMA) hydrogel micro-spheres, both formed via high-throughput fabrication techniques. No significant difference in viability between the construct assembled utilising the automated bioassembly system and manually assembled construct was observed. Bioassembly of pre-differentiated micro-tissues as well as chondrocyte-laden hydrogel micro-spheres demonstrated the flexibility of the platform while supporting tissue fusion, long-term cell viability, and deposition of cartilage-specific extracellular matrix proteins. This technology provides an automated and scalable pathway for bioassembly of both simple and complex 3D tissue constructs of clinically relevant shape and size, with demonstrated capability to facilitate direct spatial organisation and hierarchical 3D assembly of micro-tissue modules, ranging from biomaterial free cell pellets to cell-laden hydrogel formulations.

A bio-inspired microstructure induced by slow injection moulding of cylindrical block copolymers.

PubMed

Stasiak, Joanna; Brubert, Jacob; Serrani, Marta; Nair, Sukumaran; de Gaetano, Francesco; Costantino, Maria Laura; Moggridge, Geoff D

2014-08-28

It is well known that block copolymers with cylindrical morphology show alignment with shear, resulting in anisotropic mechanical properties. Here we show that well-ordered bi-directional orientation can be achieved in such materials by slow injection moulding. This results in a microstructure, and anisotropic mechanical properties, similar to many natural tissues, making this method attractive for engineering prosthetic fibrous tissues. An application of particular interest to us is prosthetic polymeric heart valve leaflets, mimicking the shape, microstructure and hence performance of the native valve. Anisotropic layers have been observed for cylinder-forming block copolymers centrally injected into thin circular discs. The skin layers exhibit orientation parallel to the flow direction, whilst the core layer shows perpendicularly oriented domains; the balance of skin to core layers can be controlled by processing parameters such as temperature and injection rate. Heart valve leaflets with a similar layered structure have been prepared by injection moulding. Numerical modelling demonstrates that such complex orientation can be explained and predicted by the balance of shear and extensional flow.

Light microscopical structure and ultrastructure of a Besnoitia sp. in the naturally infected lizard Ameiva ameiva (Teiidae) from north Brazil, and in experimentally infected mice.

PubMed

Paperna, I; Son, R L

2001-09-01

A Besnoitia species of the teiid lizard Ameiva ameiva (L.), from north Brazil was established in laboratory mice and hamster by the intraperitoneal inoculation of bradyzoites in the tissue cysts. In the lizards all the cyst wall layers were closely apposed. In the mice the layers of the wall were distinguishable, and ultrastructurally the inner cytoplasmic layer contained either a tight network of endoplasmic reticulum or packed mitochondria or both. These components were less frequent or sparse in the inner cytoplasmic layer of cysts in the lizard. The only animals available for experiments in attempts to indicate the definitive host of the parasite were 3 kittens of the domestic cat and a juvenile specimen of the snake Boa constrictor raised in captivity. No evidence of infection could be detected in these animals after feeding them with the tissues of mice harbouring cysts with very large number of bradyzoites.

Diagnostic efficiency of Mueller-matrix polarization reconstruction system of the phase structure of liver tissue

NASA Astrophysics Data System (ADS)

Zabolotna, Natalia I.; Pavlov, Sergii V.; Radchenko, Kostiantyn O.; Stasenko, Vladyslav A.; Wójcik, Waldemar; Kussambayeva, Nazym

2015-12-01

The application field of using the Mueller-matrix polarizing reconstruction system of phase structure of biological layer for optical-anisotropic parameters differentiation of histological sections of healthy and rat's liver with hepatitis were investigated. Comparison of system informativity with known systems on indexes of sensitivity, specificity and balanced accuracy were performed.

Photosynthetic Acclimation of Symbiodinium in hospite Depends on Vertical Position in the Tissue of the Scleractinian Coral Montastrea curta

PubMed Central

Lichtenberg, Mads; Larkum, Anthony W. D.; Kühl, Michael

2016-01-01

Coral photophysiology has been studied intensively from the colony scale down to the scale of single fluorescent pigment granules as light is one of the key determinants for coral health. We studied the photophysiology of the oral and aboral symbiont band of scleractinian coral Montastrea curta to investigate if different acclimation to light exist in hospite on a polyp scale. By combined use of electrochemical and fiber-optic microsensors for O2, scalar irradiance and variable chlorophyll fluorescence, we could characterize the physical and chemical microenvironment experienced by the symbionts and, for the first time, estimate effective quantum yields of PSII photochemistry and rates of electron transport at the position of the zooxanthellae corrected for the in-tissue gradient of scalar irradiance. The oral- and aboral Symbiodinium layers received ∼71% and ∼33% of surface scalar irradiance, respectively, and the two symbiont layers experience considerable differences in light exposure. Rates of gross photosynthesis did not differ markedly between the oral- and aboral layer and curves of PSII electron transport rates corrected for scalar irradiance in hospite, showed that the light use efficiency under sub-saturating light conditions were similar between the two layers. However, the aboral Symbiodinium band did not experience photosynthetic saturation, even at the highest investigated irradiance where the oral layer was clearly saturated. We thus found a different light acclimation response for the oral and aboral symbiont bands in hospite, and discuss whether such response could be shaped by spectral shifts caused by tissue gradients of scalar irradiance. Based on our experimental finding, combined with previous knowledge, we present a conceptual model on the photophysiology of Symbiodinium residing inside living coral tissue under natural gradients of light and chemical parameters. PMID:26955372

Photosynthetic Acclimation of Symbiodinium in hospite Depends on Vertical Position in the Tissue of the Scleractinian Coral Montastrea curta.

PubMed

Lichtenberg, Mads; Larkum, Anthony W D; Kühl, Michael

2016-01-01

Coral photophysiology has been studied intensively from the colony scale down to the scale of single fluorescent pigment granules as light is one of the key determinants for coral health. We studied the photophysiology of the oral and aboral symbiont band of scleractinian coral Montastrea curta to investigate if different acclimation to light exist in hospite on a polyp scale. By combined use of electrochemical and fiber-optic microsensors for O2, scalar irradiance and variable chlorophyll fluorescence, we could characterize the physical and chemical microenvironment experienced by the symbionts and, for the first time, estimate effective quantum yields of PSII photochemistry and rates of electron transport at the position of the zooxanthellae corrected for the in-tissue gradient of scalar irradiance. The oral- and aboral Symbiodinium layers received ∼71% and ∼33% of surface scalar irradiance, respectively, and the two symbiont layers experience considerable differences in light exposure. Rates of gross photosynthesis did not differ markedly between the oral- and aboral layer and curves of PSII electron transport rates corrected for scalar irradiance in hospite, showed that the light use efficiency under sub-saturating light conditions were similar between the two layers. However, the aboral Symbiodinium band did not experience photosynthetic saturation, even at the highest investigated irradiance where the oral layer was clearly saturated. We thus found a different light acclimation response for the oral and aboral symbiont bands in hospite, and discuss whether such response could be shaped by spectral shifts caused by tissue gradients of scalar irradiance. Based on our experimental finding, combined with previous knowledge, we present a conceptual model on the photophysiology of Symbiodinium residing inside living coral tissue under natural gradients of light and chemical parameters.

Stem Cell Research: A Novel Boulevard towards Improved Bovine Mastitis Management

PubMed Central

Sharma, Neelesh; Jeong, Dong Kee

2013-01-01

The dairy industry is a multi-billion dollar industry catering the nutritional needs of all age groups globally through the supply of milk. Clinical mastitis has a severe impact on udder tissue and is also an animal welfare issue. Moreover, it significantly reduces animal value and milk production. Mammary tissue damage reduces the number and activity of epithelial cells and consequently contributes to decreased milk production. The high incidence, low cure rate of this highly economic and sometimes deadly disease is an alarming for dairy sector as well as policy makers. Bovine mammary epithelial cells (MECs) and their stem cells are very important in milk production and bioengineering. The adult mammary epithelium consists of two main cell types; an inner layer of luminal epithelial cells, which produce the milk during lactation, and an outer layer of myoepithelial cells resting on a basement membrane, which are responsible for pushing the milk through the ductal network to the teat cistern. Inner layer of columner/luminal cells of bovine MECs, is characterized by cytokeratin18, 19 (CK18, CK19) and outer layer such as myoepithelial cells which are characterized by CK14, α-smooth muscle actin (α-SMA) and p63. Much work has been done in mouse and human, on mammary gland stem cell research, particularly in cancer therapy, but stem cell research in bovine is still in its infancy. Such stem/progenitor cell discoveries in human and mouse mammary gland bring some hope for application in bovines. These progenitors may be therapeutically adopted to correct the structural/cytological defects in the bovine udder due to mastitis. In the present review we focused on various kinds of stem/progenitor cells which can have therapeutic utility and their possibilities to use as a potential stem cell therapy in the management of bovine post-mastitis damage in orders to restore milk production. The possibilities of bovine mammary stem cell therapy offers significant potential for regeneration of tissues that can potentially replace/repair diseased and damaged tissue through differentiation into epithelial, myoepithelial and/or cuboidal/columnar cells in the udder with minimal risk of rejection and side effects. PMID:23983615

Stem cell research: a novel boulevard towards improved bovine mastitis management.

PubMed

Sharma, Neelesh; Jeong, Dong Kee

2013-01-01

The dairy industry is a multi-billion dollar industry catering the nutritional needs of all age groups globally through the supply of milk. Clinical mastitis has a severe impact on udder tissue and is also an animal welfare issue. Moreover, it significantly reduces animal value and milk production. Mammary tissue damage reduces the number and activity of epithelial cells and consequently contributes to decreased milk production. The high incidence, low cure rate of this highly economic and sometimes deadly disease is an alarming for dairy sector as well as policy makers. Bovine mammary epithelial cells (MECs) and their stem cells are very important in milk production and bioengineering. The adult mammary epithelium consists of two main cell types; an inner layer of luminal epithelial cells, which produce the milk during lactation, and an outer layer of myoepithelial cells resting on a basement membrane, which are responsible for pushing the milk through the ductal network to the teat cistern. Inner layer of columner/luminal cells of bovine MECs, is characterized by cytokeratin18, 19 (CK18, CK19) and outer layer such as myoepithelial cells which are characterized by CK14, α-smooth muscle actin (α-SMA) and p63. Much work has been done in mouse and human, on mammary gland stem cell research, particularly in cancer therapy, but stem cell research in bovine is still in its infancy. Such stem/progenitor cell discoveries in human and mouse mammary gland bring some hope for application in bovines. These progenitors may be therapeutically adopted to correct the structural/cytological defects in the bovine udder due to mastitis. In the present review we focused on various kinds of stem/progenitor cells which can have therapeutic utility and their possibilities to use as a potential stem cell therapy in the management of bovine post-mastitis damage in orders to restore milk production. The possibilities of bovine mammary stem cell therapy offers significant potential for regeneration of tissues that can potentially replace/repair diseased and damaged tissue through differentiation into epithelial, myoepithelial and/or cuboidal/columnar cells in the udder with minimal risk of rejection and side effects.

Gene evolution and functions of extracellular matrix proteins in teeth

PubMed Central

Yoshizaki, Keigo; Yamada, Yoshihiko

2013-01-01

The extracellular matrix (ECM) not only provides physical support for tissues, but it is also critical for tissue development, homeostasis and disease. Over 300 ECM molecules have been defined as comprising the “core matrisome” in mammals through the analysis of whole genome sequences. During tooth development, the structure and functions of the ECM dynamically change. In the early stages, basement membranes (BMs) separate two cell layers of the dental epithelium and the mesenchyme. Later in the differentiation stages, the BM layer is replaced with the enamel matrix and the dentin matrix, which are secreted by ameloblasts and odontoblasts, respectively. The enamel matrix genes and the dentin matrix genes are each clustered in two closed regions located on human chromosome 4 (mouse chromosome 5), except for the gene coded for amelogenin, the major enamel matrix protein, which is located on the sex chromosomes. These genes for enamel and dentin matrix proteins are derived from a common ancestral gene, but as a result of evolution, they diverged in terms of their specific functions. These matrix proteins play important roles in cell adhesion, polarity, and differentiation and mineralization of enamel and dentin matrices. Mutations of these genes cause diseases such as odontogenesis imperfect (OI) and amelogenesis imperfect (AI). In this review, we discuss the recently defined terms matrisome and matrixome for ECMs, as well as focus on genes and functions of enamel and dentin matrix proteins. PMID:23539364

Ballistic impacts on an anatomically correct synthetic skull with a surrogate skin/soft tissue layer.

PubMed

Mahoney, Peter; Carr, Debra; Arm, Richard; Gibb, Iain; Hunt, Nicholas; Delaney, Russ J

2018-03-01

The aim of this work was to further develop a synthetic model of ballistic head injury by the addition of skin and soft tissue layers to an anatomically correct polyurethane skull filled with gelatine 10% by mass. Six head models were impacted with 7.62 x 39 mm full metal jacket mild steel core (FMJ MSC) bullets with a mean velocity of 652 m/s. The impact events were filmed with high-speed cameras. The models were imaged pre- and post-impact using computed tomography. The models were assessed post impact by two experienced Home Office pathologists and the images assessed by an experienced military radiologist. The findings were scored against real injuries. The entry wounds, exit wounds and fracture patterns were scored positively, but the synthetic skin and soft tissue layer was felt to be too extendable. Further work is ongoing to address this.

Functional characterization of a novel 3D model of the epithelial-mesenchymal trophic unit.

PubMed

Bucchieri, Fabio; Pitruzzella, Alessandro; Fucarino, Alberto; Gammazza, Antonella Marino; Bavisotto, Celeste Caruso; Marcianò, Vito; Cajozzo, Massimo; Lo Iacono, Giorgio; Marchese, Roberto; Zummo, Giovanni; Holgate, Stephen T; Davies, Donna E

2017-03-01

Epithelial-mesenchymal communication plays a key role in tissue homeostasis and abnormal signaling contributes to chronic airways disease such as COPD. Most in vitro models are limited in complexity and poorly represent this epithelial-mesenchymal trophic unit. We postulated that cellular outgrowth from bronchial tissue would enable development of a mucosal structure that recapitulates better in vivo tissue architecture. Bronchial tissue was embedded in Matrigel and outgrowth cultures monitored using time-lapse microscopy, electrical resistance, light and electron microscopy. Cultures were challenged repetitively with cigarette smoke extract (CSE). The outgrowths formed as a multicellular sheet with motile cilia becoming evident as the Matrigel was remodeled to provide an air interface; cultures were viable for more than one year. Immunofluorescence and electron microscopy (EM) identified an upper layer of mucociliary epithelium and a lower layer of highly organized extracellular matrix (ECM) interspersed with fibroblastic cells separated by a basement membrane. EM analysis of the mucosal construct after repetitive exposure to CSE revealed epithelial damage, loss of cilia, and ECM remodeling, as occurs in vivo. We have developed a robust bronchial mucosal model. The structural changes observed following CSE exposure suggest the model should have utility for drug discovery and preclinical testing, especially those targeting airway remodeling.

Electrical structure in two thunderstorm anvil clouds

NASA Technical Reports Server (NTRS)

Marshall, Thomas C.; Rust, W. David; Winn, William P.; Gilbert, Kenneth E.

1989-01-01

Electrical structures in two thunderstorm anvil clouds (or 'anvils'), one in New Mexico, the other in Oklahoma, were investigated, using measurements of electric field by balloon-carried instruments and a one-dimensional model to calculate the time and spatial variations of electrical parameters in the clear air below the anvil. The electric field soundings through the two thunderstorm anvils showed similar charge structures; namely, negatively charged screening layers on the top and the bottom surfaces, a layer of positive charge in the interior, and one or two layers of zero charge. It is suggested that the positive charge originated in the main positive charge region normally found at high altitudes in the core of thunderclouds, and the negatively charged layers probably formed as screening layers, resulting from the discontinuity in the electrical conductivity at the cloud boundaries.

Microstructure and Mechanical Property of Glutaraldehyde-Treated Porcine Pulmonary Ligament.

PubMed

Chen, Huan; Zhao, Xuefeng; Berwick, Zachary C; Krieger, Joshua F; Chambers, Sean; Kassab, Ghassan S

2016-06-01

There is a significant need for fixed biological tissues with desired structural and material constituents for tissue engineering applications. Here, we introduce the lung ligament as a fixed biological material that may have clinical utility for tissue engineering. To characterize the lung tissue for potential clinical applications, we studied glutaraldehyde-treated porcine pulmonary ligament (n = 11) with multiphoton microscopy (MPM) and conducted biaxial planar experiments to characterize the mechanical property of the tissue. The MPM imaging revealed that there are generally two families of collagen fibers distributed in two distinct layers: The first family largely aligns along the longitudinal direction with a mean angle of θ = 10.7 ± 9.3 deg, while the second one exhibits a random distribution with a mean θ = 36.6 ± 27.4. Elastin fibers appear in some intermediate sublayers with a random orientation distribution with a mean θ = 39.6 ± 23 deg. Based on the microstructural observation, a microstructure-based constitutive law was proposed to model the elastic property of the tissue. The material parameters were identified by fitting the model to the biaxial stress-strain data of specimens, and good fitting quality was achieved. The parameter e0 (which denotes the strain beyond which the collagen can withstand tension) of glutaraldehyde-treated tissues demonstrated low variability implying a relatively consistent collagen undulation in different samples, while the stiffness parameters for elastin and collagen fibers showed relatively greater variability. The fixed tissues presented a smaller e0 than that of fresh specimen, confirming that glutaraldehyde crosslinking increases the mechanical strength of collagen-based biomaterials. The present study sheds light on the biomechanics of glutaraldehyde-treated porcine pulmonary ligament that may be a candidate for tissue engineering.

Quantitative assessment of optical properties in healthy cartilage and repair tissue by optical coherence tomography and histology (Conference Presentation)

NASA Astrophysics Data System (ADS)

Jansen, Sanne M. A.; Cernohorsky, Paul; de Bruin, Daniel M.; van der Pol, Edwin; Savci-Heijink, Cemile D.; Strackee, Simon D.; Faber, Dirk J.; van Leeuwen, Ton G.

2016-02-01

Quantification of the OCT signal is an important step toward clinical implementation of a diagnostic tool in cartilage imaging. Discrimination of structural cartilage differences in patients with osteoarthritis is critical, yet challenging. This study assesses the variation in the optical attenuation coefficient (μOCT) between healthy cartilage, repair tissue, bone and layers within repair tissue in a controlled setting. OCT and histology was used to assess goat talus articular surfaces in which central osteochondral defects were created. Exact matches of OCT and histology were selected for research. μOCT measurements were taken from healthy cartilage, repair tissue and bone. Measured μOCT in healthy cartilage was higher compared to both repair tissue and bone tissue. Two possible mechanisms for the difference in attenuation were investigated. We studied morphological parameters in terms of nucleus count, nucleus size and inter-nucleus distance. Collagen content in healthy cartilage and repair tissue was assessed using polarization microscopy. Quantitative analysis of the nuclei did not demonstrate a difference in nucleus size and count between healthy cartilage and repair tissue. In healthy cartilage, cells were spaced farther apart and had a lower variation in local nuclear density compared to repair tissue. Polarization microscopy suggested higher collagen content in healthy cartilage compared to repair tissue. μOCT measurements can distinguish between healthy cartilage, repair tissue and bone. Results suggest that cartilage OCT attenuation measurements could be of great impact in clinical diagnostics of osteoarthritis.

3D printing of tissue-simulating phantoms for calibration of biomedical optical devices

NASA Astrophysics Data System (ADS)

Zhao, Zuhua; Zhou, Ximing; Shen, Shuwei; Liu, Guangli; Yuan, Li; Meng, Yuquan; Lv, Xiang; Shao, Pengfei; Dong, Erbao; Xu, Ronald X.

2016-10-01

Clinical utility of many biomedical optical devices is limited by the lack of effective and traceable calibration methods. Optical phantoms that simulate biological tissues used for optical device calibration have been explored. However, these phantoms can hardly simulate both structural and optical properties of multi-layered biological tissue. To address this limitation, we develop a 3D printing production line that integrates spin coating, light-cured 3D printing and Fused Deposition Modeling (FDM) for freeform fabrication of optical phantoms with mechanical and optical heterogeneities. With the gel wax Polydimethylsiloxane (PDMS), and colorless light-curable ink as matrix materials, titanium dioxide (TiO2) powder as the scattering ingredient, graphite powder and black carbon as the absorption ingredient, a multilayer phantom with high-precision is fabricated. The absorption and scattering coefficients of each layer are measured by a double integrating sphere system. The results demonstrate that the system has the potential to fabricate reliable tissue-simulating phantoms to calibrate optical imaging devices.

Engineering Functional Epithelium for Regenerative Medicine and In Vitro Organ Models: A Review

PubMed Central

Vrana, Nihal E.; Lavalle, Philippe; Dokmeci, Mehmet R.; Dehghani, Fariba; Ghaemmaghami, Amir M.

2013-01-01

Recent advances in the fields of microfabrication, biomaterials, and tissue engineering have provided new opportunities for developing biomimetic and functional tissues with potential applications in disease modeling, drug discovery, and replacing damaged tissues. An intact epithelium plays an indispensable role in the functionality of several organs such as the trachea, esophagus, and cornea. Furthermore, the integrity of the epithelial barrier and its degree of differentiation would define the level of success in tissue engineering of other organs such as the bladder and the skin. In this review, we focus on the challenges and requirements associated with engineering of epithelial layers in different tissues. Functional epithelial layers can be achieved by methods such as cell sheets, cell homing, and in situ epithelialization. However, for organs composed of several tissues, other important factors such as (1) in vivo epithelial cell migration, (2) multicell-type differentiation within the epithelium, and (3) epithelial cell interactions with the underlying mesenchymal cells should also be considered. Recent successful clinical trials in tissue engineering of the trachea have highlighted the importance of a functional epithelium for long-term success and survival of tissue replacements. Hence, using the trachea as a model tissue in clinical use, we describe the optimal structure of an artificial epithelium as well as challenges of obtaining a fully functional epithelium in macroscale. One of the possible remedies to address such challenges is the use of bottom-up fabrication methods to obtain a functional epithelium. Modular approaches for the generation of functional epithelial layers are reviewed and other emerging applications of microscale epithelial tissue models for studying epithelial/mesenchymal interactions in healthy and diseased (e.g., cancer) tissues are described. These models can elucidate the epithelial/mesenchymal tissue interactions at the microscale and provide the necessary tools for the next generation of multicellular engineered tissues and organ-on-a-chip systems. PMID:23705900

Engineering functional epithelium for regenerative medicine and in vitro organ models: a review.

PubMed

Vrana, Nihal E; Lavalle, Philippe; Dokmeci, Mehmet R; Dehghani, Fariba; Ghaemmaghami, Amir M; Khademhosseini, Ali

2013-12-01

Recent advances in the fields of microfabrication, biomaterials, and tissue engineering have provided new opportunities for developing biomimetic and functional tissues with potential applications in disease modeling, drug discovery, and replacing damaged tissues. An intact epithelium plays an indispensable role in the functionality of several organs such as the trachea, esophagus, and cornea. Furthermore, the integrity of the epithelial barrier and its degree of differentiation would define the level of success in tissue engineering of other organs such as the bladder and the skin. In this review, we focus on the challenges and requirements associated with engineering of epithelial layers in different tissues. Functional epithelial layers can be achieved by methods such as cell sheets, cell homing, and in situ epithelialization. However, for organs composed of several tissues, other important factors such as (1) in vivo epithelial cell migration, (2) multicell-type differentiation within the epithelium, and (3) epithelial cell interactions with the underlying mesenchymal cells should also be considered. Recent successful clinical trials in tissue engineering of the trachea have highlighted the importance of a functional epithelium for long-term success and survival of tissue replacements. Hence, using the trachea as a model tissue in clinical use, we describe the optimal structure of an artificial epithelium as well as challenges of obtaining a fully functional epithelium in macroscale. One of the possible remedies to address such challenges is the use of bottom-up fabrication methods to obtain a functional epithelium. Modular approaches for the generation of functional epithelial layers are reviewed and other emerging applications of microscale epithelial tissue models for studying epithelial/mesenchymal interactions in healthy and diseased (e.g., cancer) tissues are described. These models can elucidate the epithelial/mesenchymal tissue interactions at the microscale and provide the necessary tools for the next generation of multicellular engineered tissues and organ-on-a-chip systems.

Bilayered, non-cross-linked collagen matrix for regeneration of facial defects after skin cancer removal: a new perspective for biomaterial-based tissue reconstruction.

PubMed

Ghanaati, Shahram; Kovács, Adorján; Barbeck, Mike; Lorenz, Jonas; Teiler, Anna; Sadeghi, Nader; Kirkpatrick, Charles James; Sader, Robert

2016-03-01

Classically skin defects are covered by split thickness skin grafts or by means of local or regional skin flaps. In the presented case series for the first time a bilayered, non-crossed-linked collagen matrix has been used in an off-label fashion in order to reconstruct facial skin defects following different types of skin cancer resection. The material is of porcine origin and consists of a spongy and a compact layer. The ratio of the two layers is 1:3 in favour of the spongy layer. The aim of the study was to investigate the potential of this matrix for skin regeneration as an alternative to the standard techniques of skin grafts or flaps. Six patients between 39 and 83 years old were included in the study based on a therapeutic trial. The collagen matrix was used in seven defects involving the nose, eyelid, forehead- and posterior scalp regions, and ranging from 1,2 to 6 cm in diameter. Two different head and neck surgeons at two different institutions performed the operations. Each used a different technique in covering the wound following surgery, i.e. with and without a latex-based sheet under the pressure dressing. In three cases cylindrical biopsies were taken after 14 days. In all cases the biomaterial application was performed without any complication and no adverse effects were observed. Clinically, the collagen matrix contributed to a tension-free skin regeneration, independent of the wound dressing used. The newly regenerated skin showed strong similarity to the adjacent normal tissue both in quality and colour. Histological analysis indicated that the spongy layer replaced the defective connective tissue, by providing stepwise integration into the surrounding implantation bed, while the compact layer was infiltrated by mononuclear cells and contributed to its epithelialization by means of a "conductive"process from the surrounding epithelial cells. The clinical and histological data demonstrate that the collagen bilayered matrix used in this series contributes to a "Guided-Integrative-Regeneration-Process", which still needs to be further understood. The biomimetic nature of this material seems to contribute to physiological matrix remodelling, which probably involves other matricellular proteins essential for soft tissue regeneration. A deeper understanding of the mechanism, involved in the tissue integration of this material and its contribution to soft tissue regeneration based on the direct and indirect effect of matricellular proteins could open new therapeutic avenues for biomaterial-based soft tissue regeneration as an alternative to traditional flap-based plastic surgery.

Excitation mechanism of surface plasmon polaritons in a double-layer wire grid structure

NASA Astrophysics Data System (ADS)

Motogaito, Atsushi; Nakajima, Tomoyasu; Miyake, Hideto; Hiramatsu, Kazumasa

2017-12-01

We characterize the optical properties of a double-layer wire grid structure and investigate in detail the excitation mechanism of surface plasmon polaritons (SPPs). Angular spectra for the transmittance of the transverse magnetic polarized light that are obtained through the experiment reveal two peaks. In addition, simulated mapping of the transmittance and the magnetic field distribution indicate that SPPs are excited in two areas of the wire grid structures: at the interface between the Au layer and the resist layer or the glass substrate and at the interface between the Au layer and air. The experimental data are consistent with the transmittance mapping result and the distribution of the magnetic field. Accordingly, we constructed a model of SPPs propagation. We consider that SPPs excited at the interface between the Au layer and the resist layer or the glass substrate strongly contribute to the extraordinary transmission observed in the wire grid structures.

A theoretical and experimental investigation of nonlinear propagation of ultrasound through tissue mimicking media

NASA Astrophysics Data System (ADS)

Rielly, Matthew Robert

An existing numerical model (known as the Bergen code) is used to investigate finite amplitude ultrasound propagation through multiple layers of tissue-like media. This model uses a finite difference method to solve the nonlinear parabolic KZK wave equation. The code is modified to include an arbitrary frequency dependence of absorption and transmission effects for wave propagation across a plane interface at normal incidence. In addition the code is adapted to calculate the total intensity loss associated with the absorption of the fundamental and nonlinearly generated harmonics. Measurements are also taken of the axial nonlinear pressure field generated from a circular focused, 2.25 MHz source, through single and multiple layered tissue mimicking fluids, for source pressures in the range from 13 kPa to 310 kPa. Two tissue mimicking fluids are developed to provide acoustic properties similar to amniotic fluid and a typical soft tissue. The values of the nonlinearity parameter, sound velocity and frequency dependence of attenuation for both fluids are presented, and the measurement procedures employed to obtain these characteristics are described in detail. These acoustic parameters, together with the measured source conditions are used as input to the numerical model, allowing the experimental conditions to be simulated. Extensive comparisons are made between the model's predictions and the axial pressure field measurements. Results are presented in the frequency domain showing the fundamental and three subsequent harmonic amplitudes on axis, as a function of axial distance. These show that significant nonlinear distortion can occur through media with characteristics typical of tissue. Time domain waveform comparisons are also made. An excellent agreement is found between theory and experiment indicating that the model can be used to predict nonlinear ultrasound propagation through multiple layers of tissue-like media. The numerical code is also used to model the intensity loss through layered tissue mimics and results are presented illustrating the effects of altering the layered medium on the magnitude and spatial distribution of intensity loss.

The Selective Centrifugation Ensures a Better In Vitro Isolation of ASCs and Restores a Soft Tissue Regeneration In Vivo

PubMed Central

De Francesco, Francesco; Guastafierro, Antonio; Nicoletti, Gianfranco; Razzano, Sergio; Riccio, Michele; Ferraro, Giuseppe A.

2017-01-01

Autologous fat grafting procedures in plastic surgery have been extensively used to reinforce soft tissue in congenital or acquired tissue impairments. With this background, the aim of this study is firstly to examine the impact of a selective centrifugation on existing adipose stem cells (ASCs) in terms of stemness profile maintenance and, secondly, to investigate the effect of restoring volume in reconstruction on patients affected by soft tissue damage. After centrifugation, the fat graft products were separated into two layers and subsequently examined in vitro for the expression of CD34, CD90, CD117, CD105, CD29, CD31, CD44, CD73, CD133, CD14 and CD45 markers by flow cytometry and gene expression analyses were performed for Sox2, WNT3A, END, CD44, FUT4, COLL1, CTNNB1, hbEGF, KRTLG, MMP2 and VIM genes. The results showed that in the middle-high density (MHD) layer there was a peak concentration of ASCs, compared to another layer obtained after centrifugation. Research carried out on patients under treatment for soft tissue regeneration using cells obtained from MHD layer selection will be fundamental in comparative analysis. These studies will lead to an adequate standardization of outcomes, provided that treatment is performed through cell selection. Therefore, a unique procedure in tissue reconstruction and regeneration through fat grafting is presented here. PMID:28498335

Beyond the single-file fluid limit using transfer matrix method: Exact results for confined parallel hard squares

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gurin, Péter; Varga, Szabolcs

2015-06-14

We extend the transfer matrix method of one-dimensional hard core fluids placed between confining walls for that case where the particles can pass each other and at most two layers can form. We derive an eigenvalue equation for a quasi-one-dimensional system of hard squares confined between two parallel walls, where the pore width is between σ and 3σ (σ is the side length of the square). The exact equation of state and the nearest neighbor distribution functions show three different structures: a fluid phase with one layer, a fluid phase with two layers, and a solid-like structure where the fluidmore » layers are strongly correlated. The structural transition between differently ordered fluids develops continuously with increasing density, i.e., no thermodynamic phase transition occurs. The high density structure of the system consists of clusters with two layers which are broken with particles staying in the middle of the pore.« less

Spatial frequency domain spectroscopy of two layer media

NASA Astrophysics Data System (ADS)

Yudovsky, Dmitry; Durkin, Anthony J.

2011-10-01

Monitoring of tissue blood volume and oxygen saturation using biomedical optics techniques has the potential to inform the assessment of tissue health, healing, and dysfunction. These quantities are typically estimated from the contribution of oxyhemoglobin and deoxyhemoglobin to the absorption spectrum of the dermis. However, estimation of blood related absorption in superficial tissue such as the skin can be confounded by the strong absorption of melanin in the epidermis. Furthermore, epidermal thickness and pigmentation varies with anatomic location, race, gender, and degree of disease progression. This study describes a technique for decoupling the effect of melanin absorption in the epidermis from blood absorption in the dermis for a large range of skin types and thicknesses. An artificial neural network was used to map input optical properties to spatial frequency domain diffuse reflectance of two layer media. Then, iterative fitting was used to determine the optical properties from simulated spatial frequency domain diffuse reflectance. Additionally, an artificial neural network was trained to directly map spatial frequency domain reflectance to sets of optical properties of a two layer medium, thus bypassing the need for iteration. In both cases, the optical thickness of the epidermis and absorption and reduced scattering coefficients of the dermis were determined independently. The accuracy and efficiency of the iterative fitting approach was compared with the direct neural network inversion.

Radiation properties of two types of luminous textile devices containing plastic optical fibers

NASA Astrophysics Data System (ADS)

Selm, Bärbel; Rothmaier, Markus

2007-05-01

Luminous textiles have the potential to satisfy a need for thin and flexible light diffusers for treatment of intraoral cancerous tissue. Plastic optical fibers (POF) with diameters of 250 microns and smaller are used to make the textiles luminous. Usually light is supplied to the optical fiber at both ends. On the textile surface light emission occurs in a woven structure via damaged straight POFs, whereas the embroidered structure radiates the light out of macroscopically bent POFs. We compared the optical properties of these two types of textile diffusers using red light laser for the embroidery and light emitting diode (LED) for the woven structure as light sources, and found efficiencies for the luminous areas of the two samples of 19 % (woven) and 32 % (embroidery), respectively. It was shown that the efficiency can be greatly improved using an aluminium backing. Additional scattering layers lower the fluence rate by around 30 %. To analyse the homogeneity we took a photo of the illuminated surface using a 3CCD camera and found, for both textiles, a slightly skewed distribution of the dark and bright pixels. The interquartile range of brightness distribution of the embroidery is more than double as the woven structure.

Stem/Progenitor Cell–Mediated De Novo Regeneration of Dental Pulp with Newly Deposited Continuous Layer of Dentin in an In Vivo Model

PubMed Central

Yamaza, Takayoshi; Shea, Lonnie D.; Djouad, Farida; Kuhn, Nastaran Z.; Tuan, Rocky S.; Shi, Songtao

2010-01-01

The ultimate goal of this study is to regenerate lost dental pulp and dentin via stem/progenitor cell–based approaches and tissue engineering technologies. In this study, we tested the possibility of regenerating vascularized human dental pulp in emptied root canal space and producing new dentin on existing dentinal walls using a stem/progenitor cell–mediated approach with a human root fragment and an immunocompromised mouse model. Stem/progenitor cells from apical papilla and dental pulp stem cells were isolated, characterized, seeded onto synthetic scaffolds consisting of poly-D,L-lactide/glycolide, inserted into the tooth fragments, and transplanted into mice. Our results showed that the root canal space was filled entirely by a pulp-like tissue with well-established vascularity. In addition, a continuous layer of dentin-like tissue was deposited onto the canal dentinal wall. This dentin-like structure appeared to be produced by a layer of newly formed odontoblast-like cells expressing dentin sialophosphoprotein, bone sialoprotein, alkaline phosphatase, and CD105. The cells in regenerated pulp-like tissue reacted positively to anti-human mitochondria antibodies, indicating their human origin. This study provides the first evidence showing that pulp-like tissue can be regenerated de novo in emptied root canal space by stem cells from apical papilla and dental pulp stem cells that give rise to odontoblast-like cells producing dentin-like tissue on existing dentinal walls. PMID:19737072

Organic electrochemical transistors for cell-based impedance sensing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rivnay, Jonathan, E-mail: rivnay@emse.fr, E-mail: owens@emse.fr; Ramuz, Marc; Hama, Adel

2015-01-26

Electrical impedance sensing of biological systems, especially cultured epithelial cell layers, is now a common technique to monitor cell motion, morphology, and cell layer/tissue integrity for high throughput toxicology screening. Existing methods to measure electrical impedance most often rely on a two electrode configuration, where low frequency signals are challenging to obtain for small devices and for tissues with high resistance, due to low current. Organic electrochemical transistors (OECTs) are conducting polymer-based devices, which have been shown to efficiently transduce and amplify low-level ionic fluxes in biological systems into electronic output signals. In this work, we combine OECT-based drain currentmore » measurements with simultaneous measurement of more traditional impedance sensing using the gate current to produce complex impedance traces, which show low error at both low and high frequencies. We apply this technique in vitro to a model epithelial tissue layer and show that the data can be fit to an equivalent circuit model yielding trans-epithelial resistance and cell layer capacitance values in agreement with literature. Importantly, the combined measurement allows for low biases across the cell layer, while still maintaining good broadband signal.« less

[Parasitological examinations of the hydatoid cysts of liver].

PubMed

Cielecka, D; Gierczak, A; Michałowicz, B; Grytner-Ziecina, B

2001-01-01

Liver cysts taken from 9 patients, operated on in the Medical Academy Clinic in 2000, have been examined. Cyst layers, their thickness and content have been analysed. We have searched for the cestoid scolexes and rostellar hooks. In 6 patients Echinococcus granulosus larvae have been found. In 4 cases there have been protoscolexes discovered, however with some signs of degeneration of the larva. In one case dead larva has been diagnosed on the basis of the presence of single hooks and in one patient one larva alive with unchanged structures has been revealed. Profound parasitological examination is a good supplement to the examination of the histopathological slides, especially in the case of degenerating larvae, when the tissues of the parasite lose their contact with the connective tissue layer of the host. Separated layer fragments, as well as protoscolexes and rostellar hooks need not be exposed to histological cutting.

Mechanical roles of apical constriction, cell elongation, and cell migration during neural tube formation in Xenopus.

PubMed

Inoue, Yasuhiro; Suzuki, Makoto; Watanabe, Tadashi; Yasue, Naoko; Tateo, Itsuki; Adachi, Taiji; Ueno, Naoto

2016-12-01

Neural tube closure is an important and necessary process during the development of the central nervous system. The formation of the neural tube structure from a flat sheet of neural epithelium requires several cell morphogenetic events and tissue dynamics to account for the mechanics of tissue deformation. Cell elongation changes cuboidal cells into columnar cells, and apical constriction then causes them to adopt apically narrow, wedge-like shapes. In addition, the neural plate in Xenopus is stratified, and the non-neural cells in the deep layer (deep cells) pull the overlying superficial cells, eventually bringing the two layers of cells to the midline. Thus, neural tube closure appears to be a complex event in which these three physical events are considered to play key mechanical roles. To test whether these three physical events are mechanically sufficient to drive neural tube formation, we employed a three-dimensional vertex model and used it to simulate the process of neural tube closure. The results suggest that apical constriction cued the bending of the neural plate by pursing the circumference of the apical surface of the neural cells. Neural cell elongation in concert with apical constriction further narrowed the apical surface of the cells and drove the rapid folding of the neural plate, but was insufficient for complete neural tube closure. Migration of the deep cells provided the additional tissue deformation necessary for closure. To validate the model, apical constriction and cell elongation were inhibited in Xenopus laevis embryos. The resulting cell and tissue shapes resembled the corresponding simulation results.

Computer-aided design of microvasculature systems for use in vascular scaffold production.

PubMed

Mondy, William Lafayette; Cameron, Don; Timmermans, Jean-Pierre; De Clerck, Nora; Sasov, Alexander; Casteleyn, Christophe; Piegl, Les A

2009-09-01

In vitro biomedical engineering of intact, functional vascular networks, which include capillary structures, is a prerequisite for adequate vascular scaffold production. Capillary structures are necessary since they provide the elements and compounds for the growth, function and maintenance of 3D tissue structures. Computer-aided modeling of stereolithographic (STL) micro-computer tomographic (micro-CT) 3D models is a technique that enables us to mimic the design of vascular tree systems containing capillary beds, found in tissues. In our first paper (Mondy et al 2009 Tissue Eng. at press), using micro-CT, we studied the possibility of using vascular tissues to produce data capable of aiding the design of vascular tree scaffolding, which would help in the reverse engineering of a complete vascular tree system including capillary bed structures. In this paper, we used STL models of large datasets of computer-aided design (CAD) data of vascular structures which contained capillary structures that mimic those in the dermal layers of rabbit skin. Using CAD software we created from 3D STL models a bio-CAD design for the development of capillary-containing vascular tree scaffolding for skin. This method is designed to enhance a variety of therapeutic protocols including, but not limited to, organ and tissue repair, systemic disease mediation and cell/tissue transplantation therapy. Our successful approach to in vitro vasculogenesis will allow the bioengineering of various other types of 3D tissue structures, and as such greatly expands the potential applications of biomedical engineering technology into the fields of biomedical research and medicine.

Photoacoustic characterization of the left atrium wall: healthy and ablated tissue (Conference Presentation)

NASA Astrophysics Data System (ADS)

Iskander-Rizk, Sophinese; Kruizinga, Pieter; van der Steen, Antonius F. W.; van Soest, Gijs

2017-03-01

Radio-frequency ablation (RFA) creates a thermal lesion in the atrial wall, with clearly recognizable optical and structural changes to the tissue. This can be detected by photoacoustic (PA) imaging, and used for monitoring of lesion depth, lesion functionality, and limiting excessive ablation. Porcine left atrium tissue can be split into three visually distinguishable regions, a thick white endocardium, pinkish myocardium and a thin gelatinous epicardium. In this study, we characterize the layered left atrium tissue in terms of the relevant photoacoustic parameters (wavelength, frequency content, imaging depth, lesion contrast). Previous studies in the literature targeted the photoacoustic characterization of fresh and ablated ventricular myocardium in the range of 650nm to 900nm. In this study we target the characterization of fresh and ablated left atrial tissue from 410nm to 1000nm, including the endocardium and epicardium. We generate the photoacoustic signals using a tunable pulsed laser source, and record those signals using either a broadband 1 mm hydrophone or a L12-3v transducer connected to the Verasonics machine for more realistic conditions. Initial experiments on fresh porcine tissue show that the presence of the endocardium and epicardium layers do affect the photoacoustic signal received. The signal recorded is representative of the difference in optical and mechanical properties between the layers. Ablated and non-ablated tissue also present differences in spectra. The determined optical contrast could be used in the PA monitoring of RFA lesion to monitor the extension of the lesion to the edge of the myocardium-epicardium border avoiding complications related to over ablation.

Techniques of noninvasive optical tomographic imaging

NASA Astrophysics Data System (ADS)

Rosen, Joseph; Abookasis, David; Gokhler, Mark

2006-01-01

Recently invented methods of optical tomographic imaging through scattering and absorbing media are presented. In one method, the three-dimensional structure of an object hidden between two biological tissues is recovered from many noisy speckle pictures obtained on the output of a multi-channeled optical imaging system. Objects are recovered from many speckled images observed by a digital camera through two stereoscopic microlens arrays. Each microlens in each array generates a speckle image of the object buried between the layers. In the computer each image is Fourier transformed jointly with an image of the speckled point-like source captured under the same conditions. A set of the squared magnitudes of the Fourier-transformed pictures is accumulated to form a single average picture. This final picture is again Fourier transformed, resulting in the three-dimensional reconstruction of the hidden object. In the other method, the effect of spatial longitudinal coherence is used for imaging through an absorbing layer with different thickness, or different index of refraction, along the layer. The technique is based on synthesis of multiple peak spatial degree of coherence. This degree of coherence enables us to scan simultaneously different sample points on different altitudes, and thus decreases the acquisition time. The same multi peak degree of coherence is also used for imaging through the absorbing layer. Our entire experiments are performed with a quasi-monochromatic light source. Therefore problems of dispersion and inhomogeneous absorption are avoided.

Preparation of a novel composite nanofiber gel-encapsulated human placental extract through layer-by-layer self-assembly

PubMed Central

LIU, GUOHUI; CHEN, XI; ZHOU, WU; YANG, SHUHUA; YE, SHUNAN; CAO, FAQI; LIU, YI; XIONG, YUAN

2016-01-01

Aqueous human placenta extract (HPE) has been previously used to treat chronic soft tissue ulcer; however, the optimal dosage of HPE has yet to be elucidated. The present study investigated a novel nanofiber gel composed through layer-by-layer (LbL) self-assembly, in which HPE was encapsulated. IKVAV, RGD, RAD16 and FGL-PA were screened and combined to produce an optimal vehicle nanofiber gel through LbL assembly. Subsequently, the aqueous HPE was encapsulated into this nanofiber at the appropriate concentration, and the morphology, particle size, drug loading efficacy, encapsulation rate, release efficiency and structure validation were detected. The encapsulation efficiency of all three HPE samples was >90%, the nanofiber gel exhibited a slow releasing profile, and the structure of HPE encapsulated in the nanofiber gel was unvaried. In conclusion, this type of novel composite nanocapsules may offer a promising delivery system for HPE. PMID:27073463

Quantitative, depth-resolved determination of particle motion using multi-exposure, spatial frequency domain laser speckle imaging.

PubMed

Rice, Tyler B; Kwan, Elliott; Hayakawa, Carole K; Durkin, Anthony J; Choi, Bernard; Tromberg, Bruce J

2013-01-01

Laser Speckle Imaging (LSI) is a simple, noninvasive technique for rapid imaging of particle motion in scattering media such as biological tissue. LSI is generally used to derive a qualitative index of relative blood flow due to unknown impact from several variables that affect speckle contrast. These variables may include optical absorption and scattering coefficients, multi-layer dynamics including static, non-ergodic regions, and systematic effects such as laser coherence length. In order to account for these effects and move toward quantitative, depth-resolved LSI, we have developed a method that combines Monte Carlo modeling, multi-exposure speckle imaging (MESI), spatial frequency domain imaging (SFDI), and careful instrument calibration. Monte Carlo models were used to generate total and layer-specific fractional momentum transfer distributions. This information was used to predict speckle contrast as a function of exposure time, spatial frequency, layer thickness, and layer dynamics. To verify with experimental data, controlled phantom experiments with characteristic tissue optical properties were performed using a structured light speckle imaging system. Three main geometries were explored: 1) diffusive dynamic layer beneath a static layer, 2) static layer beneath a diffuse dynamic layer, and 3) directed flow (tube) submerged in a dynamic scattering layer. Data fits were performed using the Monte Carlo model, which accurately reconstructed the type of particle flow (diffusive or directed) in each layer, the layer thickness, and absolute flow speeds to within 15% or better.

Development of human nervous tissue upon differentiation of embryonic stem cells in three-dimensional culture.

PubMed

Preynat-Seauve, Olivier; Suter, David M; Tirefort, Diderik; Turchi, Laurent; Virolle, Thierry; Chneiweiss, Herve; Foti, Michelangelo; Lobrinus, Johannes-Alexander; Stoppini, Luc; Feki, Anis; Dubois-Dauphin, Michel; Krause, Karl Heinz

2009-03-01

Researches on neural differentiation using embryonic stem cells (ESC) require analysis of neurogenesis in conditions mimicking physiological cellular interactions as closely as possible. In this study, we report an air-liquid interface-based culture of human ESC. This culture system allows three-dimensional cell expansion and neural differentiation in the absence of added growth factors. Over a 3-month period, a macroscopically visible, compact tissue developed. Histological coloration revealed a dense neural-like neural tissue including immature tubular structures. Electron microscopy, immunochemistry, and electrophysiological recordings demonstrated a dense network of neurons, astrocytes, and oligodendrocytes able to propagate signals. Within this tissue, tubular structures were niches of cells resembling germinal layers of human fetal brain. Indeed, the tissue contained abundant proliferating cells expressing markers of neural progenitors. Finally, the capacity to generate neural tissues on air-liquid interface differed for different ESC lines, confirming variations of their neurogenic potential. In conclusion, this study demonstrates in vitro engineering of a human neural-like tissue with an organization that bears resemblance to early developing brain. As opposed to previously described methods, this differentiation (a) allows three-dimensional organization, (b) yields dense interconnected neural tissue with structurally and functionally distinct areas, and (c) is spontaneously guided by endogenous developmental cues.

Structural and material approaches to bone tissue engineering in powder-based three-dimensional printing.

PubMed

Butscher, A; Bohner, M; Hofmann, S; Gauckler, L; Müller, R

2011-03-01

This article reviews the current state of knowledge concerning the use of powder-based three-dimensional printing (3DP) for the synthesis of bone tissue engineering scaffolds. 3DP is a solid free-form fabrication (SFF) technique building up complex open porous 3D structures layer by layer (a bottom-up approach). In contrast to traditional fabrication techniques generally subtracting material step by step (a top-down approach), SFF approaches allow nearly unlimited designs and a large variety of materials to be used for scaffold engineering. Today's state of the art materials, as well as the mechanical and structural requirements for bone scaffolds, are summarized and discussed in relation to the technical feasibility of their use in 3DP. Advances in the field of 3DP are presented and compared with other SFF methods. Existing strategies on material and design control of scaffolds are reviewed. Finally, the possibilities and limiting factors are addressed and potential strategies to improve 3DP for scaffold engineering are proposed. Copyright © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

In vitro developmental model of the gastrointestinal tract from mouse embryonic stem cells.

PubMed

Torihashi, Shigeko; Kuwahara, Masaki; Kurahashi, Masaaki

2007-10-01

Mouse embryonic stem (ES) cells are pluripotent and retain their potential to form cells, tissues and organs originated from three embryonic germ layers. Recently, we developed in vitro organ--gut-like structures--from mouse ES cells. They had basically similar morphological features to a mouse gastrointestinal tract in vivo composed of three distinct layers (i.e., epithelium, connective tissue and musculature). Gut-like structures showed spontaneous contractions derived from pacemaker cells (interstitial cells of Cajal) in the musculature. We also examined their formation process and expression pattern of transcription factors crucial for gut organogenesis such as Id2, Sox17, HNF3beta/Foxa2 and GATA4. We found that they mimic the development of embryonic gut in vivo and showed a similar expression pattern of common transcription factors. They also maintain their developmental potential after transplantation to a renal capsule. Therefore, gut-like structures are suitable for in vitro models of gastrointestinal tracts and their development. In addition, we pointed out several unique features different from gut in vivo that provide useful and advantageous tools to investigate the developmental mechanism of the gastrointestinal tract.

Morphology of the copulatory apparatus of the spotted tinamou Nothura maculosa (Aves: Tinamiformes).

PubMed

de Oliveira, C A; Mahecha, G A

2000-03-01

The components of the copulatory apparatus of Nothura maculosa were identified and studied by means of dissections, parenteral latex injections and standard histological and histochemical techniques. N. maculosa possesses an intromittent phallus with a tubular cavity, within which a fixed base and a tubular portion can be distinguished. An ejaculatory groove, which is supported by a fibrous body, runs dorsally at the base of the phallus. The tubular portion of the phallus inserts into the fibrous body, the former possessing two continuous but morphologically distinct parts, one of which is eversible and is extruded during erection while the other is a fixed non-eversible glandular portion. The walls of both parts are formed by the mucosa, an intermediate layer of connective tissue containing a wide lymphatic space which surrounds the whole perimeter of the tube, and an external layer of dense connective tissue. The mucosa of the eversible portion, which lodges the phallic groove, is lined by a non-keratinized stratified squamous epithelium. In the fixed tubular portion, the tubular lumen is reduced in size, the phallic groove disappears, the mucosa becomes folded and there occurs an abrupt change to a pseudostratified columnar secretory epithelium. The copulatory apparatus of N. maculosa includes the vascular bodies of the phallus, which show morphofunctional continuity with the phallic structures, thus forming a single system involved in erection. On erection, the eversible portion of the phallus evaginates and emerges from the cloacal opening as a spiral shaped structure directed towards the left.

Nanosized CaP-silk fibroin-PCL-PEG-PCL/PCL based bilayer membranes for guided bone regeneration.

PubMed

Türkkan, Sibel; Pazarçeviren, A Engin; Keskin, Dilek; Machin, Nesrin E; Duygulu, Özgür; Tezcaner, Ayşen

2017-11-01

Guided bone regeneration (GBR) concept has been developed to prevent the formation of non-functional scar tissue layer on defect site by undertaking barrier role. In this study, a new bilayer membrane which consisted of one layer of electrospun silk fibroin/PCL-PEG-PCL incorporating nanocalcium phosphate (SPCA) 1 and one layer of PCL membrane was developed for GBR. To improve the osteoconductivity of membranes, nanosized calcium phosphate particles synthesized by Flame Spray Pyrolysis method were incorporated into membranes at 10% (wt) (SPCA10) and 20% (wt) (SPCA20) of the polymer content. The structural and chemical analyses revealed the well-integrated two layers of membranes with a total thickness of ca 100μm. In the regenerative layer, the highly porous mesh structure had a thickness of 12.6μm with randomly oriented fibers having diameters around 760nm, and nanoparticles dispersed homogenously. The mechanical test results showed remarkable improvement on the tensile strength of membranes with incorporation of nanoparticles. Higher water affinity of nanoCaP included membranes was proved by lower contact angle values and higher percent water uptake capacity. Biomineralization assay revealed that nucleation and growth of apatites around fibers of SPCA10 and SPCA20 were apparent while on SPCA0 apatite minerals were barely detected after 10days. Human dental pulp stem cells (DPSC) were seeded on electrospun layer of the bilayer membranes for biocompatibility and osteo-compatibility study. Increasing nanoCaP amount resulted in higher cell adhesion, proliferation, ALP activity and calcium deposition on membranes. These overall results confirmed the biocompatibility and potential applicability of proposed membranes for GBR treatments. Copyright © 2017 Elsevier B.V. All rights reserved.

Do Skeletal Density Changes Within the Tissue Layer of Corals Affect Paleoclimate Reconstructions?

NASA Astrophysics Data System (ADS)

Griffiths, J. S.; DeLong, K. L.; Quinn, T.; Taylor, F. W.; Kilbourne, K. H.; Wagner, A. J.

2016-02-01

Sea surface temperature (SST) reconstructions from coral geochemistry provide information on past climate variability; however, not all coral studies agree on a common calibration slope. Therefore, understanding the impacts of coral skeletal growth on strontium-to-calcium ratios (Sr/Ca) and oxygen isotopic ratios (δ18O) is necessary to ensure accurate calibrations. The study of Gagan et al. (2012) suggests that for the Pacific coral genera Porites, SST calibrations for coral Sr/Ca and δ18O need to be adjusted to account for skeletal density changes in the tissue layer, which may attenuate the seasonal cycle in coral geochemistry. We attempt to duplicate those results and density patterns in several Porites lutea colonies from two locations, yet our results do not show an increase in density in the tissue layer. Another study with Montastraea faveolata reveals reduced seasonality in coral Sr/Ca compared to slower-growing Siderastrea siderea in close proximity and same water depth, suggesting the faster growing M. faveolata geochemistry may be attenuated. By measuring skeletal density changes by micromilling a standard volume throughout the tissue layer and immediately below, we find no pattern of skeletal accumulation in the tissue layer of multiple colonies of M. faveolata and S. siderea from different locations. We conclude that these species lay down all of their skeletal material at the skeleton surface, thus skeletal density changes in the tissue layer do not account for reduced seasonality. We propose that time averaging occurs in M. faveolata as a result of the coral polyp's deep calyces mixing time intervals in the adjacent thecal wall in which micromilling for geochemical analysis produces a sample area that contains several growth increments. Our results show that skeletal density growth effects cannot be applied to all coral genera and paves the way for new research on calyx depth as an alternative explanation for differences in coral calibration slopes.

Modeling of Surface Thermodynamics and Damage Thresholds in the IR and THz Regime

DTIC Science & Technology

2007-01-01

Conference on Optical Interactions with Tissue and Cells [18th] Held in San Jose, California on January 22-24, 2007 To order the complete compilation report...United States; c Air Force Reasearch Lab, Human Effectivness Directorate Optical Branch, 2624 Louis Bauer Drive, San Antonio, TX, United States...equation (radial and axial) in a biological system construct. Tissues are represented as multi-layer structures, with optical and thermal properties

The double layers in the plasma sheet boundary layer during magnetic reconnection

NASA Astrophysics Data System (ADS)

Guo, J.; Yu, B.

2014-11-01

We studied the evolutions of double layers which appear after the magnetic reconnection through two-dimensional electromagnetic particle-in-cell simulation. The simulation results show that the double layers are formed in the plasma sheet boundary layer after magnetic reconnection. At first, the double layers which have unipolar structures are formed. And then the double layers turn into bipolar structures, which will couple with another new weak bipolar structure. Thus a new double layer or tripolar structure comes into being. The double layers found in our work are about several ten Debye lengths, which accords with the observation results. It is suggested that the electron beam formed during the magnetic reconnection is responsible for the production of the double layers.

Development and characterization of non-resonant multiphoton photoacoustic spectroscopy (NMPPAS) for brain tumor margining

NASA Astrophysics Data System (ADS)

Dahal, Sudhir

During tumor removal surgery, due to the problems associated with obtaining high-resolution, real-time chemical images of where exactly the tumor ends and healthy tissue begins (tumor margining), it is often necessary to remove a much larger volume of tissue than the tumor itself. In the case of brain tumor surgery, however, it is extremely unsafe to remove excess tissue. Therefore, without an accurate image of the tumor margins, some of the tumor's finger-like projections are inevitably left behind in the surrounding parenchyma to grow again. For this reason, the development of techniques capable of providing high-resolution real-time images of tumor margins up to centimeters below the surface of a tissue is ideal for the diagnosis and treatment of tumors, as well as surgical guidance during brain tumor excision. A novel spectroscopic technique, non-resonant multiphoton photoacoustic spectroscopy (NMPPAS), is being developed with the capabilities of obtaining high-resolution subsurface chemical-based images of underlying tumors. This novel technique combines the strengths of multiphoton tissue spectroscopy and photoacoustic spectroscopy into a diagnostic methodology that will, ultimately, provide unparalleled chemical information and images to provide the state of sub-surface tissues. The NMPPAS technique employs near-infrared light (in the diagnostic window) to excite ultraviolet and/or visible light absorbing species deep below the tissue's surface. Once a multiphoton absorption event occurs, non-radiative relaxation processes generates a localized thermal expansion and subsequent acoustic wave that can be detected using a piezoelectric transducer. Since NMPPAS employs an acoustic detection modality, much deeper diagnoses can be performed than that is possible using current state of the art high-resolution chemical imaging techniques such as multiphoton fluorescence spectroscopy. NMPPAS was employed to differentiate between excised brain tumors (astrocytoma III) and healthy tissue with over 99% accuracy. NMPPAS spectral features showed evident differences between tumor and healthy tissues, and ratiometric analysis ensured that only a few wavelengths could be used for excitation instead of using numerous wavelength excitations to create spectra. This process would significantly reduce the analysis time while maintaining the same degree of accuracy. Tissue phantoms were fabricated in order to characterize the properties of NMPPAS. Scattering particles were doped into the phantoms to simulate their light scattering properties to real tissues. This allowed for better control over shape, size, reproducibility and doping in the sample while maintaining the light-tissue interaction properties of real tissue. To make NMPPAS viable for clinical applications, the technique was characterized to determine the spatial (lateral and longitudinal) resolution, depth of penetration and its ability to image in three-dimension through layers of tissue. Both resolutions were determined to be near-cellular level resolution (50-70 microm), obtained initially with the aid of the technique of multiphoton fluorescence, and later verified using NMPPAS imaging. Additionally, the maximum depth of penetration and detection was determined to be about 1.4cm, making the technique extremely suitable to margin tumors from underlying tissues in the brain. The capability of NMPPAS to detect and image layers that lie beneath other structures and blood vessels was also investigated. Three-dimensional images were obtained for the first time using NMPPAS. The images were obtained from different depths and structures were imaged through other layers of existing structures in the sample. This verified that NMPPAS was capable of detecting and imaging structures that lie embedded within the tissues. NMPPAS images of embedded structures were also obtained with the presence of hemoglobin, which is potentially the largest source of background in blood-perfused tissues, thus showing that the technique is capable of detecting and differentiating in blood-perfused samples.

Structural stability and sustained release of protein from a multilayer nanofiber/nanoparticle composite.

PubMed

Vakilian, Saeid; Mashayekhan, Shohreh; Shabani, Iman; Khorashadizadeh, Mohsen; Fallah, Ali; Soleimani, Masoud

2015-04-01

The cellular microenvironment can be engineered through the utilization of various nano-patterns and matrix-loaded bioactive molecules. In this study, a multilayer system of electrospun scaffold containing chitosan nanoparticles was introduced to overcome the common problems of instability and burst release of proteins from nanofibrous scaffolds. Bovine serum albumin (BSA)-loaded chitosan nanoparticles was fabricated based on ionic gelation interaction between chitosan and sodium tripolyphosphate. Suspension electrospinning was employed to fabricate poly-ɛ-caprolacton (PCL) containing protein-loaded chitosan nanoparticles with a core-shell structure. To obtain the desired scaffold mechanical properties with enough elasticity for expansion and contraction, a hybrid mono and multilayer electrospun scaffold was fabricated using PCL containing protein-loaded chitosan nanoparticles and poly-L-lactic acid (PLLA). According to the BSA release profile, the multi-layered structure of nanofibers with two barrier layers provided a programmable release pattern of the loaded protein. Moreover, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and circular dichroism spectra results showed that the electrospinning process had no significant effect on the primary and secondary structure of the protein. The results indicated a desirable biocompatibility and mechanical cues of the multilayer nanofibrous scaffolds supporting structural stability and controlled release of the protein, which can offer diverse applications in hollow organ tissue engineering. Copyright © 2015 Elsevier B.V. All rights reserved.

Fourier transform Raman spectroscopic studies of human and animal skins

NASA Astrophysics Data System (ADS)

Barry, Brian W.; Edwards, Howell G.; Williams, Adrian C.

1994-01-01

The stratum corneum is the outermost layer of the skin and provides the principal barrier for the ingress of chemicals and environmental toxins into human and animal tissues. However, human skin has several advantages for the administration of therapeutic agents (transdermal drug delivery), but problems occur with the supply, storage, and biohazardous nature of human tissue. Hence, alternative animal tissues have been prepared to model drug diffusion across human skin but the molecular basis for comparison is lacking. Here, FT-Raman spectra of mammalian (human and pig) and reptilian (snake) skins have been obtained and the structural dissimilarities are correlated with drug diffusion studies across the tissues.

Gross anatomy of superficial fascia and future localised fat deposit areas of the abdomen in foetus.

PubMed

Kumar, Pramod; Pandey, Arvind Kumar; Kumar, Brijesh; Aithal, K S; Dsouza, Antony Sylvan

2013-09-01

The development and popularity of body contouring procedures such as liposuction and abdominoplasty has renewed interest in the anatomy of the superficial fascia and subcutaneous fat deposits of the abdomen. The study of anatomy of fascia and fetal adipose tissue was proposed as it may be of value in understanding the possible programing of prevention of obesity. The present study was undertaken to understand the gross anatomy of superficial fascia of abdomen and to study the gross anatomy of future localized fat deposits (LFDs) area of abdomen in fetus. Four fetus (two male & two female) of four month of intrauterine life were dissected. Attachments & layers of superficial fascia and future subcutaneous fat deposit area of upper and lower abdomen were noted. Superficial fascia of the abdomen was multi layered in mid line and number of layers reduced laterally as in adult. The future abdominal LFD (localized fat deposits) area in fetus shows brownish-white blubbary tissue without well-defined adult fat lobules. The attachment and gross anatomy of superficial fascia of the fetus was similar to that in adults. The future LFD areas showed brownish white blubbary tissue with ill-defined fat lobules.

Optical Coherence Tomography for Quantitative Assessment of Microstructural and Microvascular Alterations in Late Oral Radiation Toxicity

NASA Astrophysics Data System (ADS)

Davoudi, Bahar

More than half of head-and-neck cancer patients undergo radiotherapy at some point during their treatment. Even though the use of conformed therapeutic beams has increased radiation dose localization to the tumor, resulting in more normal tissue sparing, still, in many head-and-neck cancer patients, the healthy tissue of the oral cavity still receives a sizeable amount of radiation. This causes acute and / or late complications in these patients. The latter occur as late as several months or even years after the completion of treatment and are typically associated with severe symptoms. Currently, the clinical method for diagnosing these complications is visual examination of the oral tissue surface. However, it has been well established that such complications originate in subsurface oral tissue layers including its microvasculature. Therefore, to better understand the mechanism of these complications and to be able to diagnose them earlier, there exists a need for subsurface monitoring of the irradiated oral tissue. Histology has been used as such a tool for research purposes; however, its use in clinical diagnosis is limited due to its invasive and hazardous nature. Therefore, in this thesis, I propose to use optical coherence tomography (OCT) as a subsurface, micron-scale resolution optical imaging tool that can provide images of oral tissue subsurface layers down to a depth of 1-2 mm (structural OCT), as well as images demonstrating vessel morphology (speckle variance OCT) and blood flow information (Doppler OCT). This thesis explains the development of an OCT setup and an oral probe to acquire images in-vivo. Moreover, it introduces a software-based quantification platform for extracting specific biologically-meaningful metrics from the structural and vascular OCT images. It then describes the application of the developed imaging and quantification platform in a feasibility clinical study that was performed on 15 late oral radiation toxicity patients and 5 age-matched healthy volunteers. The results of this clinical study show that: 1) the value of the developed metrics (such as epithelium to lamina propria thickness, vessel diameter, and blood velocity) is significantly different between the two cohorts; 2) radiation-induced abnormalities in structural OCT images are primarily observed in the regions where the total radiation dose exceeded ˜50 Gy. The results of this study demonstrate the promising ability of the developed OCT imaging and quantification platform to highlight biologically meaningful differences between late oral radiation toxicity patients and healthy volunteers in the structural and vascular images. This methodology may also be used in prognostic studies to monitor the efficacy of the medication prescribed to late oral radiation toxicity patients.

Cytokeratin expression of engrafted three-dimensional culture tissues using epithelial cells derived from porcine periodontal ligaments.

PubMed

Yamada, Rie; Kitajima, Kayoko; Arai, Kyoko; Igarashi, Masaru

2014-09-01

This study investigated the differentiation and proliferation of epithelial cells derived from periodontal ligaments after three-dimensional culture using collagen gel with fibroblasts in vitro and in vivo. Epithelial cells and fibroblasts were derived from porcine periodontal ligaments. Epithelial cells were labeled using a fluorescent red membrane marker (PKH-26GL) and were seeded onto collagen gel with fibroblasts, followed by incubation in an air-liquid interface for 7 days. Three-dimensional cultures were grafted onto the backs of nude mice and removed at 1, 7, and 14 days after surgery (in vivo model). Unfixed sections (5 μm) were used to detect the presence of red fluorescent cells. Paraffin sections were analyzed histologically and immunohistochemically. Specimens were compared with three-dimensional culture tissues at 8, 14 and 21 days (in vitro model). Grafted three-dimensional cultures formed a stratified epithelial structure similar to skin in vivo. Epithelial cells were sequenced in basal-layer-like structures at 14 days in vivo. Immunohistochemical findings showed that the expression of cytokeratin was detected in the epithelial layer in in vitro and in vivo models. Ck8 + 18 + 19 was expressed in the upper epithelial layer in the in vitro model at 14 and 21 days, but not in vivo. Involucrin was expressed in the certified layers in vitro at 14 days, but not in vivo. Laminin was detected at the dermo-epidermal junction in vivo at 7 and 14 days, but not in vitro. These results suggest that differentiation of three-dimensional culture tissues differs in vivo and in vitro. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Constitutive modeling of the passive inflation-extension behavior of the swine colon.

PubMed

Patel, Bhavesh; Chen, Huan; Ahuja, Aashish; Krieger, Joshua F; Noblet, Jillian; Chambers, Sean; Kassab, Ghassan S

2018-01-01

In the present work, we propose the first structural constitutive model of the passive mechanical behavior of the swine colon that is validated against physiological inflation-extension tests, and accounts for residual strains. Sections from the spiral colon and the descending colon were considered to investigate potential regional variability. We found that the proposed constitutive model accurately captures the passive inflation-extension behavior of both regions of the swine colon (coefficient of determination R 2 =0.94±0.02). The model revealed that the circumferential muscle layer does not provide significant mechanical support under passive conditions and the circumferential load is actually carried by the submucosa layer. The stress analysis permitted by the model showed that the colon tissue can distend up to 30% radially without significant increase in the wall stresses suggesting a highly compliant behavior of the tissue. This is in-line with the requirement for the tissue to easily accommodate variable quantities of fecal matter. The analysis also showed that the descending colon is significantly more compliant than the spiral colon, which is relevant to the storage function of the descending colon. Histological analysis showed that the swine colon possesses a four-layer structure similar to the human colon, where the longitudinal muscle layer is organized into bands called taeniae, a typical feature of the human colon. The model and the estimated parameters can be used in a Finite Element framework to conduct simulations with realistic geometry of the swine colon. The resulting computational model will provide a foundation for virtual assessment of safe and effective devices for the treatment of colonic diseases. Copyright © 2017 Elsevier Ltd. All rights reserved.

Additive manufacturing and mechanical characterization of graded porosity scaffolds designed based on triply periodic minimal surface architectures.

PubMed

Afshar, M; Anaraki, A Pourkamali; Montazerian, H; Kadkhodapour, J

2016-09-01

Since the advent of additive manufacturing techniques, triply periodic minimal surfaces have emerged as a novel tool for designing porous scaffolds. Whereas scaffolds are expected to provide multifunctional performance, spatially changing pore patterns have been a promising approach to integrate mechanical characteristics of different architectures into a unique scaffold. Smooth morphological variations are also frequently seen in nature particularly in bone and cartilage structures and can be inspiring for designing of artificial tissues. In this study, we carried out experimental and numerical procedures to uncover the mechanical properties and deformation mechanisms of linearly graded porosity scaffolds for two different mathematically defined pore structures. Among TPMS-based scaffolds, P and D surfaces were subjected to gradient modeling to explore the mechanical responses for stretching and bending dominated deformations, respectively. Moreover, the results were compared to their corresponding uniform porosity structures. Mechanical properties were found to be by far greater for the stretching dominated structure (P-Surface). For bending dominated architecture (D-Surface), although there was no global fracture for uniform structures, graded structure showed a brittle fracture at 0.08 strain. A layer by layer deformation mechanism for stretching dominated structure was observed. For bending dominated scaffolds, deformation was accompanied by development of 45° shearing bands. Finite element simulations were also performed and the results showed a good agreement with the experimental observations. Copyright © 2016 Elsevier Ltd. All rights reserved.

A two-ply polymer-based flexible tactile sensor sheet using electric capacitance.

PubMed

Guo, Shijie; Shiraoka, Takahisa; Inada, Seisho; Mukai, Toshiharu

2014-01-29

Traditional capacitive tactile sensor sheets usually have a three-layered structure, with a dielectric layer sandwiched by two electrode layers. Each electrode layer has a number of parallel ribbon-like electrodes. The electrodes on the two electrode layers are oriented orthogonally and each crossing point of the two perpendicular electrode arrays makes up a capacitive sensor cell on the sheet. It is well known that compatibility between measuring precision and resolution is difficult, since decreasing the width of the electrodes is required to obtain a high resolution, however, this may lead to reduction of the area of the sensor cells, and as a result, lead to a low Signal/Noise (S/N) ratio. To overcome this problem, a new multilayered structure and related calculation procedure are proposed. This new structure stacks two or more sensor sheets with shifts in position. Both a high precision and a high resolution can be obtained by combining the signals of the stacked sensor sheets. Trial production was made and the effect was confirmed.

Quantifying the ultrastructure of carotid arteries using high-resolution micro-diffusion tensor imaging—comparison of intact versus open cut tissue

NASA Astrophysics Data System (ADS)

Salman Shahid, Syed; Gaul, Robert T.; Kerskens, Christian; Flamini, Vittoria; Lally, Caitríona

2017-12-01

Diffusion magnetic resonance imaging (dMRI) can provide insights into the microstructure of intact arterial tissue. The current study employed high magnetic field MRI to obtain ultra-high resolution dMRI at an isotropic voxel resolution of 117 µm3 in less than 2 h of scan time. A parameter selective single shell (128 directions) diffusion-encoding scheme based on Stejskel-Tanner sequence with echo-planar imaging (EPI) readout was used. EPI segmentation was used to reduce the echo time (TE) and to minimise the susceptibility-induced artefacts. The study utilised the dMRI analysis with diffusion tensor imaging (DTI) framework to investigate structural heterogeneity in intact arterial tissue and to quantify variations in tissue composition when the tissue is cut open and flattened. For intact arterial samples, the region of interest base comparison showed significant differences in fractional anisotropy and mean diffusivity across the media layer (p  <  0.05). For open cut flat samples, DTI based directionally invariant indices did not show significant differences across the media layer. For intact samples, fibre tractography based indices such as calculated helical angle and fibre dispersion showed near circumferential alignment and a high degree of fibre dispersion, respectively. This study demonstrates the feasibility of fast dMRI acquisition with ultra-high spatial and angular resolution at 7 T. Using the optimised sequence parameters, this study shows that DTI based markers are sensitive to local structural changes in intact arterial tissue samples and these markers may have clinical relevance in the diagnosis of atherosclerosis and aneurysm.

Three-dimensional Organization of Layered Apical Cytoskeletal Networks Associated with Mouse Airway Tissue Development

NASA Astrophysics Data System (ADS)

Tateishi, Kazuhiro; Nishida, Tomoki; Inoue, Kanako; Tsukita, Sachiko

2017-03-01

The cytoskeleton is an essential cellular component that enables various sophisticated functions of epithelial cells by forming specialized subcellular compartments. However, the functional and structural roles of cytoskeletons in subcellular compartmentalization are still not fully understood. Here we identified a novel network structure consisting of actin filaments, intermediate filaments, and microtubules directly beneath the apical membrane in mouse airway multiciliated cells and in cultured epithelial cells. Three-dimensional imaging by ultra-high voltage electron microscopy and immunofluorescence revealed that the morphological features of each network depended on the cell type and were spatiotemporally integrated in association with tissue development. Detailed analyses using Odf2 mutant mice, which lack ciliary basal feet and apical microtubules, suggested a novel contribution of the intermediate filaments to coordinated ciliary beating. These findings provide a new perspective for viewing epithelial cell differentiation and tissue morphogenesis through the structure and function of apical cytoskeletal networks.

Fabrication and characterization of gels with integrated channels using 3D printing with microfluidic nozzle for tissue engineering applications.

PubMed

Attalla, R; Ling, C; Selvaganapathy, P

2016-02-01

The lack of a simple and effective method to integrate vascular network with engineered scaffolds and tissue constructs remains one of the biggest challenges in true 3D tissue engineering. Here, we detail the use of a commercially available, low-cost, open-source 3D printer modified with a microfluidic print-head in order to develop a method for the generation of instantly perfusable vascular network integrated with gel scaffolds seeded with cells. The print-head features an integrated coaxial nozzle that allows the fabrication of hollow, calcium-polymerized alginate tubes that can be easily patterned using 3D printing techniques. The diameter of the hollow channel can be precisely controlled and varied between 500 μm - 2 mm by changing applied flow rates or print-head speed. These channels are integrated into gel layers with a thickness of 800 μm - 2.5 mm. The structural rigidity of these constructs allows the fabrication of multi-layered structures without causing the collapse of hollow channels in lower layers. The 3D printing method was fully characterized at a range of operating speeds (0-40 m/min) and corresponding flow rates (1-30 mL/min) were identified to produce precise definition. This microfluidic design also allows the incorporation of a wide range of scaffold materials as well as biological constituents such as cells, growth factors, and ECM material. Media perfusion of the channels causes a significant viability increase in the bulk of cell-laden structures over the long-term. With this setup, gel constructs with embedded arrays of hollow channels can be created and used as a potential substitute for blood vessel networks.

Simulation of plasma double-layer structures

NASA Technical Reports Server (NTRS)

Borovsky, J. E.; Joyce, G.

1982-01-01

Electrostatic plasma double layers are numerically simulated by means of a magnetized 2 1/2 dimensional particle in cell method. The investigation of planar double layers indicates that these one dimensional potential structures are susceptible to periodic disruption by instabilities in the low potential plasmas. Only a slight increase in the double layer thickness with an increase in its obliqueness to the magnetic field is observed. Weak magnetization results in the double layer electric field alignment of accelerated particles and strong magnetization results in their magnetic field alignment. The numerical simulations of spatially periodic two dimensional double layers also exhibit cyclical instability. A morphological invariance in two dimensional double layers with respect to the degree of magnetization implies that the potential structures scale with Debye lengths rather than with gyroradii. Electron beam excited electrostatic electron cyclotron waves and (ion beam driven) solitary waves are present in the plasmas adjacent to the double layers.

High precision position sensor based on CPA in a composite multi-layered system.

PubMed

Dey, Sanjeeb; Singh, Suneel; Rao, Desai Narayana

2018-04-16

We propose a scheme for high precision position sensing based on coherent perfect absorption (CPA) in a five-layered structure comprising three layers of metal-dielectric composites and two spacer (air) layers. Both the outermost interfaces of the five layered medium are irradiated by two identical coherent light waves at the same angle of incidence. We first investigate the occurrence of CPA in a symmetric layered structure as a function of different system parameters for oblique incidence. Thereafter, by shifting the middle layer, beginning from one end of the structure to the other, we observe the periodic occurrence of extremely narrow CPA resonances at several positions of the middle layer. Moreover this phenomenon is seen to recur even at many other wavelengths. We discuss how the position sensitivity of this phenomenon can be utilized for designing a CPA based high precision position sensing device.

[Analysis and comprehensive evaluation on cold resistance of six varieties of Michelia].

PubMed

Li, Rui Xue; Jin, Xiao Ling; Hu, Xi Jun; Chai, Yi Xia; Cai, Meng Ying; Luo, Feng; Zhang, Fang Jing

2017-05-18

Taking six varieties of Michelia as test materials, their responses under cold situation in the field were investigated and the semilethal low temperatures were calculated by fitting Logistic equation. The nine structure indexes of leaf tissue were observed by paraffin section, and a comprehensive evaluation on cold resistance of different varieties was given according to subordinate function value analysis. The results showed that the relative electrical conductivity of six varieties of Michelia was significantly positively correlated with the semilethal low temperature (LT 50 ) of 3 h 0-25 ℃ treatment. From high to low, the order of LT 50 , which ranged between -20.48 ℃and -8.67 ℃, was M. maudiae ＞ M. maudiae var. rubicunda ＞ M. wilsonii ＞ M. 'liubanhanxiao' ♀× M. shiluensis ♂ ＞ M. platypetala ＞ M. 'liubanhanxiao'. The epidermal anticlinal walls of six varieties of Michelia leaves had 1-2 layers and showed slightly sinuated shape. The leaf had 1-3 layers of palisade tissue cells and the differences among the indexes of nine anatomical structures were extremely significant. The thickness of palisade tissue, ratio between palisade tissue and spongy tissue, and thickness of the vein were the key factors affecting cold resistance. The order of cold resistance of six varieties of Michelia, from the strong to the weak, was M. 'liubanhanxiao' ＞ M. platypetala ＞ M. 'liubanhanxiao' ♀× M. shiluensis ♂＞M. wilsonii ＞ M. maudiae var. rubicunda ＞ M. maudiae, which was basically consistent with the result of field investigation.

Characterization of Canopy Layering in Forested Ecosystems Using Full Waveform Lidar

NASA Technical Reports Server (NTRS)

Whitehurst, Amanda S.; Swatantran, Anu; Blair, J. Bryan; Hofton, Michelle A.; Dubayah, Ralph

2013-01-01

Canopy structure, the vertical distribution of canopy material, is an important element of forest ecosystem dynamics and habitat preference. Although vertical stratification, or "canopy layering," is a basic characterization of canopy structure for research and forest management, it is difficult to quantify at landscape scales. In this paper we describe canopy structure and develop methodologies to map forest vertical stratification in a mixed temperate forest using full-waveform lidar. Two definitions-one categorical and one continuous-are used to map canopy layering over Hubbard Brook Experimental Forest, New Hampshire with lidar data collected in 2009 by NASA's Laser Vegetation Imaging Sensor (LVIS). The two resulting canopy layering datasets describe variation of canopy layering throughout the forest and show that layering varies with terrain elevation and canopy height. This information should provide increased understanding of vertical structure variability and aid habitat characterization and other forest management activities.

Adhesion of Dental Materials to Tooth Structure

NASA Astrophysics Data System (ADS)

Mitra, Sumita B.

2000-03-01

The understanding and proper application of the principles of adhesion has brought forth a new paradigm in the realm of esthetic dentistry. Modern restorative tooth procedures can now conserve the remaining tooth-structure and also provide for the strengthening of the tooth. Adhesive restorative techniques call for the application and curing of the dental adhesive at the interface between the tooth tissue and the filling material. Hence the success of the restoration depends largely on the integrity of this interface. The mechanism of adhesion of the bonding materials to the dental hard tissue will be discussed in this paper. There are four main steps that occur during the application of the dental adhesive to the oral hard tissues: 1) The first step is the creation of a microstructure in the tooth enamel or dentin by means of an acidic material. This can be through the application of a separate etchant or can be accomplished in situ by the adhesive/primer. This agent has to be effective in removing or modifying the proteinaceous “smear” layer, which would otherwise act as a weak boundary layer on the surface to be bonded. 2) The primer/adhesive must then be able to wet and penetrate the microstructure created in the tooth. Since the surface energies of etched enamel and that of etched dentin are different finding one material to prime both types of dental tissues can be quite challenging. 3) The ionomer types of materials, particularly those that are carboxylate ion-containing, can chemically bond with the calcium ions of the hydroxyapatite mineral. 4) Polymerization in situ allows for micromechanical interlocking of the adhesive. The importance of having the right mechanical properties of the cured adhesive layer and its role in absorbing and dissipating stresses encountered by a restored tooth will also be discussed.

Use of chemical-mechanical polishing for fabricating photonic bandgap structures

DOEpatents

Fleming, James G.; Lin, Shawn-Yu; Hetherington, Dale L.; Smith, Bradley K.

1999-01-01

A method is disclosed for fabricating a two- or three-dimensional photonic bandgap structure (also termed a photonic crystal, photonic lattice, or photonic dielectric structure). The method uses microelectronic integrated circuit (IC) processes to fabricate the photonic bandgap structure directly upon a silicon substrate. One or more layers of arrayed elements used to form the structure are deposited and patterned, with chemical-mechanical polishing being used to planarize each layer for uniformity and a precise vertical tolerancing of the layer. The use of chemical-mechanical planarization allows the photonic bandgap structure to be formed over a large area with a layer uniformity of about two-percent. Air-gap photonic bandgap structures can also be formed by removing a spacer material separating the arrayed elements by selective etching. The method is useful for fabricating photonic bandgap structures including Fabry-Perot resonators and optical filters for use at wavelengths in the range of about 0.2-20 .mu.m.

Modeling marine boundary-layer clouds with a two-layer model: A one-dimensional simulation

NASA Technical Reports Server (NTRS)

Wang, Shouping

1993-01-01

A two-layer model of the marine boundary layer is described. The model is used to simulate both stratocumulus and shallow cumulus clouds in downstream simulations. Over cold sea surfaces, the model predicts a relatively uniform structure in the boundary layer with 90%-100% cloud fraction. Over warm sea surfaces, the model predicts a relatively strong decoupled and conditionally unstable structure with a cloud fraction between 30% and 60%. A strong large-scale divergence considerably limits the height of the boundary layer and decreases relative humidity in the upper part of the cloud layer; thus, a low cloud fraction results. The efffects of drizzle on the boundary-layer structure and cloud fraction are also studied with downstream simulations. It is found that drizzle dries and stabilizes the cloud layer and tends to decouple the cloud from the subcloud layer. Consequently, solid stratocumulus clouds may break up and the cloud fraction may decrease because of drizzle.

Drug nano-reservoirs synthesized using layer-by-layer technologies.

PubMed

Costa, Rui R; Alatorre-Meda, Manuel; Mano, João F

2015-11-01

The pharmaceutical industry has been able to tackle the emergence of new microorganisms and diseases by synthesizing new specialized drugs to counter them. Their administration must ensure that a drug is effectively encapsulated and protected until it reaches its target, and that it is released in a controlled way. Herein, the potential of layer-by-layer (LbL) structures to act as drug reservoirs is presented with an emphasis to "nano"-devices of various geometries, from planar coatings to fibers and capsules. The inherent versatile nature of this technique allows producing carriers resorting to distinct classes of materials, variable geometry and customized release profiles that fit within adequate criteria required for disease treatment or for novel applications in the tissue engineering field. The production methods of LbL reservoirs are varied and allow for different kinds of molecules to be incorporated, such as antibiotics, growth factors and biosensing substances, not limited to water-soluble molecules but including hydrophobic drugs. We will also debate the future of LbL in the pharmaceutical industry. Currently, multilayered structures are yet to be covered by the regulatory guidelines that govern the fabrication of nanotechnology products. However, as they stand now, LbL nanodevices have already shown usefulness for antifouling applications, gene therapy, nanovaccines and the formation of de novo tissues. Copyright © 2015 Elsevier Inc. All rights reserved.

Preparation of TiO(2) layers on cp-Ti and Ti6Al4V by thermal and anodic oxidation and by sol-gel coating techniques and their characterization.

PubMed

Velten, D; Biehl, V; Aubertin, F; Valeske, B; Possart, W; Breme, J

2002-01-01

The excellent biocompatibility of titanium and its alloys used, for example, for medical devices, is associated with the properties of their surface oxide. For a better understanding of the tissue reaction in contact with the oxide layer, knowledge of the chemical and physical properties of this layer is of increasing interest. In this study, titania films were produced on cp-Ti and Ti6Al4V substrates by thermal oxidation, anodic oxidation, and by the sol-gel process. The thickness and structure of the films produced under different conditions were determined by ellipsometry, infrared spectroscopy, and X-ray diffraction measurements. The corrosion properties of these layers were investigated by current density-potential curves under physiological conditions. The oxide layers produced on cp-Ti and Ti6Al4V by thermal oxidation consist of TiO(2) in the rutile structure. For the anodized samples the structure of TiO(2) is a mixture of amorphous phase and anatase. The structure of the coatings produced by the sol-gel process for a constant annealing time depends on the annealing temperature, and with increasing temperature successively amorphous, anatase, and rutile structure is observed. Compared to the uncoated, polished substrate with a natural oxide layer, the corrosion resistance of cp-Ti and Ti6Al4V is increased for the samples with an oxide layer thickness of about 100 nm, independent of the oxidation procedure. Copyright 2001 John Wiley & Sons, Inc.

Study of ion beam sputtered Fe/Si interfaces as a function of Si layer thickness

NASA Astrophysics Data System (ADS)

Kumar, Anil; Brajpuriya, Ranjeet; Singh, Priti

2018-01-01

The exchange interaction in metal/semiconductor interfaces is far from being completely understood. Therefore, in this paper, we have investigated the nature of silicon on the Fe interface in the ion beam deposited Fe/Si/Fe trilayers keeping the thickness of the Fe layers fixed at 3 nm and varying the thickness of the silicon sandwich layer from 1.5 nm to 4 nm. Grazing incidence x-ray diffraction and atomic force microscopy techniques were used, respectively, to study the structural and morphological changes in the deposited films as a function of layer thickness. The structural studies show silicide formation at the interfaces during deposition and better crystalline structure of Fe layers at a lower spacer layer thickness. The magnetization behavior was investigated using magneto-optical Kerr effect, which clearly shows that coupling between the ferromagnetic layers is highly influenced by the semiconductor spacer layer thickness. A strong antiferromagnetic coupling was observed for a value of tSi = 2.5 nm but above this value an unexpected behavior of hysteresis loop (step like) with two coercivity values is recorded. For spacer layer thickness greater than 2.5 nm, an elemental amorphous Si layer starts to appear in the spacer layer in addition to the silicide layer at the interfaces. It is observed that in the trilayer structure, Fe layers consist of various stacks, viz., Si doped Fe layers, ferromagnetic silicide layer, and nonmagnetic silicide layer at the interfaces. The two phase hysteresis loop is explained on the basis of magnetization reversal of two ferromagnetic layers, independent of each other, with different coercivities. X-ray photo electron spectroscopy technique was also used to study interfaces characteristics as a function of tSi.

Delivery of Plasmid DNA to Vascular Tissue in vivo using Catheter Balloons Coated with Polyelectrolyte Multilayers

PubMed Central

Saurer, Eric M.; Yamanouchi, Dai; Liu, Bo; Lynn, David M.

2010-01-01

We report an approach for the localized delivery of plasmid DNA to vascular tissue from the surfaces of inflatable embolectomy catheter balloons. Using a layer-by-layer approach, ultrathin multilayered polyelectrolyte films were fabricated on embolectomy catheter balloons by alternately adsorbing layers of a hydrolytically degradable poly(β-amino ester) and plasmid DNA. Fluorescence microscopy revealed that the films coated the surfaces of the balloons uniformly. Coated balloons that were incubated in phosphate-buffered saline at 37 °C released ~25 μg DNA/cm2 over 24 hours. Analysis of the DNA by gel electrophoresis showed that the DNA was released in open-circular (‘nicked’) and supercoiled conformations, and in vitro cell transfection assays confirmed that the released DNA was transcriptionally active. Arterial injury was induced in the internal carotid arteries of Sprague-Dawley rats using uncoated balloons, followed by treatment with film-coated balloons for 20 minutes. X-gal, immunohistochemical, and immunofluorescence staining of sectioned arteries indicated high levels of β-galactosidase or enhanced green fluorescent protein (EGFP) expression in arteries treated with film-coated balloons. β-galactosidase and EGFP expression were observed throughout the medial layers of arterial tissue, and around approximately two-thirds of the circumference of the treated arteries. The layer-by-layer approach reported here provides a general platform for the balloon-mediated delivery of DNA to vascular tissue. Our results suggest the potential of this approach to deliver therapeutically relevant DNA to prevent complications such as intimal hyperplasia that arise after vascular interventions. PMID:20933275

The Nanomechanics of Biomineralized Soft-Tissues and Organic Matrices

NASA Astrophysics Data System (ADS)

Bezares-Chavez, Jiddu

The research reported on in this dissertation is concerned with the macro-molecular constitution and geometrical organization of the soft-tissue comprising the matrix of the nacreous portion of the shell of Haliotis rufescens, the Red abalone. Nacre is one of literally legions of intricate biomineralized structures that exist in nature and has long served as a paradigm for elegant and optimized structural de-sign. Biomineralization involves, inter alia, the uptake and synthesis of elements and compounds from the environment and their incorporation into highly optimized functional structures. Nacre has a structure described as a brick wall like with a matrix of biopolymer layers that are preformed and serve as a template into which nanocrystalline tiles of CaCO3 precipitate. The matrix, or what are known as inter-lamellar layers, are of particular interest as they impart both toughness and strength to the composite ceramic nacre structure. The work first involved a histochemical mapping of the macromolecular structure of the interlamellar layers; this revealed the locations of proteins and functional molecular groups that serve as nucleation sites for the ceramic tiles. Parallel studies on the nacre of Nautilus pompilius, the Chambered Nautilus, revealed the generality of the findings. Of particular interest was determining both the content and layout of chitin within these layers. In fact it was determined that chitin was organized as mostly unidirectional architecture of fibrils, with a certain fraction of fibrils laying at cross directions. Most remarkably, it was found that the fibrils possessed a very long range connectivity that spanned many tiles. This was determined by systematic atomic force (afm) and analytical optical histochemical microscopy. These findings were further verified by a unique form of mechanical testing whereby tensile testing was conducted on groups of interlamellar layers extracted from nacre. Mechanical testing led to a quantitative visco-elastic constitutive model for these layers and, in turn, to a complete mechanical/structural model for the complete nacre composite. Further verification was obtained via micro- and nano-indentation experiments which were modeled via detailed FEM numerical simulations. Nano-indentation also allowed a detailed assessment of the nano-structure and properties of the ceramic tiles which are best described as nano-scale composites composed of protein infiltrated CaCO3 nano-grains within a biopolymer matrix. The role of water content, i.e. moisture content, was also determined via, in part experiments conducted on dehydrated nacre. These findings lead to a pathway for specifying optimal bio-mimicked or bio-inspired synthetic materials.

Analysing photonic structures in plants

PubMed Central

Vignolini, Silvia; Moyroud, Edwige; Glover, Beverley J.; Steiner, Ullrich

2013-01-01

The outer layers of a range of plant tissues, including flower petals, leaves and fruits, exhibit an intriguing variation of microscopic structures. Some of these structures include ordered periodic multilayers and diffraction gratings that give rise to interesting optical appearances. The colour arising from such structures is generally brighter than pigment-based colour. Here, we describe the main types of photonic structures found in plants and discuss the experimental approaches that can be used to analyse them. These experimental approaches allow identification of the physical mechanisms producing structural colours with a high degree of confidence. PMID:23883949

Model for the ultrasound reflection from micro-beads and cells distributed in layers on a uniform surface

NASA Astrophysics Data System (ADS)

Couture, O.; Cherin, E.; Foster, F. S.

2007-07-01

A model predicting the reflection of ultrasound from multiple layers of small scattering spheres is developed. Predictions of the reflection coefficient, which takes into account the interferences between the different sphere layers, are compared to measurements performed in the 10-80 MHz and 15-35 MHz frequency range with layers of glass beads and spherical acute myeloid leukemia (AML) cells, respectively. For both types of scatterers, the reflection coefficient increases as a function of their density on the surface for less than three superimposed layers, at which point it saturates at 0.38 for glass beads and 0.02 for AML cells. Above three layers, oscillations of the reflection coefficient due to constructive or destructive interference between layers are observed experimentally and are accurately predicted by the model. The use of such a model could lead to a better understanding of the structures observed in layered tissue images.

Two-Dimensional Layered Oxide Structures Tailored by Self-Assembled Layer Stacking via Interfacial Strain.

PubMed

Zhang, Wenrui; Li, Mingtao; Chen, Aiping; Li, Leigang; Zhu, Yuanyuan; Xia, Zhenhai; Lu, Ping; Boullay, Philippe; Wu, Lijun; Zhu, Yimei; MacManus-Driscoll, Judith L; Jia, Quanxi; Zhou, Honghui; Narayan, Jagdish; Zhang, Xinghang; Wang, Haiyan

2016-07-06

Study of layered complex oxides emerge as one of leading topics in fundamental materials science because of the strong interplay among intrinsic charge, spin, orbital, and lattice. As a fundamental basis of heteroepitaxial thin film growth, interfacial strain can be used to design materials that exhibit new phenomena beyond their conventional forms. Here, we report a strain-driven self-assembly of bismuth-based supercell (SC) with a two-dimensional (2D) layered structure. With combined experimental analysis and first-principles calculations, we investigated the full SC structure and elucidated the fundamental growth mechanism achieved by the strain-enabled self-assembled atomic layer stacking. The unique SC structure exhibits room-temperature ferroelectricity, enhanced magnetic responses, and a distinct optical bandgap from the conventional double perovskite structure. This study reveals the important role of interfacial strain modulation and atomic rearrangement in self-assembling a layered singe-phase multiferroic thin film, which opens up a promising avenue in the search for and design of novel 2D layered complex oxides with enormous promise.

[Strategies to choose scaffold materials for tissue engineering].

PubMed

Gao, Qingdong; Zhu, Xulong; Xiang, Junxi; Lü, Yi; Li, Jianhui

2016-02-01

Current therapies of organ failure or a wide range of tissue defect are often not ideal. Transplantation is the only effective way for long time survival. But it is hard to meet huge patients demands because of donor shortage, immune rejection and other problems. Tissue engineering could be a potential option. Choosing a suitable scaffold material is an essential part of it. According to different sources, tissue engineering scaffold materials could be divided into three types which are natural and its modified materials, artificial and composite ones. The purpose of tissue engineering scaffold is to repair the tissues or organs damage, so could reach the ideal recovery in its function and structure aspect. Therefore, tissue engineering scaffold should even be as close as much to the original tissue or organs in function and structure. We call it "organic scaffold" and this strategy might be the drastic perfect substitute for the tissues or organs in concern. Optimized organization with each kind scaffold materials could make up for biomimetic structure and function of the tissue or organs. Scaffold material surface modification, optimized preparation procedure and cytosine sustained-release microsphere addition should be considered together. This strategy is expected to open new perspectives for tissue engineering. Multidisciplinary approach including material science, molecular biology, and engineering might find the most ideal tissue engineering scaffold. Using the strategy of drawing on each other strength and optimized organization with each kind scaffold material to prepare a multifunctional biomimetic tissue engineering scaffold might be a good method for choosing tissue engineering scaffold materials. Our research group had differentiated bone marrow mesenchymal stem cells into bile canaliculi like cells. We prepared poly(L-lactic acid)/poly(ε-caprolactone) biliary stent. The scaffold's internal played a part in the long-term release of cytokines which mixed with sustained-release nano-microsphere containing growth factors. What's more, the stent internal surface coated with glue/collagen matrix mixing layer containing bFGF and EGF so could supplying the early release of the two cytokines. Finally, combining the poly(L-lactic acid)/poly(ε-caprolactone) biliary stent with the induced cells was the last step for preparing tissue-engineered bile duct. This literature reviewed a variety of the existing tissue engineering scaffold materials and briefly introduced the impact factors on the characteristics of tissue engineering scaffold materials such as preparation procedure, surface modification of scaffold, and so on. We explored the choosing strategy of desired tissue engineering scaffold materials.

Alginate/nanohydroxyapatite scaffolds with designed core/shell structures fabricated by 3D plotting and in situ mineralization for bone tissue engineering.

PubMed

Luo, Yongxiang; Lode, Anja; Wu, Chengtie; Chang, Jiang; Gelinsky, Michael

2015-04-01

Composite scaffolds, especially polymer/hydroxyapatite (HAP) composite scaffolds with predesigned structures, are promising materials for bone tissue engineering. Various methods including direct mixing of HAP powder with polymers or incubating polymer scaffolds in simulated body fluid for preparing polymer/HAP composite scaffolds are either uncontrolled or require long times of incubation. In this work, alginate/nano-HAP composite scaffolds with designed pore parameters and core/shell structures were fabricated using 3D plotting technique and in situ mineralization under mild conditions (at room temperature and without the use of any organic solvents). Light microscopy, scanning electron microscopy, microcomputer tomography, X-ray diffraction, and Fourier transform infrared spectroscopy were applied to characterize the fabricated scaffolds. Mechanical properties and protein delivery of the scaffolds were evaluated, as well as the cell response to the scaffolds by culturing human bone-marrow-derived mesenchymal stem cells (hBMSC). The obtained data indicate that this method is suitable to fabricate alginate/nano-HAP composite scaffolds with a layer of nano-HAP, coating the surface of the alginate strands homogeneously and completely. The surface mineralization enhanced the mechanical properties and improved the cell attachment and spreading, as well as supported sustaining protein release, compared to pure alginate scaffolds without nano-HAP shell layer. The results demonstrated that the method provides an interesting option for bone tissue engineering application.

A new Pb{sup II}(ethylenediaminetetraacetate) coordination polymer with a two-dimensional layer structure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhao, D., E-mail: iamzd@hpu.edu.cn; Zhang, R. H.; Li, F. F.

2016-12-15

A new Pb{sup II}−edta{sup 4–} coordination polymer, Pb{sub 2}(edta)(H{sub 2}O){sub 0.76} (edta{sup 4–} = ethylenediaminetetraacetate) was synthesized under hydrothermal condition. Single crystal X-ray analysis reveals that it represents a novel two-dimensional (2D) Pb{sup 2+}–edta{sup 4–} layer structure with a (4,8{sup 2})-topology. Each edta{sup 4–} ligand employs its four carboxylate O and two N atoms to chelate one Pb{sup II} atom (hexa-coordinated) and connects five Pb{sup II} atoms (ennea-coordinated) via its four carboxylate groups to form 2D layer framework. Adjacent layers are packed into the overall structure through vander Waals interactions.

Blockface histology with optical coherence tomography: a comparison with Nissl staining.

PubMed

Magnain, Caroline; Augustinack, Jean C; Reuter, Martin; Wachinger, Christian; Frosch, Matthew P; Ragan, Timothy; Akkin, Taner; Wedeen, Van J; Boas, David A; Fischl, Bruce

2014-01-01

Spectral domain optical coherence tomography (SD-OCT) is a high resolution imaging technique that generates excellent contrast based on intrinsic optical properties of the tissue, such as neurons and fibers. The SD-OCT data acquisition is performed directly on the tissue block, diminishing the need for cutting, mounting and staining. We utilized SD-OCT to visualize the laminar structure of the isocortex and compared cortical cytoarchitecture with the gold standard Nissl staining, both qualitatively and quantitatively. In histological processing, distortions routinely affect registration to the blockface image and prevent accurate 3D reconstruction of regions of tissue. We compared blockface registration to SD-OCT and Nissl, respectively, and found that SD-OCT-blockface registration was significantly more accurate than Nissl-blockface registration. Two independent observers manually labeled cortical laminae (e.g. III, IV and V) in SD-OCT images and Nissl stained sections. Our results show that OCT images exhibit sufficient contrast in the cortex to reliably differentiate the cortical layers. Furthermore, the modalities were compared with regard to cortical laminar organization and showed good agreement. Taken together, these SD-OCT results suggest that SD-OCT contains information comparable to standard histological stains such as Nissl in terms of distinguishing cortical layers and architectonic areas. Given these data, we propose that SD-OCT can be used to reliably generate 3D reconstructions of multiple cubic centimeters of cortex that can be used to accurately and semi-automatically perform standard histological analyses. © 2013.

Blockface Histology with Optical Coherence Tomography: A Comparison with Nissl Staining

PubMed Central

Magnain, Caroline; Augustinack, Jean C.; Reuter, Martin; Wachinger, Christian; Frosch, Matthew P.; Ragan, Timothy; Akkin, Taner; Wedeen, Van J.; Boas, David A.; Fischl, Bruce

2015-01-01

Spectral domain optical coherence tomography (SD-OCT) is a high resolution imaging technique that generates excellent contrast based on intrinsic optical properties of the tissue, such as neurons and fibers. The SD-OCT data acquisition is performed directly on the tissue block, diminishing the need for cutting, mounting and staining. We utilized SD-OCT to visualize the laminar structure of the isocortex and compared cortical cytoarchitecture with the gold standard Nissl staining, both qualitatively and quantitatively. In histological processing, distortions routinely affect registration to the blockface image and prevent accurate 3D reconstruction of regions of tissue. We compared blockface registration to SD-OCT and Nissl, respectively, and found that SD-OCT-blockface registration was significantly more accurate than Nissl-blockface registration. Two independent observers manually labeled cortical laminae (e.g. III, IV and V) in SD-OCT images and Nissl stained sections. Our results show that OCT images exhibit sufficient contrast in the cortex to reliably differentiate the cortical layers. Furthermore, the modalities were compared with regard to cortical laminar organization and showed good agreement. Taken together, these SD-OCT results suggest that SD-OCT contains information comparable to standard histological stains such as Nissl in terms of distinguishing cortical layers and architectonic areas. Given these data, we propose that SD-OCT can be used to reliably generate 3D reconstructions of multiple cubic centimeters of cortex that can be used to accurately and semi-automatically perform standard histological analyses. PMID:24041872

Topology-Scaling Identification of Layered Solids and Stable Exfoliated 2D Materials.

PubMed

Ashton, Michael; Paul, Joshua; Sinnott, Susan B; Hennig, Richard G

2017-03-10

The Materials Project crystal structure database has been searched for materials possessing layered motifs in their crystal structures using a topology-scaling algorithm. The algorithm identifies and measures the sizes of bonded atomic clusters in a structure's unit cell, and determines their scaling with cell size. The search yielded 826 stable layered materials that are considered as candidates for the formation of two-dimensional monolayers via exfoliation. Density-functional theory was used to calculate the exfoliation energy of each material and 680 monolayers emerge with exfoliation energies below those of already-existent two-dimensional materials. The crystal structures of these two-dimensional materials provide templates for future theoretical searches of stable two-dimensional materials. The optimized structures and other calculated data for all 826 monolayers are provided at our database (https://materialsweb.org).

Long-term biopersistence of tangled oxidized carbon nanotubes inside and outside macrophages in rat subcutaneous tissue

NASA Astrophysics Data System (ADS)

Sato, Yoshinori; Yokoyama, Atsuro; Nodasaka, Yoshinobu; Kohgo, Takao; Motomiya, Kenichi; Matsumoto, Hiroaki; Nakazawa, Eiko; Numata, Tomoko; Zhang, Minfang; Yudasaka, Masako; Hara, Hideyuki; Araki, Rikita; Tsukamoto, Osamu; Saito, Hiroaki; Kamino, Takeo; Watari, Fumio; Tohji, Kazuyuki

2013-08-01

Because of their mechanical strength, chemical stability, and low molecular weight, carbon nanotubes (CNTs) are attractive biological implant materials. Biomaterials are typically implanted into subcutaneous tissue or bone; however, the long-term biopersistence of CNTs in these tissues is unknown. Here, tangled oxidized multi-walled CNTs (t-ox-MWCNTs) were implanted into rat subcutaneous tissues and structural changes in the t-ox-MWCNTs located inside and outside of macrophages were studied for 2 years post-implantation. The majority of the large agglomerates were present in the intercellular space, maintained a layered structure, and did not undergo degradation. By contrast, small agglomerates were found inside macrophages, where they were gradually degraded in lysosomes. None of the rats displayed symptoms of cancer or severe inflammatory reactions such as necrosis. These results indicate that t-ox-MWCNTs have high biopersistence and do not evoke adverse events in rat subcutaneous tissue in vivo, demonstrating their potential utility as implantable biomaterials.

Multizone Paper Platform for 3D Cell Cultures

PubMed Central

Derda, Ratmir; Hong, Estrella; Mwangi, Martin; Mammoto, Akiko; Ingber, Donald E.; Whitesides, George M.

2011-01-01

In vitro 3D culture is an important model for tissues in vivo. Cells in different locations of 3D tissues are physiologically different, because they are exposed to different concentrations of oxygen, nutrients, and signaling molecules, and to other environmental factors (temperature, mechanical stress, etc). The majority of high-throughput assays based on 3D cultures, however, can only detect the average behavior of cells in the whole 3D construct. Isolation of cells from specific regions of 3D cultures is possible, but relies on low-throughput techniques such as tissue sectioning and micromanipulation. Based on a procedure reported previously (“cells-in-gels-in-paper” or CiGiP), this paper describes a simple method for culture of arrays of thin planar sections of tissues, either alone or stacked to create more complex 3D tissue structures. This procedure starts with sheets of paper patterned with hydrophobic regions that form 96 hydrophilic zones. Serial spotting of cells suspended in extracellular matrix (ECM) gel onto the patterned paper creates an array of 200 micron-thick slabs of ECM gel (supported mechanically by cellulose fibers) containing cells. Stacking the sheets with zones aligned on top of one another assembles 96 3D multilayer constructs. De-stacking the layers of the 3D culture, by peeling apart the sheets of paper, “sections” all 96 cultures at once. It is, thus, simple to isolate 200-micron-thick cell-containing slabs from each 3D culture in the 96-zone array. Because the 3D cultures are assembled from multiple layers, the number of cells plated initially in each layer determines the spatial distribution of cells in the stacked 3D cultures. This capability made it possible to compare the growth of 3D tumor models of different spatial composition, and to examine the migration of cells in these structures. PMID:21573103

Determination of the Dynamics of Healing at the Tissue-Implant Interface by Means of Microcomputed Tomography and Functional Apparent Moduli

PubMed Central

Chang, Po-Chun; Seol, Yang-Jo; Goldstein, Steven A.; Giannobile, William V.

2014-01-01

Purpose It is currently a challenge to determine the biomechanical properties of the hard tissue–dental implant interface. Recent advances in intraoral imaging and tomographic methods, such as microcomputed tomography (micro-CT), provide three-dimensional details, offering significant potential to evaluate the bone-implant interface, but yield limited information regarding osseointegration because of physical scattering effects emanating from metallic implant surfaces. In the present study, it was hypothesized that functional apparent moduli (FAM), generated from functional incorporation of the peri-implant structure, would eliminate the radiographic artifact–affected layer and serve as a feasible means to evaluate the biomechanical dynamics of tissue-implant integration in vivo. Materials and Methods Cylindric titanium mini-implants were placed in osteotomies and osteotomies with defects in rodent maxillae. The layers affected by radiographic artifacts were identified, and the pattern of tissue-implant integration was evaluated from histology and micro-CT images over a 21-day observation period. Analyses of structural information, FAM, and the relationship between FAM and interfacial stiffness (IS) were done before and after eliminating artifacts. Results Physical artifacts were present within a zone of about 100 to 150 μm around the implant in both experimental defect situations (osteotomy alone and osteotomy + defect). All correlations were evaluated before and after eliminating the artifact-affected layers, most notably during the maturation period of osseointegration. A strong correlation existed between functional bone apparent modulus and IS within 300 μm at the osteotomy defects (r > 0.9) and functional composite tissue apparent modulus in the osteotomy defects (r > 0.75). Conclusion Micro-CT imaging and FAM were of value in measuring the temporal process of tissue-implant integration in vivo. This approach will be useful to complement imaging technologies for longitudinal monitoring of osseointegration. PMID:23377049

Robust Formation and Maintenance of Continuous Stratified Cortical Neuroepithelium by Laminin-Containing Matrix in Mouse ES Cell Culture

PubMed Central

Nasu, Makoto; Takata, Nozomu; Danjo, Teruko; Sakaguchi, Hideya; Kadoshima, Taisuke; Futaki, Sugiko; Sekiguchi, Kiyotoshi; Eiraku, Mototsugu; Sasai, Yoshiki

2012-01-01

In the mammalian cortex, the dorsal telencephalon exhibits a characteristic stratified structure. We previously reported that three-dimensional (3D) culture of mouse ES cells (mESCs) can efficiently generate cortical neuroepithelium (NE) and layer-specific cortical neurons. However, the cortical NE generated in this mESC culture was structurally unstable and broke into small neural rosettes by culture day 7, suggesting that some factors for reinforcing the structural integrity were missing. Here we report substantial supporting effects of the extracellular matrix (ECM) protein laminin on the continuous formation of properly polarized cortical NE in floating aggregate culture of mESCs. The addition of purified laminin and entactin (a laminin-associated protein), even at low concentrations, stabilized the formation of continuous cortical NE as well as the maintenance of basement membrane and prevented rosette formation. Treatment with the neutralizing ß1-integrin antibody impaired the continuous NE formation. The stabilized cortical NE exhibited typical interkinetic nuclear migration of cortical progenitors, as seen in the embryonic cortex. The laminin-treated cortical NE maintained a continuous structure even on culture days 12 and 15, and contained ventricular, basal-progenitor, cortical-plate and Cajal-Retzius cell layers. The cortical NE in this culture was flanked by cortical hem-like tissue. Furthermore, when Shh was added, ventral telencephalic structures such as lateral ganglionic eminence–like tissue formed in the region adjacent to the cortical NE. Thus, our results indicate that laminin-entactin ECM promotes the formation of structurally stable telencephalic tissues in 3D ESC culture, and supports the morphogenetic recapitulation of cortical development. PMID:23300850

Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf

NASA Astrophysics Data System (ADS)

Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; An Nguyen, Thien; Alfano, Robert R.

2014-06-01

Two-photon (2P) excitation of the second singlet (S) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S2 state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf.

PubMed

Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; Nguyen, Thien An; Alfano, Robert R

2014-06-01

Two-photon (2P) excitation of the second singlet (S₂) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S₂ state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S₂ state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

Opinion competition dynamics on multiplex networks

NASA Astrophysics Data System (ADS)

Amato, R.; Kouvaris, N. E.; San Miguel, M.; Díaz-Guilera, A.

2017-12-01

Multilayer and multiplex networks represent a good proxy for the description of social phenomena where social structure is important and can have different origins. Here, we propose a model of opinion competition where individuals are organized according to two different structures in two layers. Agents exchange opinions according to the Abrams-Strogatz model in each layer separately and opinions can be copied across layers by the same individual. In each layer a different opinion is dominant, so each layer has a different absorbing state. Consensus in one opinion is not the only possible stable solution because of the interaction between the two layers. A new mean field solution has been found where both opinions coexist. In a finite system there is a long transient time for the dynamical coexistence of both opinions. However, the system ends in a consensus state due to finite size effects. We analyze sparse topologies in the two layers and the existence of positive correlations between them, which enables the coexistence of inter-layer groups of agents sharing the same opinion.

Patterns of oriented cell division during the steady-state morphogenesis of the body column in hydra.

PubMed

Shimizu, H; Bode, P M; Bode, H R

1995-12-01

In an adult hydra, the tissue of the body column is in a dynamic state. The epithelial cells of both layers are constantly in the mitotic cycle. As the tissue expands, it is continuously displaced along the body axis in either an apical or basal direction, but not in a circumferential direction. Using a modified whole mount method we examined the orientation of mitotic spindles to determine what role the direction of cell division plays in axial displacement. Surprisingly, the direction of cell division was found to differ in the two epithelial layers. In the ectoderm it was somewhat biased in an axial direction. In the endoderm it was strongly biased in a circumferential direction. For both layers, the directional biases occurred throughout the length of the body column, with some regional variation in its extent. As buds developed into adults, the bias in each layer increased from an almost random distribution to the distinctly different orientations of the adult. Thus, to maintain the observed axial direction of tissue displacement, rearrangement of the epithelial cells of both layers must occur continuously in the adult as well as in developing animals. How the locomotory and contractile behavior of the muscle processes of the epithelial cells may effect changes in cell shape, and thereby influence the direction of cell division in each layer, is discussed.

Tissue engineering and regenerative medicine as applied to the gastrointestinal tract.

PubMed

Bitar, Khalil N; Zakhem, Elie

2013-10-01

The gastrointestinal (GI) tract is a complex system characterized by multiple cell types with a determined architectural arrangement. Tissue engineering of the GI tract aims to reinstate the architecture and function of all structural layers. The key point for successful tissue regeneration includes the use of cells/biomaterials that elucidate minimal immune response after implantation. Different biomaterial choices and cell sources have been proposed to engineer the GI tract. This review summarizes the recent advances in bioengineering the GI tract with emphasis on cell sources and scaffolding biomaterials. Copyright © 2013 Elsevier Ltd. All rights reserved.

Sound transmission through finite lightweight multilayered structures with thin air layers.

PubMed

Dijckmans, A; Vermeir, G; Lauriks, W

2010-12-01

The sound transmission loss (STL) of finite lightweight multilayered structures with thin air layers is studied in this paper. Two types of models are used to describe the vibro-acoustic behavior of these structures. Standard transfer matrix method assumes infinite layers and represents the plane wave propagation in the layers. A wave based model describes the direct sound transmission through a rectangular structure placed between two reverberant rooms. Full vibro-acoustic coupling between rooms, plates, and air cavities is taken into account. Comparison with double glazing measurements shows that this effect of vibro-acoustic coupling is important in lightweight double walls. For infinite structures, structural damping has no significant influence on STL below the coincidence frequency. In this frequency region, the non-resonant transmission or so-called mass-law behavior dominates sound transmission. Modal simulations suggest a large influence of structural damping on STL. This is confirmed by experiments with double fiberboard partitions and sandwich structures. The results show that for thin air layers, the damping induced by friction and viscous effects at the air gap surfaces can largely influence and improve the sound transmission characteristics.

Al{sub 2}O{sub 3} multi-density layer structure as a moisture permeation barrier deposited by radio frequency remote plasma atomic layer deposition

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jung, Hyunsoo; Samsung Display Co. Ltd., Tangjeong, Chungcheongnam-Do 336-741; Jeon, Heeyoung

2014-02-21

Al{sub 2}O{sub 3} films deposited by remote plasma atomic layer deposition have been used for thin film encapsulation of organic light emitting diode. In this study, a multi-density layer structure consisting of two Al{sub 2}O{sub 3} layers with different densities are deposited with different deposition conditions of O{sub 2} plasma reactant time. This structure improves moisture permeation barrier characteristics, as confirmed by a water vapor transmission rate (WVTR) test. The lowest WVTR of the multi-density layer structure was 4.7 × 10{sup −5} gm{sup −2} day{sup −1}, which is one order of magnitude less than WVTR for the reference single-density Al{submore » 2}O{sub 3} layer. This improvement is attributed to the location mismatch of paths for atmospheric gases, such as O{sub 2} and H{sub 2}O, in the film due to different densities in the layers. This mechanism is analyzed by high resolution transmission electron microscopy, elastic recoil detection, and angle resolved X-ray photoelectron spectroscopy. These results confirmed that the multi-density layer structure exhibits very good characteristics as an encapsulation layer via location mismatch of paths for H{sub 2}O and O{sub 2} between the two layers.« less

Plasmonic enhancement of second-harmonic generation of dielectric layer embedded in metal-dielectric-metal structure

NASA Astrophysics Data System (ADS)

Kang, Byungjun; Imakita, Kenji; Fujii, Minoru; Hayashi, Shinji

2018-03-01

The enhancement of second-harmonic generation from a dielectric layer embedded in a metal-dielectric-metal structure upon excitation of surface plasmon polaritons is demonstrated experimentally. The metal-dielectric-metal structure consisting of a Gex(SiO2)1-x layer sandwiched by two Ag layers was prepared, and the surface plasmon polaritons were excited in an attenuated total reflection geometry. The measured attenuated total reflection spectra exhibited two reflection dips corresponding to the excitation of two different surface plasmon polariton modes. Strong second-harmonic signals were observed under the excitation of these surface plasmon polariton modes. The results demonstrate that the second-harmonic intensity of the Gex(SiO2)1-x layer is highly enhanced relative to that of the single layer deposited on a substrate. Under the excitation of one of the two surface plasmon polariton modes, the estimated enhancement factor falls in a range between 39.9 and 171, while under the excitation of the other surface plasmon polariton mode, it falls in a range between 3.96 and 84.6.

A Novel Actuator for Simulation of Epidural Anesthesia and Other Needle Insertion Procedures

PubMed Central

Magill, John C.; Byl, Marten F.; Hinds, Michael F.; Agassounon, William; Pratt, Stephen D.; Hess, Philip E.

2010-01-01

Introduction When navigating a needle from skin to epidural space, a skilled clinician maintains a mental model of the anatomy and uses the various forms of haptic and visual feedback to track the location of the needle tip. Simulating the procedure requires an actuator that can produce the feel of tissue layers even as the needle direction changes from the ideal path. Methods A new actuator and algorithm architecture simulate forces associated with passing a needle through varying tissue layers. The actuator uses a set of cables to suspend a needle holder. The cables are wound onto spools controlled by brushless motors. An electromagnetic tracker is used to monitor the position of the needle tip. Results Novice and expert clinicians simulated epidural insertion with the simulator. Preliminary depth-time curves show that the user responds to changes in tissue properties as the needle is advanced. Some discrepancy in clinician response indicates that the feel of the simulator is sensitive to technique, thus perfect tissue property simulation has not been achieved. Conclusions The new simulator is able to approximately reproduce properties of complex multilayer tissue structures, including fine-scale texture. Methods for improving fidelity of the simulation are identified. PMID:20651481

A novel actuator for simulation of epidural anesthesia and other needle insertion procedures.

PubMed

Magill, John C; Byl, Marten F; Hinds, Michael F; Agassounon, William; Pratt, Stephen D; Hess, Philip E

2010-06-01

When navigating a needle from skin to epidural space, a skilled clinician maintains a mental model of the anatomy and uses the various forms of haptic and visual feedback to track the location of the needle tip. Simulating the procedure requires an actuator that can produce the feel of tissue layers even as the needle direction changes from the ideal path. A new actuator and algorithm architecture simulate forces associated with passing a needle through varying tissue layers. The actuator uses a set of cables to suspend a needle holder. The cables are wound onto spools controlled by brushless motors. An electromagnetic tracker is used to monitor the position of the needle tip. Novice and expert clinicians simulated epidural insertion with the simulator. Preliminary depth-time curves show that the user responds to changes in tissue properties as the needle is advanced. Some discrepancy in clinician response indicates that the feel of the simulator is sensitive to technique, thus perfect tissue property simulation has not been achieved. The new simulator is able to approximately reproduce properties of complex multilayer tissue structures, including fine-scale texture. Methods for improving fidelity of the simulation are identified.

Two dimensional disorder in black phosphorus and layered monochalcogenides

NASA Astrophysics Data System (ADS)

Barraza-Lopez, Salvador; Mehboudi, Mehrshad; Kumar, Pradeep; Harriss, Edmund O.; Churchill, Hugh O. H.; Dorio, Alex M.; Zhu, Wenjuan; van der Zande, Arend; Pacheco Sanjuan, Alejandro A.

The degeneracies of the structural ground state of materials with a layered orthorhombic structure such as black phosphorus and layered monochalcogenides GeS, GeSe, SnS, and SnSe, lead to an order/disorder transition in two dimensions at finite temperature. This transition has consequences on applications based on these materials requiring a crystalline two-dimensional structure. Details including a Potts model that explains the two-dimensional transition, among other results, will be given in this talk. References: M. Mehboudi, A.M. Dorio, W. Zhu, A. van der Zande, H.O.H. Churchill, A.A. Pacheco Sanjuan, E.O.H. Harris, P. Kumar, and S. Barraza-Lopez. arXiv:1510.09153.

Reconsideration of F-layer seismic model in the south polar region

NASA Astrophysics Data System (ADS)

Ohtaki, T.; Kaneshima, S.

2017-12-01

Previously, we analyzed the seismic structure near the inner core boundary beneath Antarctica (Ohtaki et al., 2012). In the study, we determined the velocity of the lowermost outer core (F-layer) using amplitude ratio observations between the inner-core phase (PKIKP) and the inner-core grazing/diffracted phase (PKPbc/c-diff). Because the observations are not so sensitive to the F-layer structure, a constant velocity is assumed in the layer to simplify the model. The obtained model (SPR) has a flat velocity zone with a 75 km thick on the inner core boundary. With this F-layer structure and using travel times of these phases as well as the phase that reflects at the boundary, we determined the seismic structure of the inner core in the south polar region. However, a constant velocity layer is unrealistic, although it is reasonable assumption.Recently, we determined F-layer velocity structures more accurately using the combined observations of PKiKP-PKPbc differential travel times and of PKPbc/c-diff dispersion (Ohtaki et al., 2015, 2016). The former observation is sensitive to average velocity in the F-layer; the latter to velocity gradient in the layer. By analyzing these two observations together, we can determine the detailed velocity structure in the F-layer. The surveyed areas are beneath the Northeast Pacific and Australia. The seismic velocity models obtained are quite different between the two regions. Thus our results require laterally heterogeneous F-layer, and show that F-layer is more complicated than we ever imagined.Then there is one question; which structure is that of the south polar region close to? Unfortunately, the seismic waveforms that we analyzed in the previous study may not have quality high enough to analyze the PKiKP-PKPbc or PKPbc dispersion. However, it would be meaningful to reanalyze the amplitude data and reconsider the F-layer velocity there. And we also estimate how large slope of velocity can be acceptable for the F-layer velocity structure in this region.

Proof-of-concept: 3D bioprinting of pigmented human skin constructs.

PubMed

Ng, Wei Long; Qi, Jovina Tan Zhi; Yeong, Wai Yee; Naing, May Win

2018-01-23

Three-dimensional (3D) pigmented human skin constructs have been fabricated using a 3D bioprinting approach. The 3D pigmented human skin constructs are obtained from using three different types of skin cells (keratinocytes, melanocytes and fibroblasts from three different skin donors) and they exhibit similar constitutive pigmentation (pale pigmentation) as the skin donors. A two-step drop-on-demand bioprinting strategy facilitates the deposition of cell droplets to emulate the epidermal melanin units (pre-defined patterning of keratinocytes and melanocytes at the desired positions) and manipulation of the microenvironment to fabricate 3D biomimetic hierarchical porous structures found in native skin tissue. The 3D bioprinted pigmented skin constructs are compared to the pigmented skin constructs fabricated by conventional a manual-casting approach; in-depth characterization of both the 3D pigmented skin constructs has indicated that the 3D bioprinted skin constructs have a higher degree of resemblance to native skin tissue in term of the presence of well-developed stratified epidermal layers and the presence of a continuous layer of basement membrane proteins as compared to the manually-cast samples. The 3D bioprinting approach facilitates the development of 3D in vitro pigmented human skin constructs for potential toxicology testing and fundamental cell biology research.

Trapped-mode-induced Fano resonance and acoustical transparency in a one-dimensional solid-fluid phononic crystal

NASA Astrophysics Data System (ADS)

Quotane, Ilyasse; El Boudouti, El Houssaine; Djafari-Rouhani, Bahram

2018-01-01

We investigate theoretically and numerically the possibility of existence of Fano and acoustic-induced transparency (AIT) resonances in a simple though realistic one-dimensional acoustic structure made of solid-fluid layers inserted between two fluids. These resonances are obtained by combining appropriately the zeros of transmission (antiresonance) induced by the solid layers and the local resonances induced by the solid or combined solid-fluid layers with surface free boundary conditions. In particular, we show the possibility of trapped modes, also called bound states in continuum, which have recently found a high renewal interest. These modes appear as resonances with zero width in the transmission spectra as well as in the density of states (DOS). We consider three different structures: (i) a single solid layer inserted between two fluids. This simple structure shows the possibility of existence of trapped modes, which are discrete modes of the solid layer that lie in the continuum modes of the surrounding fluids. We give explicit analytical expressions of the dispersion relation of these eigenmodes of the solid layer which are found independent of the nature of the surrounding fluids. By slightly detuning the angle of incidence from that associated to the trapped mode, we get a well-defined Fano resonance characterized by an asymmetric Fano profile in the transmission spectra. (ii) The second structure consists of a solid-fluid-solid triple layer embedded between two fluids. This structure is found more appropriate to show both Fano and acoustic-induced transparency resonances. We provide detailed analytical expressions for the transmission and reflection coefficients that enable us to deduce a closed-form expression of the dispersion relation giving the trapped modes. Two situations can be distinguished in the triple-layer system: in the case of a symmetric structure (i.e., the same solid layers) we show, by detuning the incidence angle θ , the possibility of existence of Fano resonances that can be fitted following a Fano-type expression. The variation of the Fano parameter that describes the asymmetry of such resonances as well as their width versus θ is studied in detail. In the case of an asymmetric structure (i.e., different solid layers), we show the existence of an incidence angle that enables to squeeze a resonance between two transmission zeros induced by the two solid layers. This resonance behaves like an AIT resonance, its position and width depend on the nature of the fluid and solid layers as well as on the difference between the thicknesses of the solid layers. (iii) In the case of a periodic structure (phononic crystal), we show that trapped modes and Fano resonances give rise, respectively, to dispersionless flat bands with zero group velocity and nearly flat bands with negative or positive group velocities. The analytical results presented here are obtained by means of the Green's function method which enables to deduce in closed form: dispersion curves, transmission and reflection coefficients, DOS, as well as the displacement fields. The proposed solid-fluid layered structures should have important applications for designing acoustic mirrors and acoustic filters as well as supersonic and subsonic materials.

Fabrication and characterization of a 3-D non-homogeneous tissue-like mouse phantom for optical imaging

NASA Astrophysics Data System (ADS)

Avtzi, Stella; Zacharopoulos, Athanasios; Psycharakis, Stylianos; Zacharakis, Giannis

2013-11-01

In vivo optical imaging of biological tissue not only requires the development of new theoretical models and experimental procedures, but also the design and construction of realistic tissue-mimicking phantoms. However, most of the phantoms available currently in literature or the market, have either simple geometrical shapes (cubes, slabs, cylinders) or when realistic in shape they use homogeneous approximations of the tissue or animal under investigation. The goal of this study is to develop a non-homogeneous realistic phantom that matches the anatomical geometry and optical characteristics of the mouse head in the visible and near-infrared spectral range. The fabrication of the phantom consisted of three stages. Initially, anatomical information extracted from either mouse head atlases or structural imaging modalities (MRI, XCT) was used to design a digital phantom comprising of the three main layers of the mouse head; the brain, skull and skin. Based on that, initial prototypes were manufactured by using accurate 3D printing, allowing complex objects to be built layer by layer with sub-millimeter resolution. During the second stage the fabrication of individual molds was performed by embedding the prototypes into a rubber-like silicone mixture. In the final stage the detailed phantom was constructed by loading the molds with epoxy resin of controlled optical properties. The optical properties of the resin were regulated by using appropriate quantities of India ink and intralipid. The final phantom consisted of 3 layers, each one with different absorption and scattering coefficient (μa,μs) to simulate the region of the mouse brain, skull and skin.

Mucin Production Dynamics at the Surface of Corneal Epithelial Cells

NASA Astrophysics Data System (ADS)

Hormel, Tristan; Bhattacharjee, Tapomoy; Pitenis, Angela; Urueã+/-A, Juan; Sawyer, Gregory; Angelini, Thomas

Mucous layers form at the apical surface of many epithelia, protecting tissues from pathogens and environmental wear and damage. Although these layers contain many materials they are primarily composed of mucin glycoproteins, the concentration of which may be physiologically controlled to maintain specific rheological properties and to provide proper lubrication. Nowhere is this truer than at the surface of the eye's corneal epithelium, where the mucous layer must additionally achieve structural integrity to withstand the stresses created by blinking, and remain transparent in order to enable vision. I will present results on the growth dynamics, concentration, and rheology of a model corneal epithelial mucous layer, all of which can be viewed as important parameters at this interface. I will also discuss modulation of the mucous layer's dynamics with variation in environmental conditions. Alcon.

Morphological and ultrastructural comparative analysis of bone tissue after Er:YAG laser and surgical drill osteotomy.

PubMed

Panduric, Dragana Gabric; Juric, Ivona Bago; Music, Svetozar; Molčanov, Krešimir; Sušic, Mato; Anic, Ivica

2014-07-01

The purpose of this study was to analyze morphological, chemical, and crystallographic changes of bone tissue after osteotomy performed with an erbium:yttrium-aluminium-garnet (Er:YAG) laser and a low speed pilot drill. Bone blocks were prepared from porcine ribs, and on each block, two tunnel preparations were performed using the Er:YAG laser (pulse energy: 1000 mJ, pulse duration: 300 μs, pulse repetition rate: 20 Hz) or the low-speed surgical pilot drill. The morphological changes of the cortical and the spongious surface of the tunnel preparations were analyzed under the field emission scanning electron microscopy (FE-SEM) at low and high resolution. The distribution and the level of chemical elements in the treated surfaces were evaluated by qualitative and semiquantitative energy dispersive x-ray analysis (SEM-EDX). Diffraction x-ray analysis was used to detect any differences and thermally induced modifications of hydroxyapatite crystals. FE-SEM revealed sharp edges of the Er:YAG preparations, with empty intertrabecular spaces and no signs of carbonization. In the drill group, the surface of the preparations was smooth, completely covered with smear layer and microcracks, and with hairy-like irregularities on the edges. SEM-EDX analysis did not reveal any differences in the number of specific chemical elements between the laser and the drill group. There were no thermally induced modifications of hydroxyapatite crystal structure in the bone tissue in either group. The Er:YAG laser ablation did not cause any chemical or crystallographic changes of the bone tissue. Compared with the drill, Er:YAG laser created well-defined edges of the preparations, and cortical bone had no smear layer.

Micro-PIXE mapping of mineral distribution in mature grain of two pearl millet cultivars

NASA Astrophysics Data System (ADS)

Minnis-Ndimba, R.; Kruger, J.; Taylor, J. R. N.; Mtshali, C.; Pineda-Vargas, C. A.

2015-11-01

Micro-proton-induced X-ray emission (micro-PIXE) was used to map the distribution of several nutritionally important minerals found in the grain tissue of two cultivars of pearl millet (Pennisetum glaucum (L.) R. Br.). The distribution maps revealed that the predominant localisation of minerals was within the germ (consisting of the scutellum and embryo) and the outer grain layers (specifically the pericarp and aleurone); whilst the bulk of the endosperm tissue featured relatively low concentrations of the surveyed minerals. Within the germ, the scutellum was revealed as a major storage tissue for P and K, whilst Ca, Mn and Zn were more prominent within the embryo. Fe was revealed to have a distinctive distribution pattern, confined to the dorsal end of the scutellum; but was also highly concentrated in the outer grain layers. Interestingly, the hilar region was also revealed as a site of high accumulation of minerals, particularly for S, Ca, Mn, Fe and Zn, which may be part of a defensive strategy against infection or damage. Differences between the two cultivars, in terms of the bulk Fe and P content obtained via inductively coupled plasma optical emission spectrometry (ICP-OES), concurred with the average concentration data determined from the analysis of micro-PIXE spectra specifically extracted from the endosperm tissue.

Six-band terahertz metamaterial absorber based on the combination of multiple-order responses of metallic patches in a dual-layer stacked resonance structure.

PubMed

Wang, Ben-Xin; Wang, Gui-Zhen; Sang, Tian; Wang, Ling-Ling

2017-01-25

This paper reports on a numerical study of the six-band metamaterial absorber composed of two alternating stack of metallic-dielectric layers on top of a continuous metallic plane. Six obvious resonance peaks with high absorption performance (average larger than 99.37%) are realized. The first, third, fifth, and the second, fourth, sixth resonance absorption bands are attributed to the multiple-order responses (i.e., the 1-, 3- and 5-order responses) of the bottom- and top-layer of the structure, respectively, and thus the absorption mechanism of six-band absorber is due to the combination of two sets of the multiple-order resonances of these two layers. Besides, the size changes of the metallic layers have the ability to tune the frequencies of the six-band absorber. Employing the results, we also present a six-band polarization tunable absorber through varying the sizes of the structure in two orthogonal polarization directions. Moreover, nine-band terahertz absorber can be achieved by using a three-layer stacked structure. Simulation results indicate that the absorber possesses nine distinct resonance bands, and average absorptivities of them are larger than 94.03%. The six-band or nine-band absorbers obtained here have potential applications in many optoelectronic and engineering technology areas.

Computing dispersion curves of elastic/viscoelastic transversely-isotropic bone plates coupled with soft tissue and marrow using semi-analytical finite element (SAFE) method.

PubMed

Nguyen, Vu-Hieu; Tran, Tho N H T; Sacchi, Mauricio D; Naili, Salah; Le, Lawrence H

2017-08-01

We present a semi-analytical finite element (SAFE) scheme for accurately computing the velocity dispersion and attenuation in a trilayered system consisting of a transversely-isotropic (TI) cortical bone plate sandwiched between the soft tissue and marrow layers. The soft tissue and marrow are mimicked by two fluid layers of finite thickness. A Kelvin-Voigt model accounts for the absorption of all three biological domains. The simulated dispersion curves are validated by the results from the commercial software DISPERSE and published literature. Finally, the algorithm is applied to a viscoelastic trilayered TI bone model to interpret the guided modes of an ex-vivo experimental data set from a bone phantom. Copyright © 2017 Elsevier Ltd. All rights reserved.

Quantitative evaluation of deep and shallow tissue layers' contribution to fNIRS signal using multi-distance optodes and independent component analysis.

PubMed

Funane, Tsukasa; Atsumori, Hirokazu; Katura, Takusige; Obata, Akiko N; Sato, Hiroki; Tanikawa, Yukari; Okada, Eiji; Kiguchi, Masashi

2014-01-15

To quantify the effect of absorption changes in the deep tissue (cerebral) and shallow tissue (scalp, skin) layers on functional near-infrared spectroscopy (fNIRS) signals, a method using multi-distance (MD) optodes and independent component analysis (ICA), referred to as the MD-ICA method, is proposed. In previous studies, when the signal from the shallow tissue layer (shallow signal) needs to be eliminated, it was often assumed that the shallow signal had no correlation with the signal from the deep tissue layer (deep signal). In this study, no relationship between the waveforms of deep and shallow signals is assumed, and instead, it is assumed that both signals are linear combinations of multiple signal sources, which allows the inclusion of a "shared component" (such as systemic signals) that is contained in both layers. The method also assumes that the partial optical path length of the shallow layer does not change, whereas that of the deep layer linearly increases along with the increase of the source-detector (S-D) distance. Deep- and shallow-layer contribution ratios of each independent component (IC) are calculated using the dependence of the weight of each IC on the S-D distance. Reconstruction of deep- and shallow-layer signals are performed by the sum of ICs weighted by the deep and shallow contribution ratio. Experimental validation of the principle of this technique was conducted using a dynamic phantom with two absorbing layers. Results showed that our method is effective for evaluating deep-layer contributions even if there are high correlations between deep and shallow signals. Next, we applied the method to fNIRS signals obtained on a human head with 5-, 15-, and 30-mm S-D distances during a verbal fluency task, a verbal working memory task (prefrontal area), a finger tapping task (motor area), and a tetrametric visual checker-board task (occipital area) and then estimated the deep-layer contribution ratio. To evaluate the signal separation performance of our method, we used the correlation coefficients of a laser-Doppler flowmetry (LDF) signal and a nearest 5-mm S-D distance channel signal with the shallow signal. We demonstrated that the shallow signals have a higher temporal correlation with the LDF signals and with the 5-mm S-D distance channel than the deep signals. These results show the MD-ICA method can discriminate between deep and shallow signals. Copyright © 2013 Elsevier Inc. All rights reserved.

Sandwiched polymer fibre in fibrin matrices for the dictation of endothelial cells undergoing angiogenesis

NASA Astrophysics Data System (ADS)

Sukmana, I.; Djuansjah, J. R. P.

2013-04-01

We present here a three-dimensional (3D) sandwich system made by poly(ethylene terephthalate) (PET) fibre and fibrin extracellular matrix (ECM) for endothelial cell dictation and angiogenesis guidance. In this three-dimensional system, Human Umbilical Vein Endothelial cells (HUVECs) were firstly cultured for 2 (two) days to cover the PET fibre before sandwiched in two layer fibrin gel containing HUVECs. After 4 (four) days of culture, cel-to-cel connection, tube-like structure and multi-cellular lumen formation were then assessed and validated. Phase contrast and fluorescence imaging using an inverted microscope were used to determine cell-to-cell and cell-ECM interactions. Laser scanning confocal microscopy and histological techniques were used to confirm the development of tube-like structure and multi-cellular lumen formation. This study shows that polymer fibres sandwiched in fibrin gel can be used to dictate endothelial cells undergoing angiogenesis with potential application in cancer and cardiovascular study and tissue engineering vascularisation.

Speckle reduction in optical coherence tomography using two-step iteration method (Conference Presentation)

NASA Astrophysics Data System (ADS)

Wang, Xianghong; Liu, Xinyu; Wang, Nanshuo; Yu, Xiaojun; Bo, En; Chen, Si; Liu, Linbo

2017-02-01

Optical coherence tomography (OCT) provides high resolution and cross-sectional images of biological tissue and is widely used for diagnosis of ocular diseases. However, OCT images suffer from speckle noise, which typically considered as multiplicative noise in nature, reducing the image resolution and contrast. In this study, we propose a two-step iteration (TSI) method to suppress those noises. We first utilize augmented Lagrange method to recover a low-rank OCT image and remove additive Gaussian noise, and then employ the simple and efficient split Bregman method to solve the Total-Variation Denoising model. We validated such proposed method using images of swine, rabbit and human retina. Results demonstrate that our TSI method outperforms the other popular methods in achieving higher peak signal-to-noise ratio (PSNR) and structure similarity (SSIM) while preserving important structural details, such as tiny capillaries and thin layers in retinal OCT images. In addition, the results of our TSI method show clearer boundaries and maintains high image contrast, which facilitates better image interpretations and analyses.

Multi-layer carbon-based coatings for field emission

DOEpatents

Sullivan, John P.; Friedmann, Thomas A.

1998-01-01

A multi-layer resistive carbon film field emitter device for cold cathode field emission applications. The multi-layered film of the present invention consists of at least two layers of a conductive carbon material, preferably amorphous-tetrahedrally coordinated carbon, where the resistivities of adjacent layers differ. For electron emission from the surface, the preferred structure can be a top layer having a lower resistivity than the bottom layer. For edge emitting structures, the preferred structure of the film can be a plurality of carbon layers, where adjacent layers have different resistivities. Through selection of deposition conditions, including the energy of the depositing carbon species, the presence or absence of certain elements such as H, N, inert gases or boron, carbon layers having desired resistivities can be produced.

Optimized capping layers for EUV multilayers

DOEpatents

Bajt, Sasa [Livermore, CA; Folta, James A [Livermore, CA; Spiller, Eberhard A [Livermore, CA

2004-08-24

A new capping multilayer structure for EUV-reflective Mo/Si multilayers consists of two layers: A top layer that protects the multilayer structure from the environment and a bottom layer that acts as a diffusion barrier between the top layer and the structure beneath. One embodiment combines a first layer of Ru with a second layer of B.sub.4 C. Another embodiment combines a first layer of Ru with a second layer of Mo. These embodiments have the additional advantage that the reflectivity is also enhanced. Ru has the best oxidation resistance of all materials investigated so far. B.sub.4 C is an excellent barrier against silicide formation while the silicide layer formed at the Si boundary is well controlled.

Development of novel layered nanoparticles for more efficient cancer treatment

NASA Astrophysics Data System (ADS)

Priest, Thomas A.

Cancer is the second-most leading cause of death in the United States, with 1.66 million new cases expected to be diagnosed and over 580,000 Americans expected to die of cancer in 2013 alone. (American Cancer Society 2013) Current treatments result in damage to the healthy tissues and incomplete resections of solid tumors, but by harnessing nanotechnology, more effective treatments can be constructed. Gold nanoshells present a promising option for targeted cancer therapy. The anatomy of tumors causes the "enhanced permeability and retention" effect, which means that nanoscale particles will extravasate from the bloodstream and accumulate in the tumors. However, small nanoparticles must still diffuse from the tumor vasculature into the tumor tissue. Due to impaired vascularization, the particles are unable to reach into the entire tumor region. The purpose of our project is to create a "two-layer" nanoshell coated with alkanethiol and phosphatidlycholine and a "three-layer" nanoshell that coats the "two-layer" system with a layer of high-density lipoprotein. It is proposed that these coatings will allow for better penetration of solid tumors compared to the standard nanoshells modified with poly(ethylene glycol) (PEG). In addition to the nanoshells, citrate-gold nanoparticles were investigated as a control. Size, zeta potential, and morphology were optimized, and the penetration of the particles into solid tumors was investigated using dark-field microscopy. It was discovered that the "two-layer" nanoshells exhibited significantly more uptake into the solid tumors compared to PEGylated nanoshells, and should be further investigated as a platform for targeted cancer therapies.

The global mechanical properties and multi-scale failure mechanics of heterogeneous human stratum corneum.

PubMed

Liu, X; Cleary, J; German, G K

2016-10-01

The outermost layer of skin, or stratum corneum, regulates water loss and protects underlying living tissue from environmental pathogens and insults. With cracking, chapping or the formation of exudative lesions, this functionality is lost. While stratum corneum exhibits well defined global mechanical properties, macroscopic mechanical testing techniques used to measure them ignore the structural heterogeneity of the tissue and cannot provide any mechanistic insight into tissue fracture. As such, a mechanistic understanding of failure in this soft tissue is lacking. This insight is critical to predicting fracture risk associated with age or disease. In this study, we first quantify previously unreported global mechanical properties of isolated stratum corneum including the Poisson's ratio and mechanical toughness. African American breast stratum corneum is used for all assessments. We show these parameters are highly dependent on the ambient humidity to which samples are equilibrated. A multi-scale investigation assessing the influence of structural heterogeneities on the microscale nucleation and propagation of cracks is then performed. At the mesoscale, spatially resolved equivalent strain fields within uniaxially stretched stratum corneum samples exhibit a striking heterogeneity, with localized peaks correlating closely with crack nucleation sites. Subsequent crack propagation pathways follow inherent topographical features in the tissue and lengthen with increased tissue hydration. At the microscale, intact corneocytes and polygonal shaped voids at crack interfaces highlight that cracks propagate in superficial cell layers primarily along intercellular junctions. Cellular fracture does occur however, but is uncommon. Human stratum corneum protects the body against harmful environmental pathogens and insults. Upon mechanical failure, this barrier function is lost. Previous studies characterizing the mechanics of stratum corneum have used macroscopic testing equipment designed for homogenous materials. Such measurements ignore the tissue's rich topography and heterogeneous structure, and cannot describe the underlying mechanistic process of tissue failure. For the first time, we establish a mechanistic insight into the failure mechanics of soft heterogeneous tissues by investigating how cracks nucleate and propagate in stratum corneum. We further quantify previously unreported values of the tissue's Poisson's ratio and toughness, and their dramatic variation with ambient humidity. To date, skin models examining drug delivery, wound healing, and ageing continue to estimate these parameters. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Novel Layered Supercell Structure from Bi 2AlMnO 6 for Multifunctionalities

DOE PAGES

Li, Leigang; Boullay, Philippe; Lu, Ping; ...

2017-10-02

Layered materials, e.g., graphene and transition metal (di)chalcogenides, holding great promises in nanoscale device applications have been extensively studied in fundamental chemistry, solid state physics and materials research areas. In parallel, layered oxides (e.g., Aurivillius and Ruddlesden–Popper phases) present an attractive class of materials both because of their rich physics behind and potential device applications. In this work, we report a novel layered oxide material with self-assembled layered supercell structure consisting of two mismatch-layered sublattices of [Bi 3O 3+δ] and [MO 2] 1.84 (M = Al/Mn, simply named BAMO), i.e., alternative layered stacking of two mutually incommensurate sublattices made ofmore » a three-layer-thick Bi–O slab and a one-layer-thick Al/Mn–O octahedra slab in the out-of-plane direction. Strong room-temperature ferromagnetic and piezoelectric responses as well as anisotropic optical property have been demonstrated with great potentials in various device applications. Furthermore, the realization of the novel BAMO layered supercell structure in this work has paved an avenue toward exploring and designing new materials with multifunctionalities.« less

Spectroscopy of Multilayered Biological Tissues for Diabetes Care

NASA Astrophysics Data System (ADS)

Yudovsky, Dmitry

Neurological and vascular complications of diabetes mellitus are known to cause foot ulceration in diabetic patients. Present clinical screening techniques enable the diabetes care provider to triage treatment by identifying diabetic patients at risk of foot ulceration. However, these techniques cannot effectively identify specific areas of the foot at risk of ulceration. This study aims to develop non-invasive optical techniques for accurate assessment of tissue health and viability with spatial resolution on the order of 1 mm². The thesis can be divided into three parts: (1) the use of hyperspectral tissue oximetry to detect microcirculatory changes prior to ulcer formation, (2) development of a two-layer tissue spectroscopy algorithm and its application to detection of callus formation or epidermal degradation prior to ulceration, and (3) multi-layered tissue fluorescence modeling for identification of bacterial growth in existing diabetic foot wounds. The first part of the dissertation describes a clinical study in which hyperspectral tissue oximetry was performed on multiple diabetic subjects at risk of ulceration. Tissue oxyhemoglobin and deoxyhemoglobin concentrations were estimated using the Modified Beer-Lambert law. Then, an ulcer prediction algorithm was developed based on retrospective analysis of oxyhemoglobin and deoxyhemoglobin concentrations in sites that were known to ulcerate. The ulcer prediction algorithm exhibited a large sensitivity but low specificity of 95 and 80%, respectively. The second part of the dissertation revisited the hyperspectral data presented in part one with a new and novel two-layer tissue spectroscopy algorithm. This algorithm was able to detect not only oxyhemoglobin and deoxyhemoglobin concentrations, but also the thickness of the epidermis, and the tissue's scattering coefficient. Specifically, change in epidermal thickness provided insight into the formation of diabetic foot ulcers over time. Indeed, callus formation or the thickening of the epidermis which preempts ulcer formation was detectable prior to ulceration. This added dimension of information increased the specificity of the ulcer prediction algorithm by 7% without reducing the sensitivity. Finally, the third part of the dissertation describes the feasibility of detecting bacteria in open ulcers. First, a semi-empirical model of multi-layered tissue fluorescence was developed. Then, an inverse method was developed and applied to simulated fluorescence emission spectra of diabetic foot wounds infected with Staphylococcus aureus and stained with indocyanine green dye (ICG). The inverse method was able to detect the blood volume fraction, oxygen saturation, and the intrinsic fluorescence spectrum of the ICG dye from simulated fluorescence emission spectra.

Thicker host tissues moderate light stress in a cnidarian endosymbiont.

PubMed

Dimond, James L; Holzman, Benjamin J; Bingham, Brian L

2012-07-01

The susceptibility of algal-cnidarian holobionts to environmental stress is dependent on attributes of both host and symbiont, but the role of the host is often unclear. We examined the influence of the host on symbiont light stress, comparing the photophysiology of the chlorophyte symbiont Elliptochloris marina in two species of sea anemones in the genus Anthopleura. After 3 months of acclimation in outdoor tanks, polyp photoprotective contraction behavior was similar between the two host species, but photochemical efficiency was 1.5 times higher in A. xanthogrammica than in A. elegantissima. Maximum relative electron transport rates, derived from rapid light curves, were 1.5 times higher in A. xanthogrammica than in A. elegantissima when symbionts were inside intact tissues, but were not significantly different between host species upon removal of outer (epidermis and mesoglea) tissue layers from symbiont-containing gastrodermal cells. Tissues of A. xanthogrammica were 1.8 times thicker than those of A. elegantissima, with outer tissue layers attenuating 1.6 times more light. We found no significant differences in light absorption properties per unit volume of tissue, confirming the direct effect of tissue thickness on light attenuation. The thicker tissues of A. xanthogrammica thus provide a favorable environment for E. marina - a relatively stress-susceptible symbiont - and may explain its higher prevalence and expanded range in A. xanthogrammica along the Pacific coast of North America. Our findings also support a photoprotective role for thicker host tissues in reef corals that has long been thought to influence variability in bleaching susceptibility among coral taxa.

Vocal Fold Epithelial Barrier in Health and Injury: A Research Review

ERIC Educational Resources Information Center

Levendoski, Elizabeth Erickson; Leydon, Ciara; Thibeault, Susan L.

2014-01-01

Purpose: Vocal fold epithelium is composed of layers of individual epithelial cells joined by junctional complexes constituting a unique interface with the external environment. This barrier provides structural stability to the vocal folds and protects underlying connective tissue from injury while being nearly continuously exposed to potentially…

Design and fabrication of a chitosan hydrogel with gradient structures via a step-by-step cross-linking process.

PubMed

Xu, Yongxiang; Yuan, Shenpo; Han, Jianmin; Lin, Hong; Zhang, Xuehui

2017-11-15

The development of scaffolds to mimic the gradient structure of natural tissue is an important consideration for effective tissue engineering. In the present study, a physical cross-linking chitosan hydrogel with gradient structures was fabricated via a step-by-step cross-linking process using sodium tripolyphosphate and sodium hydroxide as sequential cross-linkers. Chitosan hydrogels with different structures (single, double, and triple layers) were prepared by modifying the gelling process. The properties of the hydrogels were further adjusted by varying the gelling conditions, such as gelling time, pH, and composition of the crosslinking solution. Slight cytotoxicity was showed in MTT assay for hydrogels with uncross-linking chitosan solution and non-cytotoxicity was showed for other hydrogels. The results suggest that step-by-step cross-linking represents a practicable method to fabricate scaffolds with gradient structures. Copyright © 2017. Published by Elsevier Ltd.

Fabrication of triple-layered bifurcated vascular scaffold with a certain degree of three-dimensional structure

NASA Astrophysics Data System (ADS)

Liu, Yuanyuan; Jiang, Weijian; Yang, Yang; Pu, Huayan; Peng, Yan; Xin, Liming; Zhang, Yi; Sun, Yu

2018-01-01

Constructing vascular scaffolds is important in tissue engineering. However, scaffolds with characteristics such as multiple layers and a certain degree of spatial morphology still cannot be readily constructed by current vascular scaffolds fabrication techniques. This paper presents a three-layered bifurcated vascular scaffold with a curved structure. The technique combines 3D printed molds and casting hydrogel and fugitive ink to create vessel-mimicking constructs with customizable structural parameters. Compared with other fabrication methods, the technique can create more native-like 3D geometries. The diameter and wall thickness of the fabricated constructs can be independently controlled, providing a feasible approach for vascular scaffold construction. Enzymatically-crosslinked gelatin was used as the scaffold material. The morphology and mechanical properties were evaluated. Human umbilical cord derived endothelial cells (HUVECs) were seeded on the scaffolds and cultured for 72 h. Cell viability and morphology were assessed. The results showed that the proposed process had good application potentials, and will hopefully provide a feasible approach for constructing vascular scaffolds.

The role of apical contractility in determining cell morphology in multilayered epithelial sheets and tubes

NASA Astrophysics Data System (ADS)

Zhen Tan, Rui; Lai, Tanny; Chiam, K.-H.

2017-08-01

A multilayered epithelium is made up of individual cells that are stratified in an orderly fashion, layer by layer. In such tissues, individual cells can adopt a wide range of shapes ranging from columnar to squamous. From histological images, we observe that, in flat epithelia such as the skin, the cells in the top layer are squamous while those in the middle and bottom layers are columnar, whereas in tubular epithelia, the cells in all layers are columnar. We develop a computational model to understand how individual cell shape is governed by the mechanical forces within multilayered flat and curved epithelia. We derive the energy function for an epithelial sheet of cells considering intercellular adhesive and intracellular contractile forces. We determine computationally the cell morphologies that minimize the energy function for a wide range of cellular parameters. Depending on the dominant adhesive and contractile forces, we find four dominant cell morphologies for the multilayered-layered flat sheet and three dominant cell morphologies for the two-layered curved sheet. We study the transitions between the dominant cell morphologies for the two-layered flat sheet and find both continuous and discontinuous transitions and also the presence of multistable states. Matching our computational results with histological images, we conclude that apical contractile forces from the actomyosin belt in the epithelial cells is the dominant force determining cell shape in multilayered epithelia. Our computational model can guide tissue engineers in designing artificial multilayered epithelia, in terms of figuring out the cellular parameters needed to achieve realistic epithelial morphologies.

Statistical and fractal analysis of autofluorescent myocardium images in posthumous diagnostics of acute coronary insufficiency

NASA Astrophysics Data System (ADS)

Boichuk, T. M.; Bachinskiy, V. T.; Vanchuliak, O. Ya.; Minzer, O. P.; Garazdiuk, M.; Motrich, A. V.

2014-08-01

This research presents the results of investigation of laser polarization fluorescence of biological layers (histological sections of the myocardium). The polarized structure of autofluorescence imaging layers of biological tissues was detected and investigated. Proposed the model of describing the formation of polarization inhomogeneous of autofluorescence imaging biological optically anisotropic layers. On this basis, analytically and experimentally tested to justify the method of laser polarimetry autofluorescent. Analyzed the effectiveness of this method in the postmortem diagnosis of infarction. The objective criteria (statistical moments) of differentiation of autofluorescent images of histological sections myocardium were defined. The operational characteristics (sensitivity, specificity, accuracy) of these technique were determined.

The competition of densification and structure ordering during crystallization of HCP-Mg in the framework of layering

NASA Astrophysics Data System (ADS)

Luo, Jie; Jiang, Yewei; Yu, Ronggang; Wu, Yongquan

2017-06-01

In this paper, we performed an NPT molecular dynamics simulation of crystallization process of HCP-Mg to probe the competition between densification and structural ordering. Two opposite layering patterns, i.e. outward and inward, were designed for analysis. From the perspective of solid-like cluster (SLC) itself, structural ordering always precedes densification; but from the perspective of SLC's precursor, structural ordering always lags behind densification; the reversion occurs at the closest two liquid layers around SLC. We call it dip-rebound phenomenon. This phenomenon is a completely new finding. It resolves, to some extent, recent debate about whether densification or structural ordering triggers crystallization.

Assessment of tissue viability by polarization spectroscopy

NASA Astrophysics Data System (ADS)

Nilsson, G.; Anderson, C.; Henricson, J.; Leahy, M.; O'Doherty, J.; Sjöberg, F.

2008-09-01

A new and versatile method for tissue viability imaging based on polarization spectroscopy of blood in superficial tissue structures such as the skin is presented in this paper. Linearly polarized light in the visible wavelength region is partly reflected directly by the skin surface and partly diffusely backscattered from the dermal tissue matrix. Most of the directly reflected light preserves its polarization state while the light returning from the deeper tissue layers is depolarized. By the use of a polarization filter positioned in front of a sensitive CCD-array, the light directly reflected from the tissue surface is blocked, while the depolarized light returning from the deeper tissue layers reaches the detector array. By separating the colour planes of the detected image, spectroscopic information about the amount of red blood cells (RBCs) in the microvascular network of the tissue under investigation can be derived. A theory that utilizes the differences in light absorption of RBCs and bloodless tissue in the red and green wavelength region forms the basis of an algorithm for displaying a colour coded map of the RBC distribution in a tissue. Using a fluid model, a linear relationship (cc. = 0.99) between RBC concentration and the output signal was demonstrated within the physiological range 0-4%. In-vivo evaluation using transepidermal application of acetylcholine by the way of iontophoresis displayed the heterogeneity pattern of the vasodilatation produced by the vasoactive agent. Applications of this novel technology are likely to be found in drug and skin care product development as well as in the assessment of skin irritation and tissue repair processes and even ultimately in a clinic case situation.

Collagen fibre characterisation in arterial tissue under load using SALS.

PubMed

Gaul, R T; Nolan, D R; Lally, C

2017-11-01

The collagen fibre architecture of arterial tissue is known to play a key role in its resultant mechanical behaviour, while maladaptive remodelling of this architecture may be linked to disease. Many of the techniques currently used to analyse collagen fibre architecture require time consuming tissue preparation procedures and are destructive in nature. The aim of this study is to fully explore Small Angle Light Scattering (SALS) as a means to non-destructively assess collagen fibre architecture in arterial tissue and subsequently gain insights into load induced reorientation. The optimised configuration of the SALS system for arterial tissue was determined using quantitative comparisons to histological analyses of porcine carotid artery as its basis. Once established, layer specific fibre orientation and the influence of tissue loading was determined for thin sections of carotid artery using SALS. This process was subsequently repeated for intact carotid artery layers. A single family of circumferentially orientated collagen fibres were found in the intima (- 0.1 ± 1.4° (5.5°)) and media (- 1.7 ± 1.9° (4.7°)) while two perpendicular families of fibres were identified in the adventitia (- 6.4 ± 0.7° (37.7°)) and (118.3 ± 2.7 (39.9°)). An increase in fibre alignment in response to a 20% circumferential strain was also identified using SALS, characterised by an increase in scattered light eccentricity. determined using SALS agreed with those found using traditional destructive techniques, however SALS has the important benefits of allowing vessel layers to remain intact, and has a fast processing time. SALS unique ability to identify load induced reorganisation in intact arterial layers offers an efficient means to gain crucial insights into arterial disease and its development over time. Copyright © 2017 Elsevier Ltd. All rights reserved.

Concentrations of dapivirine in the rhesus macaque and rabbit following once daily intravaginal administration of a gel formulation of [14C]dapivirine for 7 days.

PubMed

Nuttall, Jeremy P; Thake, Daryl C; Lewis, Mark G; Ferkany, John W; Romano, Joseph W; Mitchnick, Mark A

2008-03-01

Dapivirine is a nonnucleoside reverse transcriptase inhibitor being developed as a topical microbicide for the prevention of human immunodeficiency virus infection. The distribution of radioactivity and drug in plasma and in vaginal, cervical, and draining lymph node tissues was investigated after daily application of a vaginal gel formulation of [14C]dapivirine to rhesus macaques. This was preceded by a preliminary study with rabbits. Following the intravaginal administration of [14C]dapivirine ( approximately 0.1 mg/ml [15 microCi/ml]) to rabbits (0.5 ml/day) and macaques (1 ml/day) for 7 days, the dapivirine levels associated with vaginal and cervical tissue samples 1 h after the final dose were high (quantities of microg/g of tissue) and remained detectable at 24 h (mean, >or=2.5 ng/g in rabbits) and 48 h (mean, >80 ng/g in macaques). Radioactivity levels were low in the plasma and very low or unquantifiable in the draining lymph nodes of the macaques. Microautoradiography identified drug-related material (DRM) on the surfaces of the vaginal and cervical tissues of the rabbits and macaques. Although DRM was primarily associated with the outermost layer of shedding cells in rabbits, two animals showed some evidence of small quantities in the mucosal epithelium of the cervix. In macaques, DRM was seen within the keratinized layer of the vaginal epithelium and and was found to extend into the superficial cellular layers, and in at least one animal it appeared to be present in the deepest (germinal) layer of the epithelium and in submucosal tissues. The persistence of biologically significant concentrations of dapivirine in vaginal and cervical tissues for >24 h supports the development of dapivirine as a microbicide for once daily application.

Nonlinear low frequency electrostatic structures in a magnetized two-component auroral plasma

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rufai, O. R., E-mail: rajirufai@gmail.com; Scientific Computing, Memorial University of Newfoundland, St John's, Newfoundland and Labrador A1C 5S7; Bharuthram, R., E-mail: rbharuthram@uwc.ac.za

2016-03-15

Finite amplitude nonlinear ion-acoustic solitons, double layers, and supersolitons in a magnetized two-component plasma composed of adiabatic warm ions fluid and energetic nonthermal electrons are studied by employing the Sagdeev pseudopotential technique and assuming the charge neutrality condition at equilibrium. The model generates supersoliton structures at supersonic Mach numbers regime in addition to solitons and double layers, whereas in the unmagnetized two-component plasma case only, soliton and double layer solutions can be obtained. Further investigation revealed that wave obliqueness plays a critical role for the evolution of supersoliton structures in magnetized two-component plasmas. In addition, the effect of ion temperaturemore » and nonthermal energetic electron tends to decrease the speed of oscillation of the nonlinear electrostatic structures. The present theoretical results are compared with Viking satellite observations.« less

Emission of Biophotons and Adjustable Sounds by the Fascial System: Review and Reflections for Manual Therapy.

PubMed

Bordoni, Bruno; Marelli, Fabiola; Morabito, Bruno; Sacconi, Beatrice

2018-01-01

Every body structure is wrapped in connective tissue or fascia, creating a structural continuity that gives form and function to every tissue and organ. The fascial tissue is uniformly distributed throughout the body, enveloping, interacting with and permeating blood vessels, nerves, viscera, meninges, bones and muscles, creating various layers at different depths and forming a tridimensional metabolic and mechanical matrix. This article reviews the literature on the emission of biophotons and adjustable sounds by the fascial system, because these biological changes could be a means of local and systemic cellular communication and become another assessment tool for manual (therapy) practitioners. This is the first article that discusses these topics in a single text, attempting to bring such information into an area of application that is beneficial to osteopaths, chiropractors, and manual therapists.

Emission of Biophotons and Adjustable Sounds by the Fascial System: Review and Reflections for Manual Therapy

PubMed Central

Marelli, Fabiola; Sacconi, Beatrice

2018-01-01

Every body structure is wrapped in connective tissue or fascia, creating a structural continuity that gives form and function to every tissue and organ. The fascial tissue is uniformly distributed throughout the body, enveloping, interacting with and permeating blood vessels, nerves, viscera, meninges, bones and muscles, creating various layers at different depths and forming a tridimensional metabolic and mechanical matrix. This article reviews the literature on the emission of biophotons and adjustable sounds by the fascial system, because these biological changes could be a means of local and systemic cellular communication and become another assessment tool for manual (therapy) practitioners. This is the first article that discusses these topics in a single text, attempting to bring such information into an area of application that is beneficial to osteopaths, chiropractors, and manual therapists. PMID:29405763

Ordered organic-organic multilayer growth

DOEpatents

Forrest, Stephen R.; Lunt, Richard R.

2016-04-05

An ordered multilayer crystalline organic thin film structure is formed by depositing at least two layers of thin film crystalline organic materials successively wherein the at least two thin film layers are selected to have their surface energies within .+-.50% of each other, and preferably within .+-.15% of each other, whereby every thin film layer within the multilayer crystalline organic thin film structure exhibit a quasi-epitaxial relationship with the adjacent crystalline organic thin film.

Ordered organic-organic multilayer growth

DOEpatents

Forrest, Stephen R; Lunt, Richard R

2015-01-13

An ordered multilayer crystalline organic thin film structure is formed by depositing at least two layers of thin film crystalline organic materials successively wherein the at least two thin film layers are selected to have their surface energies within .+-.50% of each other, and preferably within .+-.15% of each other, whereby every thin film layer within the multilayer crystalline organic thin film structure exhibit a quasi-epitaxial relationship with the adjacent crystalline organic thin film.

Corrosion processes of physical vapor deposition-coated metallic implants.

PubMed

Antunes, Renato Altobelli; de Oliveira, Mara Cristina Lopes

2009-01-01

Protecting metallic implants from the harsh environment of physiological fluids is essential to guaranteeing successful long-term use in a patient's body. Chemical degradation may lead to the failure of an implant device in two different ways. First, metal ions may cause inflammatory reactions in the tissues surrounding the implant and, in extreme cases, these reactions may inflict acute pain on the patient and lead to loosening of the device. Therefore, increasing wear strength is beneficial to the performance of the metallic implant. Second, localized corrosion processes contribute to the nucleation of fatigue cracks, and corrosion fatigue is the main reason for the mechanical failure of metallic implants. Common biomedical alloys such as stainless steel, cobalt-chrome alloys, and titanium alloys are prone to at least one of these problems. Vapor-deposited hard coatings act directly to improve corrosion, wear, and fatigue resistances of metallic materials. The effectiveness of the corrosion protection is strongly related to the structure of the physical vapor deposition layer. The aim of this paper is to present a comprehensive review of the correlation between the structure of physical vapor deposition layers and the corrosion properties of metallic implants.

Structure of a magnetic flux annihilation layer formed by the collision of supersonic, magnetized plasma flows

DOE PAGES

Suttle, L. G.; Hare, J. D.; Lebedev, S. V.; ...

2016-05-31

We present experiments characterizing the detailed structure of a current layer, generated by the collision of two counter-streaming, supersonic and magnetized aluminum plasma flows. The anti parallel magnetic fields advected by the flows are found to be mutually annihilated inside the layer, giving rise to a bifurcated current structure—two narrow current sheets running along the outside surfaces of the layer. Measurements with Thomson scattering show a fast outflow of plasma along the layer and a high ion temperature (T i~¯ZT e, with average ionization ¯Z=7). Lastly, analysis of the spatially resolved plasma parameters indicates that the advection and subsequent annihilationmore » of the in-flowing magnetic flux determines the structure of the layer, while the ion heating could be due to the development of kinetic, current-driven instabilities.« less

Structure of a magnetic flux annihilation layer formed by the collision of supersonic, magnetized plasma flows

DOE Office of Scientific and Technical Information (OSTI.GOV)

Suttle, L. G.; Hare, J. D.; Lebedev, S. V.

We present experiments characterizing the detailed structure of a current layer, generated by the collision of two counter-streaming, supersonic and magnetized aluminum plasma flows. The anti parallel magnetic fields advected by the flows are found to be mutually annihilated inside the layer, giving rise to a bifurcated current structure—two narrow current sheets running along the outside surfaces of the layer. Measurements with Thomson scattering show a fast outflow of plasma along the layer and a high ion temperature (T i~¯ZT e, with average ionization ¯Z=7). Lastly, analysis of the spatially resolved plasma parameters indicates that the advection and subsequent annihilationmore » of the in-flowing magnetic flux determines the structure of the layer, while the ion heating could be due to the development of kinetic, current-driven instabilities.« less

Preparation and Characterization of a Novel Decellularized Fibrocartilage "Book" Scaffold for Use in Tissue Engineering.

PubMed

Guo, Liyun; Qu, Jin; Zheng, Cheng; Cao, Yong; Zhang, Tao; Lu, Hongbin; Hu, Jianzhong

2015-01-01

At the tendon-to-bone insertion, there is a unique transitional structure: tendon, non-calcified fibrocartilage, calcified fibrocartilage, and bone. The reconstruction of this special graded structure after defects or damage is an important but challenging task in orthopedics. In particular, reconstruction of the fibrocartilage zone has yet to be successfully achieved. In this study, the development of a novel book-shape scaffold derived from the extracellular matrix of fibrocartilage was reported. Specifically, fibrocartilage from the pubic symphysis was obtained from rabbits and sliced into the shape of a book (dimensions: 10 mm × 3 mm × 1 mm) with 10 layers, each layer (akin to a page of a book) with a thickness of 100-μm. These fibrocartilage "book" scaffolds were decellularized using sequentially 3 freeze-thaw cycles, 0.1% Triton X-100 with 1.5 M KCl, 0.25% trypsin, and a nuclease. Histology and DNA quantification analysis confirmed substantial removal of cells from the fibrocartilage scaffolds. Furthermore, the quantities of DNA, collagen, and glycosaminoglycan in the fibrocartilage were markedly reduced following decellularization. Scanning electron microscopy confirmed that the intrinsic ultrastructure of the fibrocartilage tissue was well preserved. Therefore, the results of this study suggest that the novel "book" fibrocartilage scaffold could have potential applications in tissue engineering.

Preparation and Characterization of a Novel Decellularized Fibrocartilage “Book” Scaffold for Use in Tissue Engineering

PubMed Central

Guo, Liyun; Qu, Jin; Zheng, Cheng; Cao, Yong; Zhang, Tao; Lu, Hongbin; Hu, Jianzhong

2015-01-01

At the tendon-to-bone insertion, there is a unique transitional structure: tendon, non-calcified fibrocartilage, calcified fibrocartilage, and bone. The reconstruction of this special graded structure after defects or damage is an important but challenging task in orthopedics. In particular, reconstruction of the fibrocartilage zone has yet to be successfully achieved. In this study, the development of a novel book-shape scaffold derived from the extracellular matrix of fibrocartilage was reported. Specifically, fibrocartilage from the pubic symphysis was obtained from rabbits and sliced into the shape of a book (dimensions: 10 mm × 3 mm × 1 mm) with 10 layers, each layer (akin to a page of a book) with a thickness of 100-μm. These fibrocartilage “book” scaffolds were decellularized using sequentially 3 freeze-thaw cycles, 0.1% Triton X-100 with 1.5 M KCl, 0.25% trypsin, and a nuclease. Histology and DNA quantification analysis confirmed substantial removal of cells from the fibrocartilage scaffolds. Furthermore, the quantities of DNA, collagen, and glycosaminoglycan in the fibrocartilage were markedly reduced following decellularization. Scanning electron microscopy confirmed that the intrinsic ultrastructure of the fibrocartilage tissue was well preserved. Therefore, the results of this study suggest that the novel “book” fibrocartilage scaffold could have potential applications in tissue engineering. PMID:26636672

Morphology and ultrastructure of the esophagus during the ontogeny of the spider crab Maja brachydactyla (Decapoda, Brachyura, Majidae).

PubMed

Castejón, Diego; Rotllant, Guiomar; Ribes, Enric; Durfort, Mercè; Guerao, Guillermo

2018-06-01

The esophagus of the eucrustaceans is known as a short tube that connects the mouth with the stomach but has generally received little attention by the carcinologists, especially during the larval stages. By this reason, the present study is focused on the morphology and ultrastructure of the esophagus in the brachyuran Maja brachydactyla during the larval development and adult stage. The esophagus shows internally four longitudinal folds. The simple columnar epithelium is covered by a thick cuticle. The epithelial cells of the adults are intensively interdigitated and show abundant apical mitochondria and bundles of filamentous structures. The cuticle surface has microspines and mutually exclusive pores. Three muscle layers surrounded by the connective tissue are reported: circular muscles forming a broad continuous band, longitudinal muscle bundles adjacent to the circular muscles, and dilator muscles crossing the connective tissue vertically toward the epithelium. The connective tissue has rosette glands. The esophagus of the larvae have epithelial cells with big vesicles but poorly developed interdigitations and filamentous structures, the cuticle is formed by a procuticle without differentiated exocuticle and endocuticle, the connective layer is thin and the rosette glands are absent. The observed features can be explained by his role in the swallowing of the food. © 2018 Wiley Periodicals, Inc.

Comparison of structural, architectural and mechanical aspects of cellular and acellular bone in two teleost fish.

PubMed

Cohen, Liat; Dean, Mason; Shipov, Anna; Atkins, Ayelet; Monsonego-Ornan, Efrat; Shahar, Ron

2012-06-01

The histological diversity of the skeletal tissues of fishes is impressive compared with that of other vertebrate groups, yet our understanding of the functional consequences of this diversity is limited. In particular, although it has been known since the mid-1800s that a large number of fish species possess acellular bones, the mechanical advantages and consequences of this structural characteristic - and therefore the nature of the evolution of this feature - remain unclear. Although several studies have examined the material properties of fish bone, these have used a variety of techniques and there have been no direct contrasts of acellular and cellular bone. We report on a comparison of the structural and mechanical properties of the ribs and opercula between two freshwater fish - the common carp Cyprinus carpio (a fish with cellular bone) and the tilapia Oreochromis aureus (a fish with acellular bone). We used light microscopy to show that the bones in both fish species exhibit poor blood supply and possess discrete tissue zones, with visible layering suggesting differences in the underlying collagen architecture. We performed identical micromechanical testing protocols on samples of the two bone types to determine the mechanical properties of the bone material of opercula and ribs. Our data support the consensus of literature values, indicating that Young's moduli of cellular and acellular bones are in the same range, and lower than Young's moduli of the bones of mammals and birds. Despite these similarities in mechanical properties between the bone tissues of the fish species tested here, cellular bone had significantly lower mineral content than acellular bone; furthermore, the percentage ash content and bone mineral density values (derived from micro-CT scans) show that the bone of these fishes is less mineralized than amniote bone. Although we cannot generalize from our data to the numerous remaining teleost species, the results presented here suggest that while cellular and acellular fish bone may perform similarly from a mechanical standpoint, there are previously unappreciated differences in the structure and composition of these bone types.

Forced free-shear layer measurements

NASA Technical Reports Server (NTRS)

Leboeuf, Richard L.

1994-01-01

Detailed three-dimensional three-component phase averaged measurements of the spanwise and streamwise vorticity formation and evolution in acoustically forced plane free-shear flows have been obtained. For the first time, phase-averaged measurements of all three velocity components have been obtained in both a mixing layer and a wake on three-dimensional grids, yielding the spanwise and streamwise vorticity distributions without invoking Taylor's hypothesis. Initially, two-frequency forcing was used to phase-lock the roll-up and first pairing of the spanwise vortical structures in a plane mixing layer. The objective of this study was to measure the near-field vortical structure morphology in a mixing layer with 'natural' laminar initial boundary layers. For the second experiment the second and third subharmonics of the fundamental roll-up frequency were added to the previous two-frequency forcing in order to phase-lock the roll-up and first three pairings of the spanwise rollers in the mixing layer. The objective of this study was to determine the details of spanwise scale changes observed in previous time-averaged measurements and flow visualization of unforced mixing layers. For the final experiment, single-frequency forcing was used to phase-lock the Karman vortex street in a plane wake developing from nominally two-dimensional laminar initial boundary layers. The objective of this study was to compare measurements of the three-dimensional structure in a wake developing from 'natural' initial boundary layers to existing models of wake vortical structure.

Dependence of air masses type on PBL vertical structure retrieved at the Mace Head station during EUCAARI campaign.

NASA Astrophysics Data System (ADS)

Milroy, Conor; Martucci, Giovanni; O'Dowd, Colin

2010-05-01

During the EUCAARI Intensive Observing Period held at the Mace Head GAW station from mid-May to mid-June, 2008, the PBL depth has been continuously measured by two ceilometers (Vaisala CL31 and Jenoptik CHM15K) and a microwave radiometer (RPG-HATPRO). The Lidar-Ceilometer, through the gradients in aerosol backscatter profiles, and the microwave profiler, through gradients in the specific humidity profiles, were used to remotely-sense the boundary layer structure. An automatic, newly developed Temporal Height-Tracking (THT) algorithm (Martucci et al., 2010) have been applied to both type of instruments data to retrieve the 2-layered structure of the local marine boundary layer. The two layers are defined as a lower, well mixed layer, i.e. the surface mixed layer, and the layer occupying the region below the free Troposphere inversion, i.e. the decoupled residual or convective layer. A categorization of the incoming air masses has been performed based on their origins and been used to asses the correlation with the PBL depths. The study confirmed the dependence of PBL vertical structure on different air masses and different type of advected aerosol.

Light dosimetry for focused and defocused beam irradiation in multi-layered tissue models

NASA Astrophysics Data System (ADS)

Petrova, Kremena S.; Stoykova, Elena V.

2006-09-01

Treatment of acupuncture points, trigger points, joint inflammations in low level laser therapy as well as various applications of lasers for treatment of soft tissues in dental medicine, require irradiation by a narrow converging laser beam. The aim of this study is to compare light delivery produced by focused or defocused narrow beam irradiation in a multi-layered skin tissue model at increasing depth of the target. The task is solved by 3-D Monte-Carlo simulation for matched and mismatched refractive indices at the tissue/ambient medium interface. The modeled light beams have a circular cross-section at the tissue entrance with uniform or Gaussian intensity distribution. Three are the tissue models used in simulation : i) a bloodless skin layer; ii) a bloodless skin layer with embedded scattering object; iii) a skin layer with small blood vessels of varying size, which are modeled as infinite cylinders parallel to the tissue surface located at different depths. Optical properties (absorption coefficient, scattering coefficient, anisotropy factor, g, and index of refraction) of different tissue constituents are chosen from the literature.

Morphometric analysis of high-intensity focused ultrasound-induced lipolysis on cadaveric abdominal and thigh skin.

PubMed

Lee, Sugun; Kim, Hee-Jin; Park, Hyun Jun; Kim, Hyoung Moon; Lee, So Hyun; Cho, Sung Bin

2017-07-01

Non-focused ultrasound and high-intensity focused ultrasound (HIFU) devices induce lipolysis by generating acoustic cavitation and coagulation necrosis in targeted tissues. We aimed to investigate the morphometric characteristics of immediate tissue reactions induced by 2 MHz, 13-mm focused HIFU via two-dimensional ultrasound images and histologic evaluation of cadaveric skin from the abdomen and thigh. Acoustic fields of a 2 MHz, 38-mm HIFU transducer were characterized by reconstruction of the fields using acoustic intensity measurement. Additionally, abdominal and thigh tissues from a fresh cadaver were treated with a HIFU device for a single, two, and three pulses at the pulse energy of 130 J/cm 2 and a penetration depth of 13 mm. Acoustic intensity measurement revealed characteristic focal zones of significant thermal injury at the depth of 38 mm. In both the abdomen and thigh tissue, round to oval ablative thermal injury zones (TIZs) were visualized in subcutaneous fat layers upon treatment with a single pulse of HIFU treatment. Two to three HIFU pulses generated larger and more remarkable ablative zones throughout subcutaneous fat layers. Finally, experimental treatment in a tumescent infiltration-like setting induced larger HIFU-induced TIZs of an oval or columnar shape, compared to non-tumescent settings. Although neither acoustic intensity measurement nor cadaveric tissue exactly reflects in vivo HIFU-induced reactions in human tissue, we believe that our data will help guide further in vivo studies in investigating the therapeutic efficacy and safety of HIFU-induced lipolysis.

Design of nano- and microfiber combined scaffolds by electrospinning of collagen onto starch-based fiber meshes: a man-made equivalent of natural extracellular matrix.

PubMed

Tuzlakoglu, Kadriye; Santos, Marina I; Neves, Nuno; Reis, Rui L

2011-02-01

Mimicking the structural organization and biologic function of natural extracellular matrix has been one of the main goals of tissue engineering. Nevertheless, the majority of scaffolding materials for bone regeneration highlights biochemical functionality in detriment of mechanical properties. In this work we present a rather innovative construct that combines in the same structure electrospun type I collagen nanofibers with starch-based microfibers. These combined structures were obtained by a two-step methodology and structurally consist in a type I collagen nano-network incorporated on a macro starch-based support. The morphology of the developed structures was assessed by several microscopy techniques and the collagenous nature of the nano-network was confirmed by immunohistochemistry. In addition, and especially regarding the requirements of large bone defects, we also successfully introduced the concept of layer by layer, as a way to produce thicker structures. In an attempt to recreate bone microenvironment, the design and biochemical composition of the combined structures also envisioned bone-forming cells and endothelial cells (ECs). The inclusion of a type I collagen nano-network induced a stretched morphology and improved the metabolic activity of osteoblasts. Regarding ECs, the presence of type I collagen on the combined structures provided adhesive support and obviated the need of precoating with fibronectin. It was also importantly observed that ECs on the nano-network organized into circular structures, a three-dimensional arrangement distinct from that observed for osteoblasts and resembling the microcappillary-like organizations formed during angiogenesis. By providing simultaneously physical and chemical cues for cells, the herein-proposed combined structures hold a great potential in bone regeneration as a man-made equivalent of extracellular matrix.

The zebrafish tailbud contains two independent populations of midline progenitor cells that maintain long-term germ layer plasticity and differentiate in response to local signaling cues

PubMed Central

Row, Richard H.; Tsotras, Steve R.; Goto, Hana; Martin, Benjamin L.

2016-01-01

Vertebrate body axis formation depends on a population of bipotential neuromesodermal cells along the posterior wall of the tailbud that make a germ layer decision after gastrulation to form spinal cord and mesoderm. Despite exhibiting germ layer plasticity, these cells never give rise to midline tissues of the notochord, floor plate and dorsal endoderm, raising the question of whether midline tissues also arise from basal posterior progenitors after gastrulation. We show in zebrafish that local posterior signals specify germ layer fate in two basal tailbud midline progenitor populations. Wnt signaling induces notochord within a population of notochord/floor plate bipotential cells through negative transcriptional regulation of sox2. Notch signaling, required for hypochord induction during gastrulation, continues to act in the tailbud to specify hypochord from a notochord/hypochord bipotential cell population. Our results lend strong support to a continuous allocation model of midline tissue formation in zebrafish, and provide an embryological basis for zebrafish and mouse bifurcated notochord phenotypes as well as the rare human congenital split notochord syndrome. We demonstrate developmental equivalency between the tailbud progenitor cell populations. Midline progenitors can be transfated from notochord to somite fate after gastrulation by ectopic expression of msgn1, a master regulator of paraxial mesoderm fate, or if transplanted into the bipotential progenitors that normally give rise to somites. Our results indicate that the entire non-epidermal posterior body is derived from discrete, basal tailbud cell populations. These cells remain receptive to extracellular cues after gastrulation and continue to make basic germ layer decisions. PMID:26674311

Anchoring TGF-β1 on biomaterial surface via affinitive interactions: Effects on spatial structures and bioactivity.

PubMed

Xiao, Meng; Xiao, Jiangwei; Wu, Gang; Ke, Yu; Fang, Liming; Deng, Chunlin; Liao, Hua

2018-06-01

Protein adsorption on biomaterial surfaces is clinically applied to increase therapeutic effects; however, this adsorption is possibly accompanied by conformational changes and results in loss of protein bioactivity or adverse reactions. In this research, a transforming growth factor β1 (TGF-β1) affinitive peptide HSNGLPL was grafted onto biopolymer surface to stabilize TGF-β1 spatial conformation after adhesion. The peptide with azide end group was combined with the propynyl pendant group on polyurethane via copper-catalyzed azide-alkyne cycloaddition (CuAAC) click reaction. The final polymer was characterized by Fourier transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy, which indicated that the affinitive peptide was introduced to the polymer. Quartz crystal microbalance with dissipation (QCM-D) was performed to monitor TGF-β1 adsorption and desorption on the surfaces coated with polyurethane with and without peptide combination. Results showed that TGF-β1 adhered on polyurethane surface and formed a compact and rigid layer. This layer showed spatial structural change but presented a loose and diffuse layer on the peptide-grafted polyurethane surface, indicating stable spatial conformation after adherence. Similar regulations were observed on the two surfaces where BSA layer was coated in advance. In vivo animal experiments revealed that immune reactions and tissue regenerations occurred earlier on peptide-modified polyurethane than on polyurethane, which did not undergo peptide grafting. This finding confirmed that affinitive interactions may preserve TGF-β1 bioactivity on polymer surface after adsorption. Copyright © 2018 Elsevier B.V. All rights reserved.

Agent-based modeling of porous scaffold degradation and vascularization: Optimal scaffold design based on architecture and degradation dynamics.

PubMed

Mehdizadeh, Hamidreza; Bayrak, Elif S; Lu, Chenlin; Somo, Sami I; Akar, Banu; Brey, Eric M; Cinar, Ali

2015-11-01

A multi-layer agent-based model (ABM) of biomaterial scaffold vascularization is extended to consider the effects of scaffold degradation kinetics on blood vessel formation. A degradation model describing the bulk disintegration of porous hydrogels is incorporated into the ABM. The combined degradation-angiogenesis model is used to investigate growing blood vessel networks in the presence of a degradable scaffold structure. Simulation results indicate that higher porosity, larger mean pore size, and rapid degradation allow faster vascularization when not considering the structural support of the scaffold. However, premature loss of structural support results in failure for the material. A strategy using multi-layer scaffold with different degradation rates in each layer was investigated as a way to address this issue. Vascularization was improved with the multi-layered scaffold model compared to the single-layer model. The ABM developed provides insight into the characteristics that influence the selection of optimal geometric parameters and degradation behavior of scaffolds, and enables easy refinement of the model as new knowledge about the underlying biological phenomena becomes available. This paper proposes a multi-layer agent-based model (ABM) of biomaterial scaffold vascularization integrated with a structural-kinetic model describing bulk degradation of porous hydrogels to consider the effects of scaffold degradation kinetics on blood vessel formation. This enables the assessment of scaffold characteristics and in particular the disintegration characteristics of the scaffold on angiogenesis. Simulation results indicate that higher porosity, larger mean pore size, and rapid degradation allow faster vascularization when not considering the structural support of the scaffold. However, premature loss of structural support by scaffold disintegration results in failure of the material and disruption of angiogenesis. A strategy using multi-layer scaffold with different degradation rates in each layer was investigated as away to address this issue. Vascularization was improved with the multi-layered scaffold model compared to the single-layer model. The ABM developed provides insight into the characteristics that influence the selection of optimal geometric and degradation characteristics of tissue engineering scaffolds. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Multi-layer carbon-based coatings for field emission

DOEpatents

Sullivan, J.P.; Friedmann, T.A.

1998-10-13

A multi-layer resistive carbon film field emitter device for cold cathode field emission applications is disclosed. The multi-layered film of the present invention consists of at least two layers of a conductive carbon material, preferably amorphous-tetrahedrally coordinated carbon, where the resistivities of adjacent layers differ. For electron emission from the surface, the preferred structure can be a top layer having a lower resistivity than the bottom layer. For edge emitting structures, the preferred structure of the film can be a plurality of carbon layers, where adjacent layers have different resistivities. Through selection of deposition conditions, including the energy of the depositing carbon species, the presence or absence of certain elements such as H, N, inert gases or boron, carbon layers having desired resistivities can be produced. 8 figs.

Cryogenic 3D printing for producing hierarchical porous and rhBMP-2-loaded Ca-P/PLLA nanocomposite scaffolds for bone tissue engineering.

PubMed

Wang, Chong; Zhao, Qilong; Wang, Min

2017-06-07

The performance of bone tissue engineering scaffolds can be assessed through cell responses to scaffolds, including cell attachment, infiltration, morphogenesis, proliferation, differentiation, etc, which are determined or heavily influenced by the composition, structure, mechanical properties, and biological properties (e.g. osteoconductivity and osteoinductivity) of scaffolds. Although some promising 3D printing techniques such as fused deposition modeling and selective laser sintering could be employed to produce biodegradable bone tissue engineering scaffolds with customized shapes and tailored interconnected pores, effective methods for fabricating scaffolds with well-designed hierarchical porous structure (both interconnected macropores and surface micropores) and tunable osteoconductivity/osteoinductivity still need to be developed. In this investigation, a novel cryogenic 3D printing technique was investigated and developed for producing hierarchical porous and recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded calcium phosphate (Ca-P) nanoparticle/poly(L-lactic acid) nanocomposite scaffolds, in which the Ca-P nanoparticle-incorporated scaffold layer and rhBMP-2-encapsulated scaffold layer were deposited alternatingly using different types of emulsions as printing inks. The mechanical properties of the as-printed scaffolds were comparable to those of human cancellous bone. Sustained releases of Ca 2+ ions and rhBMP-2 were achieved and the biological activity of rhBMP-2 was well-preserved. Scaffolds with a desirable hierarchical porous structure and dual delivery of Ca 2+ ions and rhBMP-2 exhibited superior performance in directing the behaviors of human bone marrow-derived mesenchymal stem cells and caused improved cell viability, attachment, proliferation, and osteogenic differentiation, which has suggested their great potential for bone tissue engineering.

Ultrastructural and immunocytochemical characterization of ameloblast-enamel adhesion at maturation stage in amelogenesis in Macaca fuscata tooth germ.

PubMed

Sawada, Takashi

2015-12-01

Maturation-stage ameloblasts are firmly bound to the tooth enamel by a basal lamina-like structure. The mechanism underlying this adhesion, however, remains to be fully clarified. The goal of this study was to investigate the mechanism underlying adhesion between the basal lamina-like structure and the enamel in monkey tooth germ. High-resolution immunogold labeling was performed to localize amelotin and laminin 332 at the interface between ameloblasts and tooth enamel. Minute, electron-dense strands were observed on the enamel side of the lamina densa, extending into the degrading enamel matrix to produce a well-developed fibrous layer (lamina fibroreticularis). In un-demineralized tissue sections, mineral crystals smaller than those in the bulk of the enamel were observed adhering to these strands where they protruded into the surface enamel. Immunogold particles reactive for amelotin were preferentially localized on these strands in the fibrous layer. On the other hand, those for laminin 332 were localized solely in the lamina densa; none were observed in the fibrous layer. These results suggest that the fibrous layer of the basal lamina-like structure is partly composed of amelotin molecules, and that these molecules facilitate ameloblast-enamel adhesion by promoting mineralization of the fibrous layer during the maturation stage of amelogenesis.

Electrospraying of microfluidic encapsulated cells for the fabrication of cell-laden electrospun hybrid tissue constructs.

PubMed

Weidenbacher, L; Abrishamkar, A; Rottmar, M; Guex, A G; Maniura-Weber, K; deMello, A J; Ferguson, S J; Rossi, R M; Fortunato, G

2017-12-01

The fabrication of functional 3D tissues is a major goal in tissue engineering. While electrospinning is a promising technique to manufacture a structure mimicking the extracellular matrix, cell infiltration into electrospun scaffolds remains challenging. The robust and in situ delivery of cells into such biomimetic scaffolds would potentially enable the design of tissue engineered constructs with spatial control over cellular distribution but often solvents employed in the spinning process are problematic due to their high cytotoxicity. Herein, microfluidic cell encapsulation is used to establish a temporary protection vehicle for the in situ delivery of cells for the development of a fibrous, cell-laden hybrid biograft. Therefore a layer-by-layer process is used by alternating fiber electrospinning and cell spraying procedures. Both encapsulation and subsequent electrospraying of capsules has no negative effect on the viability and myogenic differentiation of murine myoblast cells. Propidium iodide positive stained cells were analyzed to quantify the amount of dead cells and the presence of myosin heavy chain positive cells after the processes was shown. Furthermore, encapsulation successfully protects cells from cytotoxic solvents (such as dimethylformamide) during in situ delivery of the cells into electrospun poly(vinylidene fluoride-co-hexafluoropropylene) scaffolds. The resulting cell-populated biografts demonstrate the clear potential of this approach in the creation of viable tissue engineering constructs. Infiltration of cells and their controlled spatial distribution within fibrous electrospun membranes is a challenging task but allows for the development of functional highly organized 3D hybrid tissues. Combining polymer electrospinning and cell electrospraying in a layer-by-layer approach is expected to overcome current limitations of reduced cell infiltration after traditional static seeding. However, organic solvents, used during the electrospinning process, impede often major issues due to their high cytotoxicity. Utilizing microfluidic encapsulation as a mean to embed cells within a protective polymer casing enables the controlled deposition of viable cells without interfering with the cellular phenotype. The presented techniques allow for novel cell manipulation approaches being significant for enhanced 3D tissue engineering based on its versatility in terms of material and cell selection. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Electrical characterization of anodic alumina substrate with via-in-pad structure

NASA Astrophysics Data System (ADS)

Kim, Moonjung

2013-10-01

An anodic alumina substrate has been developed as a package substrate for dynamic random access memory devices. Unlike the conventional package substrates commonly made by laminating an epoxy-based core and cladding with copper, this substrate is fabricated using aluminum anodization technology. The anodization process produces a thick aluminum oxide layer on the aluminum substrate to be used as a dielectric layer. Placing copper patterns on the anodic aluminum oxide layer forms a new substrate structure that consists of a layered structure of aluminum, anodic aluminum oxide, and copper. Using selective anodization in the fabrication process, a via structure connecting the top copper layer and bottom aluminum layer is demonstrated. Additionally, by putting vias directly in the bond and ball pads in the substrate design, the via-in-pad structure is applied in this work. These two-layer metal structures and via-in-pad arrangements make routing easier and thus provide more design flexibility. Additionally, this new package substrate has improved the power distribution network impedance given the characteristics of these structures.

Mechanical characterization of stomach tissue under uniaxial tensile action.

PubMed

Jia, Z G; Li, W; Zhou, Z R

2015-02-26

In this article, the tensile properties of gastric wall were investigated by using biomechanical test and theoretical analysis. The samples of porcine stomach strips from smaller and greater curvature of the stomach were cut in longitudinal and circumferential direction, respectively. The loading-unloading, stress relaxation, strain creep, tensile fracture tests were performed at mucosa-submucosa, serosa-muscle and intact layer, respectively. Results showed that the biomechanical properties of the porcine stomach depended on the layers, orientations and locations of the gastric wall and presented typical viscoelastic, nonlinear and anisotropic mechanical properties. During loading-unloading test, the stress of serosa-muscle layer in the longitudinal direction was 15-20% more than that in the circumferential direction at 12% stretch ratio, while it could reach about 40% for the intact layer and 50% for the mucosa-submucosa layer. The results of stress relaxation and strain creep showed that the variation degree was obviously faster in the circumferential direction than that in the longitudinal direction, and the ultimate residual values were also different for the different layers, orientations and locations. In the process of fracture test, the serosa-muscle layer fractured firstly followed by the mucosa-submucosa layer when the intact layer was tested, the longitudinal strips firstly began to fracture and the required stress value was about twice as much as that in the circumferential strips. The anisotropy and heterogeneity of mechanical characterization of the porcine stomach were related to its complicated geometry, structure and functions. The results would help us to understand the biomechanics of soft organ tissue. Copyright © 2015 Elsevier Ltd. All rights reserved.

Measuring temperature induced phase change kinetics in subcutaneous fatty tissues using near infrared spectroscopy, magnetic resonance imaging and optical coherence tomography (Conference Presentation)

NASA Astrophysics Data System (ADS)

Sajjadi, Amir Y.; Carp, Stefan A.; Manstein, Dieter

2017-02-01

Monitoring phase transition in adipose tissue and formation of lipid crystals is important in Cryo-procedures such as cryosurgery or Selective Cryolipolysis (SC). In this work, we exploited a Near-Infrared Spectroscopy (NIRS) method to monitor the onset of fat freezing/melting. Concurrent measurements using frequency domain NIRS and MR Spectroscopy during cooling/heating were performed on an in vitro porcine skin sample with a thick subcutaneous fat layer in a human MR scanner. The NIRS probe was placed on the skin measuring the average optical scattering of the fatty layer. Two fiber optic temperature probes were inserted in the area of the MRS and NIRS measurements. To further investigate the microscopic features of the phase-transition, an identical cooling/heating procedure was replicated on the same fat tissue while being imaged by Optical Coherence Tomography. The temperature relationships of optical scattering, MRS peak characteristics and OCT reflection intensity were analyzed to find signatures related to the onset of phase transition. The optical scattering in the fatty tissues decreases during the heating and increases by cooling. However, there is an inflexion in the rate of change of the scattering while the phase transition happens in the fatty layer. The methylene fat peaks on the MR Spectrum are also shown to be broadened during the cooling. OCT intensity displays a sharp increase at the transition temperature. The results from multiple samples show two transition points around 5-10 ˚C (cooling) and 15-20 ˚C (heating) through all three methods, demonstrating that adipose tissue phase change can be monitored non-invasively.

Asymptotic structure and similarity solutions for three-dimensional turbulent boundary layers

NASA Technical Reports Server (NTRS)

Degani, A. T.; Walker, J. D. A.

1989-01-01

The asymptotic structure of the three-dimensional turbulent boundary layer is investigated in the limit of large Reynolds numbers. A self-consistent, but relatively complex, two-layer structure exists and the simplest situation, corresponding to a plane of symmetry, is considered in this paper as a first step. The adjustment of the streamwise velocity to relative rest, through an outer defect layer and then an inner wall layer, is similar to that in two-dimensional flow. The adjustment of the cross-streamwise velocity is more complicated and it is shown that two terms in the expansion are required to obtain useful results, and in particular to obtain the velocity skew angle at the wall near the symmetry plane. The conditions under which self-similarity is achieved near a plane of symmetry are investigated. A set of ordinary differential equations is developed which describe the streamwise and cross-streamwise velocities near a plane of symmetry in a self-similar flow through two orders of magnitude. Calculated numerical solutions of these equations yield trends which are consistent with experimental observations.

Production and emission of volatile compounds by petal cells.

PubMed

Baudino, Sylvie; Caissard, Jean-Claude; Bergougnoux, Véronique; Jullien, Frédéric; Magnard, Jean-Louis; Scalliet, Gabriel; Cock, J Mark; Hugueney, Philippe

2007-11-01

We localized the tissues and cells that contribute to scent biosynthesis in scented and non-scented Rosa x hybrida cultivars as part of a detailed cytological analysis of the rose petal. Adaxial petal epidermal cells have a typical conical, papillate shape whereas abaxial petal epidermal cells are flat. Using two different techniques, solid/liquid phase extraction and headspace collection of volatiles, we showed that, in roses, both epidermal layers are capable of producing and emitting scent volatiles, despite the different morphologies of the cells of these two tissues. Moreover, OOMT, an enzyme involved in scent molecule biosynthesis, was localized in both epidermal layers. These results are discussed in view of results found in others species such as Antirrhinum majus, where it has been shown that the adaxial epidermis is the preferential site of scent production and emission.

Production and Emission of Volatile Compounds by Petal Cells

PubMed Central

Caissard, Jean-Claude; Bergougnoux, Véronique; Jullien, Frédéric; Magnard, Jean-Louis; Scalliet, Gabriel; Cock, J Mark; Hugueney, Philippe

2007-01-01

We localized the tissues and cells that contribute to scent biosynthesis in scented and non-scented Rosa × hybrida cultivars as part of a detailed cytological analysis of the rose petal. Adaxial petal epidermal cells have a typical conical, papillate shape whereas abaxial petal epidermal cells are flat. Using two different techniques, solid/liquid phase extraction and headspace collection of volatiles, we showed that, in roses, both epidermal layers are capable of producing and emitting scent volatiles, despite the different morphologies of the cells of these two tissues. Moreover, OOMT, an enzyme involved in scent molecule biosynthesis, was localized in both epidermal layers. These results are discussed in view of results found in others species such as Antirrhinum majus, where it has been shown that the adaxial epidermis is the preferential site of scent production and emission. PMID:19704548

Geometry and Mechanics of Chiral Pod Opening

NASA Astrophysics Data System (ADS)

Sharon, Eran; Armon, Shahaf; Efrati, Efi; Kupferman, Raz

2012-02-01

We study the geometry and mechanics that drive the opening of Bauhinia seeds pods. The pod valve wall consists of two fibrous layers oriented at ± 45^o with respect to the pod axis. Upon drying, each of the layers shrinks uniaxially, perpendicularly to the fibers orientation. This active deformation turn the valve into an incompatible sheet with reference saddle-like curvature tensor and a flat (Euclidean) reference metric. These two intrinsic properties are incompatible. The shape is, therefore, selected by a stretching-bending competition. Strips cut from the valve tissue and from synthetic model material adopt various helical configurations. We provide analytical expressions for these configurations in the bending and stretching dominated regimes. Surface measurements show the transition from minimal surfaces (narrow limit) to cylindrical ones (wide limit). Finally, we show how plants use these mechanical principles using different tissue architectures.

Experimental high-frequency ultrasound can detect graft rejection after small bowel transplantation.

PubMed

Yang, R; Liu, Q; Wu, E X; Pescovitz, M D; Collins, M H; Kopecky, K K; Grosfeld, J L

1994-02-01

Early diagnosis of graft rejection after small bowel transplantation (SBT) can allow prompt institution of vigorous immunosuppressive therapy, with resultant reversal of the rejection process. The current method for graft monitoring is random mucosal biopsy from a stomal site or through an endoscope. However, because early rejection often has a patchy distribution, it could be missed by random biopsy. We hypothesized that the pathological process of rejection would alter acoustic impedance of the tissue and thus change the ultrasonic patterns of the graft intestinal wall. If this hypothesis is correct, then high-frequency endoscopic ultrasound (US) could be used to monitor the entire transplanted bowel and guide the biopsy, with improved yields. This hypothesis was tested in a rat orthotopic SBT model. Sixty-two intestinal specimens (9 isografts, 12 allografts treated with cyclosporine A [CsA], 22 untreated allografts, and 19 intestines from normal rats) were collected for in vitro transluminal US imaging (30 MHz) and histopathologic study. The echo pattern of normal rat intestinal wall consisted of five echo layers that correlated spatially with the histological layers: the innermost hyperechoic layer 1, plus hypoechoic layer 2, corresponded to the mucosa; hyperechoic layer 3, the submucosa; anechoic layer 4, the muscularis propria; and hyperechoic layer 5, the serosa. The isografts and CsA-treated allografts were identical histologically and ultrasonically to normal intestine. However, the echo patterns of the untreated allografts had progressive loss of architectural stratification, with worsening rejection. The change began with patchy indistinctness and disruption of hyperechoic layers 1, 3 and 5, and progressed to total obliteration of the layers, with the intestinal wall becoming a nonstratified hypoechoic structure.(ABSTRACT TRUNCATED AT 250 WORDS)

Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye.

PubMed

Sharma, Robin; Williams, David R; Palczewska, Grazyna; Palczewski, Krzysztof; Hunter, Jennifer J

2016-02-01

Although extrinsic fluorophores can be introduced to label specific cell types in the retina, endogenous fluorophores, such as NAD(P)H, FAD, collagen, and others, are present in all retinal layers. These molecules are a potential source of optical contrast and can enable noninvasive visualization of all cellular layers. We used a two-photon fluorescence adaptive optics scanning light ophthalmoscope (TPF-AOSLO) to explore the native autofluorescence of various cell classes spanning several layers in the unlabeled retina of a living primate eye. Three macaques were imaged on separate occasions using a custom TPF-AOSLO. Two-photon fluorescence was evoked by pulsed light at 730 and 920 nm excitation wavelengths, while fluorescence emission was collected in the visible range from several retinal layers and different locations. Backscattered light was recorded simultaneously in confocal modality and images were postprocessed to remove eye motion. All retinal layers yielded two-photon signals and the heterogeneous distribution of fluorophores provided optical contrast. Several structural features were observed, such as autofluorescence from vessel walls, Müller cell processes in the nerve fibers, mosaics of cells in the ganglion cell and other nuclear layers of the inner retina, as well as photoreceptor and RPE layers in the outer retina. This in vivo survey of two-photon autofluorescence throughout the primate retina demonstrates a wider variety of structural detail in the living eye than is available through conventional imaging methods, and broadens the use of two-photon imaging of normal and diseased eyes.

Low resolution crystal structure of Arenicola erythrocruorin: influence of coiled coils on the architecture of a megadalton respiratory protein.

PubMed

Royer, William E; Omartian, Michael N; Knapp, James E

2007-01-05

Annelid erythrocruorins are extracellular respiratory complexes assembled from 180 subunits into hexagonal bilayers. Cryo-electron microscopic experiments have identified two different architectural classes. In one, designated type I, the vertices of the two hexagonal layers are partially staggered, with one hexagonal layer rotated by about 16 degrees relative to the other layer, whereas in the other class, termed type II, the vertices are essentially eclipsed. We report here the first crystal structure of a type II erythrocruorin, that from Arenicola marina, at 6.2 A resolution. The structure reveals the presence of long continuous triple-stranded coiled-coil "spokes" projecting towards the molecular center from each one-twelfth unit; interdigitation of these spokes provides the only contacts between the two hexagonal layers of the complex. This arrangement contrasts with that of a type I erythrocruorin from Lumbricus terrestris in which the spokes are broken into two triple-stranded coiled coils with a disjointed connection. The disjointed connection allows formation of a more compact structure in the type I architecture, with the two hexagonal layers closer together and additional extensive contacts between the layers. Comparison of sequences of the coiled-coil regions of various linker subunits shows that the linker subunits from type II erythrocruorins possess continuous heptad repeats, whereas a sequence gap places these repeats out of register in the type I linker subunits, consistent with a disjointed coiled-coil arrangement.

Histopathologic study of human vocal fold mucosa unphonated over a decade.

PubMed

Sato, Kiminori; Umeno, Hirohito; Ono, Takeharu; Nakashima, Tadashi

2011-12-01

Mechanotransduction caused by vocal fold vibration could possibly be an important factor in the maintenance of extracellular matrices and layered structure of the human adult vocal fold mucosa as a vibrating tissue after the layered structure has been completed. Vocal fold stellate cells (VFSCs) in the human maculae flavae of the vocal fold mucosa are inferred to be involved in the metabolism of extracellular matrices of the vocal fold mucosa. Maculae flavae are also considered to be an important structure in the growth and development of the human vocal fold mucosa. Tension caused by phonation (vocal fold vibration) is hypothesized to stimulate the VFSCs to accelerate production of extracellular matrices. A human adult vocal fold mucosa unphonated over a decade was investigated histopathologically. Vocal fold mucosa unphonated for 11 years and 2 months of a 64-year-old male with cerebral hemorrhage was investigated by light and electron microscopy. The vocal fold mucosae (including maculae flavae) were atrophic. The vocal fold mucosa did not have a vocal ligament, Reinke's space or a layered structure. The lamina propria appeared as a uniform structure. Morphologically, the VFSCs synthesized fewer extracellular matrices, such as fibrous protein and glycosaminoglycan. Consequently, VFSCs appeared to decrease their level of activity.

Morphologie und Ultrastruktur der Koralle Cornularia cornucopiae (Anthozoa, Octocorallia)

NASA Astrophysics Data System (ADS)

Benke, H.; Hündgen, M.

1984-03-01

Cornularia cornucopiae is a colonial coral whose polyps arise singly from stolons. In contrast with other octocrallia C. cornucopiae lacks calcareous spicules. Therefore, tissue preparation for electron microscopic investigations can be performed. The presence of a calyx such as the theca of hydroids, in which the polyps may be completely retracted, is conspicuous. The calyx consists of three layers. The structure of the basal layer suggests massive collagen. The body wall is connected with the calyx by living desmocytes. The histology of the oral disc and the actinopharynx is identical. The ventral side of the polyps bears the siphonoglyph. Below the pharynx the inner edges of the mesenteries are free and form the mesenterial filaments. The two ventral mesenteries differ from the others; the one is long and exhibits a large and heavily flagellated filament, the other is short and lacks a filament. The muscular system is represented by gastrodermal circular fibres in the body wall and by radial and longitudinal fibres in the septa; a large septal retractor muscle is missing.

Study of non-linear deformation of vocal folds in simulations of human phonation

NASA Astrophysics Data System (ADS)

Saurabh, Shakti; Bodony, Daniel

2014-11-01

Direct numerical simulation is performed on a two-dimensional compressible, viscous fluid interacting with a non-linear, viscoelastic solid as a model for the generation of the human voice. The vocal fold (VF) tissues are modeled as multi-layered with varying stiffness in each layer and using a finite-strain Standard Linear Solid (SLS) constitutive model implemented in a quadratic finite element code and coupled to a high-order compressible Navier-Stokes solver through a boundary-fitted fluid-solid interface. The large non-linear mesh deformation is handled using an elliptic/poisson smoothening technique. Supra-glottal flow shows asymmetry in the flow, which in turn has a coupling effect on the motion of the VF. The fully compressible simulations gives direct insight into the sound produced as pressure distributions and the vocal fold deformation helps study the unsteady vortical flow resulting from the fluid-structure interaction along the full phonation cycle. Supported by the National Science Foundation (CAREER Award Number 1150439).

Laser optoacoustic tomography for medical diagnostics: principles

NASA Astrophysics Data System (ADS)

Oraevsky, Alexander A.; Esenaliev, Rinat O.; Jacques, Steven L.; Tittel, Frank K.

1996-04-01

This paper is to describe principles of laser optoacoustic tomography for medical diagnostics. Two types of imaging modes are presented. The first is the tomography in transmission mode, which utilizes detection of stress transients transmitted from the laser-excited volume toward the depth through thick layers of tissue. The second is the tomography in reflection mode which utilizes detection of stress transients generated in superficial tissue layer and reflected back toward tissue surface. To distinguish the two modes, we have abbreviated them as (1) laser optoacoustic tomography in transmission mode, LOATT, and (2) time-resolved stress detection tomography of light absorption, TRSDTLA, in reflection mode where emphasis is made on high spatial resolution of images. The basis for laser optoacoustic tomography is the time-resolved detection of laser-induced transient stress waves, selectively generated in absorbing tissues of diagnostic interest. Such a technique allows one to visualize absorbed light distribution in turbid biological tissues irradiated by short laser pulses. Laser optoacoustic tomography can be used for detection of tissue pathological changes that result in either increased concentration of various tissue chromophores such as hemoglobin or in development of enhanced microcirculation in diseased tissue. Potential areas of applications are diagnosis of cancer, brain hemorrhages, arterial atherosclerotic plaques, and other diseased tissues. In addition, it can provide feedback information during medical treatments. Both LOATT and TRSDTLA utilize laser excitation of biological tissues and sensitive detection of laser-induced stress waves. Optical selectivity is based upon differences in optical properties of pathologically different tissues. Sensitivity comes from stress generation under irradiation conditions of temporal stress confinement. The use of sensitive wide-band lithium niobate acoustic transducers expands limits of laser optoacoustic tomography. The technology allows us to determine directly temperature distributions in tissues and locate tissues volumes with different absorption. To demonstrate principles of TRSDTLA, experiments were conducted in vivo with mice-model for breast cancer using specially designed front-surface transducers- reflectometers. To present advantages and limitation of LOATT, experiments were performed in phantoms made of gel with polystyrene spheres colored with copper sulfate. Our experimental results and theoretical calculations show that TRSDTLA can be applied for non- invasive histology of layered tissues with in-depth resolution of up to 2 microns. TRSDTLA in acoustic reflection mode is promising for diagnostics of skin and ocular diseases. LOATT in acoustic transmission mode can be applied for detection of small tissue volumes with enhanced absorption located inside organs at the depth of up to 10 cm.

Fabrication of a multi-layer three-dimensional scaffold with controlled porous micro-architecture for application in small intestine tissue engineering.

PubMed

Knight, Toyin; Basu, Joydeep; Rivera, Elias A; Spencer, Thomas; Jain, Deepak; Payne, Richard

2013-01-01

Various methods can be employed to fabricate scaffolds with characteristics that promote cell-to-material interaction. This report examines the use of a novel technique combining compression molding with particulate leaching to create a unique multi-layered scaffold with differential porosities and pore sizes that provides a high level of control to influence cell behavior. These cell behavioral responses were primarily characterized by bridging and penetration of two cell types (epithelial and smooth muscle cells) on the scaffold in vitro. Larger pore sizes corresponded to an increase in pore penetration, and a decrease in pore bridging. In addition, smaller cells (epithelial) penetrated further into the scaffold than larger cells (smooth muscle cells). In vivo evaluation of a multi-layered scaffold was well tolerated for 75 d in a rodent model. This data shows the ability of the components of multi-layered scaffolds to influence cell behavior, and demonstrates the potential for these scaffolds to promote desired tissue outcomes in vivo.

Cornea Transplant

MedlinePlus

... transplants may be used that remove only certain layers of cornea tissue, or only tissue affected by ... procedure removes diseased tissue from the back corneal layers, including the endothelium, along with the Descemet membrane, ...

Hybrid method to estimate two-layered superficial tissue optical properties from simulated data of diffuse reflectance spectroscopy.

PubMed

Hsieh, Hong-Po; Ko, Fan-Hua; Sung, Kung-Bin

2018-04-20

An iterative curve fitting method has been applied in both simulation [J. Biomed. Opt.17, 107003 (2012)JBOPFO1083-366810.1117/1.JBO.17.10.107003] and phantom [J. Biomed. Opt.19, 077002 (2014)JBOPFO1083-366810.1117/1.JBO.19.7.077002] studies to accurately extract optical properties and the top layer thickness of a two-layered superficial tissue model from diffuse reflectance spectroscopy (DRS) data. This paper describes a hybrid two-step parameter estimation procedure to address two main issues of the previous method, including (1) high computational intensity and (2) converging to local minima. The parameter estimation procedure contained a novel initial estimation step to obtain an initial guess, which was used by a subsequent iterative fitting step to optimize the parameter estimation. A lookup table was used in both steps to quickly obtain reflectance spectra and reduce computational intensity. On simulated DRS data, the proposed parameter estimation procedure achieved high estimation accuracy and a 95% reduction of computational time compared to previous studies. Furthermore, the proposed initial estimation step led to better convergence of the following fitting step. Strategies used in the proposed procedure could benefit both the modeling and experimental data processing of not only DRS but also related approaches such as near-infrared spectroscopy.

Shear wave elastography using Wigner-Ville distribution: a simulated multilayer media study.

PubMed

Bidari, Pooya Sobhe; Alirezaie, Javad; Tavakkoli, Jahan

2016-08-01

Shear Wave Elastography (SWE) is a quantitative ultrasound-based imaging modality for distinguishing normal and abnormal tissue types by estimating the local viscoelastic properties of the tissue. These properties have been estimated in many studies by propagating ultrasound shear wave within the tissue and estimating parameters such as speed of wave. Vast majority of the proposed techniques are based on the cross-correlation of consecutive ultrasound images. In this study, we propose a new method of wave detection based on time-frequency (TF) analysis of the ultrasound signal. The proposed method is a modified version of the Wigner-Ville Distribution (WVD) technique. The TF components of the wave are detected in a propagating ultrasound wave within a simulated multilayer tissue and the local properties are estimated based on the detected waves. Image processing techniques such as Alternative Sequential Filters (ASF) and Circular Hough Transform (CHT) have been utilized to improve the estimation of TF components. This method has been applied to a simulated data from Wave3000™ software (CyberLogic Inc., New York, NY). This data simulates the propagation of an acoustic radiation force impulse within a two-layer tissue with slightly different viscoelastic properties between the layers. By analyzing the local TF components of the wave, we estimate the longitudinal and shear elasticities and viscosities of the media. This work shows that our proposed method is capable of distinguishing between different layers of a tissue.

Fabrication of micro-alginate gel tubes utilizing micro-gelatin fibers

NASA Astrophysics Data System (ADS)

Sakaguchi, Katsuhisa; Arai, Takafumi; Shimizu, Tatsuya; Umezu, Shinjiro

2017-05-01

Tissues engineered utilizing biofabrication techniques have recently been the focus of much attention, because these bioengineered tissues have great potential to improve the quality of life of patients with various hard-to-treat diseases. Most tissues contain micro-tubular structures including blood vessels, lymphatic vessels, and bile canaliculus. Therefore, we bioengineered a micro diameter tube using alginate gel to coat the core gelatin gel. Micro-gelatin fibers were fabricated by the coacervation method and then coated with a very thin alginate gel layer by dipping. A micro diameter alginate tube was produced by dissolving the core gelatin gel. Consequently, these procedures led to the formation of micro-alginate gel tubes of various shapes and sizes. This biofabrication technique should contribute to tissue engineering research fields.

Two-photon Microscopy and Polarimetry for Assessment of Myocardial Tissue Organization

NASA Astrophysics Data System (ADS)

Archambault-Wallenburg, Marika

Optical methods can provide useful tissue characterization tools. For this project, two-photon microscopy and polarized light examinations (polarimetry) were used to assess the organizational state of myocardium in healthy, infarcted, and stem-cell regenerated states. Two-photon microscopy visualizes collagen through second-harmonic generation and myocytes through two-photon excitation autofluorescence, providing information on the composition and structure/organization of the tissue. Polarimetry measurements yield a value of linear retardance that can serve as an indicator of tissue anisotropy, and with a dual-projection method, information about the anisotropy axis orientation can also be extracted. Two-photon microscopy results reveal that stem-cell treated tissue retains more myocytes and structure than infarcted myocardium, while polarimetry findings suggest that the injury caused by temporary ligation of a coronary artery is less severe and more diffuse that than caused by a permanent ligation. Both these methods show potential for tissue characterization.

Design of a dual band metamaterial absorber for Wi-Fi bands

NASA Astrophysics Data System (ADS)

Alkurt, Fatih Özkan; Baǧmancı, Mehmet; Karaaslan, Muharrem; Bakır, Mehmet; Altıntaş, Olcay; Karadaǧ, Faruk; Akgöl, Oǧuzhan; Ünal, Emin

2018-02-01

The goal of this work is to design and fabrication of a dual band metamaterial based absorber for Wireless Fidelity (Wi-Fi) bands. Wi-Fi has two different operating frequencies such as 2.45 GHz and 5 GHz. A dual band absorber is proposed and the proposed structure consists of two layered unit cells, and different sized square split ring (SSR) resonators located on each layers. Copper is used for metal layer and resonator structure, FR-4 is used as substrate layer in the proposed structure. This designed dual band metamaterial absorber is used in the wireless frequency bands which has two center frequencies such as 2.45 GHz and 5 GHz. Finite Integration Technique (FIT) based simulation software used and according to FIT based simulation results, the absorption peak in the 2.45 GHz is about 90% and the another frequency 5 GHz has absorption peak near 99%. In addition, this proposed structure has a potential for energy harvesting applications in future works.

I-III-VI.sub.2 based solar cell utilizing the structure CuInGaSe.sub.2 CdZnS/ZnO

DOEpatents

Chen, Wen S.; Stewart, John M.

1992-01-07

A thin film I-III-VI.sub.2 based solar cell having a first layer of copper indium gallium selenide, a second layer of cadmium zinc sulfide, a double layer of zinc oxide, and a metallization structure comprised of a layer of nickel covered by a layer of aluminum. An optional antireflective coating may be placed on said metallization structure. The cadmium zinc sulfide layer is deposited by means of an aqueous solution growth deposition process and may actually consist of two layers: a low zinc content layer and a high zinc content layer. Photovoltaic efficiencies of 12.5% at Air Mass 1.5 illumination conditions and 10.4% under AMO illumination can be achieved.

Enhanced adherence of mouse fibroblast and vascular cells to plasma modified polyethylene.

PubMed

Reznickova, Alena; Novotna, Zdenka; Kolska, Zdenka; Kasalkova, Nikola Slepickova; Rimpelova, Silvie; Svorcik, Vaclav

2015-01-01

Since the last decade, tissue engineering has shown a sensational promise in providing more viable alternatives to surgical procedures for harvested tissues, implants and prostheses. Biomedical polymers, such as low-density polyethylene (LDPE), high-density polyethylene (HDPE) and ultra-high molecular weight polyethylene (UHMWPE), were activated by Ar plasma discharge. Degradation of polymer chains was examined by determination of the thickness of ablated layer. The amount of an ablated polymer layer was measured by gravimetry. Contact angle, measured by goniometry, was studied as a function of plasma exposure and post-exposure aging times. Chemical structure of modified polymers was characterized by angle resolved X-ray photoelectron spectroscopy. Surface chemistry and polarity of the samples were investigated by electrokinetic analysis. Changes in surface morphology were followed using atomic force microscopy. Cytocompatibility of plasma activated polyethylene foils was studied using two distinct model cell lines; VSMCs (vascular smooth muscle cells) as a model for vascular graft testing and connective tissue cells L929 (mouse fibroblasts) approved for standardized material cytotoxicity testing. Specifically, the cell number, morphology, and metabolic activity of the adhered and proliferated cells on the polyethylene matrices were studied in vitro. It was found that the plasma treatment caused ablation of the polymers, resulting in dramatic changes in their surface morphology and roughness. ARXPS and electrokinetic measurements revealed oxidation of the polymer surface. It was found that plasma activation has a positive effect on the adhesion and proliferation of VSMCs and L929 cells. Copyright © 2015 Elsevier B.V. All rights reserved.

Nanostructured 3D constructs based on chitosan and chondroitin sulphate multilayers for cartilage tissue engineering.

PubMed

Silva, Joana M; Georgi, Nicole; Costa, Rui; Sher, Praveen; Reis, Rui L; Van Blitterswijk, Clemens A; Karperien, Marcel; Mano, João F

2013-01-01

Nanostructured three-dimensional constructs combining layer-by-layer technology (LbL) and template leaching were processed and evaluated as possible support structures for cartilage tissue engineering. Multilayered constructs were formed by depositing the polyelectrolytes chitosan (CHT) and chondroitin sulphate (CS) on either bidimensional glass surfaces or 3D packet of paraffin spheres. 2D CHT/CS multi-layered constructs proved to support the attachment and proliferation of bovine chondrocytes (BCH). The technology was transposed to 3D level and CHT/CS multi-layered hierarchical scaffolds were retrieved after paraffin leaching. The obtained nanostructured 3D constructs had a high porosity and water uptake capacity of about 300%. Dynamical mechanical analysis (DMA) showed the viscoelastic nature of the scaffolds. Cellular tests were performed with the culture of BCH and multipotent bone marrow derived stromal cells (hMSCs) up to 21 days in chondrogenic differentiation media. Together with scanning electronic microscopy analysis, viability tests and DNA quantification, our results clearly showed that cells attached, proliferated and were metabolically active over the entire scaffold. Cartilaginous extracellular matrix (ECM) formation was further assessed and results showed that GAG secretion occurred indicating the maintenance of the chondrogenic phenotype and the chondrogenic differentiation of hMSCs.

An anatomical, histopathological, and molecular biological function study of the fascias posterior to the interperitoneal colon and its associated mesocolon: their relevance to colonic surgery

PubMed Central

Gao, Zhidong; Ye, Yingjiang; Zhang, Weiguang; Shen, Danhua; Zhong, Yanfeng; Jiang, Kewei; Yang, Xiaodong; Yin, Mujun; Liang, Bin; Tian, Long; Wang, Shan

2013-01-01

The study aim was to explore the anatomy, histopathology, and molecular biological function of the fascias posterior to the interperitoneal colon and its mesocolon to provide information for improving complete mesocolic excision. To accomplish this aim, we performed intraoperative observations in 60 interperitoneal colon-cancer patients accepted for complete mesocolic excision and conducted local anatomy observations for five embalmed cadavers. An additional two embalmed child cadaver specimens were studied with large slices and paraffin sections. Ten of the 60 patients were examined with a lymph node tracer technique in vivo, while fresh specimens from these patients were assessed by histopathological examination and transwell cell migration assays in vitro. The anatomical and histopathological findings showed that the fascias posterior to the interperitoneal colon and its associated mesocolon were composed of two independent layers: the visceral and parietal fascias. These two fascias were primarily composed of collagen fibers, with the parietal fascia containing a small amount of muscle fiber. The in vivo test showed that the visceral fascia surrounded the colon and its associated mesocolon, including vessels and lymphatics, and that it had no lymphatic flow through it into the rear tissues. Moreover, the in vitro assays showed the visceral fascia was able to block tumor cell migration. Although many surgical scholars have known of the existence of fascia tissue posterior to the intraperitoneal colon, the detailed structure has been ignored and been unclear. As shown by our findings, the visceral and parietal fascias are truly formed structures that have not been previously reported. A thorough understanding of fascial structures and the function of the visceral fascia barrier in blocking tumor cells will facilitate surgeons when performing high-quality complete mesocolic excision procedures. PMID:23721400

WE-AB-303-04: A Tissue Model of Cherenkov Emission From the Skin Surface During Megavoltage X-Ray Radiotherapy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wiles, A. N.; Loyalka, S. K.; Izaguirre, E. W.

Purpose: To develop a tissue model of Cherenkov radiation emitted from the skin surface during external beam radiotherapy. Imaging Cherenkov radiation emitted from human skin allows visualization of the beam position and potentially surface dose estimates, and our goal is to characterize the optical properties of these emissions. Methods: We developed a Monte Carlo model of Cherenkov radiation generated in a semi-infinite tissue slab by megavoltage x-ray beams with optical transmission properties determined by a two-layered skin model. We separate the skin into a dermal and an epidermal layer in our model, where distinct molecular absorbers modify the Cherenkov intensitymore » spectrum in each layer while we approximate the scattering properties with Mie and Rayleigh scattering from the highly structured molecular organization found in human skin. Results: We report on the estimated distributions of the Cherenkov wavelength spectrum, emission angles, and surface distribution for the modeled irradiated skin surface. The expected intensity distribution of Cherenkov radiation emitted from skin shows a distinct intensity peak around 475 nm, the blue region of the visible spectrum, between a pair of optical absorption bands in hemoglobin and a broad plateau beginning near 600 nm and extending to at least 700 nm where melanin and hemoglobin absorption are both low. We also find that the Cherenkov intensity decreases with increasing angle from the surface normal, the majority being emitted within 20 degrees of the surface normal. Conclusion: Our estimate of the spectral distribution of Cherenkov radiation emitted from skin indicates an advantage to using imaging devices with long wavelength spectral responsivity. We also expect the most efficient imaging to be near the surface normal where the intensity is greatest; although for contoured surfaces, the relative intensity across the surface may appear to vary due to decreasing Cherenkov intensity with increased angle from the skin normal. This research was supported in part by a GAANN Fellowship from the Department of Education.« less

A structural review of foliar glands in Passiflora L. (Passifloraceae)

PubMed Central

da Costa Silva, Delmira; Flavia de Albuquerque Melo-de-Pinna, Gladys

2017-01-01

Extrafloral glands in Passifloraceae species have aroused the interest of many researchers because of their wide morphological diversity. The present work analyzed the foliar glands on 34 species of Passiflora from samples containing glands in the petiole and foliar blade fixed in 50% solution of formaldehyde-ethanol-acetic acid and stored in a 70% ethanol solution. For anatomical analyses, part of the material was embedded in Paraplast, longitudinally sectioned and double stained with safranin and astra blue. Scanning electron microscopy analysis was also carried out. To analyze the presence of sugars in the secretion of foliar glands, a glucose strip test was used. Based on the results of morphological, anatomical and glucose strip tests, the foliar secretory glands in Passiflora can be grouped into two categories: Type I glands, defined as nectaries, can be elevated or flattened, and can have a sugar content high enough to be detected by the glucose strip test analysis. Type II glands are elevated and did not show a positive reaction to the glucose strip test. From an anatomical viewpoint, glands characterized as extrafloral nectaries show a multistratified secretory epidermis, typically followed by two flat layers of nectariferous parenchyma with dense content. Internal to these layers, vascular bundles are immersed in the subsecretory parenchyma and terminate in phloem cells. On the other hand, type II glands show a single layer of elongated secretory epidermal cells. Internal to this single layer, parenchyma and vascular tissue with both phloem and xylem elements can be observed. The analyzed species show a wide diversity of gland shape and distribution, and the combined analysis of morphology, anatomy and preliminary tests for the presence of glucose in the exudate in different Passiflora subgenera suggests the occurrence of two categories of glands: nectaries and resin glands. PMID:29136029

A Solvent-Free Surface Suspension Melt Technique for Making Biodegradable PCL Membrane Scaffolds for Tissue Engineering Applications.

PubMed

Suntornnond, Ratima; An, Jia; Tijore, Ajay; Leong, Kah Fai; Chua, Chee Kai; Tan, Lay Poh

2016-03-21

In tissue engineering, there is limited availability of a simple, fast and solvent-free process for fabricating micro-porous thin membrane scaffolds. This paper presents the first report of a novel surface suspension melt technique to fabricate a micro-porous thin membrane scaffolds without using any organic solvent. Briefly, a layer of polycaprolactone (PCL) particles is directly spread on top of water in the form of a suspension. After that, with the use of heat, the powder layer is transformed into a melted layer, and following cooling, a thin membrane is obtained. Two different sizes of PCL powder particles (100 µm and 500 µm) are used. Results show that membranes made from 100 µm powders have lower thickness, smaller pore size, smoother surface, higher value of stiffness but lower ultimate tensile load compared to membranes made from 500 µm powder. C2C12 cell culture results indicate that the membrane supports cell growth and differentiation. Thus, this novel membrane generation method holds great promise for tissue engineering.

Micro-arc oxidation treatment to improve the hard-tissue compatibility of Ti-29Nb-13Ta-4.6Zr alloy

NASA Astrophysics Data System (ADS)

Tsutsumi, Yusuke; Niinomi, Mitsuo; Nakai, Masaaki; Tsutsumi, Harumi; Doi, Hisashi; Nomura, Naoyuki; Hanawa, Takao

2012-12-01

Micro-arc oxidation (MAO) was performed on a β-type Ti-29Nb-13Ta-4.6Zr alloy (TNTZ) in this study to improve its bioactivity in a body fluid and its hard-tissue compatibility. The surface oxide layer formed on TNTZ by MAO treatment in a mixture of calcium glycerophosphate and magnesium acetate was characterized using various surface analyses. The oxide layer was mainly composed of two types of TiO2 (rutile and anatase), and it also contained Ca, P, and Mg, which were incorporated from the electrolyte during the treatment. The calcium phosphate formation on the surface of the specimens after immersion in Hanks' solution was evaluated to determine the bioactivity of TNTZ with and without MAO treatment. As a result, thick calcium phosphate layers formed on the TNTZ specimen that underwent MAO treatment, whereas only a small amount of precipitate was observed on TNTZ without treatment. Thus, the MAO treatment is a promising method to improve the bioactivity and hard-tissue compatibility of TNTZ.

Largely Tunable Band Structures of Few-Layer InSe by Uniaxial Strain.

PubMed

Song, Chaoyu; Fan, Fengren; Xuan, Ningning; Huang, Shenyang; Zhang, Guowei; Wang, Chong; Sun, Zhengzong; Wu, Hua; Yan, Hugen

2018-01-31

Because of the strong quantum confinement effect, few-layer γ-InSe exhibits a layer-dependent band gap, spanning the visible and near infrared regions, and thus recently has been drawing tremendous attention. As a two-dimensional material, the mechanical flexibility provides an additional tuning knob for the electronic structures. Here, for the first time, we engineer the band structures of few-layer and bulk-like InSe by uniaxial tensile strain and observe a salient shift of photoluminescence peaks. The shift rate of the optical gap is approximately 90-100 meV per 1% strain for four- to eight-layer samples, which is much larger than that for the widely studied MoS 2 monolayer. Density functional theory calculations well reproduce the observed layer-dependent band gaps and the strain effect and reveal that the shift rate decreases with the increasing layer number for few-layer InSe. Our study demonstrates that InSe is a very versatile two-dimensional electronic and optoelectronic material, which is suitable for tunable light emitters, photodetectors, and other optoelectronic devices.

In Vitro Engineering of Vascularized Tissue Surrogates

PubMed Central

Sakaguchi, Katsuhisa; Shimizu, Tatsuya; Horaguchi, Shigeto; Sekine, Hidekazu; Yamato, Masayuki; Umezu, Mitsuo; Okano, Teruo

2013-01-01

In vitro scaling up of bioengineered tissues is known to be limited by diffusion issues, specifically a lack of vasculature. Here, we report a new strategy for preserving cell viability in three-dimensional tissues using cell sheet technology and a perfusion bioreactor having collagen-based microchannels. When triple-layer cardiac cell sheets are incubated within this bioreactor, endothelial cells in the cell sheets migrate to vascularize in the collagen gel, and finally connect with the microchannels. Medium readily flows into the cell sheets through the microchannels and the newly developed capillaries, while the cardiac construct shows simultaneous beating. When additional triple-layer cell sheets are repeatedly layered, new multi-layer construct spontaneously integrates and the resulting construct becomes a vascularized thick tissue. These results confirmed our method to fabricate in vitro vascularized tissue surrogates that overcomes engineered-tissue thickness limitations. The surrogates promise new therapies for damaged organs as well as new in vitro tissue models. PMID:23419835

The two-dimensional hybrid surface plasma micro-cavity

NASA Astrophysics Data System (ADS)

Kai, Tong; Mei-yu, Wang; Fu-cheng, Wang; Jia, Guo

2018-07-01

A hybrid surface plasma micro-cavity structure with a defect cavity is formed based on the two-dimensional surface plasmon resonance photonic crystal waveguide structure. A cell defect is introduced in the centre of the photonic crystal layer to build the hybrid surface plasma micro-cavity structure. This work is numerical based on the finite-difference time-domain method. The photon energy is confined to the micro-cavity and the photon energy is strongest at the interface between the insulating layer and the metal layer. The micro-cavity structure has a very small mode volume of sub-wavelength scale in the 1550 nm communication band. The value of Q/V is up to 7132.08 λ/n-3.

Long-Distance Translocation of Protein during Morphogenesis of the Fruiting Body in the Filamentous Fungus, Agaricus bisporus

PubMed Central

Woolston, Benjamin M.; Schlagnhaufer, Carl; Wilkinson, Jack; Larsen, Jeffrey; Shi, Zhixin; Mayer, Kimberly M.; Walters, Donald S.; Curtis, Wayne R.; Romaine, C. Peter

2011-01-01

Commercial cultivation of the mushroom fungus, Agaricus bisporus, utilizes a substrate consisting of a lower layer of compost and upper layer of peat. Typically, the two layers are seeded with individual mycelial inoculants representing a single genotype of A. bisporus. Studies aimed at examining the potential of this fungal species as a heterologous protein expression system have revealed unexpected contributions of the mycelial inoculants in the morphogenesis of the fruiting body. These contributions were elucidated using a dual-inoculant method whereby the two layers were differientially inoculated with transgenic β-glucuronidase (GUS) and wild-type (WT) lines. Surprisingly, use of a transgenic GUS line in the lower substrate and a WT line in the upper substrate yielded fruiting bodies expressing GUS activity while lacking the GUS transgene. Results of PCR and RT-PCR analyses for the GUS transgene and RNA transcript, respectively, suggested translocation of the GUS protein from the transgenic mycelium colonizing the lower layer into the fruiting body that developed exclusively from WT mycelium colonizing the upper layer. Effective translocation of the GUS protein depended on the use of a transgenic line in the lower layer in which the GUS gene was controlled by a vegetative mycelium-active promoter (laccase 2 and β-actin), rather than a fruiting body-active promoter (hydrophobin A). GUS-expressing fruiting bodies lacking the GUS gene had a bonafide WT genotype, confirmed by the absence of stably inherited GUS and hygromycin phosphotransferase selectable marker activities in their derived basidiospores and mycelial tissue cultures. Differientially inoculating the two substrate layers with individual lines carrying the GUS gene controlled by different tissue-preferred promoters resulted in up to a ∼3.5-fold increase in GUS activity over that obtained with a single inoculant. Our findings support the existence of a previously undescribed phenomenon of long-distance protein translocation in A. bisporus that has potential application in recombinant protein expression and biotechnological approaches for crop improvement. PMID:22163014

A composites-based hyperelastic constitutive model for soft tissue with application to the human annulus fibrosus

NASA Astrophysics Data System (ADS)

Guo, Z. Y.; Peng, X. Q.; Moran, B.

2006-09-01

This paper presents a composites-based hyperelastic constitutive model for soft tissue. Well organized soft tissue is treated as a composite in which the matrix material is embedded with a single family of aligned fibers. The fiber is modeled as a generalized neo-Hookean material in which the stiffness depends on fiber stretch. The deformation gradient is decomposed multiplicatively into two parts: a uniaxial deformation along the fiber direction and a subsequent shear deformation. This permits the fiber-matrix interaction caused by inhomogeneous deformation to be estimated by using effective properties from conventional composites theory based on small strain linear elasticity and suitably generalized to the present large deformation case. A transversely isotropic hyperelastic model is proposed to describe the mechanical behavior of fiber-reinforced soft tissue. This model is then applied to the human annulus fibrosus. Because of the layered anatomical structure of the annulus fibrosus, an orthotropic hyperelastic model of the annulus fibrosus is developed. Simulations show that the model reproduces the stress-strain response of the human annulus fibrosus accurately. We also show that the expression for the fiber-matrix shear interaction energy used in a previous phenomenological model is compatible with that derived in the present paper.

Fluctuations and differential contraction during regeneration of Hydra vulgaris tissue toroids

NASA Astrophysics Data System (ADS)

Krahe, Michael; Wenzel, Iris; Lin, Kao-Nung; Fischer, Julia; Goldmann, Joseph; Kästner, Markus; Fütterer, Claus

2013-03-01

We studied regenerating bilayered tissue toroids dissected from Hydra vulgaris polyps and relate our macroscopic observations to the dynamics of force-generating mesoscopic cytoskeletal structures. Tissue fragments undergo a specific toroid-spheroid folding process leading to complete regeneration towards a new organism. The time scale of folding is too fast for biochemical signalling or morphogenetic gradients, which forced us to assume purely mechanical self-organization. The initial pattern selection dynamics was studied by embedding toroids into hydro-gels, allowing us to observe the deformation modes over longer periods of time. We found increasing mechanical fluctuations which break the toroidal symmetry, and discuss the evolution of their power spectra for various gel stiffnesses. Our observations are related to single-cell studies which explain the mechanical feasibility of the folding process. In addition, we observed switching of cells from a tissue bound to a migrating state after folding failure as well as in tissue injury. We found a supra-cellular actin ring assembled along the toroid's inner edge. Its contraction can lead to the observed folding dynamics as we could confirm by finite element simulations. This actin ring in the inner cell layer is assembled by myosin-driven length fluctuations of supra-cellular F-actin bundles (myonemes) in the outer cell layer. This paper is dedicated to Malcolm Steinberg.

Predicting multicellular function through multi-layer tissue networks

PubMed Central

Zitnik, Marinka; Leskovec, Jure

2017-01-01

Abstract Motivation: Understanding functions of proteins in specific human tissues is essential for insights into disease diagnostics and therapeutics, yet prediction of tissue-specific cellular function remains a critical challenge for biomedicine. Results: Here, we present OhmNet, a hierarchy-aware unsupervised node feature learning approach for multi-layer networks. We build a multi-layer network, where each layer represents molecular interactions in a different human tissue. OhmNet then automatically learns a mapping of proteins, represented as nodes, to a neural embedding-based low-dimensional space of features. OhmNet encourages sharing of similar features among proteins with similar network neighborhoods and among proteins activated in similar tissues. The algorithm generalizes prior work, which generally ignores relationships between tissues, by modeling tissue organization with a rich multiscale tissue hierarchy. We use OhmNet to study multicellular function in a multi-layer protein interaction network of 107 human tissues. In 48 tissues with known tissue-specific cellular functions, OhmNet provides more accurate predictions of cellular function than alternative approaches, and also generates more accurate hypotheses about tissue-specific protein actions. We show that taking into account the tissue hierarchy leads to improved predictive power. Remarkably, we also demonstrate that it is possible to leverage the tissue hierarchy in order to effectively transfer cellular functions to a functionally uncharacterized tissue. Overall, OhmNet moves from flat networks to multiscale models able to predict a range of phenotypes spanning cellular subsystems. Availability and implementation: Source code and datasets are available at http://snap.stanford.edu/ohmnet. Contact: jure@cs.stanford.edu PMID:28881986

Structuring of composite hydrogel bioadhesives and its effect on properties and bonding mechanism.

PubMed

Pinkas, Oded; Goder, Daniella; Noyvirt, Roni; Peleg, Sivan; Kahlon, Maayan; Zilberman, Meital

2017-03-15

Bioadhesives are polymeric hydrogels that can adhere to a tissue after crosslinking and are an essential element in nearly all surgeries worldwide. Several bioadhesives are commercially available. However, none of them are ideal. The main limitation of current tissue adhesives is the tradeoff between biocompatibility and mechanical strength, especially in wet hemorrhagic environments. Our novel bioadhesives are based on the natural polymers gelatin (coldwater fish) and alginate, crosslinked by carbodiimide (EDC). Two types of hemostatic agents with a layered silicate structure, montmorillonite (MMT) and kaolin, were loaded in order to improve the sealing ability in a hemorrhagic environment. The effect of the adhesive's components on its mechanical strength was studied by three different methods - burst strength, lap shear and compression. The viscosity, gelation time and structural features of the adhesive were also studied. A qualitative model that describes the effect of the bioadhesive's parameters on the cohesive and adhesive strength was developed. A formulation based on 400mg/mL gelatin, 10mg/mL alginate and 20mg/mL EDC was found as optimal, enabling a burst strength of 387mmHg. Incorporation of kaolin increased the burst strength by 25% due to microcomposite structuring, whereas MMT increased the burst strength by 50% although loaded in a smaller concentration, due to nano-structuring effects. This research clearly shows that the incorporation of kaolin and MMT in gelatin-alginate surgical sealants is a very promising novel approach for improving the bonding strength and physical properties of surgical sealants for use in hemorrhagic environments. The current manuscript focuses on novel bioadhesives, based on natural polymers and loaded with hemostatic agents with a layered silicate structure, in order to improve the sealing ability in hemorrhagic environment. Such composite bioadhesives have not been developed and studied before. The effect of the adhesive's components on its mechanical strength was studied by three different methods, as well as the physical properties and structural features. Thorough understanding of these unique biomaterials resulted in a qualitative model which describes the effect of the bioadhesive's parameters on the cohesive and adhesive strength. Thus, structure-property-function relationships are presented. Structuring of the composite bioadhesives and its effect of the properties and bonding mechanism, are expected to be of high interest to Acta readership. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

New Synthesis of Ocean Crust Velocity Structure From Two-Dimensional Profiles

NASA Astrophysics Data System (ADS)

Christeson, G. L.; Goff, J.; Carlson, R. L.; Reece, R.

2017-12-01

The velocity structure of typical oceanic crust consists of Layer 2, where velocities increase rapidly with depth from seafloor, and Layer 3, which is thicker and has a lower velocity gradient. Previous syntheses have found no correlation of velocity structure with spreading rate, even though we know that magmatic processes differ between slow-spreading and fast-spreading crust. We present a new synthesis of ocean crust velocity structure, compiling observations from two-dimensional studies in the Atlantic, Pacific, and Indian ocean basins. The Layer 2/3 boundary was picked from each publication at a change in gradient either on velocity-depth functions or contour plots (with at least 0.5 km/s contour interval), or from the appropriate layer boundary for layered models. We picked multiple locations at each seismic refraction profile if warranted by model variability. Preliminary results show statistically significant differences in average Layer 2 and Layer 3 thicknesses between slow-spreading and superfast-spreading crust, with Layer 2 thinner and Layer 3 thicker for the higher spreading rate crust. The thickness changes are about equivalent, resulting in no change in mean crustal thickness. The Layer 2/3 boundary is often interpreted as the top of the gabbros; however, a comparison with mapped magma lens depths at the ridge axis shows that the boundary is typically deeper than average axial melt lens depth at superfast-spreading crust, and shallower at intermediate-spreading crust.

Superficial Macromolecular Arrays on the Cell Wall of Spirillum putridiconchylium

PubMed Central

Beveridge, T. J.; Murray, R. G. E.

1974-01-01

Electron microscopy of the cell envelope of Spirillum putridiconchylium, using negatively stained, thin-sectioned, and replicated freeze-etched preparations, showed two superficial wall layers forming a complex macromolecular pattern on the external surface. The outer structured layer was a linear array of particles overlying an inner tetragonal array of larger subunits. They were associated in a very regular fashion, and the complex was bonded to the outer, pitted surface of the lipopolysaccharide tripartite layer of the cell wall. The relationship of the components of the two structured layers was resolved with the aid of optical diffraction, combined with image filtering and reconstruction and linear and rotary integration techniques. The outer structural layer consisted of spherical 1.5-nm units set in double lines determined by the size and arrangement of 6- by 3-nm inner structural layer subunits, which bore one outer structural layer unit on each outer corner. The total effect of this arrangement was a double-ridged linear structure that was evident in surface replicas and negatively stained fragments of the whole wall. The packing of these units was not square but skewed by 2° off the perpendicular so that the “unit array” described by optical diffraction and linear integration appeared to be a deformed tetragon. The verity of the model was checked by using a photographically reduced image to produce an optical diffraction pattern for comparison with that of the actual layers. The correspondence was nearly perfect. Images PMID:4137219

Silicon Alignment Pins: An Easy Way to Realize a Wafer-To-Wafer Alignment

NASA Technical Reports Server (NTRS)

Peralta, Alejandro (Inventor); Gill, John J. (Inventor); Toda, Risaku (Inventor); Lin, Robert H. (Inventor); Jung-Kubiak, Cecile (Inventor); Reck, Theodore (Inventor); Thomas, Bertrand (Inventor); Siles, Jose V. (Inventor); Lee, Choonsup (Inventor); Chattopadhyay, Goutam (Inventor)

2016-01-01

A silicon alignment pin is used to align successive layers of components made in semiconductor chips and/or metallic components to make easier the assembly of devices having a layered structure. The pin is made as a compressible structure which can be squeezed to reduce its outer diameter, have one end fit into a corresponding alignment pocket or cavity defined in a layer of material to be assembled into a layered structure, and then allowed to expand to produce an interference fit with the cavity. The other end can then be inserted into a corresponding cavity defined in a surface of a second layer of material that mates with the first layer. The two layers are in registry when the pin is mated to both. Multiple layers can be assembled to create a multilayer structure. Examples of such devices are presented.

Rich interfacial chemistry and properties of carbon-doped hexagonal boron nitride nanosheets revealed by electronic structure calculations

NASA Astrophysics Data System (ADS)

Xie, Wei; Tamura, Takahiro; Yanase, Takashi; Nagahama, Taro; Shimada, Toshihiro

2018-04-01

The effect of C doping to hexagonal boron nitride (h-BN) to its electronic structure is examined by first principles calculations using the association from π-electron systems of organic molecules embedded in a two-dimensional insulator. In a monolayered carbon-doped structure, odd-number doping with carbon atoms confers metallic properties with different work functions. Various electronic interactions occur between two layers with odd-number carbon substitution. A direct sp3 covalent chemical bond is formed when C replaces adjacent B and N in different layers. A charge transfer complex between layers is found when C replaces B and N in the next-neighboring region, which results in narrower band gaps (e.g., 0.37 eV). Direct bonding between C and B atoms is found when two C atoms in different layers are at a certain distance.

The oesophageal zero-stress state and mucosal folding from a GIOME perspective

PubMed Central

Liao, Donghua; Zhao, Jingbo; Yang, Jian; Gregersen, Hans

2007-01-01

The oesophagus is a cylindrical organ with a collapsed lumen and mucosal folds. The mucosal folding may serve to advance the function of the oesophagus, i.e. the folds have a major influence on the flow of air and bolus through the oesophagus. Experimental studies have demonstrated oesophageal mucosal folds in the no-load state. This indicates that mucosal buckling must be considered in the analysis of the mechanical reference state since the material stiffness drops dramatically after tissue collapse. Most previous work on the oesophageal zero-stress state and mucosal folding has been experimental. However, numerical analysis offers a promising alternative approach, with the additional ability to predict the mucosal buckling behaviour and to calculate the regional stress and strain in complex structures. A numerical model used for describing the mechanical behaviour of the mucosal-folded, three-layered, two-dimensional oesophageal model is reviewed. GIOME models can be used in the future to predict the tissue function physiologically and pathologically. PMID:17457964

Forward and inverse models of electromagnetic scattering from layered media with rough interfaces

NASA Astrophysics Data System (ADS)

Tabatabaeenejad, Seyed Alireza

This work addresses the problem of electromagnetic scattering from layered dielectric structures with rough boundaries and the associated inverse problem of retrieving the subsurface parameters of the structure using the scattered field. To this end, a forward scattering model based on the Small Perturbation Method (SPM) is developed to calculate the first-order spectral-domain bistatic scattering coefficients of a two-layer rough surface structure. SPM requires the boundaries to be slightly rough compared to the wavelength, but to understand the range of applicability of this method in scattering from two-layer rough surfaces, its region of validity is investigated by comparing its output with that of a first principle solver that does not impose roughness restrictions. The Method of Moments (MoM) is used for this purpose. Finally, for retrieval of the model parameters of the layered structure using scattered field, an inversion scheme based on the Simulated Annealing method is investigated and a strategy is proposed to address convergence to local minimum.

Multiple-Step Injection Molding for Fibrin-Based Tissue-Engineered Heart Valves

PubMed Central

Weber, Miriam; Gonzalez de Torre, Israel; Moreira, Ricardo; Frese, Julia; Oedekoven, Caroline; Alonso, Matilde; Rodriguez Cabello, Carlos J.

2015-01-01

Heart valves are elaborate and highly heterogeneous structures of the circulatory system. Despite the well accepted relationship between the structural and mechanical anisotropy and the optimal function of the valves, most approaches to create tissue-engineered heart valves (TEHVs) do not try to mimic this complexity and rely on one homogenous combination of cells and materials for the whole construct. The aim of this study was to establish an easy and versatile method to introduce spatial diversity into a heart valve fibrin scaffold. We developed a multiple-step injection molding process that enables the fabrication of TEHVs with heterogeneous composition (cell/scaffold material) of wall and leaflets without the need of gluing or suturing components together, with the leaflets firmly connected to the wall. The integrity of the valves and their functionality was proved by either opening/closing cycles in a bioreactor (proof of principle without cells) or with continuous stimulation over 2 weeks. We demonstrated the potential of the method by the two-step molding of the wall and the leaflets containing different cell lines. Immunohistology after stimulation confirmed tissue formation and demonstrated the localization of the different cell types. Furthermore, we showed the proof of principle fabrication of valves using different materials for wall (fibrin) and leaflets (hybrid gel of fibrin/elastin-like recombinamer) and with layered leaflets. The method is easy to implement, does not require special facilities, and can be reproduced in any tissue-engineering lab. While it has been demonstrated here with fibrin, it can easily be extended to other hydrogels. PMID:25654448

Multiple-Step Injection Molding for Fibrin-Based Tissue-Engineered Heart Valves.

PubMed

Weber, Miriam; Gonzalez de Torre, Israel; Moreira, Ricardo; Frese, Julia; Oedekoven, Caroline; Alonso, Matilde; Rodriguez Cabello, Carlos J; Jockenhoevel, Stefan; Mela, Petra

2015-08-01

Heart valves are elaborate and highly heterogeneous structures of the circulatory system. Despite the well accepted relationship between the structural and mechanical anisotropy and the optimal function of the valves, most approaches to create tissue-engineered heart valves (TEHVs) do not try to mimic this complexity and rely on one homogenous combination of cells and materials for the whole construct. The aim of this study was to establish an easy and versatile method to introduce spatial diversity into a heart valve fibrin scaffold. We developed a multiple-step injection molding process that enables the fabrication of TEHVs with heterogeneous composition (cell/scaffold material) of wall and leaflets without the need of gluing or suturing components together, with the leaflets firmly connected to the wall. The integrity of the valves and their functionality was proved by either opening/closing cycles in a bioreactor (proof of principle without cells) or with continuous stimulation over 2 weeks. We demonstrated the potential of the method by the two-step molding of the wall and the leaflets containing different cell lines. Immunohistology after stimulation confirmed tissue formation and demonstrated the localization of the different cell types. Furthermore, we showed the proof of principle fabrication of valves using different materials for wall (fibrin) and leaflets (hybrid gel of fibrin/elastin-like recombinamer) and with layered leaflets. The method is easy to implement, does not require special facilities, and can be reproduced in any tissue-engineering lab. While it has been demonstrated here with fibrin, it can easily be extended to other hydrogels.

Voluntary exercise improves murine dermal connective tissue status in high-fat diet-induced obesity.

PubMed

Lőrincz, Kende; Haluszka, Dóra; Kiss, Norbert; Gyöngyösi, Nóra; Bánvölgyi, András; Szipőcs, Róbert; Wikonkál, Norbert M

2017-04-01

Obesity is a risk factor for several cardiovascular and metabolic diseases. Its influence on the skin is less obvious, yet certain negative effects of adipose tissue inflammation on the dermis have been suggested. Excess weight is closely associated with sedentary behavior, so any increase in physical activity is considered beneficial against obesity. To investigate the effects of obesity and physical exercise on the skin, we established a mouse model in which mice were kept either on a high-fat diet or received standard chow. After the two groups achieved a significant weight difference, physical exercise was introduced to both. Animals were given the opportunity to perform voluntary exercise for 40 min daily in a hamster wheel for a period of 8 weeks. We evaluated the status of the dermis at the beginning and at the end of the exercise period by in vivo nonlinear microscopy. Obese mice kept on high-fat diet lost weight steadily after they started to exercise. In the high-fat diet group, we could detect significantly larger adipocytes and a thicker layer of subcutaneous tissue; both changes started to normalize after exercise. Nonlinear microscopy revealed an impaired collagen structure in obese mice that improved considerably after physical activity was introduced. With the ability to detect damage on collagen structure, we set out to address the question whether this process is reversible. With the use of a novel imaging method, we were able to show the reversibility of connective tissue deterioration as a benefit of physical exercise.

Method for estimating optimal spectral and energy parameters of laser irradiation in photodynamic therapy of biological tissue

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lisenko, S A; Kugeiko, M M

We have solved the problem of layer-by-layer laser-light dosimetry in biological tissues and of selecting an individual therapeutic dose in laser therapy. A method is proposed for real-time monitoring of the radiation density in tissue layers in vivo, concentrations of its endogenous (natural) and exogenous (specially administered) chromophores, as well as in-depth distributions of the spectrum of light action on these chromophores. As the background information use is made of the spectrum of diffuse light reflected from a patient's tissue, measured by a fibre-optic spectrophotometer. The measured spectrum is quantitatively analysed by the method of approximating functions for fluxes ofmore » light multiply scattered in tissue and by a semi-analytical method for calculating the in-depth distribution of the light flux in a multi-layered medium. We have shown the possibility of employing the developed method for monitoring photosensitizer and oxyhaemoglobin concentrations in tissue, light power absorbed by chromophores in tissue layers at different depths and laser-induced changes in the tissue morphology (vascular volume content and ratios of various forms of haemoglobin) during photodynamic therapy. (biophotonics)« less

Characterization of a Honeycomb-Like Scaffold With Dielectrophoresis-Based Patterning for Tissue Engineering.

PubMed

Huan, Zhijie; Chu, Henry K; Yang, Jie; Sun, Dong

2017-04-01

Seeding and patterning of cells with an engineered scaffold is a critical process in artificial tissue construction and regeneration. To date, many engineered scaffolds exhibit simple intrinsic designs, which fail to mimic the geometrical complexity of native tissues. In this study, a novel scaffold that can automatically seed cells into multilayer honeycomb patterns for bone tissue engineering application was designed and examined. The scaffold incorporated dielectrophoresis for noncontact manipulation of cells and intrinsic honeycomb architectures were integrated in each scaffold layer. When a voltage was supplied to the stacked scaffold layers, three-dimensional electric fields were generated, thereby manipulating cells to form into honeycomb-like cellular patterns for subsequent culture. The biocompatibility of the scaffold material was confirmed through the cell viability test. Experiments were conducted to evaluate the cell viability during DEP patterning at different voltage amplitudes, frequencies, and manipulating time. Three different mammalian cells were examined and the effects of the cell size and the cell concentration on the resultant cellular patterns were evaluated. Results showed that the proposed scaffold structure was able to construct multilayer honeycomb cellular patterns in a manner similar to the natural tissue. This honeycomb-like scaffold and the dielectrophoresis-based patterning technique examined in this study could provide the field with a promising tool to enhance seeding and patterning of a wide range of cells for the development of high-quality artificial tissues.

Analytical and numerical calculations of optimum design frequency for focused ultrasound therapy and acoustic radiation force.

PubMed

Ergün, A Sanlı

2011-10-01

Focused ultrasound therapy relies on acoustic power absorption by tissue. The stronger the absorption the higher the temperature increase is. However, strong acoustic absorption also means faster attenuation and limited penetration depth. Hence, there is a trade-off between heat generation efficacy and penetration depth. In this paper, we formulated the acoustic power absorption as a function of frequency and attenuation coefficient, and defined two figures of merit to measure the power absorption: spatial peak of the acoustic power absorption density, and the acoustic power absorbed within the focal area. Then, we derived "rule of thumb" expressions for the optimum frequencies that maximized these figures of merit given the target depth and homogeneous tissue type. We also formulated a method to calculate the optimum frequency for inhomogeneous tissue given the tissue composition for situations where the tissue structure can be assumed to be made of parallel layers of homogeneous tissue. We checked the validity of the rules using linear acoustic field simulations. For a one-dimensional array of 4cm acoustic aperture, and for a two-dimensional array of 4×4cm(2) acoustic aperture, we found that the power absorbed within the focal area is maximized at 0.86MHz, and 0.79MHz, respectively, when the target depth is 4cm in muscle tissue. The rules on the other hand predicted the optimum frequencies for acoustic power absorption as 0.9MHz and 0.86MHz, respectively for the 1D and 2D array case, which are within 6% and 9% of the field simulation results. Because radiation force generated by an acoustic wave in a lossy propagation medium is approximately proportional to the acoustic power absorption, these rules can be used to maximize acoustic radiation force generated in tissue as well. Copyright © 2011 Elsevier B.V. All rights reserved.

3D bio-etching of a complex composite-like embryonic tissue.

PubMed

Hazar, Melis; Kim, Yong Tae; Song, Jiho; LeDuc, Philip R; Davidson, Lance A; Messner, William C

2015-08-21

Morphogenesis involves a complex series of cell signaling, migration and differentiation events that are coordinated as tissues self-assemble during embryonic development. Collective cell movements such as those that occur during morphogenesis have typically been studied in 2D with single layers of cultured cells adhering to rigid substrates such as glass or plastic. In vivo, the intricacies of the 3D microenvironment and complex 3D responses are pivotal in the formation of functional tissues. To study such processes as collective cell movements within 3D multilayered tissues, we developed a microfluidic technique capable of producing complex 3D laminar multicellular structures. We call this technique "3D tissue-etching" because it is analogous to techniques used in the microelectromechanics (MEMS) field where complex 3D structures are built by successively removing material from a monolithic solid through subtractive manufacturing. We use a custom-designed microfluidic control system to deliver a range of tissue etching reagents (detergents, chelators, proteases, etc.) to specific regions of multilayered tissues. These tissues were previously isolated by microsurgical excision from embryos of the African claw-toed frog, Xenopus laevis. The ability to shape the 3D form of multicellular tissues and to control 3D stimulation will have a high impact on tissue engineering and regeneration applications in bioengineering and medicine as well as provide significant improvements in the synthesis of highly complex 3D integrated multicellular biosystems.

Interaction of intraluminal tissue and coronary sinus lead stabilized with stent placement.

PubMed

Balázs, Tibor; Merkely, Béla; Bognár, Eszter; Zima, Endre

2013-04-01

The aim of our investigation was to examine the intraluminal interaction of the vascular tissue and the implanted coronary sinus lead stabilized with stent on two human hearts removed before transplantation. The coronary sinus lumen was sectioned under operational microscope and opened carefully. The leads and stents were found separately positioned beside each other completely covered by an intact intimal tissue layer. No sign of occluding proliferative tissue was observed. Stent fixation technique and extraction of the CS lead in our cases did not have any particular damaging effect on the vascular system. © 2012 Wiley Periodicals, Inc.

The muscular force transmission system: role of the intramuscular connective tissue.

PubMed

Turrina, Andrea; Martínez-González, Miguel Antonio; Stecco, Carla

2013-01-01

The objective of this review is to analyze in detail the microscopic structure and relations among muscular fibers, endomysium, perimysium, epimysium and deep fasciae. In particular, the multilayer organization and the collagen fiber orientation of these elements are reported. The endomysium, perimysium, epimysium and deep fasciae have not just a role of containment, limiting the expansion of the muscle with the disposition in concentric layers of the collagen tissue, but are fundamental elements for the transmission of muscular force, each one with a specific role. From this review it appears that the muscular fibers should not be studied as isolated elements, but as a complex inseparable from their fibrous components. The force expressed by a muscle depends not only on its anatomical structure, but also the angle at which its fibers are attached to the intramuscular connective tissue and the relation with the epimysium and deep fasciae. Copyright © 2012 Elsevier Ltd. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Carneiro, Karina M. M.; Zhai, Halei; Zhu, Li

We report that enamel, the outermost layer of teeth, is an acellular mineralized tissue that cannot regenerate; the mature tissue is composed of high aspect ratio apatite nanocrystals organized into rods and inter-rod regions. Amelogenin constitutes 90% of the protein matrix in developing enamel and plays a central role in guiding the hierarchical organization of apatite crystals observed in mature enamel. To date, a convincing link between amelogenin supramolecular structures and mature enamel has yet to be described, in part because the protein matrix is degraded during tissue maturation. Here we show compelling evidence that amelogenin self-assembles into an amyloid-likemore » structure in vitro and in vivo. We show that enamel matrices stain positive for amyloids and we identify a specific region within amelogenin that self-assembles into β-sheets. Lastly, we propose that amelogenin nanoribbons template the growth of apatite mineral in human enamel. This is a paradigm shift from the current model of enamel development.« less

Design of a thermal waist-pad

NASA Astrophysics Data System (ADS)

Kursun Bahadir, S.; Sahin, U. K.; Acikgoz Tufan, H.

2017-10-01

The objective of the current study is designing a thermal waist-pad for people who have backaches with a sandwich-like multi-layered structure. Two model is developed; one is three-layered and second is five-layered with waterproof woven outer layer fabric, Thermolite® knitted fabric (for five-layered structures), wool knitted, polyester nonwoven fabric, polypropylene nonwoven fabric and viscose nonwoven fabric for mid-layer. 10 different structures are designed and produced. All samples are tested for thermal comfort properties of waist-pad. Multi-layer structures were tested, and according to their thermal performance and thermal comfort criteria, all results are evaluated for identifying the best product. These three factors are examined by analysis of thermal conductivity, thermal resistance, thermal absorptivity, relative water vapour/air permeability, water absorption. Highest thermal resistance test result, 150,42 mK/Wm2, is achieved in five-layered sandwich structure with waterproof fabric, Thermolite® fabric, wool based knitted fabric, Thermolite® fabric and waterproof fabric, respectively. Thermal conductivity result of this structure is 46,2 mW/mK, which is one of the lowest results among the alternative structures. Structures with Thermolite® fabric show higher thermal comfort when compared to others.

EFFECTS OF CYCLIC FLEXURAL FATIGUE ON PORCINE BIOPROSTHETIC HEART VALVE HETEROGRAFT BIOMATERIALS

PubMed Central

Mirnajafi, Ali; Zubiate, Brett; Sacks, Michael S.

2009-01-01

While bioprosthetic heart valves (BHV) remain the primary treatment modality for adult heart valve replacement, continued problems with durability remain. Several studies have implicated flexure as a major damage mode in porcine-derived heterograft biomaterials used in BHV fabrication. While conventional accelerated wear testing can provide valuable insights into BHV damage phenomena, the constituent tissues are subjected to complex, time-varying deformation modes (i.e. tension and flexure), that do not allow for the control of the amount, direction, and location of flexure. Thus, in the present study customized fatigue testing devices were developed to subject circumferentially oriented porcine BHV tissue strips to controlled cyclic flexural loading. By using this approach, we were able to study layer-specific structural damage induced by cyclic flexural tensile and compressive stresses alone. 10×106, 25×106 and 50×106 cycle levels were used, with resulting changes in flexural stiffness and collagen structure assessed. Results indicated that flexural rigidity was markedly reduced after only 10×106 cycles, and progressively decayed at a lower rate with cycle number thereafter. Moreover, the against-curvature fatigue direction induced the most damage, suggesting that the ventricularis and fibrosa layers have low resistance to cyclic flexural compressive and tensile loads, respectively. The histological analyses indicated progressive collagen fiber delamination as early as 10×106 cycles, but otherwise no change in gross collagen orientation. Our results underscore that porcine-derived heterograft biomaterials are very sensitive to flexural fatigue, with delamination of the tissue layers the primary underlying mechanism. This appears to be in contrast to pericardial BHV, wherein high tensile stresses are considered to be the major cause of structural failure. These findings point towards the need for the development of chemical fixation technologies that minimize flexure induced damage to extend porcine heterograft biomaterial durability. PMID:20166221

Reconstructing in-vivo reflectance spectrum of pigmented skin lesion by Monte Carlo simulation

NASA Astrophysics Data System (ADS)

Wang, Shuang; He, Qingli; Zhao, Jianhua; Lui, Harvey; Zeng, Haishan

2012-03-01

In dermatology applications, diffuse reflectance spectroscopy has been extensively investigated as a promising tool for the noninvasive method to distinguish melanoma from benign pigmented skin lesion (nevus), which is concentrated with the skin chromophores like melanin and hemoglobin. We carried out a theoretical study to examine melanin distribution in human skin tissue and establish a practical optical model for further pigmented skin investigation. The theoretical simulation was using junctional nevus as an example. A multiple layer skin optical model was developed on established anatomy structures of skin, the published optical parameters of different skin layers, blood and melanin. Monte Carlo simulation was used to model the interaction between excitation light and skin tissue and rebuild the diffuse reflectance process from skin tissue. A testified methodology was adopted to determine melanin contents in human skin based on in vivo diffuse reflectance spectra. The rebuild diffuse reflectance spectra were investigated by adding melanin into different layers of the theoretical model. One of in vivo reflectance spectra from Junctional nevi and their surrounding normal skin was studied by compare the ratio between nevus and normal skin tissue in both the experimental and simulated diffuse reflectance spectra. The simulation result showed a good agreement with our clinical measurements, which indicated that our research method, including the spectral ratio method, skin optical model and modifying the melanin content in the model, could be applied in further theoretical simulation of pigmented skin lesions.

Hydrogel-laden paper scaffold system for origami-based tissue engineering

PubMed Central

Kim, Su-Hwan; Lee, Hak Rae; Yu, Seung Jung; Han, Min-Eui; Lee, Doh Young; Kim, Soo Yeon; Ahn, Hee-Jin; Han, Mi-Jung; Lee, Tae-Ik; Kim, Taek-Soo; Kwon, Seong Keun; Im, Sung Gap; Hwang, Nathaniel S.

2015-01-01

In this study, we present a method for assembling biofunctionalized paper into a multiform structured scaffold system for reliable tissue regeneration using an origami-based approach. The surface of a paper was conformally modified with a poly(styrene-co-maleic anhydride) layer via initiated chemical vapor deposition followed by the immobilization of poly-l-lysine (PLL) and deposition of Ca2+. This procedure ensures the formation of alginate hydrogel on the paper due to Ca2+ diffusion. Furthermore, strong adhesion of the alginate hydrogel on the paper onto the paper substrate was achieved due to an electrostatic interaction between the alginate and PLL. The developed scaffold system was versatile and allowed area-selective cell seeding. Also, the hydrogel-laden paper could be folded freely into 3D tissue-like structures using a simple origami-based method. The cylindrically constructed paper scaffold system with chondrocytes was applied into a three-ring defect trachea in rabbits. The transplanted engineered tissues replaced the native trachea without stenosis after 4 wks. As for the custom-built scaffold system, the hydrogel-laden paper system will provide a robust and facile method for the formation of tissues mimicking native tissue constructs. PMID:26621717

Hydrogel-laden paper scaffold system for origami-based tissue engineering.

PubMed

Kim, Su-Hwan; Lee, Hak Rae; Yu, Seung Jung; Han, Min-Eui; Lee, Doh Young; Kim, Soo Yeon; Ahn, Hee-Jin; Han, Mi-Jung; Lee, Tae-Ik; Kim, Taek-Soo; Kwon, Seong Keun; Im, Sung Gap; Hwang, Nathaniel S

2015-12-15

In this study, we present a method for assembling biofunctionalized paper into a multiform structured scaffold system for reliable tissue regeneration using an origami-based approach. The surface of a paper was conformally modified with a poly(styrene-co-maleic anhydride) layer via initiated chemical vapor deposition followed by the immobilization of poly-l-lysine (PLL) and deposition of Ca(2+). This procedure ensures the formation of alginate hydrogel on the paper due to Ca(2+) diffusion. Furthermore, strong adhesion of the alginate hydrogel on the paper onto the paper substrate was achieved due to an electrostatic interaction between the alginate and PLL. The developed scaffold system was versatile and allowed area-selective cell seeding. Also, the hydrogel-laden paper could be folded freely into 3D tissue-like structures using a simple origami-based method. The cylindrically constructed paper scaffold system with chondrocytes was applied into a three-ring defect trachea in rabbits. The transplanted engineered tissues replaced the native trachea without stenosis after 4 wks. As for the custom-built scaffold system, the hydrogel-laden paper system will provide a robust and facile method for the formation of tissues mimicking native tissue constructs.

The in vitro and in vivo influence of 4-META/MMA-TBB resin components on dental pulp tissues.

PubMed

Inoue, T; Miyakoshi, S; Shimono, M

2001-08-01

The purpose of this study was to qualitate the penetration of the major components of 4-META/MMA-TBB adhesive resin (4-META resin) and to characterize their influence on the in vitro and in vivo wound healing of dental pulp tissues. Fresh 4-META resin was applied to rabbit mesentery; its components penetrated the mesentery to form three of layers, depending on the amounts of monomer components in the tissue. The superficial layer was a soft-tissue hybrid layer (STHL), the intermediate layer contained small particles of polymerized 4-META resin, while the deepest layer contained unpolymerized monomer components including MMA and butanol, which were detected by gas chromatography (GC). To characterize the in vivo effects of the deepest layer, we immersed the pulp tissue in MMA or in 5% 4-META/MMA and autotransplanted it to placement beneath a rabbit kidney capsule. The MMA-immersed pulp was positive for osteocalcin and presented osteodentin formation at 7 days, as did the untreated control pulp tissue. In contrast, the 5% 4-META/MMA-immersed pulp collapsed into the cell-deficient fibrous connective tissue, with slight calcification by 7 days and less osteodentin formation at 14 days. Analysis of these data suggests that MMA does not inhibit osteogenic activity of pulp tissue, while 5% 4-META/MMA does inhibit osteogenic activity to some extent.

3D topography of biologic tissue by multiview imaging and structured light illumination

NASA Astrophysics Data System (ADS)

Liu, Peng; Zhang, Shiwu; Xu, Ronald

2014-02-01

Obtaining three-dimensional (3D) information of biologic tissue is important in many medical applications. This paper presents two methods for reconstructing 3D topography of biologic tissue: multiview imaging and structured light illumination. For each method, the working principle is introduced, followed by experimental validation on a diabetic foot model. To compare the performance characteristics of these two imaging methods, a coordinate measuring machine (CMM) is used as a standard control. The wound surface topography of the diabetic foot model is measured by multiview imaging and structured light illumination methods respectively and compared with the CMM measurements. The comparison results show that the structured light illumination method is a promising technique for 3D topographic imaging of biologic tissue.

Modeling ultrasonic transient scattering from biological tissues including their dispersive properties directly in the time domain.

PubMed

Norton, G V; Novarini, J C

2007-06-01

Ultrasonic imaging in medical applications involves propagation and scattering of acoustic waves within and by biological tissues that are intrinsically dispersive. Analytical approaches for modeling propagation and scattering in inhomogeneous media are difficult and often require extremely simplifying approximations in order to achieve a solution. To avoid such approximations, the direct numerical solution of the wave equation via the method of finite differences offers the most direct tool, which takes into account diffraction and refraction. It also allows for detailed modeling of the real anatomic structure and combination/layering of tissues. In all cases the correct inclusion of the dispersive properties of the tissues can make the difference in the interpretation of the results. However, the inclusion of dispersion directly in the time domain proved until recently to be an elusive problem. In order to model the transient signal a convolution operator that takes into account the dispersive characteristics of the medium is introduced to the linear wave equation. To test the ability of this operator to handle scattering from localized scatterers, in this work, two-dimensional numerical modeling of scattering from an infinite cylinder with physical properties associated with biological tissue is calculated. The numerical solutions are compared with the exact solution synthesized from the frequency domain for a variety of tissues having distinct dispersive properties. It is shown that in all cases, the use of the convolutional propagation operator leads to the correct solution for the scattered field.

Seeking: Accurate Measurement Techniques for Deep-Bone Density and Structure

NASA Technical Reports Server (NTRS)

Sibonga, Jean

2009-01-01

We are seeking a clinically-useful technology with enough sensitivity to assess the microstructure of "spongy" bone that is found in the marrow cavities of whole bones. However, this technology must be for skeletal sites surrounded by layers of soft tissues, such as the spine and the hip. Soft tissue interferes with conventional imaging and using a more accessible area -- for example, the wrist or the ankle of limbs-- as a proxy for the less accessible skeletal regions, will not be accurate. A non-radioactive technology is strongly preferred.

Ordering transitions of weakly anisotropic hard rods in narrow slitlike pores.

PubMed

Aliabadi, Roohollah; Gurin, Péter; Velasco, Enrique; Varga, Szabolcs

2018-01-01

The effect of strong confinement on the positional and orientational ordering is examined in a system of hard rectangular rods with length L and diameter D (L>D) using the Parsons-Lee modification of the second virial density-functional theory. The rods are nonmesogenic (L/D<3) and confined between two parallel hard walls, where the width of the pore (H) is chosen in such a way that both planar (particle's long axis parallel to the walls) and homeotropic (particle's long axis perpendicular to the walls) orderings are possible and a maximum of two layers is allowed to form in the pore. In the extreme confinement limit of H≤2D, where only one-layer structures appear, we observe a structural transition from a planar to a homeotropic fluid layer with increasing density, which becomes sharper as L→H. In wider pores (2D

Calculation of metamorphic two-dimensional quantum energy system: Application to wetting layer states in InAs/InGaAs metamorphic quantum dot nanostructures

DOE Office of Scientific and Technical Information (OSTI.GOV)

Seravalli, L.; Trevisi, G.; Frigeri, P.

In this work, we calculate the two-dimensional quantum energy system of the In(Ga)As wetting layer that arises in InAs/InGaAs/GaAs metamorphic quantum dot structures. Model calculations were carried on the basis of realistic material parameters taking in consideration their dependence on the strain relaxation of the metamorphic buffer; results of the calculations were validated against available literature data. Model results confirmed previous hypothesis on the extrinsic nature of the disappearance of wetting layer emission in metamorphic structures with high In composition. We also show how, by adjusting InGaAs metamorphic buffer parameters, it could be possible: (i) to spatially separate carriers confinedmore » in quantum dots from wetting layer carriers, (ii) to create an hybrid 0D-2D system, by tuning quantum dot and wetting layer levels. These results are interesting not only for the engineering of quantum dot structures but also for other applications of metamorphic structures, as the two design parameters of the metamorphic InGaAs buffer (thickness and composition) provide additional degrees of freedom to control properties of interest.« less

[Feasibility of using connective tissue prosthesis for autoplastic repair of urinary bladder wall defects (an experimental study)].

PubMed

Tyumentseva, N V; Yushkov, B G; Medvedeva, S Y; Kovalenko, R Y; Uzbekov, O K; Zhuravlev, V N

2016-12-01

Experiments on laboratory rats have shown the feasibility of autoplastic repair of urinary bladder wall defects using a connective-tissue capsule formed as the result of an inflammatory response to the presence of a foreign body. The formation of connective tissue prosthesis is characterized by developing fibrous connective tissue, ordering of collagen fibers, reducing the number of cells per unit area with a predominance of more mature cells - fibroblasts. With increasing time of observation, connective tissue prostheses were found to acquire a morphological structure similar to that of the urinary bladder wall. By month 12, the mucosa, the longitudinal and circular muscle layers were formed. The proposed method of partial autoplastic repair of urinary bladder wall is promising, has good long-term results, but requires further experimental studies.

A novel whole tooth-in-jaw-bone culture of rat molars: morphological, immunohistochemical, and laser capture microdissection analysis.

PubMed

Chokechanachaisakul, Uraiwan; Kaneko, Tomoatsu; Yamanaka, Yusuke; Okiji, Takashi; Suda, Hideaki

2012-10-01

In conventional whole-tooth culture systems, limitation exists regarding maintenance of the vitality of the dental pulp, because this tissue is encased in rigid dentin walls that hinder nutrition supply. We here report a whole tooth-in-jaw-bone culture system of rat mandibular first molars, where transcardiac perfusion with culture medium was carried out before placement of the jaw bone into culture medium, aiming to facilitate longer time preservation of the dental pulp tissue. Following 7 days of culture, the pulp tissues were analyzed by histology and immunohistochemistry to ED2 (antiresident macrophage). ED2-positive macrophages were also analyzed for their Class II MHC, interleukin-6 (IL-6), and p53 mRNA expression levels by means of immune-laser capture microdissection (immune-LCM). Dentin sialophosphoprotein (DSPP) mRNA expression in odontobalstic layer was also examined by LCM. Teeth cultured following saline-perfusion and nonperfusion served as cultured controls. Normal teeth also served as noncultured controls. Histological examination demonstrated that the structure of the pulp tissue was well preserved in the medium-perfused explants in contrast to the cultured control groups. The Class II MHC, IL-6, and p53 mRNA expression levels of ED2-positive cells and DSPP expression levels of odontoblastic layer tissues in the pulp of medium-perfused explants were not significantly different from those in the noncultured normal teeth. In conclusion, the structural integrity and mRNA expression in the pulp were maintained at the in vivo level in the ex vivo whole tooth-in-jaw-bone culture system. The system may lay the foundation for studies aiming at defining further histological and molecular mechanism of the pulp. Copyright © 2012 Wiley Periodicals, Inc.

Two-dimensional protein crystals (S-layers): fundamentals and applications.

PubMed

Sleytr, U B; Sára, M; Messner, P; Pum, D

1994-10-01

Two-dimensional crystalline surface layers (S-layers) composed of protein or glycoprotein subunits are one of the most commonly observed prokaryotic cell envelope structures. Isolated S-layer subunits are endowed with the ability to assemble into monomolecular arrays in suspension, on surfaces or interfaces by an entropy-driven process. S-layer lattices are isoporous structures with functional groups located on the surface in an identical position and orientation. These characteristic features have already led to applications of S-layers as (1) ultrafiltration membranes with well-defined molecular weight cut-offs and excellent antifouling characteristics, (2) immobilization matrices for functional molecules as required for affinity and enzyme membranes, affinity microcarriers and biosensors, (3) conjugate vaccines, (4) carriers for Langmuir-Blodgett films and reconstituted biological membranes, and (5) patterning elements in molecular nanotechnology.

Understanding the Biological Basis of Autofluorescence Imaging for Oral Cancer Detection: High-Resolution Fluorescence Microscopy in Viable Tissue

PubMed Central

Pavlova, Ina; Williams, Michelle; El-Naggar, Adel; Richards-Kortum, Rebecca; Gillenwater, Ann

2009-01-01

Purpose Autofluorescence imaging is increasingly used to noninvasively identify neoplastic oral cavity lesions. Improving the diagnostic accuracy of these techniques requires a better understanding of the biological basis for optical changes associated with neoplastic transformation in oral tissue. Experimental Design A total of 49 oral biopsies were considered in this study. The autofluorescence patterns of viable normal, benign, and neoplastic oral tissue were imaged using high-resolution confocal fluorescence microscopy. Results The autofluorescence properties of oral tissue vary significantly based on anatomic site and pathologic diagnosis. In normal oral tissue, most of the epithelial autofluorescence originates from the cytoplasm of cells in the basal and intermediate regions, whereas structural fibers are responsible for most of the stromal fluorescence. A strongly fluorescent superficial layer was observed in tissues from the palate and the gingiva, which contrasts with the weakly fluorescent superficial layer found in other oral sites. Upon UV excitation, benign inflammation shows decreased epithelial fluorescence, whereas dysplasia displays increased epithelial fluorescence compared with normal oral tissue. Stromal fluorescence in both benign inflammation and dysplasia drops significantly at UV and 488 nm excitation. Conclusion Imaging oral lesions with optical devices/probes that sample mostly stromal fluorescence may result in a similar loss of fluorescence intensity and may fail to distinguish benign from precancerous lesions. Improved diagnostic accuracy may be achieved by designing optical probes/devices that distinguish epithelial fluorescence from stromal fluorescence and by using excitation wavelengths in the UV range. PMID:18413830

Interfacial ionic 'liquids': connecting static and dynamic structures

DOE PAGES

Uysal, Ahmet; Zhou, Hua; Feng, Guang; ...

2014-12-05

It is well known that room temperature ionic liquids (RTILs) often adopt a charge-separated layered structure, i.e. with alternating cation- and anion-rich layers, at electrified interfaces. However, the dynamic response of the layered structure to temporal variations in applied potential is not well understood. For this research, we used in situ, real-time x-ray reflectivity to study the potential-dependent electric double layer (EDL) structure of an imidazolium-based RTIL on charged epitaxial graphene during potential cycling as a function of temperature. The results suggest that the graphene–RTIL interfacial structure is bistable in which the EDL structure at any intermediate potential can bemore » described by the combination of two extreme-potential structures whose proportions vary depending on the polarity and magnitude of the applied potential. This picture is supported by the EDL structures obtained by fully atomistic molecular dynamics simulations at various static potentials. Lastly, the potential-driven transition between the two structures is characterized by an increasing width but with an approximately fixed hysteresis magnitude as a function of temperature. The results are consistent with the coexistence of distinct anion- and cation-adsorbed structures separated by an energy barrier (~0.15 eV).« less

Interfacial ionic 'liquids': connecting static and dynamic structures.

PubMed

Uysal, Ahmet; Zhou, Hua; Feng, Guang; Lee, Sang Soo; Li, Song; Cummings, Peter T; Fulvio, Pasquale F; Dai, Sheng; McDonough, John K; Gogotsi, Yury; Fenter, Paul

2015-01-28

It is well known that room temperature ionic liquids (RTILs) often adopt a charge-separated layered structure, i.e. with alternating cation- and anion-rich layers, at electrified interfaces. However, the dynamic response of the layered structure to temporal variations in applied potential is not well understood. We used in situ, real-time x-ray reflectivity to study the potential-dependent electric double layer (EDL) structure of an imidazolium-based RTIL on charged epitaxial graphene during potential cycling as a function of temperature. The results suggest that the graphene-RTIL interfacial structure is bistable in which the EDL structure at any intermediate potential can be described by the combination of two extreme-potential structures whose proportions vary depending on the polarity and magnitude of the applied potential. This picture is supported by the EDL structures obtained by fully atomistic molecular dynamics simulations at various static potentials. The potential-driven transition between the two structures is characterized by an increasing width but with an approximately fixed hysteresis magnitude as a function of temperature. The results are consistent with the coexistence of distinct anion- and cation-adsorbed structures separated by an energy barrier (∼0.15 eV).

Triangular Black Phosphorus Atomic Layers by Liquid Exfoliation.

PubMed

Seo, Soonjoo; Lee, Hyun Uk; Lee, Soon Chang; Kim, Yooseok; Kim, Hyeran; Bang, Junhyeok; Won, Jonghan; Kim, Youngjun; Park, Byoungnam; Lee, Jouhahn

2016-03-30

Few-layer black phosphorus (BP) is the most promising material among the two-dimensional materials due to its layered structure and the excellent semiconductor properties. Currently, thin BP atomic layers are obtained mostly by mechanical exfoliation of bulk BP, which limits applications in thin-film based electronics due to a scaling process. Here we report highly crystalline few-layer black phosphorus thin films produced by liquid exfoliation. We demonstrate that the liquid-exfoliated BP forms a triangular crystalline structure on SiO2/Si (001) and amorphous carbon. The highly crystalline BP layers are faceted with a preferred orientation of the (010) plane on the sharp edge, which is an energetically most favorable facet according to the density functional theory calculations. Our results can be useful in understanding the triangular BP structure for large-area applications in electronic devices using two-dimensional materials. The sensitivity and selectivity of liquid-exfoliated BP to gas vapor demonstrate great potential for practical applications as sensors.

Triangular Black Phosphorus Atomic Layers by Liquid Exfoliation

PubMed Central

Seo, Soonjoo; Lee, Hyun Uk; Lee, Soon Chang; Kim, Yooseok; Kim, Hyeran; Bang, Junhyeok; Won, Jonghan; Kim, Youngjun; Park, Byoungnam; Lee, Jouhahn

2016-01-01

Few-layer black phosphorus (BP) is the most promising material among the two-dimensional materials due to its layered structure and the excellent semiconductor properties. Currently, thin BP atomic layers are obtained mostly by mechanical exfoliation of bulk BP, which limits applications in thin-film based electronics due to a scaling process. Here we report highly crystalline few-layer black phosphorus thin films produced by liquid exfoliation. We demonstrate that the liquid-exfoliated BP forms a triangular crystalline structure on SiO2/Si (001) and amorphous carbon. The highly crystalline BP layers are faceted with a preferred orientation of the (010) plane on the sharp edge, which is an energetically most favorable facet according to the density functional theory calculations. Our results can be useful in understanding the triangular BP structure for large-area applications in electronic devices using two-dimensional materials. The sensitivity and selectivity of liquid-exfoliated BP to gas vapor demonstrate great potential for practical applications as sensors. PMID:27026070

Rapid Characterization of Molecular Chemistry, Nutrient Make-Up and Microlocation of Internal Seed Tissue

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yu,P.; Block, H.; Niu, Z.

2007-01-01

Wheat differs from corn in biodegradation kinetics and fermentation characteristics. Wheat exhibits a relatively high rate (23% h{sup 01}) and extent (78% DM) of biodegradation, which can lead to metabolic problems such as acidosis and bloat in ruminants. The objective of this study was to rapidly characterize the molecular chemistry of the internal structure of wheat (cv. AC Barrie) and reveal both its structural chemical make-up and nutrient component matrix by analyzing the intensity and spatial distribution of molecular functional groups within the intact seed using advanced synchrotron-powered Fourier transform infrared (FTIR) microspectroscopy. The experiment was performed at the U2Bmore » station of the National Synchrotron Light Source at Brookhaven National Laboratory, New York, USA. The wheat tissue was imaged systematically from the pericarp, seed coat, aleurone layer and endosperm under the peaks at {approx}1732 (carbonyl C{double_bond}O ester), 1515 (aromatic compound of lignin), 1650 (amide I), 1025 (non-structural CHO), 1550 (amide II), 1246 (cellulosic material), 1160, 1150, 1080, 930, 860 (all CHO), 3350 (OH and NH stretching), 2928 (CH{sub 2} stretching band) and 2885 cm{sup -1} (CH{sub 3} stretching band). Hierarchical cluster analysis and principal component analysis were applied to analyze the molecular FTIR spectra obtained from the different inherent structures within the intact wheat tissues. The results showed that, with synchrotron-powered FTIR microspectroscopy, images of the molecular chemistry of wheat could be generated at an ultra-spatial resolution. The features of aromatic lignin, structural and non-structural carbohydrates, as well as nutrient make-up and interactions in the seeds, could be revealed. Both principal component analysis and hierarchical cluster analysis methods are conclusive in showing that they can discriminate and classify the different inherent structures within the seed tissue. The wheat exhibited distinguishable differences in the structural and nutrient make-up among the pericarp, seed coat, aleurone layer and endosperm. Such information on the molecular chemistry can be used for grain-breeding programs for selecting a superior variety of wheat targeted for food and feed purposes and for predicting wheat quality and nutritive value in humans and animals. Thus advanced synchrotron-powered FTIR technology can provide a greater understanding of the plant-animal interface.« less

Electrostatic repulsive out-of-plane actuator using conductive substrate.

PubMed

Wang, Weimin; Wang, Qiang; Ren, Hao; Ma, Wenying; Qiu, Chuankai; Chen, Zexiang; Fan, Bin

2016-10-07

A pseudo-three-layer electrostatic repulsive out-of-plane actuator is proposed. It combines the advantages of two-layer and three-layer repulsive actuators, i.e., fabrication requirements and fill factor. A theoretical model for the proposed actuator is developed and solved through the numerical calculation of Schwarz-Christoffel mapping. Theoretical and simulated results show that the pseudo-three-layer actuator offers higher performance than the two-layer and three-layer actuators with regard to the two most important characteristics of actuators, namely, driving force and theoretical stroke. Given that the pseudo-three-layer actuator structure is compatible with both the parallel-plate actuators and these two types of repulsive actuators, a 19-element two-layer repulsive actuated deformable mirror is operated in pseudo-three-layer electrical connection mode. Theoretical and experimental results demonstrate that the pseudo-three-layer mode produces a larger displacement of 0-4.5 μm for a dc driving voltage of 0-100 V, when compared with that in two-layer mode.

Electrostatic repulsive out-of-plane actuator using conductive substrate

PubMed Central

Wang, Weimin; Wang, Qiang; Ren, Hao; Ma, Wenying; Qiu, Chuankai; Chen, Zexiang; Fan, Bin

2016-01-01

A pseudo-three-layer electrostatic repulsive out-of-plane actuator is proposed. It combines the advantages of two-layer and three-layer repulsive actuators, i.e., fabrication requirements and fill factor. A theoretical model for the proposed actuator is developed and solved through the numerical calculation of Schwarz-Christoffel mapping. Theoretical and simulated results show that the pseudo-three-layer actuator offers higher performance than the two-layer and three-layer actuators with regard to the two most important characteristics of actuators, namely, driving force and theoretical stroke. Given that the pseudo-three-layer actuator structure is compatible with both the parallel-plate actuators and these two types of repulsive actuators, a 19-element two-layer repulsive actuated deformable mirror is operated in pseudo-three-layer electrical connection mode. Theoretical and experimental results demonstrate that the pseudo-three-layer mode produces a larger displacement of 0–4.5 μm for a dc driving voltage of 0–100 V, when compared with that in two-layer mode. PMID:27713542

Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye

PubMed Central

Sharma, Robin; Williams, David R.; Palczewska, Grazyna; Palczewski, Krzysztof; Hunter, Jennifer J.

2016-01-01

Purpose Although extrinsic fluorophores can be introduced to label specific cell types in the retina, endogenous fluorophores, such as NAD(P)H, FAD, collagen, and others, are present in all retinal layers. These molecules are a potential source of optical contrast and can enable noninvasive visualization of all cellular layers. We used a two-photon fluorescence adaptive optics scanning light ophthalmoscope (TPF-AOSLO) to explore the native autofluorescence of various cell classes spanning several layers in the unlabeled retina of a living primate eye. Methods Three macaques were imaged on separate occasions using a custom TPF-AOSLO. Two-photon fluorescence was evoked by pulsed light at 730 and 920 nm excitation wavelengths, while fluorescence emission was collected in the visible range from several retinal layers and different locations. Backscattered light was recorded simultaneously in confocal modality and images were postprocessed to remove eye motion. Results All retinal layers yielded two-photon signals and the heterogeneous distribution of fluorophores provided optical contrast. Several structural features were observed, such as autofluorescence from vessel walls, Müller cell processes in the nerve fibers, mosaics of cells in the ganglion cell and other nuclear layers of the inner retina, as well as photoreceptor and RPE layers in the outer retina. Conclusions This in vivo survey of two-photon autofluorescence throughout the primate retina demonstrates a wider variety of structural detail in the living eye than is available through conventional imaging methods, and broadens the use of two-photon imaging of normal and diseased eyes. PMID:26903224

Monte Carlo modeling of time-resolved fluorescence for depth-selective interrogation of layered tissue.

PubMed

Pfefer, T Joshua; Wang, Quanzeng; Drezek, Rebekah A

2011-11-01

Computational approaches for simulation of light-tissue interactions have provided extensive insight into biophotonic procedures for diagnosis and therapy. However, few studies have addressed simulation of time-resolved fluorescence (TRF) in tissue and none have combined Monte Carlo simulations with standard TRF processing algorithms to elucidate approaches for cancer detection in layered biological tissue. In this study, we investigate how illumination-collection parameters (e.g., collection angle and source-detector separation) influence the ability to measure fluorophore lifetime and tissue layer thickness. Decay curves are simulated with a Monte Carlo TRF light propagation model. Multi-exponential iterative deconvolution is used to determine lifetimes and fractional signal contributions. The ability to detect changes in mucosal thickness is optimized by probes that selectively interrogate regions superficial to the mucosal-submucosal boundary. Optimal accuracy in simultaneous determination of lifetimes in both layers is achieved when each layer contributes 40-60% of the signal. These results indicate that depth-selective approaches to TRF have the potential to enhance disease detection in layered biological tissue and that modeling can play an important role in probe design optimization. Published by Elsevier Ireland Ltd.

Permeability of nanonet structures constructed on the basis of carbon tubes

NASA Astrophysics Data System (ADS)

Bubenchikov, M. A.; Nikipelova, T. I.; Tsyrenova, V. B.; Chelnokova, A. S.

2017-11-01

An approximate integration of a potential of paired molecular interactions over the circumferential coordinate is used to find the potential of the interaction between a molecule and an infinite carbon nanotube (CNT). Based on the obtained energy of interaction between the tube and the molecules, local effective radii of the investigated carbon structures with respect to the molecules of the separated gas mixture are found. This makes it possible to calculate permeability of a single-layer, along with a two-layer CNT packing. The conducted research allowed calculating permeability of a nanonet structure as permeability of a two-layer packing of carbon nanotubes for the first time.

Is the stripping technique a tissue-sparing procedure in large simple ovarian cysts in children?

PubMed

Arena, Francesco; Romeo, Carmelo; Castagnetti, Marco; Scalfari, GianFranco; Cimador, Marcello; Impellizzeri, Pietro; Villari, Daniela; Zimbaro, Fabrizio; DeGrazia, Enrico

2008-07-01

Stripping of the cystic wall is performed by gynecologists to treat large ovarian cysts. Information in the pediatric population is poor. We prospectively evaluated the pathologic specimens of large ovarian cyst to determine whether the stripping technique is a tissue-sparing procedure even in this age. We evaluated 5 patients. Samples were taken from the intermediate part of the cystic wall and from the layer covering the cyst during excision. The presence of ovarian tissue adjacent to the cyst wall, and the morphological features of the surrounding tissue were both evaluated. Pelvic ultrasound follow-up was also performed. Patients' mean age was 4.5 years (7 days to 12 years). All cysts were removed because all were symptomatic. The mean diameter was 86.6 mm (74-100 mm). Cysts were follicular in 2 cases, serous in other two, and endometriotic in 1 case. Adjacent ovarian tissue was present in 1 of 5 specimens and was approximately 1 to 2 mm in thickness. The layer adjacent to the cystic wall always appeared as normal ovarian tissue. Ultrasound scans at follow-up revealed presence of ovarian tissue. The stripping procedure for large ovarian cyst excision allows to spare the adjacent normal ovarian tissue even in pediatric age because ovarian tissue is rarely excised with the cyst wall during the procedure.

Application of structured support vector machine backpropagation to a convolutional neural network for human pose estimation.

PubMed

Witoonchart, Peerajak; Chongstitvatana, Prabhas

2017-08-01

In this study, for the first time, we show how to formulate a structured support vector machine (SSVM) as two layers in a convolutional neural network, where the top layer is a loss augmented inference layer and the bottom layer is the normal convolutional layer. We show that a deformable part model can be learned with the proposed structured SVM neural network by backpropagating the error of the deformable part model to the convolutional neural network. The forward propagation calculates the loss augmented inference and the backpropagation calculates the gradient from the loss augmented inference layer to the convolutional layer. Thus, we obtain a new type of convolutional neural network called an Structured SVM convolutional neural network, which we applied to the human pose estimation problem. This new neural network can be used as the final layers in deep learning. Our method jointly learns the structural model parameters and the appearance model parameters. We implemented our method as a new layer in the existing Caffe library. Copyright © 2017 Elsevier Ltd. All rights reserved.

Development of the α-IGZO/Ag/α-IGZO Triple-Layer Structure Films for the Application of Transparent Electrode.

PubMed

Chen, Kun-Neng; Yang, Cheng-Fu; Wu, Chia-Ching; Chen, Yu-Hsin

2017-02-24

We investigated the structural, optical, and electrical properties of amorphous IGZO/silver/amorphous IGZO (α-IGZO/Ag/α-IGZO) triple-layer structures that were deposited at room temperature on Eagle XG glass and flexible polyethylene terephthalate substrates through the sputtering method. Thin Ag layers with different thicknesses were inserted between two IGZO layers to form a triple-layer structure. Ag was used because of its lower absorption and resistivity. Field emission scanning electron microscopy measurements of the triple-layer structures revealed that the thicknesses of the Ag layers ranged from 13 to 41 nm. The thickness of the Ag layer had a large effect on the electrical and optical properties of the electrodes. The optimum thickness of the Ag metal thin film could be evaluated according to the optical transmittance, electrical conductivity, and figure of merit of the electrode. This study demonstrates that the α-IGZO/Ag/α-IGZO triple-layer transparent electrode can be fabricated with low sheet resistance (4.2 Ω/□) and high optical transmittance (88.1%) at room temperature without postannealing processing on the deposited thin films.

The three-dimensional turbulent boundary layer near a plane of symmetry

NASA Technical Reports Server (NTRS)

Degani, A. T.; Smith, F. T.; Walker, J. D. A.

1992-01-01

The asymptotic structure of the three-dimensional turbulent boundary layer near a plane of symmetry is considered in the limit of large Reynolds number. A self-consistent two-layer structure is shown to exist wherein the streamwise velocity is brought to rest through an outer defect layer and an inner wall layer in a manner similar to that in two-dimensional boundary layers. The cross-stream velocity distribution is more complex and two terms in the asymptotic expansion are required to yield a complete profile which is shown to exhibit a logarithmic region. The flow in the inner wall layer is demonstrated to be collateral to leading order; pressure-gradient effects are formally of higher order but can cause the velocity profile to skew substantially near the wall at the large but finite Reynolds numbers encountered in practice. The governing set of ordinary differential equations describing a self-similar flow is derived. The calculated numerical solutions of these equations are matched asymptotically to an inner wall-layer solution and the results show trends that are consistent with experimental observations.

Weaving for heart valve tissue engineering.

PubMed

Liberski, Albert; Ayad, Nadia; Wojciechowska, Dorota; Kot, Radoslaw; Vo, Duy M P; Aibibu, Dilibaier; Hoffmann, Gerald; Cherif, Chokri; Grobelny-Mayer, Katharina; Snycerski, Marek; Goldmann, Helmut

2017-11-01

Weaving is a resourceful technology which offers a large selection of solutions that are readily adaptable for tissue engineering (TE) of artificial heart valves (HV). The different ways that the yarns are interlaced in this technique could be used to produce complex architectures, such as the three-layer architecture of the leaflets. Once the assembly is complete, growth of cells in the scaffold would occur in the orientation of the yarn, enabling the deposition of extra cellular matrixes proteins in an oriented manner. Weaving technology is a rapidly evolving field that, first, needs to be understood, and then explored by tissue engineers, so that it could be used to create efficient scaffolds. Similarly, the textile engineers need to gain a basic understanding of key structural and mechanical aspects of the heart valve. The aim of this review is to provide the platform for joining these two fields and to enable cooperative research efforts. Moreover, examples of woven medical products and patents as well as related publication are discussed in this review, nevertheless due to the large, and continuously growing volume of data, only the aspects strictly associated with HVTE lay in the scope of this paper. Copyright © 2017 Elsevier Inc. All rights reserved.

Evaluation of blood flow in human exercising muscle by diffuse correlation spectroscopy: a phantom model study

NASA Astrophysics Data System (ADS)

Nakabayashi, Mikie; Ono, Yumie; Ichinose, Masashi

2018-02-01

Diffuse correlation spectroscopy (DCS) has a potential to noninvasively and quantitatively measure the blood flow in the exercising muscle that could contribute to the fields of sports physiology and medicine. However, the blood flow index (BFI) measured from skin surface by DCS reflects hemodynamic signals from both superficial tissue and muscle layer. Thus, an appropriate calibration technology is required to quantify the absolute blood flow in the muscle layer. We therefore fabricated a realistic two-layer phantom model consisted of a static silicon layer imitating superficial tissue and a dynamic flow layer imitating the muscle blood flow and investigated the relationship between the simulated blood flow rate in the muscle layer and the BFI measured from the surface of the phantom. The absorption coefficient and the reduced scattering coefficient of the forearm were measured from 25 healthy young adults using a time-resolved nearinfrared spectroscopy. The depths of the superficial and muscle layers of forearm were also determined by ultrasound tomography images from 25 healthy young adults. The phantoms were fabricated to satisfy these optical coefficients and anatomical constraints. The simulated blood flow rate were set from 0 mL/ min to 68.7 mL/ min in ten steps, which is considered to cover a physiological range of mean blood flow of the forearm between per 100g of muscle tissue at rest to heavy dynamic handgrip exercise. We found a proportional relationship between the flow rates and BFIs with significant correlation coefficient of R = 0.986. Our results suggest that the absolute exercising muscle blood flow could be estimated by DCS with optimal calibration using phantom models.

Method of depositing multi-layer carbon-based coatings for field emission

DOEpatents

Sullivan, John P.; Friedmann, Thomas A.

1999-01-01

A novel field emitter device for cold cathode field emission applications, comprising a multi-layer resistive carbon film. The multi-layered film of the present invention is comprised of at least two layers of a resistive carbon material, preferably amorphous-tetrahedrally coordinated carbon, such that the resistivities of adjacent layers differ. For electron emission from the surface, the preferred structure comprises a top layer having a lower resistivity than the bottom layer. For edge emitting structures, the preferred structure of the film comprises a plurality of carbon layers, wherein adjacent layers have different resistivities. Through selection of deposition conditions, including the energy of the depositing carbon species, the presence or absence of certain elements such as H, N, inert gases or boron, carbon layers having desired resistivities can be produced. Field emitters made according the present invention display improved electron emission characteristics in comparison to conventional field emitter materials.