Lee, Sung-Il; Ko, Youngkyung; Park, Jun-Beom
2017-09-01
Three-dimensional cell culture systems provide a convenient in vitro model for the study of complex cell-cell and cell-matrix interactions in the absence of exogenous substrates. The current study aimed to evaluate the osteogenic differentiation potential of gingiva-derived stem cells cultured in two-dimensional or three-dimensional systems. To the best of our knowledge, the present study is the first to compare the growth of gingiva-derived stem cells in monolayer culture to a three-dimensional culture system with microwells. For three-dimensional culture, gingiva-derived stem cells were isolated and seeded into polydimethylsiloxane-based concave micromolds. Alkaline phosphatase activity and alizarin red S staining assays were then performed to evaluate osteogenesis and the degree of mineralization, respectively. Stem cell spheroids had a significantly increased level of alkaline phosphatase activity and mineralization compared with cells from the two-dimensional culture. In addition, an increase in mineralized deposits was observed with an increase in the loading cell number. The results of present study indicate that gingiva-derived stem cell spheroids exhibit an increased osteogenic potential compared with stem cells from two-dimensional culture. This highlights the potential of three-dimensional culture systems using gingiva-derived stem cells for regenerative medicine applications requiring stem cells with osteogenic potential.
NASA Technical Reports Server (NTRS)
Parker, Peter A. (Inventor)
2003-01-01
A single vector calibration system is provided which facilitates the calibration of multi-axis load cells, including wind tunnel force balances. The single vector system provides the capability to calibrate a multi-axis load cell using a single directional load, for example loading solely in the gravitational direction. The system manipulates the load cell in three-dimensional space, while keeping the uni-directional calibration load aligned. The use of a single vector calibration load reduces the set-up time for the multi-axis load combinations needed to generate a complete calibration mathematical model. The system also reduces load application inaccuracies caused by the conventional requirement to generate multiple force vectors. The simplicity of the system reduces calibration time and cost, while simultaneously increasing calibration accuracy.
MacDonald, Cristin; Barbee, Kenneth; Polyak, Boris
2012-05-01
To investigate the kinetics, mechanism and extent of MNP loading into endothelial cells and the effect of this loading on cell function. MNP uptake was examined under field on/off conditions, utilizing varying magnetite concentration MNPs. MNP-loaded cell viability and functional integrity was assessed using metabolic respiration, cell proliferation and migration assays. MNP uptake in endothelial cells significantly increased under the influence of a magnetic field versus non-magnetic conditions. Larger magnetite density of the MNPs led to a higher MNP internalization by cells under application of a magnetic field without compromising cellular respiration activity. Two-dimensional migration assays at no field showed that higher magnetite loading resulted in greater cell migration rates. In a three-dimensional migration assay under magnetic field, the migration rate of MNP-loaded cells was more than twice that of unloaded cells and was comparable to migration stimulated by a serum gradient. Our results suggest that endothelial cell uptake of MNPs is a force dependent process. The in vitro assays determined that cell health is not adversely affected by high MNP loadings, allowing these highly magnetically responsive cells to be potentially beneficial therapy (gene, drug or cell) delivery systems.
MacDonald, Cristin; Barbee, Kenneth
2015-01-01
Purpose To investigate the kinetics, mechanism and extent of MNP loading into endothelial cells and the effect of this loading on cell function. Methods MNP uptake was examined under field on/off conditions, utilizing varying magnetite concentration MNPs. MNP-loaded cell viability and functional integrity was assessed using metabolic respiration, cell proliferation and migration assays. Results MNP uptake in endothelial cells significantly increased under the influence of a magnetic field versus non-magnetic conditions. Larger magnetite density of the MNPs led to a higher MNP internalization by cells under application of a magnetic field without compromising cellular respiration activity. Two-dimensional migration assays at no field showed that higher magnetite loading resulted in greater cell migration rates. In a three-dimensional migration assay under magnetic field, the migration rate of MNP-loaded cells was more than twice that of unloaded cells and was comparable to migration stimulated by a serum gradient. Conclusions Our results suggest that endothelial cell uptake of MNPs is a force dependent process. The in vitro assays determined that cell health is not adversely affected by high MNP loadings, allowing these highly magnetically responsive cells to be potentially beneficial therapy (gene, drug or cell) delivery systems. PMID:22234617
Zeng, Yang; Feng, Siyu; Liu, Wei; Fu, Qinyouen; Li, Yaqian; Li, Xiaokang; Chen, Chun; Huang, Chenyu; Ge, Zigang; Du, Yanan
2017-04-01
To precondition mesenchymal stromal/stem cells (MSCs) with mechanical stimulation may enhance cell survival and functions following implantation in load bearing environment such as nucleus pulposus (NP) in intervertebral disc (IVD). In this study, preconditioning of MSCs toward NP-like cells was achieved in previously developed poly (ethylene glycol) diacrylate (PEGDA) microcryogels (PMs) within a syringe-based three-dimensional (3D) culture system which provided a facile and cost-effective pressure loading approach. PMs loaded with alginate and MSCs could be incubated in a sealable syringe which could be air-compressed to apply pressure loading through a programmable syringe pump. Expression levels of chondrogenic marker genes SOX9, COL II, and ACAN were significantly upregulated in MSCs when pressure loading of 0.2 MPa or 0.8 MPa was implemented. Expression levels of COL I and COL X were downregulated when pressure loading was applied. In a nude mouse model, MSCs loaded in PMs mechanically stimulated for three days were subcutaneously injected using the same culture syringe. Three weeks postinjection, more proteoglycans (PGs) were deposited and more SOX9 and COL II but less COL I and COL X were stained in 0.2 MPa group. Furthermore, injectable MSCs-loaded PMs were utilized in an ex vivo rabbit IVD organ culture model that demonstrated the leak-proof function and enhanced cell retention of PMs assisted cell delivery to a load bearing environment for potential NP regeneration. This microcryogels-based 3D cell culture and syringe-based pressure loading system represents a novel method for 3D cell culture with mechanical stimulation for better function. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 507-520, 2017. © 2015 Wiley Periodicals, Inc.
Hydrodynamic compression of young and adult rat osteoblast-like cells on titanium fiber mesh.
Walboomers, X F; Elder, S E; Bumgardner, J D; Jansen, J A
2006-01-01
Living bone cells are responsive to mechanical loading. Consequently, numerous in vitro models have been developed to examine the application of loading to cells. However, not all systems are suitable for the fibrous and porous three-dimensional materials, which are preferable for tissue repair purposes, or for the production of tissue engineering scaffolds. For three-dimensional applications, mechanical loading of cells with either fluid flow systems or hydrodynamic pressure systems has to be considered. Here, we aimed to evaluate the response of osteoblast-like cells to hydrodynamic compression, while growing in a three-dimensional titanium fiber mesh scaffolding material. For this purpose, a custom hydrodynamic compression chamber was built. Bone marrow cells were obtained from the femora of young (12-day-old) or old (1-year-old) rats, and precultured in the presence of dexamethasone and beta-glycerophosphate to achieve an osteoblast-like phenotype. Subsequently, cells were seeded onto the titanium mesh scaffolds, and subjected to hydrodynamic pressure, alternating between 0.3 to 5.0 MPa at 1 Hz, at 15-min intervals for a total of 60 min per day for up to 3 days. After pressurization, cell viability was checked. Afterward, DNA levels, alkaline phosphatase (ALP) activity, and extracellular calcium content were measured. Finally, all specimens were observed with scanning electron microscopy. Cell viability studies showed that the applied pressure was not harmful to the cells. Furthermore, we found that cells were able to detect the compression forces, because we did see evident effects on the cell numbers of the cells derived from old animals. However, there were no other changes in the cells under pressure. Finally, it was also noticeable that cells from old animals did not express ALP activity, but did show similar calcified extracellular matrix formation to the cells from young animals. In conclusion, the difference in DNA levels as reaction toward pressure, and the difference in ALP levels, suggest that the osteogenic properties of bone marrow-derived osteoblast-like cells are different with respect to the age of the donor. (c) 2005 Wiley Periodicals, Inc
NASA Astrophysics Data System (ADS)
Astolfo, Alberto; Arfelli, Fulvia; Schültke, Elisabeth; James, Simon; Mancini, Lucia; Menk, Ralf-Hendrik
2013-03-01
In the present study complementary high-resolution imaging techniques on different length scales are applied to elucidate a cellular loading protocol of gold nanoparticles and subsequently its impact on long term and high-resolution cell-tracking utilizing X-ray technology. Although demonstrated for malignant cell lines the results can be applied to non-malignant cell lines as well. In particular the accumulation of the gold marker per cell has been assessed quantitatively by virtue of electron microscopy, two-dimensional X-ray fluorescence imaging techniques and X-ray CT with micrometric and sub-micrometric resolution. Moreover, utilizing these techniques the three dimensional distribution of the incorporated nanoparticles, which are sequestered in lysosomes as a permanent marker, could be determined. The latter allowed elucidation of the gold partition during mitosis and the cell size, which subsequently enabled us to define the optimal instrument settings of a compact microCT system to visualize gold loaded cells. The results obtained demonstrate the feasibility of cell-tracking using X-ray CT with compact sources.
Bouet, G; Cruel, M; Laurent, C; Vico, L; Malaval, L; Marchat, D
2015-05-15
An engineered three dimensional (3D) in vitro cell culture system was designed with the goal of inducing and controlling in vitro osteogenesis in a reproducible manner under conditions more similar to the in vivo bone microenvironment than traditional two-dimensional (2D) models. This bioreactor allows efficient mechanical loading and perfusion of an original cubic calcium phosphate bioceramic of highly controlled composition and structure. This bioceramic comprises an internal portion containing homogeneously interconnected macropores surrounded by a dense layer, which minimises fluid flow bypass around the scaffold. This dense and flat layer permits the application of a homogeneous loading on the bioceramic while also enhancing its mechanical strength. Numerical modelling of constraints shows that the system provides direct mechanical stimulation of cells within the scaffold. Experimental results establish that under perfusion at a steady flow of 2 µL/min, corresponding to 3 ≤ Medium velocity ≤ 23 µm/s, mouse calvarial cells grow and differentiate as osteoblasts in a reproducible manner, and lay down a mineralised matrix. Moreover, cells respond to mechanical loading by increasing C-fos expression, which demonstrates the effective mechanical stimulation of the culture within the scaffold. In summary, we provide a "proof-of-concept" for osteoblastic cell culture in a controlled 3D culture system under perfusion and mechanical loading. This model will be a tool to analyse bone cell functions in vivo, and will provide a bench testing system for the clinical assessment of bioactive bone-targeting molecules under load.
[Three-dimensional finite element analysis on cell culture membrane under mechanical load].
Guo, Xin; Fan, Yubo; Song, Jinlin; Chen, Junkai
2002-01-01
A three-dimensional finite element model of the cell culture membrane was developed in the culture device under tension state made by us. The magnitude of tension and the displacement distribution in the membrane made of silicon rubber under different hydrostatic load were obtained by use of FEM analysis. A comparative study was made between the numerical and the experimental results. These results can serve as guides to the related cellular mechanical research.
Mittal, Nitika; Xia, Zeyang; Chen, Jie; Stewart, Kelton T; Liu, Sean Shih-Yao
2013-05-01
To quantify the three-dimensional moments and forces produced by pretorqued nickel-titanium (NiTi) rectangular archwires fully engaged in 0.018- and 0.022-inch slots of central incisor and molar edgewise and prescription brackets. Ten identical acrylic dental models with retroclined maxillary incisors were fabricated for bonding with various bracket-wire combinations. Edgewise, Roth, and MBT brackets with 0.018- and 0.022-inch slots were bonded in a simulated 2 × 4 clinical scenario. The left central incisor and molar were sectioned and attached to load cells. Correspondingly sized straight and pretorqued NiTi archwires were ligated to the brackets using 0.010-inch ligatures. Each load cell simultaneously measured three force (Fx, Fy, Fz) and three moment (Mx, My, Mz) components. The faciolingual, mesiodistal, and inciso-occluso/apical axes of the teeth corresponded to the x, y, and z axes of the load cells, respectively. Each wire was removed and retested seven times. Three-way analysis of variance (ANOVA) examined the effects of wire type, wire size, and bracket type on the measured orthodontic load systems. Interactions among the three effects were examined and pair-wise comparisons between significant combinations were performed. The force and moment components on each tooth were quantified according to their local coordinate axes. The three-way ANOVA interaction terms were significant for all force and moment measurements (P < .05), except for Fy (P > .05). The pretorqued wire generates a significantly larger incisor facial crown torquing moment in the MBT prescription compared to Roth, edgewise, and the straight NiTi wire.
Transient Response of a PEM Fuel Cell Representing Variable Load for a Moving Vehicle on Urban Roads
DOT National Transportation Integrated Search
2001-01-01
Three-dimensional numerical simulation of transient response of a Polymer Electrolyte Membrane (PEM) fuel cell subjected to a variable load is developed. The model parameters are typical of experimental cell for a 10-cm2 reactive area with serpentine...
Gajda, Steven; Chen, Jie
2012-03-01
To experimentally quantify the effects of the loop design on three-dimensional orthodontic load systems of two types of commercial closing loop archwires: Teardrop and Keyhole. An orthodontic force tester and custom-made dentoform were used to measure the load systems produced on two teeth during simulated space closure. The system included three force components along and three moment components about three clinically defined axes on two target teeth: the left maxillary canine and the lateral incisor. The archwires were attached to the dentoform and were activated following a standard clinical procedure. The resulting six load components produced by the two archwires were reported and compared. The results were also compared with those of the T-loop archwire published previously. The three designs deliver similar loading patterns; however, the component magnitudes are dependent on the design. All of the designs result in lingual tipping of the teeth, canine lingual-mesial displacement, canine crown-mesial-in rotation, and incisor crown-distal-in rotation.
Stasuk, Alexander
2017-01-01
Adipose-derived mesenchymal stem cells have become a popular cell choice for tendon repair strategies due to their relative abundance, ease of isolation, and ability to differentiate into tenocytes. In this study, we investigated the solo effect of different uniaxial tensile strains and loading frequencies on the matrix directionality and tenogenic differentiation of adipose-derived stem cells encapsulated within three-dimensional collagen scaffolds. Samples loaded at 0%, 2%, 4%, and 6% strains and 0.1 Hz and 1 Hz frequencies for 2 hours/day over a 7-day period using a custom-built uniaxial tensile strain bioreactor were characterized in terms of matrix organization, cell viability, and musculoskeletal gene expression profiles. The results displayed that the collagen fibers of the loaded samples exhibited increased matrix directionality with an increase in strain values. Gene expression analyses demonstrated that ASC-encapsulated collagen scaffolds loaded at 2% strain and 0.1 Hz frequency showed significant increases in extracellular matrix genes and tenogenic differentiation markers. Importantly, no cross-differentiation potential to osteogenic, chondrogenic, and myogenic lineages was observed at 2% strain and 0.1 Hz frequency loading condition. Thus, 2% strain and 0.1 Hz frequency were identified as the appropriate mechanical loading regime to induce tenogenic differentiation of adipose-derived stem cells cultured in a three-dimensional environment. PMID:29375625
Gajda, Steven; Chen, Jie
2014-01-01
Objective To experimentally quantify the effects of the loop design on three-dimensional orthodontic load systems of two types of commercial closing loop archwires: Teardrop and Keyhole. Materials and Methods An orthodontic force tester and custom-made dentoform were used to measure the load systems produced on two teeth during simulated space closure. The system included three force components along and three moment components about three clinically defined axes on two target teeth: the left maxillary canine and the lateral incisor. The archwires were attached to the dentoform and were activated following a standard clinical procedure. Results The resulting six load components produced by the two archwires were reported and compared. The results were also compared with those of the T-loop archwire published previously. Conclusions The three designs deliver similar loading patterns; however, the component magnitudes are dependent on the design. All of the designs result in lingual tipping of the teeth, canine lingual-mesial displacement, canine crown-mesial-in rotation, and incisor crown-distal-in rotation. PMID:21879793
NASA Technical Reports Server (NTRS)
Dorsey, D. R., Jr.
1975-01-01
A mathematical model was developed of the three-dimensional dynamics of a high-altitude scientific research balloon system perturbed from its equilibrium configuration by an arbitrary gust loading. The platform is modelled as a system of four coupled pendula, and the equations of motion were developed in the Lagrangian formalism assuming a small-angle approximation. Three-dimensional pendulation, torsion, and precessional motion due to Coriolis forces are considered. Aerodynamic and viscous damping effects on the pendulatory and torsional motions are included. A general model of the gust field incident upon the balloon system was developed. The digital computer simulation program is described, and a guide to its use is given.
Advances in solid polymer electrolyte fuel cell technology with low-platinum-loading electrodes
NASA Technical Reports Server (NTRS)
Srinivasan, Supramaniam; Ticianelli, E. A.; Derouin, C. R.; Redondo, A.
1987-01-01
The Gemini Space program demonstrated the first major application of fuel cell systems. Solid polymer electrolyte fuel cells were used as auxiliary power sources in the spacecraft. There has been considerable progress in this technology since then, particularly with the substitution of Nafion for the polystyrene sulfonate membrane as the electrolyte. Until recently the performance was good only with high platinum loading (4 mg/sq cm) electrodes. Methods are presented to advance the technology by (1) use of low platinum loading (0.35 mg/sq cm) electrodes; (2) optimization of anode/membrane/cathode interfaces by hot pressing; (3) pressurization of reactant gases, which is most important when air is used as cathodic reactant; and (4) adequate humidification of reactant gases to overcome the water management problem. The high performance of the fuel cell with the low loading of platinum appears to be due to the extension of the three dimensional reaction zone by introduction of a proton conductor, Nafion. This was confirmed by cyclic voltammetry.
Pierini, Michela; Bevilacqua, Alessandro; Torre, Maria Luisa; Lucarelli, Enrico
2017-01-01
Cell interaction with biomaterials is one of the keystones to developing medical devices for tissue engineering applications. Biomaterials are the scaffolds that give three-dimensional support to the cells, and are vectors that deliver the cells to the injured tissue requiring repair. Features of biomaterials can influence the behaviour of the cells and consequently the efficacy of the tissue-engineered product. The adhesion, distribution and motility of the seeded cells onto the scaffold represent key aspects, and must be evaluated in vitro during the product development, especially when the efficacy of a specific tissue-engineered product depends on viable and functional cell loading. In this work, we propose a non-invasive and non-destructive imaging analysis for investigating motility, viability and distribution of Mesenchymal Stem Cells (MSCs) on silk fibroin-based alginate microcarriers, to test the adhesion capacity of the fibroin coating onto alginate which is known to be unsuitable for cell adhesion. However, in depth characterization of the biomaterial is beyond the scope of this paper. Scaffold-loaded MSCs were stained with Calcein-AM and Ethidium homodimer-1 to detect live and dead cells, respectively, and counterstained with Hoechst to label cell nuclei. Time-lapse Light Sheet Fluorescent Microscopy (LSFM) was then used to produce three-dimensional images of the entire cells-loaded fibroin/alginate microcarriers. In order to quantitatively track the cell motility over time, we also developed an open source user friendly software tool called Fluorescent Cell Tracker in Three-Dimensions (F-Tracker3D). Combining LSFM with F-Tracker3D we were able for the first time to assess the distribution and motility of stem cells in a non-invasive, non-destructive, quantitative, and three-dimensional analysis of the entire surface of the cell-loaded scaffold. We therefore propose this imaging technique as an innovative holistic tool for monitoring cell-biomaterial interactions, and as a tool for the design, fabrication and functionalization of a scaffold as a medical device. PMID:28817694
Assembly of multiple cell gradients directed by three-dimensional microfluidic channels.
Li, Yiwei; Feng, Xiaojun; Wang, Yachao; Du, Wei; Chen, Peng; Liu, Chao; Liu, Bi-Feng
2015-08-07
Active control over the cell gradient is essential for understanding biological systems and the reconstitution of the functionality of many types of tissues, particularly for organ-on-a-chip. Here, we propose a three-dimensional (3D) microfluidic strategy for generating controllable cell gradients. In this approach, a homogeneous cell suspension is loaded into a 3D stair-shaped PDMS microchannel to generate a cell gradient within 10 min by sedimentation. We demonstrate that cell gradients of various profiles (exponential and piecewise linear) can be achieved by precisely controlling the height of each layer during the fabrication. With sequential seeding, we further demonstrate the generation of two overlapping cell gradients on the same glass substrate with pre-defined designs. The cell gradient-based QD cytotoxicity assay also demonstrated that cell behaviors and resistances were regulated by the changes in cell density. These results reveal that the proposed 3D microfluidic strategy provides a simple and versatile means for establishing controllable gradients in cell density, opening up a new avenue for reconstructing functional tissues.
Lynch, Maureen E; Chiou, Aaron E; Lee, Min Joon; Marcott, Stephen C; Polamraju, Praveen V; Lee, Yeonkyung; Fischbach, Claudia
2016-08-01
Dynamic mechanical loading is a strong anabolic signal in the skeleton, increasing osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BM-MSCs) and increasing the bone-forming activity of osteoblasts, but its role in bone metastatic cancer is relatively unknown. In this study, we integrated a hydroxyapatite-containing three-dimensional (3D) scaffold platform with controlled mechanical stimulation to investigate the effects of cyclic compression on the interplay between breast cancer cells and BM-MSCs as it pertains to bone metastasis. BM-MSCs cultured within mineral-containing 3D poly(lactide-co-glycolide) (PLG) scaffolds differentiated into mature osteoblasts, and exposure to tumor-derived soluble factors promoted this process. When BM-MSCs undergoing osteogenic differentiation were exposed to conditioned media collected from mechanically loaded breast cancer cells, their gene expression of osteopontin was increased. This was further enhanced when mechanical compression was simultaneously applied to BM-MSCs, leading to more uniformly deposited osteopontin within scaffold pores. These results suggest that mechanical loading of 3D scaffold-based culture models may be utilized to evaluate the role of physiologically relevant physical cues on bone metastatic breast cancer. Furthermore, our data imply that cyclic mechanical stimuli within the bone microenvironment modulate interactions between tumor cells and BM-MSCs that are relevant to bone metastasis.
Engineering three-dimensional cell mechanical microenvironment with hydrogels.
Huang, Guoyou; Wang, Lin; Wang, Shuqi; Han, Yulong; Wu, Jinhui; Zhang, Qiancheng; Xu, Feng; Lu, Tian Jian
2012-12-01
Cell mechanical microenvironment (CMM) significantly affects cell behaviors such as spreading, migration, proliferation and differentiation. However, most studies on cell response to mechanical stimulation are based on two-dimensional (2D) planar substrates, which cannot mimic native three-dimensional (3D) CMM. Accumulating evidence has shown that there is a significant difference in cell behavior in 2D and 3D microenvironments. Among the materials used for engineering 3D CMM, hydrogels have gained increasing attention due to their tunable properties (e.g. chemical and mechanical properties). In this paper, we provide an overview of recent advances in engineering hydrogel-based 3D CMM. Effects of mechanical cues (e.g. hydrogel stiffness and externally induced stress/strain in hydrogels) on cell behaviors are described. A variety of approaches to load mechanical stimuli in 3D hydrogel-based constructs are also discussed.
Godugu, Chandraiah; Singh, Mandip
2016-01-01
Routinely used two-dimensional cell culture-based models often fail while translating the observations into in vivo models. This setback is more common in cancer research, due to several reasons. The extracellular matrix and cell-to-cell interactions are not present in two-dimensional (2D) cell culture models. Diffusion of drug molecules into cancer cells is hindered by barriers of extracellular components in in vivo conditions, these barriers are absent in 2D cell culture models. To better mimic or simulate the in vivo conditions present in tumors, the current study used the alginate based three-dimensional cell culture (AlgiMatrix™) model, which resembles close to the in vivo tumor models. The current study explains the detailed protocols involved in AlgiMatrix™ based in vitro non-small-cell lung cancer (NSCLC) models. The suitability of this model was studied by evaluating, cytotoxicity, apoptosis, and penetration of nanoparticles into the in vitro tumor spheroids. This study also demonstrated the effect of EphA2 receptor targeted docetaxel-loaded nanoparticles on MDA-MB-468 TNBC cell lines. The methods section is subdivided into three subsections such as (1) preparation of AlgiMatrix™-based 3D in vitro tumor models and cytotoxicity assays, (2) free drug and nanoparticle uptake into spheroid studies, and (3) western blot, IHC, and RT-PCR studies.
Microfabrication of Cell-Laden Hydrogels for Engineering Mineralized and Load Bearing Tissues.
Li, Chia-Cheng; Kharaziha, Mahshid; Min, Christine; Maas, Richard; Nikkhah, Mehdi
2015-01-01
Microengineering technologies and advanced biomaterials have extensive applications in the field of regenerative medicine. In this chapter, we review the integration of microfabrication techniques and hydrogel-based biomaterials in the field of dental, bone, and cartilage tissue engineering. We primarily discuss the major features that make hydrogels attractive candidates to mimic extracellular matrix (ECM), and we consider the benefits of three-dimensional (3D) culture systems for tissue engineering applications. We then focus on the fundamental principles of microfabrication techniques including photolithography, soft lithography and bioprinting approaches. Lastly, we summarize recent research on microengineering cell-laden hydrogel constructs for dental, bone and cartilage regeneration, and discuss future applications of microfabrication techniques for load-bearing tissue engineering.
Thermal Analysis on Cryogenic Liquid Hydrogen Tank on an Unmanned Aerial Vehicle System
NASA Technical Reports Server (NTRS)
Wang, Xiao-Yen; Harpster, George; Hunter, James
2007-01-01
Thermal analyses are performed on the liquid hydrogen (LH2) tank designed for an unmanned aerial vehicle (UAV) powered by solar arrays and a regenerative proton-exchange membrane (PEM) fuel cell. A 14-day cruise mission at a 65,000 ft altitude is considered. Thermal analysis provides the thermal loads on the tank system and the boiling-off rates of LH2. Different approaches are being considered to minimize the boiling-off rates of the LH2. It includes an evacuated multilayer insulation (MLI) versus aerogel insulation on the LH2 tank and aluminum versus stainless steel spacer rings between the inner and outer tank. The resulting boil-off rates of LH2 provided by the one-dimensional model and three-dimensional finite element analysis (FEA) on the tank system are presented and compared to validate the results of the three-dimensional FEA. It concludes that heat flux through penetrations by conduction is as significant as that through insulation around the tank. The tank system with MLI insulation and stainless steel spacer rings result in the lowest boiling-off rate of LH2.
NASA Astrophysics Data System (ADS)
Kasperska, Kamila; Wieczorowski, Michał; Krolczyk, Jolanta B.
2017-10-01
Three-dimensional scanning is used in many fields: medicine, architecture, industry, reverse engineering. The aim of the article was to analyze the changes in the shape of the limbs under the influence of a mechanical external load using the method of three-dimensional scanner uses white light technology. The paper presents a system of human movement, passive part - skeleton and active part - the muscles, and principles of their interaction, which results in a change of the position of the body. Furthermore, by using the 3D scan, the differences in appearance of the arm and leg depending on the size of the external load in different positions have been presented. The paper shows that with increasing load, which muscles must prevent, increases the volume of certain parts of the legs, while another parts of them will be reduced. Results of the research using three-dimensional scanner allow determining what impact on changing the legs shape has an external mechanical load.
Lynch, Maureen E.; Chiou, Aaron E.; Lee, Min Joon; Marcott, Stephen C.; Polamraju, Praveen V.; Lee, Yeonkyung
2016-01-01
Dynamic mechanical loading is a strong anabolic signal in the skeleton, increasing osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BM-MSCs) and increasing the bone-forming activity of osteoblasts, but its role in bone metastatic cancer is relatively unknown. In this study, we integrated a hydroxyapatite-containing three-dimensional (3D) scaffold platform with controlled mechanical stimulation to investigate the effects of cyclic compression on the interplay between breast cancer cells and BM-MSCs as it pertains to bone metastasis. BM-MSCs cultured within mineral-containing 3D poly(lactide-co-glycolide) (PLG) scaffolds differentiated into mature osteoblasts, and exposure to tumor-derived soluble factors promoted this process. When BM-MSCs undergoing osteogenic differentiation were exposed to conditioned media collected from mechanically loaded breast cancer cells, their gene expression of osteopontin was increased. This was further enhanced when mechanical compression was simultaneously applied to BM-MSCs, leading to more uniformly deposited osteopontin within scaffold pores. These results suggest that mechanical loading of 3D scaffold-based culture models may be utilized to evaluate the role of physiologically relevant physical cues on bone metastatic breast cancer. Furthermore, our data imply that cyclic mechanical stimuli within the bone microenvironment modulate interactions between tumor cells and BM-MSCs that are relevant to bone metastasis. PMID:27401765
Transient Three-Dimensional Analysis of Side Load in Liquid Rocket Engine Nozzles
NASA Technical Reports Server (NTRS)
Wang, Ten-See
2004-01-01
Three-dimensional numerical investigations on the nozzle start-up side load physics were performed. The objective of this study is to identify the three-dimensional side load physics and to compute the associated aerodynamic side load using an anchored computational methodology. The computational methodology is based on an unstructured-grid, and pressure-based computational fluid dynamics formulation, and a simulated inlet condition based on a system calculation. Finite-rate chemistry was used throughout the study so that combustion effect is always included, and the effect of wall cooling on side load physics is studied. The side load physics captured include the afterburning wave, transition from free- shock to restricted-shock separation, and lip Lambda shock oscillation. With the adiabatic nozzle, free-shock separation reappears after the transition from free-shock separation to restricted-shock separation, and the subsequent flow pattern of the simultaneous free-shock and restricted-shock separations creates a very asymmetric Mach disk flow. With the cooled nozzle, the more symmetric restricted-shock separation persisted throughout the start-up transient after the transition, leading to an overall lower side load than that of the adiabatic nozzle. The tepee structures corresponding to the maximum side load were addressed.
Lewis, Natasha S; Lewis, Emily EL; Mullin, Margaret; Wheadon, Helen; Dalby, Matthew J; Berry, Catherine C
2017-01-01
Multicellular spheroids are an established system for three-dimensional cell culture. Spheroids are typically generated using hanging drop or non-adherent culture; however, an emerging technique is to use magnetic levitation. Herein, mesenchymal stem cell spheroids were generated using magnetic nanoparticles and subsequently cultured within a type I collagen gel, with a view towards developing a bone marrow niche environment. Cells were loaded with magnetic nanoparticles, and suspended beneath an external magnet, inducing self-assembly of multicellular spheroids. Cells in spheroids were viable and compared to corresponding monolayer controls, maintained stem cell phenotype and were quiescent. Interestingly, core spheroid necrosis was not observed, even with increasing spheroid size, in contrast to other commonly used spheroid systems. This mesenchymal stem cell spheroid culture presents a potential platform for modelling in vitro bone marrow stem cell niches, elucidating interactions between cells, as well as a useful model for drug delivery studies. PMID:28616152
Transient Three-Dimensional Side Load Analysis of a Film Cooled Nozzle
NASA Technical Reports Server (NTRS)
Wang, Ten-See; Guidos, Mike
2008-01-01
Transient three-dimensional numerical investigations on the side load physics for an engine encompassing a film cooled nozzle extension and a regeneratively cooled thrust chamber, were performed. The objectives of this study are to identify the three-dimensional side load physics and to compute the associated aerodynamic side load using an anchored computational methodology. The computational methodology is based on an unstructured-grid, pressure-based computational fluid dynamics formulation, and a transient inlet history based on an engine system simulation. Ultimately, the computational results will be provided to the nozzle designers for estimating of effect of the peak side load on the nozzle structure. Computations simulating engine startup at ambient pressures corresponding to sea level and three high altitudes were performed. In addition, computations for both engine startup and shutdown transients were also performed for a stub nozzle, operating at sea level. For engine with the full nozzle extension, computational result shows starting up at sea level, the peak side load occurs when the lambda shock steps into the turbine exhaust flow, while the side load caused by the transition from free-shock separation to restricted-shock separation comes at second; and the side loads decreasing rapidly and progressively as the ambient pressure decreases. For the stub nozzle operating at sea level, the computed side loads during both startup and shutdown becomes very small due to the much reduced flow area.
Susilo, Monica E; Bell, Brett J; Roeder, Blayne A; Voytik-Harbin, Sherry L; Kokini, Klod; Nauman, Eric A
2013-03-01
Mechanical signals are important factors in determining cell fate. Therefore, insights as to how mechanical signals are transferred between the cell and its surrounding three-dimensional collagen fibril network will provide a basis for designing the optimum extracellular matrix (ECM) microenvironment for tissue regeneration. Previously we described a cellular solid model to predict fibril microstructure-mechanical relationships of reconstituted collagen matrices due to unidirectional loads (Acta Biomater 2010;6:1471-86). The model consisted of representative volume elements made up of an interconnected network of flexible struts. The present study extends this work by adapting the model to account for microstructural anisotropy of the collagen fibrils and a biaxial loading environment. The model was calibrated based on uniaxial tensile data and used to predict the equibiaxial tensile stress-stretch relationship. Modifications to the model significantly improved its predictive capacity for equibiaxial loading data. With a comparable fibril length (model 5.9-8μm, measured 7.5μm) and appropriate fibril anisotropy the anisotropic model provides a better representation of the collagen fibril microstructure. Such models are important tools for tissue engineering because they facilitate prediction of microstructure-mechanical relationships for collagen matrices over a wide range of microstructures and provide a framework for predicting cell-ECM interactions. Copyright © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
2014-01-01
A three-dimensional finite element model was developed to investigate dynamic response of track-embankment-ground system subjected to moving loads caused by high speed trains. The track-embankment-ground systems such as the sleepers, the ballast, the embankment, and the ground are represented by 8-noded solid elements. The infinite elements are used to represent the infinite boundary condition to absorb vibration waves induced by the passing of train load at the boundary. The loads were applied on the rails directly to simulate the real moving loads of trains. The effects of train speed on dynamic response of the system are considered. The effect of material parameters, especially the modulus changes of ballast and embankment, is taken into account to demonstrate the effectiveness of strengthening the ballast, embankment, and ground for mitigating system vibration in detail. The numerical results show that the model is reliable for predicting the amplitude of vibrations produced in the track-embankment-ground system by high-speed trains. Stiffening of fill under the embankment can reduce the vibration level, on the other hand, it can be realized by installing a concrete slab under the embankment. The influence of axle load on the vibration of the system is obviously lower than that of train speed. PMID:24723838
Fu, Qiang; Zheng, Changjie
2014-01-01
A three-dimensional finite element model was developed to investigate dynamic response of track-embankment-ground system subjected to moving loads caused by high speed trains. The track-embankment-ground systems such as the sleepers, the ballast, the embankment, and the ground are represented by 8-noded solid elements. The infinite elements are used to represent the infinite boundary condition to absorb vibration waves induced by the passing of train load at the boundary. The loads were applied on the rails directly to simulate the real moving loads of trains. The effects of train speed on dynamic response of the system are considered. The effect of material parameters, especially the modulus changes of ballast and embankment, is taken into account to demonstrate the effectiveness of strengthening the ballast, embankment, and ground for mitigating system vibration in detail. The numerical results show that the model is reliable for predicting the amplitude of vibrations produced in the track-embankment-ground system by high-speed trains. Stiffening of fill under the embankment can reduce the vibration level, on the other hand, it can be realized by installing a concrete slab under the embankment. The influence of axle load on the vibration of the system is obviously lower than that of train speed.
Effect of load transients on SOFC operation—current reversal on loss of load
NASA Astrophysics Data System (ADS)
Gemmen, Randall S.; Johnson, Christopher D.
The dynamics of solid oxide fuel cell (SOFC) operation have been considered previously, but mainly through the use of one-dimensional codes applied to co-flow fuel cell systems. In this paper several geometries are considered, including cross-flow, co-flow, and counter-flow. The details of the model are provided, and the model is compared with some initial experimental data. For parameters typical of SOFC operation, a variety of transient cases are investigated, including representative load increase and decrease and system shutdown. Of particular note for large load decrease conditions (e.g., shutdown) is the occurrence of reverse current over significant portions of the cell, starting from the moment of load loss up to the point where equilibrated conditions again provide positive current. Consideration is given as to when such reverse current conditions might most significantly impact the reliability of the cell.
Controllable load sharing for soft adhesive interfaces on three-dimensional surfaces.
Song, Sukho; Drotlef, Dirk-Michael; Majidi, Carmel; Sitti, Metin
2017-05-30
For adhering to three-dimensional (3D) surfaces or objects, current adhesion systems are limited by a fundamental trade-off between 3D surface conformability and high adhesion strength. This limitation arises from the need for a soft, mechanically compliant interface, which enables conformability to nonflat and irregularly shaped surfaces but significantly reduces the interfacial fracture strength. In this work, we overcome this trade-off with an adhesion-based soft-gripping system that exhibits enhanced fracture strength without sacrificing conformability to nonplanar 3D surfaces. Composed of a gecko-inspired elastomeric microfibrillar adhesive membrane supported by a pressure-controlled deformable gripper body, the proposed soft-gripping system controls the bonding strength by changing its internal pressure and exploiting the mechanics of interfacial equal load sharing. The soft adhesion system can use up to ∼26% of the maximum adhesion of the fibrillar membrane, which is 14× higher than the adhering membrane without load sharing. Our proposed load-sharing method suggests a paradigm for soft adhesion-based gripping and transfer-printing systems that achieves area scaling similar to that of a natural gecko footpad.
Controllable load sharing for soft adhesive interfaces on three-dimensional surfaces
NASA Astrophysics Data System (ADS)
Song, Sukho; Drotlef, Dirk-Michael; Majidi, Carmel; Sitti, Metin
2017-05-01
For adhering to three-dimensional (3D) surfaces or objects, current adhesion systems are limited by a fundamental trade-off between 3D surface conformability and high adhesion strength. This limitation arises from the need for a soft, mechanically compliant interface, which enables conformability to nonflat and irregularly shaped surfaces but significantly reduces the interfacial fracture strength. In this work, we overcome this trade-off with an adhesion-based soft-gripping system that exhibits enhanced fracture strength without sacrificing conformability to nonplanar 3D surfaces. Composed of a gecko-inspired elastomeric microfibrillar adhesive membrane supported by a pressure-controlled deformable gripper body, the proposed soft-gripping system controls the bonding strength by changing its internal pressure and exploiting the mechanics of interfacial equal load sharing. The soft adhesion system can use up to ˜26% of the maximum adhesion of the fibrillar membrane, which is 14× higher than the adhering membrane without load sharing. Our proposed load-sharing method suggests a paradigm for soft adhesion-based gripping and transfer-printing systems that achieves area scaling similar to that of a natural gecko footpad.
Controllable load sharing for soft adhesive interfaces on three-dimensional surfaces
Song, Sukho; Drotlef, Dirk-Michael; Majidi, Carmel; Sitti, Metin
2017-01-01
For adhering to three-dimensional (3D) surfaces or objects, current adhesion systems are limited by a fundamental trade-off between 3D surface conformability and high adhesion strength. This limitation arises from the need for a soft, mechanically compliant interface, which enables conformability to nonflat and irregularly shaped surfaces but significantly reduces the interfacial fracture strength. In this work, we overcome this trade-off with an adhesion-based soft-gripping system that exhibits enhanced fracture strength without sacrificing conformability to nonplanar 3D surfaces. Composed of a gecko-inspired elastomeric microfibrillar adhesive membrane supported by a pressure-controlled deformable gripper body, the proposed soft-gripping system controls the bonding strength by changing its internal pressure and exploiting the mechanics of interfacial equal load sharing. The soft adhesion system can use up to ∼26% of the maximum adhesion of the fibrillar membrane, which is 14× higher than the adhering membrane without load sharing. Our proposed load-sharing method suggests a paradigm for soft adhesion-based gripping and transfer-printing systems that achieves area scaling similar to that of a natural gecko footpad. PMID:28507143
An Energy Model of Place Cell Network in Three Dimensional Space.
Wang, Yihong; Xu, Xuying; Wang, Rubin
2018-01-01
Place cells are important elements in the spatial representation system of the brain. A considerable amount of experimental data and classical models are achieved in this area. However, an important question has not been addressed, which is how the three dimensional space is represented by the place cells. This question is preliminarily surveyed by energy coding method in this research. Energy coding method argues that neural information can be expressed by neural energy and it is convenient to model and compute for neural systems due to the global and linearly addable properties of neural energy. Nevertheless, the models of functional neural networks based on energy coding method have not been established. In this work, we construct a place cell network model to represent three dimensional space on an energy level. Then we define the place field and place field center and test the locating performance in three dimensional space. The results imply that the model successfully simulates the basic properties of place cells. The individual place cell obtains unique spatial selectivity. The place fields in three dimensional space vary in size and energy consumption. Furthermore, the locating error is limited to a certain level and the simulated place field agrees to the experimental results. In conclusion, this is an effective model to represent three dimensional space by energy method. The research verifies the energy efficiency principle of the brain during the neural coding for three dimensional spatial information. It is the first step to complete the three dimensional spatial representing system of the brain, and helps us further understand how the energy efficiency principle directs the locating, navigating, and path planning function of the brain.
Three-dimensional finite element analysis of a newly designed onplant miniplate anchorage system.
Liu, Lin; Qu, Yin-Ying; Jiang, Li-Jun; Zhou, Qian; Tang, Tian-Qi
2016-06-01
The purpose of this research was to evaluate the structural stress and deformation of a newly designed onplant miniplate anchorage system compared to a standard anchorage system. A bone block integrated with a novel miniplate and fixation screw system was simulated in a three-dimensional model and subjected to force at different directions. The stress distribution and deformation of the miniplate system and cortical bone were evaluated using the three-dimensional finite element method. The results showed that the stress on the plate system and bone was linearly proportional to the force magnitude and was higher when the force was in a vertical direction (Y-axis). Stress and deformation values of the two screws (screw 1 and 2) were asymmetric when the force was added along Y-axis and was greater in screw 1. The highest deformation value of the screws was 7.5148 μm, much smaller than the limit value. The load was decreased for each single miniscrew, and the ability of the new anchorage system to bear the load was also enhanced to some degree. It was suggested that the newly designed onplant miniplate anchorage system is effective, easily implanted and minimally invasive.
Aerodynamic and heat transfer analysis of the low aspect ratio turbine
NASA Astrophysics Data System (ADS)
Sharma, O. P.; Nguyen, P.; Ni, R. H.; Rhie, C. M.; White, J. A.
1987-06-01
The available two- and three-dimensional codes are used to estimate external heat loads and aerodynamic characteristics of a highly loaded turbine stage in order to demonstrate state-of-the-art methodologies in turbine design. By using data for a low aspect ratio turbine, it is found that a three-dimensional multistage Euler code gives good averall predictions for the turbine stage, yielding good estimates of the stage pressure ratio, mass flow, and exit gas angles. The nozzle vane loading distribution is well predicted by both the three-dimensional multistage Euler and three-dimensional Navier-Stokes codes. The vane airfoil surface Stanton number distributions, however, are underpredicted by both two- and three-dimensional boundary value analysis.
Transient Three-Dimensional Analysis of Nozzle Side Load in Regeneratively Cooled Engines
NASA Technical Reports Server (NTRS)
Wang, Ten-See
2005-01-01
Three-dimensional numerical investigations on the start-up side load physics for a regeneratively cooled, high-aspect-ratio nozzle were performed. The objectives of this study are to identify the three-dimensional side load physics and to compute the associated aerodynamic side load using an anchored computational methodology. The computational methodology is based on an unstructured-grid, pressure-based computational fluid dynamics formulation, and a transient inlet condition based on an engine system simulation. Computations were performed for both the adiabatic and cooled walls in order to understand the effect of boundary conditions. Finite-rate chemistry was used throughout the study so that combustion effect is always included. The results show that three types of shock evolution are responsible for side loads: generation of combustion wave; transitions among free-shock separation, restricted-shock separation, and simultaneous free-shock and restricted shock separations; along with oscillation of shocks across the lip. Wall boundary conditions drastically affect the computed side load physics: the adiabatic nozzle prefers free-shock separation while the cooled nozzle favors restricted-shock separation, resulting in higher peak side load for the cooled nozzle than that of the adiabatic nozzle. By comparing the computed physics with those of test observations, it is concluded that cooled wall is a more realistic boundary condition, and the oscillation of the restricted-shock separation flow pattern across the lip along with its associated tangential shock motion are the dominant side load physics for a regeneratively cooled, high aspect-ratio rocket engine.
Powers, Christopher M; Chen, Yu-Jen; Scher, Irving; Lee, Thay Q
2006-01-01
The purpose of this study was to determine the influence of patellofemoral joint contact geometry on the modeling of three-dimensional patellofemoral joint forces. To achieve this goal, patellofemoral joint reaction forces (PFJRFs) that were measured from an in-vitro cadaveric set-up were compared to PFJRFs estimated from a computer model that did not consider patellofemoral joint contact geometry. Ten cadaver knees were used in this study. Each was mounted on a custom jig that was fixed to an Instron frame. Quadriceps muscle loads were accomplished using a pulley system and weights. The force in the patellar ligament was obtained using a buckle transducer. To quantify the magnitude and direction of the PFJRF, a six-axis load cell was incorporated into the femoral fixation system so that a rigid body assumption could be made. PFJRF data were obtained at 0 degrees , 20 degrees , 40 degrees and 60 degrees of knee flexion. Following in vitro testing, SIMM modeling software was used to develop computational models based on the three-dimensional coordinates (Microscribe digitizer) of individual muscle and patellar ligament force vectors obtained from the cadaver knees. The overall magnitude of the PFJRF estimated from the computer generated models closely matched the direct measurements from the in vitro set-up (Pearson's correlation coefficient, R(2)=0.91, p<0.001). Although the computational model accurately estimated the posteriorly directed forces acting on the joint, some discrepancies were noted in the forces acting in the superior and lateral directions. These differences however, were relatively small when expressed as a total of the overall PFJRF magnitude.
Overview of aerothermodynamic loads definition study
NASA Technical Reports Server (NTRS)
Gaugler, Raymond E.
1991-01-01
The objective of the Aerothermodynamic Loads Definition Study is to develop methods of accurately predicting the operating environment in advanced Earth-to-Orbit (ETO) propulsion systems, such as the Space Shuttle Main Engine (SSME) powerhead. Development of time averaged and time dependent three dimensional viscous computer codes as well as experimental verification and engine diagnostic testing are considered to be essential in achieving that objective. Time-averaged, nonsteady, and transient operating loads must all be well defined in order to accurately predict powerhead life. Described here is work in unsteady heat flow analysis, improved modeling of preburner flow, turbulence modeling for turbomachinery, computation of three dimensional flow with heat transfer, and unsteady viscous multi-blade row turbine analysis.
A multi scale multi-dimensional thermo electrochemical modelling of high capacity lithium-ion cells
NASA Astrophysics Data System (ADS)
Tourani, Abbas; White, Peter; Ivey, Paul
2014-06-01
Lithium iron phosphate (LFP) and lithium manganese oxide (LMO) are competitive and complementary to each other as cathode materials for lithium-ion batteries, especially for use in electric vehicles. A multi scale multi-dimensional physic-based model is proposed in this paper to study the thermal behaviour of the two lithium-ion chemistries. The model consists of two sub models, a one dimensional (1D) electrochemical sub model and a two dimensional (2D) thermo-electric sub model, which are coupled and solved concurrently. The 1D model predicts the heat generation rate (Qh) and voltage (V) of the battery cell through different load cycles. The 2D model of the battery cell accounts for temperature distribution and current distribution across the surface of the battery cell. The two cells are examined experimentally through 90 h load cycles including high/low charge/discharge rates. The experimental results are compared with the model results and they are in good agreement. The presented results in this paper verify the cells temperature behaviour at different operating conditions which will lead to the design of a cost effective thermal management system for the battery pack.
NASA Astrophysics Data System (ADS)
Santoro, Carlo; Kodali, Mounika; Kabir, Sadia; Soavi, Francesca; Serov, Alexey; Atanassov, Plamen
2017-07-01
Three-dimensional graphene nanosheets (3D-GNS) were used as cathode catalysts for microbial fuel cells (MFCs) operating in neutral conditions. 3D-GNS catalysts showed high performance towards oxygen electroreduction in neutral media with high current densities and low hydrogen peroxide generation compared to activated carbon (AC). 3D-GNS was incorporated into air-breathing cathodes based on AC with three different loadings (2, 6 and 10 mgcm-2). Performances in MFCs showed that 3D-GNS had the highest performances with power densities of 2.059 ± 0.003 Wm-2, 1.855 ± 0.007 Wm-2 and 1.503 ± 0.005 Wm-2 for loading of 10, 6 and 2 mgcm-2 respectively. Plain AC had the lowest performances (1.017 ± 0.009 Wm-2). The different cathodes were also investigated in supercapacitive MFCs (SC-MFCs). The addition of 3D-GNS decreased the ohmic losses by 14-25%. The decrease in ohmic losses allowed the SC-MFC with 3D-GNS (loading 10 mgcm-2) to have the maximum power (Pmax) of 5.746 ± 0.186 Wm-2. At 5 mA, the SC-MFC featured an "apparent" capacitive response that increased from 0.027 ± 0.007 F with AC to 0.213 ± 0.026 F with 3D-GNS (loading 2 mgcm-2) and further to 1.817 ± 0.040 F with 3D-GNS (loading 10 mgcm-2).
NASA Technical Reports Server (NTRS)
Sharobeam, Monir H.
1994-01-01
Load separation is the representation of the load in the test records of geometries containing cracks as a multiplication of two separate functions: a crack geometry function and a material deformation function. Load separation is demonstrated in the test records of several two-dimensional geometries such as compact tension geometry, single edge notched bend geometry, and center cracked tension geometry and three-dimensional geometries such as semi-elliptical surface crack. The role of load separation in the evaluation of the fracture parameter J-integral and the associated factor eta for two-dimensional geometries is discussed. The paper also discusses the theoretical basis and the procedure for using load separation as a simplified yet accurate approach for plastic J evaluation in semi-elliptical surface crack which is a three-dimensional geometry. The experimental evaluation of J, and particularly J(sub pl), for three-dimensional geometries is very challenging. A few approaches have been developed in this regard and they are either complex or very approximate. The paper also presents the load separation as a mean to identify the blunting and crack growth regions in the experimental test records of precracked specimens. Finally, load separation as a methodology in elastic-plastic fracture mechanics is presented.
Transient Two-Dimensional Analysis of Side Load in Liquid Rocket Engine Nozzles
NASA Technical Reports Server (NTRS)
Wang, Ten-See
2004-01-01
Two-dimensional planar and axisymmetric numerical investigations on the nozzle start-up side load physics were performed. The objective of this study is to develop a computational methodology to identify nozzle side load physics using simplified two-dimensional geometries, in order to come up with a computational strategy to eventually predict the three-dimensional side loads. The computational methodology is based on a multidimensional, finite-volume, viscous, chemically reacting, unstructured-grid, and pressure-based computational fluid dynamics formulation, and a transient inlet condition based on an engine system modeling. The side load physics captured in the low aspect-ratio, two-dimensional planar nozzle include the Coanda effect, afterburning wave, and the associated lip free-shock oscillation. Results of parametric studies indicate that equivalence ratio, combustion and ramp rate affect the side load physics. The side load physics inferred in the high aspect-ratio, axisymmetric nozzle study include the afterburning wave; transition from free-shock to restricted-shock separation, reverting back to free-shock separation, and transforming to restricted-shock separation again; and lip restricted-shock oscillation. The Mach disk loci and wall pressure history studies reconfirm that combustion and the associated thermodynamic properties affect the formation and duration of the asymmetric flow.
Haider, Mansoor A.; Guilak, Farshid
2009-01-01
Articular cartilage exhibits viscoelasticity in response to mechanical loading that is well described using biphasic or poroelastic continuum models. To date, boundary element methods (BEMs) have not been employed in modeling biphasic tissue mechanics. A three dimensional direct poroelastic BEM, formulated in the Laplace transform domain, is applied to modeling stress relaxation in cartilage. Macroscopic stress relaxation of a poroelastic cylinder in uni-axial confined compression is simulated and validated against a theoretical solution. Microscopic cell deformation due to poroelastic stress relaxation is also modeled. An extended Laplace inversion method is employed to accurately represent mechanical responses in the time domain. PMID:19851478
Haider, Mansoor A; Guilak, Farshid
2007-06-15
Articular cartilage exhibits viscoelasticity in response to mechanical loading that is well described using biphasic or poroelastic continuum models. To date, boundary element methods (BEMs) have not been employed in modeling biphasic tissue mechanics. A three dimensional direct poroelastic BEM, formulated in the Laplace transform domain, is applied to modeling stress relaxation in cartilage. Macroscopic stress relaxation of a poroelastic cylinder in uni-axial confined compression is simulated and validated against a theoretical solution. Microscopic cell deformation due to poroelastic stress relaxation is also modeled. An extended Laplace inversion method is employed to accurately represent mechanical responses in the time domain.
NASA Technical Reports Server (NTRS)
Juran, C. M.; Blaber, E. A.; Almeida, E. A. C.
2016-01-01
Cell and animal studies conducted onboard the International Space Station and formerly on Shuttle flights have provided groundbreaking data illuminating the deleterious biological response of bone to mechanical unloading. However the intercellular communicative mechanisms associated with the regulation of bone synthesis and bone resorption cells are still largely unknown. Connexin-43 (CX43), a gap junction protein, is hypothesized to play a significant role in osteoblast and osteocyte signaling. The purpose of this investigation was to evaluate within a novel three-dimensional microenvironment how the osteocyte-osteoblast gap-junction expression changes when cultures are exposed to exaggerated mechanical load. MLO-Y4 osteocyte-like cells were cultured on a 3D-Biotek polystyrene insert and placed in direct contact with an MC3T3-E1 pre-osteoblast co-cultured monolayer and exposed to 48 h of mechanical stimulation (pulsatile fluid flow (PFF) or monolayer cyclic stretch (MCS)) then evaluated for viability, proliferation, metabolism, and CX43 expression. Mono-cultured MLO-Y4 and MC3T3-E1 control experiments were conducted under PFF and MCS stimulation to observe how strain application stimuli (PFF cell membrane shear or MCS cell focal adhesion/attachment loading) initiates different signaling pathways or downstream regulatory controls. TotalLive cell count, viability and metabolic reduction (Trypan Blue, LIVEDead and Alamar Blue analysis respectively) indicate that mechanical activation of MC3T3-E1 cells inhibits proliferation while maintaining an average 1.04E4 reductioncell metabolic rate, *p0.05 n4. MLO-Y4s in monolayer culture increase in number when exposed to MCS loading but the percent of live cells within the population is low (46.3 total count, *p0.05 n4), these results may indicate an apoptotic signaling cascade. PFF stimulation of the three-dimensional co-cultures elicits a universal increase in CX43 in MLO-Y4 and MC3T3-E1 cells, illustrated by immunohistological observation. Increased CX43 expression is also observed with the three-dimensional co-cultures with MC3T3-E1 MCS stimulation but the increased gap-junction protein presence was limited to the osteoblast-osteocyte interface region. Previously reported PCR evaluation of osteogenic markers further corroborate that the co-cultured populations communicative networks play a role in translating mechanical signals to molecular messaging. These findings suggests an osteocyte-osteoblast gap-junction signaling feedback mechanism may regulate mechanotransduction of apoptosis initiation and transcription of cytokine signaling proteins responsible for stem cell niche recruitment much more directly than previously believed.
A three-dimensional meso-macroscopic model for Li-Ion intercalation batteries
Allu, S.; Kalnaus, S.; Simunovic, S.; ...
2016-06-09
Through this study, we present a three-dimensional computational formulation for electrode-electrolyte-electrode system of Li-Ion batteries. The physical consistency between electrical, thermal and chemical equations is enforced at each time increment by driving the residual of the resulting coupled system of nonlinear equations to zero. The formulation utilizes a rigorous volume averaging approach typical of multiphase formulations used in other fields and recently extended to modeling of supercapacitors [1]. Unlike existing battery modeling methods which use segregated solution of conservation equations and idealized geometries, our unified approach can model arbitrary battery and electrode configurations. The consistency of multi-physics solution also allowsmore » for consideration of a wide array of initial conditions and load cases. The formulation accounts for spatio-temporal variations of material and state properties such as electrode/void volume fractions and anisotropic conductivities. The governing differential equations are discretized using the finite element method and solved using a nonlinearly consistent approach that provides robust stability and convergence. The new formulation was validated for standard Li-ion cells and compared against experiments. Finally, its scope and ability to capture spatio-temporal variations of potential and lithium distribution is demonstrated on a prototypical three-dimensional electrode problem.« less
Validation of in vitro assays in three-dimensional human dermal constructs.
Idrees, Ayesha; Chiono, Valeria; Ciardelli, Gianluca; Shah, Siegfried; Viebahn, Richard; Zhang, Xiang; Salber, Jochen
2018-05-01
Three-dimensional cell culture systems are urgently needed for cytocompatibility testing of biomaterials. This work aimed at the development of three-dimensional in vitro dermal skin models and their optimization for cytocompatibility evaluation. Initially "murine in vitro dermal construct" based on L929 cells was generated, leading to the development of "human in vitro dermal construct" consisting of normal human dermal fibroblasts in rat tail tendon collagen type I. To assess the viability of the cells, different assays CellTiter-Blue ® , RealTime-Glo ™ MT, and CellTiter-Glo ® (Promega) were evaluated to optimize the best-suited assay to the respective cell type and three-dimensional system. Z-stack imaging (Live/Dead and Phalloidin/DAPI-Promokine) was performed to visualize normal human dermal fibroblasts inside matrix revealing filopodia-like morphology and a uniform distribution of normal human dermal fibroblasts in matrix. CellTiter-Glo was found to be the optimal cell viability assay among those analyzed. CellTiter-Blue reagent affected the cell morphology of normal human dermal fibroblasts (unlike L929), suggesting an interference with cell biological activity, resulting in less reliable viability data. On the other hand, RealTime-Glo provided a linear signal only with a very low cell density, which made this assay unsuitable for this system. CellTiter-Glo adapted to three-dimensional dermal construct by optimizing the "shaking time" to enhance the reagent penetration and maximum adenosine triphosphate release, indicating 2.4 times higher viability value by shaking for 60 min than for 5 min. In addition, viability results showed that cells were viable inside the matrix. This model would be further advanced with more layers of skin to make a full thickness model.
Edmondson, Rasheena; Broglie, Jessica Jenkins; Adcock, Audrey F.
2014-01-01
Abstract Three-dimensional (3D) cell culture systems have gained increasing interest in drug discovery and tissue engineering due to their evident advantages in providing more physiologically relevant information and more predictive data for in vivo tests. In this review, we discuss the characteristics of 3D cell culture systems in comparison to the two-dimensional (2D) monolayer culture, focusing on cell growth conditions, cell proliferation, population, and gene and protein expression profiles. The innovations and development in 3D culture systems for drug discovery over the past 5 years are also reviewed in the article, emphasizing the cellular response to different classes of anticancer drugs, focusing particularly on similarities and differences between 3D and 2D models across the field. The progression and advancement in the application of 3D cell cultures in cell-based biosensors is another focal point of this review. PMID:24831787
Research on the printability of hydrogels in 3D bioprinting
He, Yong; Yang, FeiFei; Zhao, HaiMing; Gao, Qing; Xia, Bing; Fu, JianZhong
2016-01-01
As the biocompatible materials, hydrogels have been widely used in three- dimensional (3D) bioprinting/organ printing to load cell for tissue engineering. It is important to precisely control hydrogels deposition during printing the mimic organ structures. However, the printability of hydrogels about printing parameters is seldom addressed. In this paper, we systemically investigated the printability of hydrogels from printing lines (one dimensional, 1D structures) to printing lattices/films (two dimensional, 2D structures) and printing 3D structures with a special attention to the accurate printing. After a series of experiments, we discovered the relationships between the important factors such as air pressure, feedrate, or even printing distance and the printing quality of the expected structures. Dumbbell shape was observed in the lattice structures printing due to the hydrogel diffuses at the intersection. Collapses and fusion of adjacent layer would result in the error accumulation at Z direction which was an important fact that could cause printing failure. Finally, we successfully demonstrated a 3D printing hydrogel scaffold through harmonize with all the parameters. The cell viability after printing was compared with the casting and the results showed that our bioprinting method almost had no extra damage to the cells. PMID:27436509
Research on the printability of hydrogels in 3D bioprinting
NASA Astrophysics Data System (ADS)
He, Yong; Yang, Feifei; Zhao, Haiming; Gao, Qing; Xia, Bing; Fu, Jianzhong
2016-07-01
As the biocompatible materials, hydrogels have been widely used in three- dimensional (3D) bioprinting/organ printing to load cell for tissue engineering. It is important to precisely control hydrogels deposition during printing the mimic organ structures. However, the printability of hydrogels about printing parameters is seldom addressed. In this paper, we systemically investigated the printability of hydrogels from printing lines (one dimensional, 1D structures) to printing lattices/films (two dimensional, 2D structures) and printing 3D structures with a special attention to the accurate printing. After a series of experiments, we discovered the relationships between the important factors such as air pressure, feedrate, or even printing distance and the printing quality of the expected structures. Dumbbell shape was observed in the lattice structures printing due to the hydrogel diffuses at the intersection. Collapses and fusion of adjacent layer would result in the error accumulation at Z direction which was an important fact that could cause printing failure. Finally, we successfully demonstrated a 3D printing hydrogel scaffold through harmonize with all the parameters. The cell viability after printing was compared with the casting and the results showed that our bioprinting method almost had no extra damage to the cells.
Kumar, Alok; Akkineni, Ashwini R; Basu, Bikramjit; Gelinsky, Michael
2016-03-01
Scaffolds for bone tissue engineering are essentially characterized by porous three-dimensional structures with interconnected pores to facilitate the exchange of nutrients and removal of waste products from cells, thereby promoting cell proliferation in such engineered scaffolds. Although hydroxyapatite is widely being considered for bone tissue engineering applications due to its occurrence in the natural extracellular matrix of this tissue, limited reports are available on additive manufacturing of hydroxyapatite-based materials. In this perspective, hydroxyapatite-based three-dimensional porous scaffolds with two different binders (maltodextrin and sodium alginate) were fabricated using the extrusion method of three-dimensional plotting and the results were compared in reference to the structural properties of scaffolds processed via chemical stabilization and sintering routes, respectively. With the optimal processing conditions regarding to pH and viscosity of binder-loaded hydroxyapatite pastes, scaffolds with parallelepiped porous architecture having up to 74% porosity were fabricated. Interestingly, sintering of the as-plotted hydroxyapatite-sodium alginate (cross-linked with CaCl2 solution) scaffolds led to the formation of chlorapatite (Ca9.54P5.98O23.8Cl1.60(OH)2.74). Both the sintered scaffolds displayed progressive deformation and delayed fracture under compressive loading, with hydroxyapatite-alginate scaffolds exhibiting a higher compressive strength (9.5 ± 0.5 MPa) than hydroxyapatite-maltodextrin scaffolds (7.0 ± 0.6 MPa). The difference in properties is explained in terms of the phase assemblage and microstructure. © The Author(s) 2015.
Three-dimensional boron particle loaded thermal neutron detector
Nikolic, Rebecca J.; Conway, Adam M.; Graff, Robert T.; Kuntz, Joshua D.; Reinhardt, Catherine; Voss, Lars F.; Cheung, Chin Li; Heineck, Daniel
2014-09-09
Three-dimensional boron particle loaded thermal neutron detectors utilize neutron sensitive conversion materials in the form of nano-powders and micro-sized particles, as opposed to thin films, suspensions, paraffin, etc. More specifically, methods to infiltrate, intersperse and embed the neutron nano-powders to form two-dimensional and/or three-dimensional charge sensitive platforms are specified. The use of nano-powders enables conformal contact with the entire charge-collecting structure regardless of its shape or configuration.
High power density proton exchange membrane fuel cells
NASA Technical Reports Server (NTRS)
Murphy, Oliver J.; Hitchens, G. Duncan; Manko, David J.
1993-01-01
Proton exchange membrane (PEM) fuel cells use a perfluorosulfonic acid solid polymer film as an electrolyte which simplifies water and electrolyte management. Their thin electrolyte layers give efficient systems of low weight, and their materials of construction show extremely long laboratory lifetimes. Their high reliability and their suitability for use in a microgravity environment makes them particularly attractive as a substitute for batteries in satellites utilizing high-power, high energy-density electrochemical energy storage systems. In this investigation, the Dow experimental PEM (XUS-13204.10) and unsupported high platinum loading electrodes yielded very high power densities, of the order of 2.5 W cm(exp -2). A platinum black loading of 5 mg per cm(exp 2) was found to be optimum. On extending the three-dimensional reaction zone of fuel cell electrodes by impregnating solid polymer electrolyte into the electrode structures, Nafion was found to give better performance than the Dow experimental PEM. The depth of penetration of the solid polymer electrolyte into electrode structures was 50-70 percent of the thickness of the platinum-catalyzed active layer. However, the degree of platinum utilization was only 16.6 percent and the roughness factor of a typical electrode was 274.
Three-dimensional canopy fuel loading predicted using upward and downward sensing LiDAR systems
Nicholas S. Skowronski; Kenneth L. Clark; Matthew Duveneck; John. Hom
2011-01-01
We calibrated upward sensing profiling and downward sensing scanning LiDAR systems to estimates of canopy fuel loading developed from field plots and allometric equations, and then used the LiDAR datasets to predict canopy bulk density (CBD) and crown fuel weight (CFW) in wildfire prone stands in the New Jersey Pinelands. LiDAR-derived height profiles were also...
Multitasking the three-dimensional transport code TORT on CRAY platforms
DOE Office of Scientific and Technical Information (OSTI.GOV)
Azmy, Y.Y.; Barnett, D.A.; Burre, C.A.
1996-04-01
The multitasking options in the three-dimensional neutral particle transport code TORT originally implemented for Cray`s CTSS operating system are revived and extended to run on Cray Y/MP and C90 computers using the UNICOS operating system. These include two coarse-grained domain decompositions; across octants, and across directions within an octant, termed Octant Parallel (OP), and Direction Parallel (DP), respectively. Parallel performance of the DP is significantly enhanced by increasing the task grain size and reducing load imbalance via dynamic scheduling of the discrete angles among the participating tasks. Substantial Wall Clock speedup factors, approaching 4.5 using 8 tasks, have been measuredmore » in a time-sharing environment, and generally depend on the test problem specifications, number of tasks, and machine loading during execution.« less
DOE Office of Scientific and Technical Information (OSTI.GOV)
Pasquariello, Vito, E-mail: vito.pasquariello@tum.de; Hammerl, Georg; Örley, Felix
2016-02-15
We present a loosely coupled approach for the solution of fluid–structure interaction problems between a compressible flow and a deformable structure. The method is based on staggered Dirichlet–Neumann partitioning. The interface motion in the Eulerian frame is accounted for by a conservative cut-cell Immersed Boundary method. The present approach enables sub-cell resolution by considering individual cut-elements within a single fluid cell, which guarantees an accurate representation of the time-varying solid interface. The cut-cell procedure inevitably leads to non-matching interfaces, demanding for a special treatment. A Mortar method is chosen in order to obtain a conservative and consistent load transfer. Wemore » validate our method by investigating two-dimensional test cases comprising a shock-loaded rigid cylinder and a deformable panel. Moreover, the aeroelastic instability of a thin plate structure is studied with a focus on the prediction of flutter onset. Finally, we propose a three-dimensional fluid–structure interaction test case of a flexible inflated thin shell interacting with a shock wave involving large and complex structural deformations.« less
Li, Zhonghua; Wang, Haiqin; Yang, Bo; Sun, Yukai; Huo, Ran
2015-12-01
The regeneration of functional skin remains elusive, due to poor engraftment, deficient vascularization, and excessive scar formation. Aiming to overcome these issues, the present study proposed the combination of a three-dimensional graphene foam (GF) scaffold loaded with bone marrow derived mesenchymal stem cells (MSCs) to improve skin wound healing. The GFs demonstrated good biocompatibility and promoted the growth and proliferation of MSCs. Meanwhile, the GFs loaded with MSCs obviously facilitated wound closure in animal model. The dermis formed in the presence of the GF structure loaded with MSCs was thicker and possessed a more complex structure at day 14 post-surgery. The transplanted MSCs correlated with upregulation of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), which may lead to neo-vascularization. Additionally, an anti-scarring effect was observed in the presence of the 3D-GF scaffold and MSCs, as evidenced by a downregulation of transforming growth factor-beta 1 (TGF-β1) and alpha-smooth muscle actin (α-SMA) together with an increase of TGF-β3. Altogether, the GF scaffold could guide the wound healing process with reduced scarring, and the MSCs were crucial to enhance vascularization and provided a better quality neo-skin. The GF scaffold loaded with MSCs possesses necessary bioactive cues to improve wound healing with reduced scarring, which may be of great clinical significance for skin wound healing. Copyright © 2015 Elsevier B.V. All rights reserved.
Liu, Yayuan; Lin, Dingchang; Jin, Yang; Liu, Kai; Tao, Xinyong; Zhang, Qiuhong; Zhang, Xiaokun; Cui, Yi
2017-01-01
Solid-state lithium (Li) metal batteries are prominent among next-generation energy storage technologies due to their significantly high energy density and reduced safety risks. Previously, solid electrolytes have been intensively studied and several materials with high ionic conductivity have been identified. However, there are still at least three obstacles before making the Li metal foil-based solid-state systems viable, namely, high interfacial resistance at the Li/electrolyte interface, low areal capacity, and poor power output. The problems are addressed by incorporating a flowable interfacial layer and three-dimensional Li into the system. The flowable interfacial layer can accommodate the interfacial fluctuation and guarantee excellent adhesion at all time, whereas the three-dimensional Li significantly reduces the interfacial fluctuation from the whole electrode level (tens of micrometers) to local scale (submicrometer) and also decreases the effective current density for high-capacity and high-power operations. As a consequence, both symmetric and full-cell configurations can achieve greatly improved electrochemical performances in comparison to the conventional Li foil, which are among the best reported values in the literature. Noticeably, solid-state full cells paired with high–mass loading LiFePO4 exhibited, at 80°C, a satisfactory specific capacity even at a rate of 5 C (110 mA·hour g−1) and a capacity retention of 93.6% after 300 cycles at a current density of 3 mA cm−2 using a composite solid electrolyte middle layer. In addition, when a ceramic electrolyte middle layer was adopted, stable cycling with greatly improved capacity could even be realized at room temperature. PMID:29062894
DOE Office of Scientific and Technical Information (OSTI.GOV)
Liu, Yayuan; Lin, Dingchang; Jin, Yang
Solid-state lithium (Li) metal batteries are prominent among next-generation energy storage technologies due to their significantly high energy density and reduced safety risks. Previously, solid electrolytes have been intensively studied and several materials with high ionic conductivity have been identified. However, there are still at least three obstacles before making the Li metal foil-based solid-state systems viable, namely, high interfacial resistance at the Li/electrolyte interface, low areal capacity, and poor power output. The problems are addressed by incorporating a flowable interfacial layer and three-dimensional Li into the system. The flowable interfacial layer can accommodate the interfacial fluctuation and guarantee excellentmore » adhesion at all time, whereas the three-dimensional Li significantly reduces the interfacial fluctuation from the whole electrode level (tens of micrometers) to local scale (submicrometer) and also decreases the effective current density for high-capacity and high-power operations. As a consequence, both symmetric and full-cell configurations can achieve greatly improved electrochemical performances in comparison to the conventional Li foil, which are among the best reported values in the literature. Noticeably, solid-state full cells paired with high–mass loading LiFePO4 exhibited, at 80°C, a satisfactory specific capacity even at a rate of 5 C (110 mA·hour g -1) and a capacity retention of 93.6% after 300 cycles at a current density of 3 mA cm -2 using a composite solid electrolyte middle layer. In addition, when a ceramic electrolyte middle layer was adopted, stable cycling with greatly improved capacity could even be realized at room temperature.« less
Liu, Yayuan; Lin, Dingchang; Jin, Yang; ...
2017-10-01
Solid-state lithium (Li) metal batteries are prominent among next-generation energy storage technologies due to their significantly high energy density and reduced safety risks. Previously, solid electrolytes have been intensively studied and several materials with high ionic conductivity have been identified. However, there are still at least three obstacles before making the Li metal foil-based solid-state systems viable, namely, high interfacial resistance at the Li/electrolyte interface, low areal capacity, and poor power output. The problems are addressed by incorporating a flowable interfacial layer and three-dimensional Li into the system. The flowable interfacial layer can accommodate the interfacial fluctuation and guarantee excellentmore » adhesion at all time, whereas the three-dimensional Li significantly reduces the interfacial fluctuation from the whole electrode level (tens of micrometers) to local scale (submicrometer) and also decreases the effective current density for high-capacity and high-power operations. As a consequence, both symmetric and full-cell configurations can achieve greatly improved electrochemical performances in comparison to the conventional Li foil, which are among the best reported values in the literature. Noticeably, solid-state full cells paired with high–mass loading LiFePO4 exhibited, at 80°C, a satisfactory specific capacity even at a rate of 5 C (110 mA·hour g -1) and a capacity retention of 93.6% after 300 cycles at a current density of 3 mA cm -2 using a composite solid electrolyte middle layer. In addition, when a ceramic electrolyte middle layer was adopted, stable cycling with greatly improved capacity could even be realized at room temperature.« less
Wang, Tao; Green, Ryan; Nair, Rajesh Ramakrishnan; Howell, Mark; Mohapatra, Subhra; Guldiken, Rasim; Mohapatra, Shyam Sundar
2015-12-19
Detection and quantification of cell viability and growth in two-dimensional (2D) and three-dimensional (3D) cell cultures commonly involve harvesting of cells and therefore requires a parallel set-up of several replicates for time-lapse or dose-response studies. Thus, developing a non-invasive and touch-free detection of cell growth in longitudinal studies of 3D tumor spheroid cultures or of stem cell regeneration remains a major unmet need. Since surface acoustic waves (SAWs) permit mass loading-based biosensing and have been touted due to their many advantages including low cost, small size and ease of assembly, we examined the potential of SAW-biosensing to detect and quantify cell growth. Herein, we demonstrate that a shear horizontal-surface acoustic waves (SH-SAW) device comprising two pairs of resonators consisting of interdigital transducers and reflecting fingers can be used to quantify mass loading by the cells in suspension as well as within a 3D cell culture platform. A 3D COMSOL model was built to simulate the mass loading response of increasing concentrations of cells in suspension in the polydimethylsiloxane (PDMS) well in order to predict the characteristics and optimize the design of the SH-SAW biosensor. The simulated relative frequency shift from the two oscillatory circuit systems (one of which functions as control) were found to be concordant to experimental data generated with RAW264.7 macrophage and A549 cancer cells. In addition, results showed that SAW measurements per se did not affect viability of cells. Further, SH-SAW biosensing was applied to A549 cells cultured on a 3D electrospun nanofiber scaffold that generate tumor spheroids (tumoroids) and the results showed the device's ability to detect changes in tumor spheroid growth over the course of eight days. Taken together, these results demonstrate the use of SH-SAW device for detection and quantification of cell growth changes over time in 2D suspension cultures and in 3D cell culture models, which may have potential applications in both longitudinal 3D cell cultures in cancer biology and in regenerative medicine.
Modelling three-dimensional cochlear micromechanics within the guinea pig organ of Corti
NASA Astrophysics Data System (ADS)
Ni, Guangjian; Elliott, Stephen J.
2018-05-01
The active amplification process in the mammalian cochlea depends on a complex interaction between cells within the organ of Corti. A three-dimensional (3D) model was developed using the finite element method based on anatomy for the apical end in the guinea pig cochlea, which is comprised of 3D discrete hair cells, 3D continuous membranes and fluid. The basilar membrane, tectorial membrane and the reticular lamina are modelled with orthotropic materials. The Y-shape structures formed by the outer hair cell (OHC), the Deiters' cell and Deiters' cell phalangeal process are also included to account for the structural longitudinal coupling. The motion within the organ of Corti was first simulated in response to a pressure difference loading on the basilar membrane, in order to calculate the passive vibration pattern. Then, the outer hair cells somatic electromotility was implemented by applying a voltage across the OHC walls to investigate its contribution to membranes motion.
Cai, Xianhua; Yu, Yang; Liu, Zhichao; Zhang, Meichao; Huang, Weibing
2014-08-01
Although there are many techniques for occipitocervical fixation, there have been no reports regarding occipitocervical fixation via the use of an anterior anatomical locking plate system. The biomechanics of this new system were analyzed by a three-dimensional finite element to provide a theoretical basis for clinical application. This was a modeling study. We studied a 27-year-old healthy male volunteer in whom cervical disease was excluded via X-ray examination. The states of stress and strain of these two internal fixation devices were analyzed. A three-dimensional finite element model of normal occiput-C2 was established based on the anatomical data from a Chinese population. An unstable model of occipital-cervical region was established by subtracting several unit structures from the normal model. An anterior occiput-to-axis locking titanium plate system was then applied and an anterior occiput-to-axis screw fixation was performed on the unstable model. Limitation of motion was performed on the surface of the fixed model, and physiological loads were imposed on the surface of the skull base. Under various loads from different directions, the peak values of displacement of the anterior occiput-to-axis locking titanium plate system decreased 15.5%, 12.5%, 14.4%, and 23.7%, respectively, under the loads of flexion, extension, lateral bending, and axial rotation. Compared with the anterior occiput-to-axis screw fixation, the peak values of stress of the anterior occiput-to-axis locking titanium plate system also decreased 3.9%, 2.9%, 9.7%, and 7.2%, respectively, under the loads of flexion, extension, lateral bending, and axial rotation. The anterior occiput-to-axis locking titanium plate system proved superior to the anterior occiput-to-axis screw system both in the stress distribution and fixation stability based on finite element analysis. It provides a new clinical option for anterior occipitocervical fixation. Copyright © 2014 Elsevier Inc. All rights reserved.
2001-05-01
Kinematic and Kinetic Data Collection Systems. Three-dimensional kinematic data were collected at 100 Hz through an Optotrak 3020 System (Northern...regardless of load. The Optotrak system unit provides an external trigger that was used to trigger the start of the force plate data collection, thereby...synchronizing the kinematic and kinetic data. Optotrak required the use of infrared light emitting diodes (IREDS), which were placed bilaterally on the
NASA Astrophysics Data System (ADS)
Evans, Conor
2015-03-01
Three dimensional, in vitro spheroid cultures offer considerable utility for the development and testing of anticancer photodynamic therapy regimens. More complex than monolayer cultures, three-dimensional spheroid systems replicate many of the important cell-cell and cell-matrix interactions that modulate treatment response in vivo. Simple enough to be grown by the thousands and small enough to be optically interrogated, spheroid cultures lend themselves to high-content and high-throughput imaging approaches. These advantages have enabled studies investigating photosensitizer uptake, spatiotemporal patterns of therapeutic response, alterations in oxygen diffusion and consumption during therapy, and the exploration of mechanisms that underlie therapeutic synergy. The use of quantitative imaging methods, in particular, has accelerated the pace of three-dimensional in vitro photodynamic therapy studies, enabling the rapid compilation of multiple treatment response parameters in a single experiment. Improvements in model cultures, the creation of new molecular probes of cell state and function, and innovations in imaging toolkits will be important for the advancement of spheroid culture systems for future photodynamic therapy studies.
Open-Porous Hydroxyapatite Scaffolds for Three-Dimensional Culture of Human Adult Liver Cells
Schmelzer, Eva; Over, Patrick; Nettleship, Ian; Gerlach, Joerg C.
2016-01-01
Liver cell culture within three-dimensional structures provides an improved culture system for various applications in basic research, pharmacological screening, and implantable or extracorporeal liver support. Biodegradable calcium-based scaffolds in such systems could enhance liver cell functionality by providing endothelial and hepatic cell support through locally elevated calcium levels, increased surface area for cell attachment, and allowing three-dimensional tissue restructuring. Open-porous hydroxyapatite scaffolds were fabricated and seeded with primary adult human liver cells, which were embedded within or without gels of extracellular matrix protein collagen-1 or hyaluronan. Metabolic functions were assessed after 5, 15, and 28 days. Longer-term cultures exhibited highest cell numbers and liver specific gene expression when cultured on hydroxyapatite scaffolds in collagen-1. Endothelial gene expression was induced in cells cultured on scaffolds without extracellular matrix proteins. Hydroxyapatite induced gene expression for cytokeratin-19 when cells were cultured in collagen-1 gel while culture in hyaluronan increased cytokeratin-19 gene expression independent of the use of scaffold in long-term culture. The implementation of hydroxyapatite composites with extracellular matrices affected liver cell cultures and cell differentiation depending on the type of matrix protein and the presence of a scaffold. The hydroxyapatite scaffolds enable scale-up of hepatic three-dimensional culture models for regenerative medicine applications. PMID:27403430
Hydrofocusing Bioreactor for Three-Dimensional Cell Culture
NASA Technical Reports Server (NTRS)
Gonda, Steve R.; Spaulding, Glenn F.; Tsao, Yow-Min D.; Flechsig, Scott; Jones, Leslie; Soehnge, Holly
2003-01-01
The hydrodynamic focusing bioreactor (HFB) is a bioreactor system designed for three-dimensional cell culture and tissue-engineering investigations on orbiting spacecraft and in laboratories on Earth. The HFB offers a unique hydrofocusing capability that enables the creation of a low-shear culture environment simultaneously with the "herding" of suspended cells, tissue assemblies, and air bubbles. Under development for use in the Biotechnology Facility on the International Space Station, the HFB has successfully grown large three-dimensional, tissuelike assemblies from anchorage-dependent cells and grown suspension hybridoma cells to high densities. The HFB, based on the principle of hydrodynamic focusing, provides the capability to control the movement of air bubbles and removes them from the bioreactor without degrading the low-shear culture environment or the suspended three-dimensional tissue assemblies. The HFB also provides unparalleled control over the locations of cells and tissues within its bioreactor vessel during operation and sampling.
NASA Astrophysics Data System (ADS)
Tsang, Alpha C. H.; Kwok, Holly Y. H.; Leung, Dennis Y. C.
2017-05-01
This manuscript presents the methodology of the production of 2D and 3D graphene based material, and their applications in fuel cell, supercapacitor, and photovoltic in recent years. Due to the uniqueness and attractive properties of graphene nanosheets, a large number of techniques have been developed for raw graphene preparation, from a chemical method to a physical deposition of carbon vapor under extreme conditions. A variety of graphene based materials were also prepared from raw graphene or graphene oxide, including the metal loaded, metal oxides loaded, to the foreign elements doped graphene. Both two-dimensional (2D) to three-dimensional (3D) structured graphene were covered. These materials included the bulk or template hybrid composite, containing graphene hydrogel, graphene aerogel, or graphene foam and its derived products. They were widely used in green energy device research, which exhibited strong activity, and developed some special usage in recent research.
Santoro, Carlo; Kodali, Mounika; Kabir, Sadia; Soavi, Francesca; Serov, Alexey; Atanassov, Plamen
2017-07-15
Three-dimensional graphene nanosheets (3D-GNS) were used as cathode catalysts for microbial fuel cells (MFCs) operating in neutral conditions. 3D-GNS catalysts showed high performance towards oxygen electroreduction in neutral media with high current densities and low hydrogen peroxide generation compared to activated carbon (AC). 3D-GNS was incorporated into air-breathing cathodes based on AC with three different loadings (2, 6 and 10 mgcm -2 ). Performances in MFCs showed that 3D-GNS had the highest performances with power densities of 2.059 ± 0.003 Wm -2 , 1.855 ± 0.007 Wm -2 and 1.503 ± 0.005 Wm -2 for loading of 10, 6 and 2 mgcm -2 respectively. Plain AC had the lowest performances (1.017 ± 0.009 Wm -2 ). The different cathodes were also investigated in supercapacitive MFCs (SC-MFCs). The addition of 3D-GNS decreased the ohmic losses by 14-25%. The decrease in ohmic losses allowed the SC-MFC with 3D-GNS (loading 10 mgcm -2 ) to have the maximum power (P max ) of 5.746 ± 0.186 Wm -2 . At 5 mA, the SC-MFC featured an "apparent" capacitive response that increased from 0.027 ± 0.007 F with AC to 0.213 ± 0.026 F with 3D-GNS (loading 2 mgcm -2 ) and further to 1.817 ± 0.040 F with 3D-GNS (loading 10 mgcm -2 ).
NASA Technical Reports Server (NTRS)
Straube, Timothy Milton
1993-01-01
The design and implementation of a vertical degree of freedom suspension system is described which provides a constant force off-load condition to counter gravity over large displacements. By accommodating motions up to one meter for structures weighing up to 100 pounds, the system is useful for experiments which simulate orbital construction events such as docking, multiple component assembly, or structural deployment. A unique aspect of this device is the combination of a large stroke passive off-load device augmented by electromotive torque actuated force feedback. The active force feedback has the effect of reducing break-away friction by a factor of twenty over the passive system alone. The thesis describes the development of the suspension hardware and the control algorithm. Experiments were performed to verify the suspensions system's effectiveness in providing a gravity off-load and simulating the motion of a structure in orbit. Additionally, a three dimensional system concept is presented as an extension of the one dimensional suspension system which was implemented.
Fracture Mechanical Analysis of Open Cell Ceramic Foams Under Thermal Shock Loading
NASA Astrophysics Data System (ADS)
Settgast, C.; Abendroth, M.; Kuna, M.
2016-11-01
Ceramic foams made by replica techniques containing sharp-edged cavities, which are potential crack initiators and therefore have to be analyzed using fracture mechanical methods. The ceramic foams made of novel carbon bonded alumina are used as filters in metal melt filtration applications, where the filters are exposed to a thermal shock. During the casting process the filters experience a complex thermo-mechanical loading, which is difficult to measure. Modern numerical methods allow the simulation of such complex processes. As a simplified foam structure an open Kelvin cell is used as a representative volume element. A three-dimensional finite element model containing realistic sharp-edged cavities and three-dimensional sub-models along these sharp edges are used to compute the transient temperature, stress and strain fields at the Kelvin foam. The sharp edges are evaluated using fracture mechanical methods like the J-integral technique. The results of this study describe the influence of the pore size, relative density of the ceramic foam, the heat transfer and selected material parameters on the fracture mechanical behaviour.
Su, Min; Fan, Chao; Gao, Sainan; Shen, Aiguo; Wang, Xiaoying; Zhang, Yuquan
2015-11-01
We investigated the expression of human chorionic gonadotropin (HCG) and its effects on vasculogenic mimicry (VM) formation in ovarian cancer cells under normoxic and hypoxic conditions in three-dimensional matrices preconditioned by an endothelial-trophoblast cell co-culture system. The co-culture model was established using human umbilical vein endothelial cells (HUVECs) and HTR-8 trophoblast cells in a three-dimensional culture system. The co-cultured cells were removed with NH4OH, and ovarian cancer cells were implanted into the preconditioned matrix. VM was identified morphologically and by detecting vascular markers expressed by cancer cells. The specificity of the effects of exogenous HCG in the microenvironment was assessed by inhibition with a neutralizing anti-HCG antibody. HCG siRNA was used to knock down endogenous HCG expression in OVCAR-3 ovarian cancer cells. HTR-8 cells 'fingerprinted' HUVECs to form capillary-like tube structures in co-cultures. In the preconditioned HCG-rich microenvironment, the number of vessel-like network structures formed by HCG receptor-positive OVCAR-3 cells and the expression levels of CD31, VEGF and factor VIII were significantly increased. The preconditioned HCG-rich microenvironment significantly increased the expression of hypoxia inducible factor-1α (HIF‑1α) and VM formation in OVCAR-3 cells under hypoxic conditions. Treatment with a neutralizing anti-HCG antibody but not HCG siRNA significantly inhibited the formation of vessel-like network structures. HCG in the microenvironment contributes to OVCAR-3 differentiation into endothelioid cells in three-dimensional matrices preconditioned with an endothelial-trophoblast cell co-culture system. HCG may synergistically enhance hypoxia-induced vascular markers and HIF-1α expression. These findings would provide perspectives on new therapeutic targets for ovarian cancer.
NASA Astrophysics Data System (ADS)
Yamauchi, Toyohiko; Kakuno, Yumi; Goto, Kentaro; Fukami, Tadashi; Sugiyama, Norikazu; Iwai, Hidenao; Mizuguchi, Yoshinori; Yamashita, Yutaka
2014-03-01
There is an increasing need for non-invasive imaging techniques in the field of stem cell research. Label-free techniques are the best choice for assessment of stem cells because the cells remain intact after imaging and can be used for further studies such as differentiation induction. To develop a high-resolution label-free imaging system, we have been working on a low-coherence quantitative phase microscope (LC-QPM). LC-QPM is a Linnik-type interference microscope equipped with nanometer-resolution optical-path-length control and capable of obtaining three-dimensional volumetric images. The lateral and vertical resolutions of our system are respectively 0.5 and 0.93 μm and this performance allows capturing sub-cellular morphological features of live cells without labeling. Utilizing LC-QPM, we reported on three-dimensional imaging of membrane fluctuations, dynamics of filopodia, and motions of intracellular organelles. In this presentation, we report three-dimensional morphological imaging of human induced pluripotent stem cells (hiPS cells). Two groups of monolayer hiPS cell cultures were prepared so that one group was cultured in a suitable culture medium that kept the cells undifferentiated, and the other group was cultured in a medium supplemented with retinoic acid, which forces the stem cells to differentiate. The volumetric images of the 2 groups show distinctive differences, especially in surface roughness. We believe that our LC-QPM system will prove useful in assessing many other stem cell conditions.
Kai, M; Aoki, O; Hiraga, A; Oki, H; Tokuriki, M
2000-08-01
To develop an instrument that could be sandwiched between the hoof and shoe of horses and that would reliably measure vertical ground reaction forces and three-dimensional acceleration at the walk, trot, and canter. 5 clinically sound Thoroughbreds. The recording instrument (weight, 350 g) consisted of 2 metal plates, 2 bolts, 4 load cells, and 3 accelerometers. It was mounted to the hoof with a glue-on shoe and devised to support as much load exerted by a limb as possible. The load cells and accelerometers were wired to a 16-channel transmitter, and transmitted signals were received and amplified with a telemetry receiver. The recording instrument could measure in real time the 4 components of the ground reaction force or their resultant force along with acceleration in 3 dimensions as horses walked, trotted, or cantered on a treadmill. Patterns of force-time curves recorded for consecutive strides were similar to each other and to those previously reported, using a force plate. The recording instrument developed for use in the present study allowed us to record vertical ground reaction force and acceleration in 3 dimensions in horses at the walk, trot, and canter.
Thermal modelling of Li-ion polymer battery for electric vehicle drive cycles
NASA Astrophysics Data System (ADS)
Chacko, Salvio; Chung, Yongmann M.
2012-09-01
Time-dependent, thermal behaviour of a lithium-ion (Li-ion) polymer cell has been modelled for electric vehicle (EV) drive cycles with a view to developing an effective battery thermal management system. The fully coupled, three-dimensional transient electro-thermal model has been implemented based on a finite volume method. To support the numerical study, a high energy density Li-ion polymer pouch cell was tested in a climatic chamber for electric load cycles consisting of various charge and discharge rates, and a good agreement was found between the model predictions and the experimental data. The cell-level thermal behaviour under stressful conditions such as high power draw and high ambient temperature was predicted with the model. A significant temperature increase was observed in the stressful condition, corresponding to a repeated acceleration and deceleration, indicating that an effective battery thermal management system would be required to maintain the optimal cell performance and also to achieve a full battery lifesapn.
Computer simulation of plasma and N-body problems
NASA Technical Reports Server (NTRS)
Harries, W. L.; Miller, J. B.
1975-01-01
The following FORTRAN language computer codes are presented: (1) efficient two- and three-dimensional central force potential solvers; (2) a three-dimensional simulator of an isolated galaxy which incorporates the potential solver; (3) a two-dimensional particle-in-cell simulator of the Jeans instability in an infinite self-gravitating compressible gas; and (4) a two-dimensional particle-in-cell simulator of a rotating self-gravitating compressible gaseous system of which rectangular coordinate and superior polar coordinate versions were written.
NASA Technical Reports Server (NTRS)
Ihrke, Chris A. (Inventor); Diftler, Myron A. (Inventor); Linn, Douglas Martin (Inventor); Platt, Robert (Inventor); Griffith, Bryan Kristian (Inventor)
2010-01-01
A tactile load cell that has particular application for measuring the load on a phalange in a dexterous robot system. The load cell includes a flexible strain element having first and second end portions that can be used to mount the load cell to the phalange and a center portion that can be used to mount a suitable contact surface to the load cell. The strain element also includes a first S-shaped member including at least three sections connected to the first end portion and the center portion and a second S-shaped member including at least three sections coupled to the second end portion and the center portion. The load cell also includes eight strain gauge pairs where each strain gauge pair is mounted to opposing surfaces of one of the sections of the S-shaped members where the strain gauge pairs provide strain measurements in six-degrees of freedom.
Chen, X C; Liu, H; Li, H; Cheng, Y; Yang, L; Liu, Y F
2016-06-27
In this study, a dynamic three-dimensional cell culture technology was used to expand and differentiate rat pancreatic duct-derived stem cells (PDSCs) into islet-like cell clusters that can secrete insulin. PDSCs were isolated from rat pancreatic tissues by in situ collagenase digestion and density gradient centrifugation. Using a dynamic three-dimensional culture technique, the cells were expanded and differentiated into functional islet-like cell clusters, which were characterized by morphological and phenotype analyses. After maintaining 1 x 108 isolated rat PDSCs in a dynamic three-dimensional cell culture for 7 days, 1.5 x 109 cells could be harvested. Passaged PDSCs expressed markers of pancreatic endocrine progenitors, including CD29 (86.17%), CD73 (90.73%), CD90 (84.13%), CD105 (78.28%), and Pdx-1. Following 14 additional days of culture in serum-free medium with nicotinamide, keratinocyte growth factor (KGF), and b fibroblast growth factor (FGF), the cells were differentiated into islet-like cell clusters (ICCs). The ICC morphology reflected that of fused cell clusters. During the late stage of differentiation, representative clusters were non-adherent and expressed insulin indicated by dithizone (DTZ)-positive staining. Insulin was detected in the extracellular fluid and cytoplasm of ICCs after 14 days of differentiation. Additionally, insulin levels were significantly higher at this time compared with the levels exhibited by PDSCs before differentiation (P < 0.01). By using a dynamic three-dimensional cell culture system, PDSCs can be expanded in vitro and can differentiate into functional islet-like cell clusters.
Improved function and growth of pancreatic cells in a three-dimensional bioreactor environment.
Samuelson, Lisa; Gerber, David A
2013-01-01
Methods of three-dimensional (3D) cell culture have made significant progress in recent years due to a better understanding of cell to cell interactions and the cell's interface with their surrounding environment. We hypothesized that a microgravity 3D culture system would improve upon the growth and function of a pancreatic progenitor cell population. We developed a rotating wall vessel bioreactor and established a culture system using a pancreatic cell line. Cells in the bioreactors showed robust proliferation, enhanced transcriptional signaling, and improved translation of pancreatic genes compared with two-dimensional static culture. Cells also gained the ability to respond to glucose stimulation, which was not observed in the control cultures. These findings suggest that a 3D microgravity bioreactor environment mimics the niche of the pancreas yielding a cell source with potential for cell-based therapy in the treatment of diabetes.
Metzger, Marco; Bareiss, Petra M; Nikolov, Ivan; Skutella, Thomas; Just, Lothar
2007-01-01
Three-dimensional intestinal cultures offer new possibilities for the examination of growth potential, analysis of time specific gene expression, and spatial cellular arrangement of enteric nervous system in an organotypical environment. We present an easy to produce in vitro model of the enteric nervous system for analysis and manipulation of cellular differentiation processes. Slice cultures of murine fetal colon were cultured on membrane inserts for up to 2 weeks without loss of autonomous contractility. After slice preparation, cultured tissue reorganized within the first days in vitro. Afterward, the culture possessed more than 35 cell layers, including high prismatic epithelial cells, smooth muscle cells, glial cells, and neurons analyzed by immunohistochemistry. The contraction frequency of intestinal slice culture could be modulated by the neurotransmitter serotonin and the sodium channel blocker tetrodotoxin. Coculture experiments with cultured neurospheres isolated from enhanced green fluorescent protein (eGFP) transgenic mice demonstrated that differentiating eGFP-positive neurons were integrated into the intestinal tissue culture. This slice culture model of enteric nervous system proved to be useful for studying cell-cell interactions, cellular signaling, and cell differentiation processes in a three-dimensional cell arrangement.
Cao, Li; Guilak, Farshid; Setton, Lori A
2011-02-01
Nucleus pulposus (NP) cells of the intervertebral disk (IVD) have unique morphological characteristics and biologic responses to mechanical stimuli that may regulate maintenance and health of the IVD. NP cells reside as single cell, paired or multiple cells in a contiguous pericellular matrix (PCM), whose structure and properties may significantly influence cell and extracellular matrix mechanics. In this study, a computational model was developed to predict the stress-strain, fluid pressure and flow fields for cells and their surrounding PCM in the NP using three-dimensional (3D) finite element models based on the in situ morphology of cell-PCM regions of the mature rat NP, measured using confocal microscopy. Three-dimensional geometries of the extracellular matrix and representative cell-matrix units were used to construct 3D finite element models of the structures as isotropic and biphasic materials. In response to compressive strain of the extracellular matrix, NP cells and PCM regions were predicted to experience volumetric strains that were 1.9-3.7 and 1.4-2.1 times greater than the extracellular matrix, respectively. Volumetric and deviatoric strain concentrations were generally found at the cell/PCM interface, while von Mises stress concentrations were associated with the PCM/extracellular matrix interface. Cell-matrix units containing greater cell numbers were associated with higher peak cell strains and lower rates of fluid pressurization upon loading. These studies provide new model predictions for micromechanics of NP cells that can contribute to an understanding of mechanotransduction in the IVD and its changes with aging and degeneration.
Microengineering methods for cell-based microarrays and high-throughput drug-screening applications.
Xu, Feng; Wu, JinHui; Wang, ShuQi; Durmus, Naside Gozde; Gurkan, Umut Atakan; Demirci, Utkan
2011-09-01
Screening for effective therapeutic agents from millions of drug candidates is costly, time consuming, and often faces concerns due to the extensive use of animals. To improve cost effectiveness, and to minimize animal testing in pharmaceutical research, in vitro monolayer cell microarrays with multiwell plate assays have been developed. Integration of cell microarrays with microfluidic systems has facilitated automated and controlled component loading, significantly reducing the consumption of the candidate compounds and the target cells. Even though these methods significantly increased the throughput compared to conventional in vitro testing systems and in vivo animal models, the cost associated with these platforms remains prohibitively high. Besides, there is a need for three-dimensional (3D) cell-based drug-screening models which can mimic the in vivo microenvironment and the functionality of the native tissues. Here, we present the state-of-the-art microengineering approaches that can be used to develop 3D cell-based drug-screening assays. We highlight the 3D in vitro cell culture systems with live cell-based arrays, microfluidic cell culture systems, and their application to high-throughput drug screening. We conclude that among the emerging microengineering approaches, bioprinting holds great potential to provide repeatable 3D cell-based constructs with high temporal, spatial control and versatility.
Microengineering Methods for Cell Based Microarrays and High-Throughput Drug Screening Applications
Xu, Feng; Wu, JinHui; Wang, ShuQi; Durmus, Naside Gozde; Gurkan, Umut Atakan; Demirci, Utkan
2011-01-01
Screening for effective therapeutic agents from millions of drug candidates is costly, time-consuming and often face ethical concerns due to extensive use of animals. To improve cost-effectiveness, and to minimize animal testing in pharmaceutical research, in vitro monolayer cell microarrays with multiwell plate assays have been developed. Integration of cell microarrays with microfluidic systems have facilitated automated and controlled component loading, significantly reducing the consumption of the candidate compounds and the target cells. Even though these methods significantly increased the throughput compared to conventional in vitro testing systems and in vivo animal models, the cost associated with these platforms remains prohibitively high. Besides, there is a need for three-dimensional (3D) cell based drug-screening models, which can mimic the in vivo microenvironment and the functionality of the native tissues. Here, we present the state-of-the-art microengineering approaches that can be used to develop 3D cell based drug screening assays. We highlight the 3D in vitro cell culture systems with live cell-based arrays, microfluidic cell culture systems, and their application to high-throughput drug screening. We conclude that among the emerging microengineering approaches, bioprinting holds a great potential to provide repeatable 3D cell based constructs with high temporal, spatial control and versatility. PMID:21725152
Platform switching: biomechanical evaluation using three-dimensional finite element analysis.
Tabata, Lucas Fernando; Rocha, Eduardo Passos; Barão, Valentim Adelino Ricardo; Assunção, Wirley Goncalves
2011-01-01
The objective of this study was to evaluate, using three-dimensional finite element analysis (3D FEA), the stress distribution in peri-implant bone tissue, implants, and prosthetic components of implant-supported single crowns with the use of the platform-switching concept. Three 3D finite element models were created to replicate an external-hexagonal implant system with peri-implant bone tissue in which three different implant-abutment configurations were represented. In the regular platform (RP) group, a regular 4.1-mm-diameter abutment (UCLA) was connected to regular 4.1-mm-diameter implant. The platform-switching (PS) group was simulated by the connection of a wide implant (5.0 mm diameter) to a regular 4.1-mm-diameter UCLA abutment. In the wide-platform (WP) group, a 5.0-mm-diameter UCLA abutment was connected to a 5.0-mm-diameter implant. An occlusal load of 100 N was applied either axially or obliquely on the models using ANSYS software. Both the increase in implant diameter and the use of platform switching played roles in stress reduction. The PS group presented lower stress values than the RP and WP groups for bone and implant. In the peri-implant area, cortical bone exhibited a higher stress concentration than the trabecular bone in all models and both loading situations. Under oblique loading, higher intensity and greater distribution of stress were observed than under axial loading. Platform switching reduced von Mises (17.5% and 9.3% for axial and oblique loads, respectively), minimum (compressive) (19.4% for axial load and 21.9% for oblique load), and maximum (tensile) principal stress values (46.6% for axial load and 26.7% for oblique load) in the peri-implant bone tissue. Platform switching led to improved biomechanical stress distribution in peri-implant bone tissue. Oblique loads resulted in higher stress concentrations than axial loads for all models. Wide-diameter implants had a large influence in reducing stress values in the implant system.
Transient Three-Dimensional Analysis of Nozzle Side Load in Regeneratively Cooled Engines
NASA Technical Reports Server (NTRS)
ng, Ten-See
2005-01-01
Nozzle side loads are potentially detrimental to the integrity and life of almost all launch vehicles. the lack of a detailed prediction capability results in reducing life and increased weight for reusable nozzle systems. A clear understanding of the mechanism that contribute to side loads during engine startup, shutdown, and steady-state operations must be established. A CFD based predictive tool must be developed to aid the understanding of side load physics and development of future reusable engine.
Aeroacoustic theory for noncompact wing-gust interaction
NASA Technical Reports Server (NTRS)
Martinez, R.; Widnall, S. E.
1981-01-01
Three aeroacoustic models for noncompact wing-gust interaction were developed for subsonic flow. The first is that for a two dimensional (infinite span) wing passing through an oblique gust. The unsteady pressure field was obtained by the Wiener-Hopf technique; the airfoil loading and the associated acoustic field were calculated, respectively, by allowing the field point down on the airfoil surface, or by letting it go to infinity. The second model is a simple spanwise superposition of two dimensional solutions to account for three dimensional acoustic effects of wing rotation (for a helicopter blade, or some other rotating planform) and of finiteness of wing span. A three dimensional theory for a single gust was applied to calculate the acoustic signature in closed form due to blade vortex interaction in helicopters. The third model is that of a quarter infinite plate with side edge through a gust at high subsonic speed. An approximate solution for the three dimensional loading and the associated three dimensional acoustic field in closed form was obtained. The results reflected the acoustic effect of satisfying the correct loading condition at the side edge.
A novel flow-perfusion bioreactor supports 3D dynamic cell culture.
Sailon, Alexander M; Allori, Alexander C; Davidson, Edward H; Reformat, Derek D; Allen, Robert J; Warren, Stephen M
2009-01-01
Bone engineering requires thicker three-dimensional constructs than the maximum thickness supported by standard cell-culture techniques (2 mm). A flow-perfusion bioreactor was developed to provide chemotransportation to thick (6 mm) scaffolds. Polyurethane scaffolds, seeded with murine preosteoblasts, were loaded into a novel bioreactor. Control scaffolds remained in static culture. Samples were harvested at days 2, 4, 6, and 8 and analyzed for cellular distribution, viability, metabolic activity, and density at the periphery and core. By day 8, static scaffolds had a periphery cell density of 67% +/- 5.0%, while in the core it was 0.3% +/- 0.3%. Flow-perfused scaffolds demonstrated peripheral cell density of 94% +/- 8.3% and core density of 76% +/- 3.1% at day 8. Flow perfusion provides chemotransportation to thick scaffolds. This system may permit high throughput study of 3D tissues in vitro and enable prefabrication of biological constructs large enough to solve clinical problems.
Engl, Tobias; Boost, Kim A; Leckel, Kerstin; Beecken, Wolf-Dietrich; Jonas, Dietger; Oppermann, Elsie; Auth, Marcus K H; Schaudt, André; Bechstein, Wolf-Otto; Blaheta, Roman A
2004-08-01
In vitro culture models that employ human liver cells could be potent tools for predictive studies on drug toxicity and metabolism in the pharmaceutical industry. However, an adequate receptor responsiveness is necessary to allow intracellular signalling and metabolic activity. We tested the ability of three-dimensionally arranged human hepatocytes to respond to the growth factors hepatocyte growth factor (HGF) or epidermal growth factor (EGF). Isolated adult human hepatocytes were cultivated within a three-dimensional collagen gel (sandwich) or on a two-dimensional collagen matrix. Cells were treated with HGF or EGF and expression and phosphorylative activity of HGF receptors (HGFr, c-met) or EGF receptors (EGFr) were measured by flow cytometry and Western blot. Increasing HGFr and EGFr levels were detected in hepatocytes growing two-dimensionally. However, both receptors were not activated in presence of growth factors. In contrast, when hepatocytes were plated within a three-dimensional matrix, HGFr and EGFr levels remained constantly low. However, both receptors became strongly phosphorylated by soluble HGF or EGF. We conclude that cultivation of human hepatocytes in a three-dimensionally arranged in vitro system allows the maintenance of specific functional activities. The necessity of cell dimensionality for HGFr and EGFr function should be considered when an adequate in vitro system has to be introduced for drug testing.
Garai, Subhadra; Sinha, Arvind
2014-03-01
An innovative biomimetic synthesis of novel three dimensional micro/macro porous carboxymethyl cellulose (CMC)-hydroxyapatite (HA) nanocomposites having four systematically different compositions has been established for its possible application as a load bearing synthetic bone graft. Our process, being in situ, involves a simple and cost effective route akin to a matrix mediated biomineralization process. Developed synthesis route not only controls the size of HA particles in the range of 15-50 nm, embedded in CMC matrix, but also assists in the formation of a mechanically strong three dimensional nanocomposite structures due to physical cross linking of HA impregnated CMC matrix. The process does not involve any toxic cross linker and works at near ambient conditions. The nanocomposites are systematically structurally and mechanically characterized using various techniques like scanning electron microscopy (SEM), atomic force microscopy (AFM), transmission electron microscopy (TEM), X-ray diffraction (XRD), Fourier transform IR (FTIR), solid state (13)C nuclear magnetic resonance ((13)C NMR), thermo-gravimetric analysis (TGA) and Universal mechanical test. It reveals that the ionic/polar or electrostatic interactions are the main driving force for formation of load bearing three dimensional nanocomposites via a process similar to matrix mediated biomineralization. Compressive strength and compressive modulus of nanocomposites, being in the range of 1.74-12 MPa and 157-330 MPa, respectively, meet the desired range of compressive strength for the synthetic grafts used in cancellous bone. An increase in the compressive strength with increase in the porosity has been an interesting observation in the present study. In vitro cytotoxicity of the synthesized nanocomposites has been evaluated using bone marrow mesenchymal stem cells (BMSC) isolated from Wistar rat. Copyright © 2013 Elsevier B.V. All rights reserved.
He, Lihui; Liu, Lijie; Gao, Bei; Gao, Shang; Chen, Yifu; Zhihui, Liu
2013-08-01
To establish three-dimensional finite element model of two-piece post crown to the mandibular first molar residual roots, and analyze the stress distribution characteristic to the residual roots with different adhesives, so as to get the best combination under different conditions. The complete mandibular first molar in vitro was selected, the crown was removed along the cemento-enamel junction, then the residual roots were scanned by CT. CT images were imported into a reverse engineering software, and the three-dimensional finite element model of the mandibular first molar residual roots was reconstructed. Titanium two-piece post crown of the mandibular first molar residual roots was produced, then was scanned by CT. The model was reconstructed and assembled by MIMICS. The stress distribution of the root canal and root section under the vertical load and lateral load with different bonding systems were analyzed. Three-dimensional finite element model of two-piece post crown to the mandibular first molar residual roots was established. With the increasing of elastic modulus of the adhesives, the maximum stress within the root canal was also increasing. Elastic modulus of zinc phosphate was the biggest, so the stress within the root canal was the biggest; elastic modulus of Superbond C&B was the smallest, so the stress within the root canal was the smallest. Lateral loading stress was much larger than the vertical load. Under vertical load, the load on the root section was even with different bonding systems. Under lateral load, the maximum stress was much larger than the vertical load. The stress on the root section was minimum using zinc phosphate binder, and the stress on the root section was maximum using Superbond C&B. In two-piece post crown restorations, there is significant difference between different adhesives on tooth protection. When the tooth structure of the root canal orifices is weak, in order to avoid the occurrence of splitting, the larger elastic modulus bonding system is the first choice, such as zinc phosphate binder. When the resistance form of the root canal orifices is good enough but the root is too weak, it is suggested that the smaller elastic modulus bonding system is the first choice, such as Superbond C&B.
Simulating Fatigue Crack Growth in Spiral Bevel Gears
NASA Technical Reports Server (NTRS)
Spievak, Lisa E.; Wawrzynek, Paul A.; Ingraffea, Anthony R.
2000-01-01
The majority of helicopter transmission systems utilize spiral bevel gears to convert the horizontal power from the engine into vertical power for the rotor. Due to the cyclical loading on a gear's tooth, fatigue crack propagation can occur. In rotorcraft applications, a crack's trajectory determines whether the gear failure will be benign or catastrophic for the aircraft. As a result, the capability to predict crack growth in gears is significant. A spiral bevel gear's complex shape requires a three dimensional model of the geometry and cracks. The boundary element method in conjunction with linear elastic fracture mechanics theories is used to predict arbitrarily shaped three dimensional fatigue crack trajectories in a spiral bevel pinion under moving load conditions. The predictions are validated by comparison to experimental results. The sensitivity of the predictions to variations in loading conditions and crack growth rate model parameters is explored. Critical areas that must be understood in greater detail prior to predicting more accurate crack trajectories and crack growth rates in three dimensions are identified.
Ingram, M; Techy, G B; Saroufeem, R; Yazan, O; Narayan, K S; Goodwin, T J; Spaulding, G F
1997-06-01
Growth patterns of a number of human tumor cell lines that from three-dimensional structures of various architectures when cultured without carrier beads in a NASA rotary cell culture system are described and illustrated. The culture system, which was designed to mimic microgravity, maintained cells in suspension under very low-shear stress throughout culture. Spheroid (particulate) production occurred within a few hours after culture was started, and spheroids increased in size by cell division and fusion of small spheroids, usually stabilizing at a spheroid diameter of about 0.5 mm. Architecture of spheroids varied with cell type. Cellular interactions that occurred in spheroids resulted in conformation and shape changes of cells, and some cell lines produced complex, epithelial-like architectures. Expression of the cell adhesion molecules, CD44 and E cadherin, was upregulated in the three-dimensional constructs. Coculture of fibroblast spheroids with PC3 prostate cancer cells induced tenascin expression by the fibroblasts underlying the adherent prostate epithelial cells. Invasion of the fibroblast spheroids by the malignant epithelium was also demonstrated.
Restrictive loads powered by separate or by common electrical sources
NASA Technical Reports Server (NTRS)
Appelbaum, J.
1989-01-01
In designing a multiple load electrical system, the designer may wish to compare the performance of two setups: a common electrical source powering all loads, or separate electrical sources powering individual loads. Three types of electrical sources: an ideal voltage source, an ideal current source, and solar cell source powering resistive loads were analyzed for their performances in separate and common source systems. A mathematical proof is given, for each case, indicating the merit of the separate or common source system. The main conclusions are: (1) identical resistive loads powered by ideal voltage sources perform the same in both system setups, (2) nonidentical resistive loads powered by ideal voltage sources perform the same in both system setups, (3) nonidentical resistive loads powered by ideal current sources have higher performance in separate source systems, and (4) nonidentical resistive loads powered by solar cells have higher performance in a common source system for a wide range of load resistances.
A new crank arm-based load cell for the 3D analysis of the force applied by a cyclist.
Balbinot, Alexandre; Milani, Cleiton; Nascimento, Jussan da Silva Bahia
2014-12-03
This report describes a new crank arm-based force platform designed to evaluate the three-dimensional force applied to the pedals by cyclists in real conditions. The force platform was designed to be fitted on a conventional competition bicycle crankset while data is transmitted wirelessly through a Bluetooth™ module and also stored on a SD card. A 3D solid model is created in the SolidWorks (Dassault Systèmes SOLIDWORKS Corp.) to analyze the static and dynamic characteristics of the crank arm by using the finite elements technique. Each crankset arm is used as a load cell based on strain gauges configured as three Wheatstone bridges. The signals are conditioned on a printed circuit board attached directly to the structure. The load cell showed a maximum nonlinearity error between 0.36% and 0.61% and a maximum uncertainty of 2.3% referred to the sensitivity of each channel. A roller trainer equipped with an optical encoder was also developed, allowing the measurement of the wheel's instantaneous velocity.
A New Crank Arm-Based Load Cell for the 3D Analysis of the Force Applied by a Cyclist
Balbinot, Alexandre; Milani, Cleiton; Nascimento, Jussan da Silva Bahia
2014-01-01
This report describes a new crank arm-based force platform designed to evaluate the three-dimensional force applied to the pedals by cyclists in real conditions. The force platform was designed to be fitted on a conventional competition bicycle crankset while data is transmitted wirelessly through a BluetoothTM module and also stored on a SD card. A 3D solid model is created in the SolidWorks (Dassault Systèmes SOLIDWORKS Corp.) to analyze the static and dynamic characteristics of the crank arm by using the finite elements technique. Each crankset arm is used as a load cell based on strain gauges configured as three Wheatstone bridges. The signals are conditioned on a printed circuit board attached directly to the structure. The load cell showed a maximum nonlinearity error between 0.36% and 0.61% and a maximum uncertainty of 2.3% referred to the sensitivity of each channel. A roller trainer equipped with an optical encoder was also developed, allowing the measurement of the wheel's instantaneous velocity. PMID:25479325
Induction of carcinoembryonic antigen expression in a three-dimensional culture system
NASA Technical Reports Server (NTRS)
Jessup, J. M.; Brown, D.; Fitzgerald, W.; Ford, R. D.; Nachman, A.; Goodwin, T. J.; Spaulding, G.
1994-01-01
MIP-101 is a poorly differentiated human colon carcinoma cell line established from ascites that produces minimal amounts of carcinoembryonic antigen (CEA), a 180 kDa glycoprotein tumor marker, and nonspecific cross-reacting antigen (NCA), a related protein that has 50 and 90 kDa isoforms, in vitro in monolayer culture. MIP-101 produces CEA when implanted into the peritoneum of nude mice but not when implanted into subcutaneous tissue. We tested whether MIP-101 cells may be induced to express CEA when cultured on microcarrier beads in three-dimensional cultures, either in static cultures as non-adherent aggregates or under dynamic conditions in a NASA-designed low shear stress bioreactor. MIP- 101 cells proliferated well under all three conditions and increased CEA and NCA production 3 - 4 fold when grown in three-dimensional cultures compared to MIP-101 cells growing logarithmically in monolayers. These results suggest that three-dimensional growth in vitro simulates tumor function in vivo and that three-dimensional growth by itself may enhance production of molecules that are associated with the metastatic process.
Orthotropic Laminated Open-cell Frameworks Retaining Strong Auxeticity under Large Uniaxial Loading
NASA Astrophysics Data System (ADS)
Tanaka, Hiro; Suga, Kaito; Iwata, Naoki; Shibutani, Yoji
2017-01-01
Anisotropic materials form inside living tissue and are widely applied in engineered structures, where sophisticated structural and functional design principles are essential to employing these materials. This paper presents a candidate laminated open-cell framework, which is an anisotropic material that shows remarkable mechanical performance. Using additive manufacturing, artificial frameworks are fabricated by lamination of in-plane orthotropic microstructures made of elbowed beam and column members; this fabricated structure features orthogonal anisotropy in three-dimensional space. Uniaxial loading tests reveal strong auxeticity (high negative Poisson’s ratios) in the out-of-plane direction, which is retained reproducibly up to the nonlinear elastic region, and is equal under tensile and compressive loading. Finite element simulations support the observed auxetic behaviors for a unit cell in the periodic framework, which preserve the theoretical elastic properties of an orthogonal solid. These findings open the possibility of conceptual materials design based on geometry.
Theory and application of a three-dimensional model of the human spine.
Belytschko, T; Schwer, L; Privitzer, E
1978-01-01
A three-dimensional, discrete model of the human spine, torso, and head was developed for the purpose of evaluating mechanical response in pilot ejection. However, it was developed in sufficient generality to be applicable to other body response problems, such as occupant response in aircraft crash and arbitrary loads on the head-spine system. The anatomy is modelled by a collection of rigid bodies, which represent skeletal segments such as the vertebrae, pelvis, head, and ribs, interconnected by deformable elements, which represent ligaments, cargilagenous joints, viscera and connective tissues. Results are presented for several conditions: different rates of onset, ejection at angles, preejection alignment, and eccentric head loadings. It is shown that slow rates of onset and angling the seat reduce both the peak axial loads and bending moments. In the presence of eccentric head masses, such as helmet-mounted devices, the reflected flexural wave is shown to be the key injury mechanism.
Multiscale strain analysis of tissue equivalents using a custom-designed biaxial testing device.
Bell, B J; Nauman, E; Voytik-Harbin, S L
2012-03-21
Mechanical signals transferred between a cell and its extracellular matrix play an important role in regulating fundamental cell behavior. To further define the complex mechanical interactions between cells and matrix from a multiscale perspective, a biaxial testing device was designed and built. Finite element analysis was used to optimize the cruciform specimen geometry so that stresses within the central region were concentrated and homogenous while minimizing shear and grip effects. This system was used to apply an equibiaxial loading and unloading regimen to fibroblast-seeded tissue equivalents. Digital image correlation and spot tracking were used to calculate three-dimensional strains and associated strain transfer ratios at macro (construct), meso, matrix (collagen fibril), cell (mitochondria), and nuclear levels. At meso and matrix levels, strains in the 1- and 2-direction were statistically similar throughout the loading-unloading cycle. Interestingly, a significant amplification of cellular and nuclear strains was observed in the direction perpendicular to the cell axis. Findings indicate that strain transfer is dependent upon local anisotropies generated by the cell-matrix force balance. Such multiscale approaches to tissue mechanics will assist in advancement of modern biomechanical theories as well as development and optimization of preconditioning regimens for functional engineered tissue constructs. Copyright © 2012 Biophysical Society. Published by Elsevier Inc. All rights reserved.
Shah, Bhranti S; Chahine, Nadeen O
2018-02-01
Dynamic hydrostatic pressure (HP) loading can modulate nucleus pulposus (NP) cell metabolism, extracellular matrix (ECM) composition, and induce transformation of notochordal NP cells into mature phenotype. However, the effects of varying cell density and dynamic HP magnitude on NP phenotype and metabolism are unknown. This study examined the effects of physiological magnitudes of HP loading applied to bovine NP cells encapsulated within three-dimensional (3D) alginate beads. Study 1: seeding density (1 M/mL versus 4 M/mL) was evaluated in unloaded and loaded (0.1 MPa, 0.1 Hz) conditions. Study 2: loading magnitude (0, 0.1, and 0.6 MPa) applied at 0.1 Hz to 1 M/mL for 7 days was evaluated. Study 1: 4 M/mL cell density had significantly lower adenosine triphosphate (ATP), glycosaminoglycan (GAG) and collagen content, and increased lactate dehydrogenase (LDH). HP loading significantly increased ATP levels, and expression of aggrecan, collagen I, keratin-19, and N-cadherin in HP loaded versus unloaded groups. Study 2: aggrecan expression increased in a dose dependent manner with HP magnitude, whereas N-cadherin and keratin-19 expression were greatest in low HP loading compared to unloaded. Overall, the findings of the current study indicate that cell seeding density within a 3D construct is a critical variable influencing the mechanobiological response of NP cells to HP loading. NP mechanobiology and phenotypic expression was also found to be dependent on the magnitude of HP loading. These findings suggest that HP loading and culture conditions of NP cells may require complex optimization for engineering an NP replacement tissue.
Quantitative volumetric Raman imaging of three dimensional cell cultures
NASA Astrophysics Data System (ADS)
Kallepitis, Charalambos; Bergholt, Mads S.; Mazo, Manuel M.; Leonardo, Vincent; Skaalure, Stacey C.; Maynard, Stephanie A.; Stevens, Molly M.
2017-03-01
The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell-material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy.
Suzuki, Teruhiko; Kazuki, Yasuhiro; Oshimura, Mitsuo; Hara, Takahiko
2014-01-01
Human artificial chromosomes (HACs) are gene-delivery vectors suitable for introducing large DNA fragments into mammalian cells. Although a HAC theoretically incorporates multiple gene expression cassettes of unlimited DNA size, its application has been limited because the conventional gene-loading system accepts only one gene-loading vector (GLV) into a HAC. We report a novel method for the simultaneous or sequential integration of multiple GLVs into a HAC vector (designated as the SIM system) via combined usage of Cre, FLP, Bxb1, and φC31 recombinase/integrase. As a proof of principle, we first attempted simultaneous integration of three GLVs encoding EGFP, Venus, and TdTomato into a gene-loading site of a HAC in CHO cells. These cells successfully expressed all three fluorescent proteins. Furthermore, microcell-mediated transfer of HACs enabled the expression of those fluorescent proteins in recipient cells. We next demonstrated that GLVs could be introduced into a HAC one-by-one via reciprocal usage of recombinase/integrase. Lastly, we introduced a fourth GLV into a HAC after simultaneous integration of three GLVs by FLP-mediated DNA recombination. The SIM system expands the applicability of HAC vectors and is useful for various biomedical studies, including cell reprogramming. PMID:25303219
Suzuki, Teruhiko; Kazuki, Yasuhiro; Oshimura, Mitsuo; Hara, Takahiko
2014-01-01
Human artificial chromosomes (HACs) are gene-delivery vectors suitable for introducing large DNA fragments into mammalian cells. Although a HAC theoretically incorporates multiple gene expression cassettes of unlimited DNA size, its application has been limited because the conventional gene-loading system accepts only one gene-loading vector (GLV) into a HAC. We report a novel method for the simultaneous or sequential integration of multiple GLVs into a HAC vector (designated as the SIM system) via combined usage of Cre, FLP, Bxb1, and φC31 recombinase/integrase. As a proof of principle, we first attempted simultaneous integration of three GLVs encoding EGFP, Venus, and TdTomato into a gene-loading site of a HAC in CHO cells. These cells successfully expressed all three fluorescent proteins. Furthermore, microcell-mediated transfer of HACs enabled the expression of those fluorescent proteins in recipient cells. We next demonstrated that GLVs could be introduced into a HAC one-by-one via reciprocal usage of recombinase/integrase. Lastly, we introduced a fourth GLV into a HAC after simultaneous integration of three GLVs by FLP-mediated DNA recombination. The SIM system expands the applicability of HAC vectors and is useful for various biomedical studies, including cell reprogramming.
Inaba, Nao; Kuroshima, Shinichiro; Uto, Yusuke; Sasaki, Muneteru; Sawase, Takashi
2017-09-01
Osteocytes play important roles in controlling bone quality as well as preferential alignment of biological apatite c -axis/collagen fibers. However, the relationship between osteocytes and mechanical stress remains unclear due to the difficulty of three-dimensional (3D) culture of osteocytes in vitro . The aim of this study was to investigate the effect of cyclic mechanical stretch on 3D-cultured osteocyte-like cells. Osteocyte-like cells were established using rat calvarial osteoblasts cultured in a 3D culture system. Cyclic mechanical stretch (8% amplitude at a rate of 2 cycles min -1 ) was applied for 24, 48 and 96 consecutive hours. Morphology, cell number and preferential cell alignment were evaluated. Apoptosis- and autophagy-related gene expression levels were measured using quantitative PCR. 3D-cultured osteoblasts became osteocyte-like cells that expressed osteocyte-specific genes such as Dmp1 , Cx43 , Sost , Fgf23 and RANKL , with morphological changes similar to osteocytes. Cell number was significantly decreased in a time-dependent manner under non-loaded conditions, whereas cyclic mechanical stretch significantly prevented decreased cell numbers with increased expression of anti-apoptosis-related genes. Moreover, cyclic mechanical stretch significantly decreased cell size and ellipticity with increased expression of autophagy-related genes, LC3b and atg7 . Interestingly, preferential cell alignment did not occur, irrespective of mechanical stretch. These findings suggest that an anti-apoptotic effect contributes to network development of osteocyte-like cells under loaded condition. Spherical change of osteocyte-like cells induced by mechanical stretch may be associated with autophagy upregulation. Preferential alignment of osteocytes induced by mechanical load in vivo may be partially predetermined before osteoblasts differentiate into osteocytes and embed into bone matrix.
Development and verification of global/local analysis techniques for laminated composites
NASA Technical Reports Server (NTRS)
Thompson, Danniella Muheim; Griffin, O. Hayden, Jr.
1991-01-01
A two-dimensional to three-dimensional global/local finite element approach was developed, verified, and applied to a laminated composite plate of finite width and length containing a central circular hole. The resulting stress fields for axial compression loads were examined for several symmetric stacking sequences and hole sizes. Verification was based on comparison of the displacements and the stress fields with those accepted trends from previous free edge investigations and a complete three-dimensional finite element solution of the plate. The laminates in the compression study included symmetric cross-ply, angle-ply and quasi-isotropic stacking sequences. The entire plate was selected as the global model and analyzed with two-dimensional finite elements. Displacements along a region identified as the global/local interface were applied in a kinematically consistent fashion to independent three-dimensional local models. Local areas of interest in the plate included a portion of the straight free edge near the hole, and the immediate area around the hole. Interlaminar stress results obtained from the global/local analyses compares well with previously reported trends, and some new conclusions about interlaminar stress fields in plates with different laminate orientations and hole sizes are presented for compressive loading. The effectiveness of the global/local procedure in reducing the computational effort required to solve these problems is clearly demonstrated through examination of the computer time required to formulate and solve the linear, static system of equations which result for the global and local analyses to those required for a complete three-dimensional formulation for a cross-ply laminate. Specific processors used during the analyses are described in general terms. The application of this global/local technique is not limited software system, and was developed and described in as general a manner as possible.
Markovic, Marica; Van Hoorick, Jasper; Hölzl, Katja; Tromayer, Maximilian; Gruber, Peter; Nürnberger, Sylvia; Dubruel, Peter; Van Vlierberghe, Sandra; Liska, Robert; Ovsianikov, Aleksandr
2015-05-01
Three-dimensional (3D) printing offers versatile possibilities for adapting the structural parameters of tissue engineering scaffolds. However, it is also essential to develop procedures allowing efficient cell seeding independent of scaffold geometry and pore size. The aim of this study was to establish a method for seeding the scaffolds using photopolymerizable cell-laden hydrogels. The latter facilitates convenient preparation, and handling of cell suspension, while distributing the hydrogel precursor throughout the pores, before it is cross-linked with light. In addition, encapsulation of living cells within hydrogels can produce constructs with high initial cell loading and intimate cell-matrix contact, similar to that of the natural extra-cellular matrix (ECM). Three dimensional scaffolds were produced from poly(lactic) acid (PLA) by means of fused deposition modeling. A solution of methacrylamide-modified gelatin (Gel-MOD) in cell culture medium containing photoinitiator Li-TPO-L was used as a hydrogel precursor. Being an enzymatically degradable derivative of natural collagen, gelatin-based matrices are biomimetic and potentially support the process of cell-induced remodeling. Preosteoblast cells MC3T3-E1 at a density of 10 × 10 6 cells per 1 mL were used for testing the seeding procedure and cell proliferation studies. Obtained results indicate that produced constructs support cell survival and proliferation over extended duration of our experiment. The established two-step approach for scaffold seeding with the cells is simple, rapid, and is shown to be highly reproducible. Furthermore, it enables precise control of the initial cell density, while yielding their uniform distribution throughout the scaffold. Such hybrid tissue engineering constructs merge the advantages of rigid 3D printed constructs with the soft hydrogel matrix, potentially mimicking the process of ECM remodeling.
Mechanical control of tissue-engineered bone.
Hung, Ben P; Hutton, Daphne L; Grayson, Warren L
2013-01-31
Bone is a load-bearing tissue and physical forces play key roles in the development and maintenance of its structure. Mechanical cues can stimulate the expression of an osteogenic phenotype, enhance matrix and mineral deposition, and influence tissue organization to improve the functional outcome of engineered bone grafts. In recent years, a number of studies have investigated the effects of biophysical forces on the bone formation properties of osteoprogenitor cells. The application of physiologically relevant stimuli to tissue-engineered bone may be determined through observation and understanding of forces to which osteoblasts, osteoclasts, and osteocytes are exposed in native bone. Subsequently, these cues may be parameterized and their effects studied in well-defined in vitro systems. The osteo-inductive effects of three specific mechanical cues - shear stress, substrate rigidity, and nanotopography - on cells cultured in monolayer or in three-dimensional biomaterial scaffolds in vitro are reviewed. Additionally, we address the time-dependent effects of mechanical cues on vascular infiltration and de novo bone formation in acellular scaffolds implanted into load-bearing sites in vivo. Recent studies employing cutting-edge advances in biomaterial fabrication and bioreactor design have provided key insights into the role of mechanical cues on cellular fate and tissue properties of engineered bone grafts. By providing mechanistic understanding, future studies may go beyond empirical approaches to rational design of engineering systems to control tissue development.
Levy, Gary; Malik, Minnie; Britten, Joy; Gilden, Melissa; Segars, James; Catherino, William H.
2014-01-01
Objective To investigate the impact of liarozole on transforming growth factor-β3 (TGF-β3) expression, TGF-β3 controlled profibrotic cytokines, and extracellular matrix formation in a three-dimensional (3D) leiomyoma model system. Design Molecular and immunohistochemical analysis in a cell line evaluated in a three-dimensional culture. Setting Laboratory study. Patient(s) None. Intervention(s) Treatment of leiomyoma and myometrial cells with liarozole and TGF-β3 in a three-dimensional culture system. Main Outcome Measure(s) Quantitative real-time reverse-transcriptase polymerase chain reaction and Western blotting to assess fold gene and protein expression of TGF-β3 and TGF-β3 regulated fibrotic cytokines: collagen 1A1 (COL1A1), fibronectin, and versican before and after treatment with liarozole, and confirmatory immunohistochemical stains of treated three-dimensional cultures. Result(s) Both TGF-β3 gene and protein expression were elevated in leiomyoma cells compared with myometrium in two-dimensional and 3D cultures. Treatment with liarozole decreased TGF-β3 gene and protein expression. Extracellular matrix components versican, COL1A1, and fibronectin were also decreased by liarozole treatment in 3D cultures. Treatment of 3D cultures with TGF-β3 increased gene expression and protein production of COL1A1, fibronectin, and versican. Conclusion(s) Liarozole decreased TGF-β3 and TGF-β3–mediated extracellular matrix expression in a 3D uterine leiomyoma culture system. PMID:24825427
Movia, Dania; Gerard, Valerie; Maguire, Ciaran Manus; Jain, Namrata; Bell, Alan P; Nicolosi, Valeria; O'Neill, Tiina; Scholz, Dimitri; Gun'ko, Yurii; Volkov, Yuri; Prina-Mello, Adriele
2014-03-01
Gold nanomaterials are currently raising a significant interest for human welfare in the field of clinical diagnosis, therapeutics for chronic pathologies, as well as of many other biomedical applications. In particular, gold nanomaterials are becoming a promising technology for developing novel approaches and treatments against widespread societal diseases such as cancer. In this study, we investigated the potential of proprietary gold nanoboxes (AuNBs) as carriers for their perspective translation into multifunctional, pre-clinical nano-enabled systems for personalized medicine approaches against lung cancer. A safe-by-design, tiered approach, with systematic tests conducted in the early phases on uncoated AuNBs and more focused testing on the coated, drug-loaded nanomaterial toward the end, was adopted. Our results showed that uncoated AuNBs could effectively penetrate into human lung adenocarcinoma (A549) cells when in simple (mono-cultures) or complex (co- and three-dimensional-cultures) in vitro microenvironments mimicking the alveolar region of human lungs. Uncoated AuNBs were biologically inert in A549 cells and demonstrated signs of biodegradability. Concurrently, preliminary data revealed that coated, drug-loaded AuNBs could efficiently deliver a chemotherapeutic agent to A549 cells, corroborating the hypothesis that AuNBs could be used in the future for the development of personalized nano-enabled systems for lung cancer treatment. Copyright © 2013 Elsevier Ltd. All rights reserved.
Development of an interactive anatomical three-dimensional eye model.
Allen, Lauren K; Bhattacharyya, Siddhartha; Wilson, Timothy D
2015-01-01
The discrete anatomy of the eye's intricate oculomotor system is conceptually difficult for novice students to grasp. This is problematic given that this group of muscles represents one of the most common sites of clinical intervention in the treatment of ocular motility disorders and other eye disorders. This project was designed to develop a digital, interactive, three-dimensional (3D) model of the muscles and cranial nerves of the oculomotor system. Development of the 3D model utilized data from the Visible Human Project (VHP) dataset that was refined using multiple forms of 3D software. The model was then paired with a virtual user interface in order to create a novel 3D learning tool for the human oculomotor system. Development of the virtual eye model was done while attempting to adhere to the principles of cognitive load theory (CLT) and the reduction of extraneous load in particular. The detailed approach, digital tools employed, and the CLT guidelines are described herein. © 2014 American Association of Anatomists.
Cellular Tug-of-War: Forces at Work and DNA Stretching in Mitosis
NASA Astrophysics Data System (ADS)
Griffin, Brian; Kilfoil, Maria L.
2013-03-01
In the microscopic world of the cell dominated by thermal noise, a cell must be able to successfully segregate its DNA with high fidelity in order to pass its genetic information on to its progeny. In this process of mitosis in eukaryotes, driving forces act on the cytoskeleton-based architecture called the mitotic spindle to promote this division. Our preliminary data demonstrates that the dynamics of this process in yeast cells is universal. Moreover, the dynamics suggest an increasing load as the chromosomes are pulled apart. To investigate this, we use three-dimensional imaging to track the dynamics of the poles of this architecture and the points of attachment to chromosomes simultaneously and with high spatial resolution. We analyze the relative motions of chromosomes as they are organized before segregation and as they are pulled apart, using this data to investigate the force-response behavior of this cytoskeleton-chromosome polymer system.
Fan, Lei; Liu, Can; Chen, Xiuxing; Zou, Yan; Zhou, Zhengnan; Lin, Chenkai; Tan, Guoxin; Zhou, Lei; Ning, Chenyun; Wang, Qiyou
2018-05-30
Current treatment approaches for spinal cord injuries (SCIs) are mainly based on cellular transplantation. Induced pluripotent stem cells (iPSCs) without supply constraints and ethical concerns have emerged as a viable treatment option for repairing neurological disorders. However, the primarily limitations in the neuroregeneration field are uncontrolled cell differentiation, and low cell viability caused by the ischemic environment. The mechanical property of three-dimensional (3D) hydrogel can be easily controlled and shared similar characteristics with nerve tissue, thus promoting cell survival and controlled cell differentiation. We propose the combination of a 3D gelatin methacrylate (GelMA) hydrogel with iPSC-derived NSCs (iNSCs) to promote regeneration after SCI. In vitro, the iNSCs photoencapsulated in the 3D GelMA hydrogel survived and differentiated well, especially in lower-moduli hydrogels. More robust neurite outgrowth and more neuronal differentiation were detected in the soft hydrogel group. To further evaluate the in vivo neuronal regeneration effect of the GelMA hydrogels, a mouse spinal cord transection model was generated. We found that GelMA/iNSC implants significantly promoted functional recovery. Further histological analysis showed that the cavity areas were significantly reduced, and less collagen was deposited in the GelMA/iNSC group. Furthermore, the GelMA and iNSC combined transplantation decreased inflammation by reducing activated macrophages/microglia (CD68-positive cells). Additionally, GelMA/iNSC implantation showed striking therapeutic effects of inhibiting GFAP-positive cells and glial scar formation while simultaneously promoting axonal regeneration. Undoubtedly, use of this 3D hydrogel stem cell-loaded system is a promising therapeutic strategy for SCI repair.
Three-dimensional micro-electrode array for recording dissociated neuronal cultures.
Musick, Katherine; Khatami, David; Wheeler, Bruce C
2009-07-21
This work demonstrates the design, fabrication, packaging, characterization, and functionality of an electrically and fluidically active three-dimensional micro-electrode array (3D MEA) for use with neuronal cell cultures. The successful function of the device implies that this basic concept-construction of a 3D array with a layered approach-can be utilized as the basis for a new family of neural electrode arrays. The 3D MEA prototype consists of a stack of individually patterned thin films that form a cell chamber conducive to maintaining and recording the electrical activity of a long-term three-dimensional network of rat cortical neurons. Silicon electrode layers contain a polymer grid for neural branching, growth, and network formation. Along the walls of these electrode layers lie exposed gold electrodes which permit recording and stimulation of the neuronal electrical activity. Silicone elastomer micro-fluidic layers provide a means for loading dissociated neurons into the structure and serve as the artificial vasculature for nutrient supply and aeration. The fluidic layers also serve as insulation for the micro-electrodes. Cells have been shown to survive in the 3D MEA for up to 28 days, with spontaneous and evoked electrical recordings performed in that time. The micro-fluidic capability was demonstrated by flowing in the drug tetrotodoxin to influence the activity of the culture.
NASA Astrophysics Data System (ADS)
Sutter, Leo; Kolbman, Dan; Wu, Mingming; Ma, Minglin; Das, Moumita
The biophysics of cell co-cultures, i.e. binary systems of cell populations, is of great interest in many biological processes including formation of embryos, and tumor progression. During these processes, different types of cells with different physical properties are mixed with each other, with important consequences for cell-cell interaction, aggregation, and migration. The role of the differences in their physical properties in their collective behavior remains poorly understood. Furthermore, until recently most theoretical studies of collective cell migration have focused on two dimensional systems. Under physiological conditions, however, cells often have to navigate three dimensional and confined micro-environments. We study a confined, three-dimensional binary system of interacting, active, and deformable particles with different physical properties such as deformability, motility, adhesion, and division rates using Langevin Dynamics simulations. Our findings may provide insights into how the differences in and interplay between cell mechanical properties, division, and motility influence emergent collective behavior such as cell aggregation and segregation experimentally observed in co-cultures of breast cancer cells and healthy breast epithelial cells. This work was partially supported by a Cottrell College Science Award.
NASA Astrophysics Data System (ADS)
Uenishi, Koji
2018-06-01
We consider stability of fracture on a three-dimensional planar interface subjected to a loading stress that is locally peaked spatially, the level of which increases quasi-statically in time. Similar to the earlier study on the two-dimensional case (Uenishi and Rice, 2003; Rice and Uenishi, 2010), as the loading stress increases, a crack, or a region of displacement discontinuity (opening gap in tension or slip for shear fracture), develops on the interface where the stress is presumed to decrease according to a displacement-weakening constitutive relation. Upon reaching the instability point at which no further quasi-static solution for the extension of the crack on the interface exists, dynamic fracture follows. For the investigation of this instability point, we employ a dimensional analysis as well as an energy approach that gives a Rayleigh-Ritz approximation for the dependence of crack size and maximum displacement discontinuity on the level and quadratic shape of the loading stress distribution. We show that, if the linear displacement-weakening law is applied and the crack may be assumed of an elliptical form, the critical crack size at instability is independent of the curvature of the loading stress distribution and it is of the same order for all two- and three-dimensional cases.
Decoupling Principle Analysis and Development of a Parallel Three-Dimensional Force Sensor
Zhao, Yanzhi; Jiao, Leihao; Weng, Dacheng; Zhang, Dan; Zheng, Rencheng
2016-01-01
In the development of the multi-dimensional force sensor, dimension coupling is the ubiquitous factor restricting the improvement of the measurement accuracy. To effectively reduce the influence of dimension coupling on the parallel multi-dimensional force sensor, a novel parallel three-dimensional force sensor is proposed using a mechanical decoupling principle, and the influence of the friction on dimension coupling is effectively reduced by making the friction rolling instead of sliding friction. In this paper, the mathematical model is established by combining with the structure model of the parallel three-dimensional force sensor, and the modeling and analysis of mechanical decoupling are carried out. The coupling degree (ε) of the designed sensor is defined and calculated, and the calculation results show that the mechanical decoupling parallel structure of the sensor possesses good decoupling performance. A prototype of the parallel three-dimensional force sensor was developed, and FEM analysis was carried out. The load calibration and data acquisition experiment system are built, and then calibration experiments were done. According to the calibration experiments, the measurement accuracy is less than 2.86% and the coupling accuracy is less than 3.02%. The experimental results show that the sensor system possesses high measuring accuracy, which provides a basis for the applied research of the parallel multi-dimensional force sensor. PMID:27649194
Flexural impact force absorption of mouthguard materials using film sensor system.
Reza, Fazal; Churei, Hiroshi; Takahashi, Hidekazu; Iwasaki, Naohiko; Ueno, Toshiaki
2014-06-01
Several methods have been used to measure the impact force absorption capacities of mouthguard materials; however, the relationships among these measurement systems have not been clearly determined. The purpose of the present study was to evaluate the impact force-absorbing capability of materials using a drop-ball system with film sensors and load cells to clarify the relationship between these two sensor systems. Disk-shaped specimens (1, 2, and 3 mm thick) were prepared using three commercial thermoplastic mouthguard materials (Bioplast, Impact Guard, MG 21) and one experimental mouthguard material [mixture of Poly (ethyl methacrylate)]. Impact force was applied by letting a stainless steel ball drop free-fall onto the specimens and then measuring the impact load under each specimen using a film sensor system and a load cell sensor system. The total load measured with the film sensor system decreased with an increase in mouthguard thickness, while almost none of the transmitted impact forces measured with the load cell system were statistically different. The film sensor system was considered to be superior to the load cell system because the maximum stress and stress area could be determined. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Load Balancing Strategies for Multiphase Flows on Structured Grids
NASA Astrophysics Data System (ADS)
Olshefski, Kristopher; Owkes, Mark
2017-11-01
The computation time required to perform large simulations of complex systems is currently one of the leading bottlenecks of computational research. Parallelization allows multiple processing cores to perform calculations simultaneously and reduces computational times. However, load imbalances between processors waste computing resources as processors wait for others to complete imbalanced tasks. In multiphase flows, these imbalances arise due to the additional computational effort required at the gas-liquid interface. However, many current load balancing schemes are only designed for unstructured grid applications. The purpose of this research is to develop a load balancing strategy while maintaining the simplicity of a structured grid. Several approaches are investigated including brute force oversubscription, node oversubscription through Message Passing Interface (MPI) commands, and shared memory load balancing using OpenMP. Each of these strategies are tested with a simple one-dimensional model prior to implementation into the three-dimensional NGA code. Current results show load balancing will reduce computational time by at least 30%.
Active Response Gravity Offload and Method
NASA Technical Reports Server (NTRS)
Dungan, Larry K. (Inventor); Lieberman, Asher P. (Inventor); Shy, Cecil (Inventor); Bankieris, Derek R. (Inventor); Valle, Paul S. (Inventor); Redden, Lee (Inventor)
2015-01-01
A variable gravity field simulator can be utilized to provide three dimensional simulations for simulated gravity fields selectively ranging from Moon, Mars, and micro-gravity environments and/or other selectable gravity fields. The gravity field simulator utilizes a horizontally moveable carriage with a cable extending from a hoist. The cable can be attached to a load which experiences the effects of the simulated gravity environment. The load can be a human being or robot that makes movements that induce swinging of the cable whereby a horizontal control system reduces swinging energy. A vertical control system uses a non-linear feedback filter to remove noise from a load sensor that is in the same frequency range as signals from the load sensor.
Preparation of a Three-Dimensional Full Thickness Skin Equivalent.
Reuter, Christian; Walles, Heike; Groeber, Florian
2017-01-01
In vitro test systems are a promising alternative to animal models. Due to the use of human cells in a three-dimensional arrangement that allows cell-cell or cell-matrix interactions these models may be more predictive for the human situation compared to animal models or two-dimensional cell culture systems. Especially for dermatological research, skin models such as epidermal or full-thickness skin equivalents (FTSE) are used for different applications. Although epidermal models provide highly standardized conditions for risk assessment, FTSE facilitate a cellular crosstalk between the dermal and epidermal layer and thus can be used as more complex models for the investigation of processes such as wound healing, skin development, or infectious diseases. In this chapter, we describe the generation and culture of an FTSE, based on a collagen type I matrix and provide troubleshooting tips for commonly encountered technical problems.
Yazdi Rouholamini, Seyede Elmira; Moghassemi, Saeid; Maharat, Zahra; Hakamivala, Amirhossien; Kashanian, Susan; Omidfar, Kobra
2018-05-01
Silibinin is a natural flavonoid with a strong antioxidant property and weak cytotoxic activity. It has demonstrated anti-tumoural activity against many types of malignancies; however, due to its hydrophobic structure, it has poor water solubility, bioavailability and permeability across intestinal epithelial cells. To improve the effect of silibinin, we have vehiculated silibinin by a highly stable niosomal nanostructure based on a Span 60/cholesterol (CH)/N-trimethyl chitosan (TMC) system in order to study its potential application for the delivery of silibinin in T47D cultured under three-dimensional (3D) and two-dimensional (2D) conditions. To study the effect of nanodrug on miRNAs expression, we evaluated quantitative expression of miRNA-21 and miRNA-15a as well as miR-141 and miR-200c which act as oncogene and tumour suppressors by real-time PCR. Results demonstrated that the mechanism of nanodrug action as well as the response of tumour cells differed in 3D culture as compared to 2D. Delivery of silibinin-loaded niosomes coated with TMC was found to be more effective in inhibiting the growth of tumour cells and inducing apoptosis than free silibinin administration. In silibinin-treated cells, death occurred in a dose- and time- dependent manner by induction of apoptosis and alteration of the cell cycle. Real-time PCR analysis revealed a decrease in miR-21, miR-15a and miR-141while increase in miR-200c expression levels was observed in silibinin-treated cells relative to the levels in the untreated cells. The results show that nanodrug delivery was more effective than free silibinin administration in changing the level of miRNAs expression in cancer cells. Therefore, niosomal nanostructure with TMC could be a suitable vehicle for hydrophobic compounds, such as silibinin, by improving their action in cancer therapy.
NASA Technical Reports Server (NTRS)
Schwarz, R. P.; Goodwin, T. J.; Wolf, D. A.
1992-01-01
High-density, three-dimensional cell cultures are difficult to grow in vitro. The rotating-wall vessel (RWV) described here has cultured BHK-21 cells to a density of 1.1 X 10(7) cells/ml. Cells on microcarriers were observed to grow with enhanced bridging in this batch culture system. The RWV is a horizontally rotated tissue culture vessel with silicon membrane oxygenation. This design results in a low-turbulence, low-shear cell culture environment with abundant oxygenation. The RWV has the potential to culture a wide variety of normal and neoplastic cells.
Magnetic resonance imaging reveals functional anatomy and biomechanics of a living dragon tree
Hesse, Linnea; Masselter, Tom; Leupold, Jochen; Spengler, Nils; Speck, Thomas; Korvink, Jan Gerrit
2016-01-01
Magnetic resonance imaging (MRI) was used to gain in vivo insight into load-induced displacements of inner plant tissues making a non-invasive and non-destructive stress and strain analysis possible. The central aim of this study was the identification of a possible load-adapted orientation of the vascular bundles and their fibre caps as the mechanically relevant tissue in branch-stem-attachments of Dracaena marginata. The complex three-dimensional deformations that occur during mechanical loading can be analysed on the basis of quasi-three-dimensional data representations of the outer surface, the inner tissue arrangement (meristem and vascular system), and the course of single vascular bundles within the branch-stem-attachment region. In addition, deformations of vascular bundles could be quantified manually and by using digital image correlation software. This combination of qualitative and quantitative stress and strain analysis leads to an improved understanding of the functional morphology and biomechanics of D. marginata, a plant that is used as a model organism for optimizing branched technical fibre-reinforced lightweight trusses in order to increase their load bearing capacity. PMID:27604526
Qin, Li; Wang, Cheng-Zheng; Fan, Hui-Jie; Zhang, Chong-Jian; Zhang, Heng-Wei; Lv, Min-Hao; Cui, Shu-DE
2014-11-01
The treatment of a brain glioma remains one of the most difficult challenges in oncology. In the present study a delivery system was developed for targeted drug delivery across the blood-brain barrier (BBB) to the brain cancer cells. A cyclic arginine-glycine-aspartic acid (RGD) peptide and transferrin (TF) were utilized as targeting ligands. Cyclic RGD peptides are specific targeting ligands of cancer cells and TFs are ligands that specifically target the BBB and cancer cells. Liposome (LP) was used to conjugate the cyclic RGD and TFs to establish the brain glioma cascade delivery system (RGD/TF-LP). The LPs were prepared by the thin film hydration method and physicochemical characterization was conducted. In vitro cell uptake and three-dimensional tumor spheroid penetration studies demonstrated that the system could target endothelial and tumor cells, as well as penetrate the tumor cells to reach the core of the tumor spheroids. The results of the in vivo imaging further demonstrated that the RGD/TF-LP provided the highest brain distribution. As a result, the paclitaxel-loaded RGD/TF-LP presents the best antiproliferative activity against C6 cells and tumor spheroids. In conclusion, the RGD/TF-LP may precisely target brain glioma, which may be valuable for glioma imaging and therapy.
Walton, Katherine D; Kolterud, Asa
2014-09-04
Most morphogenetic processes in the fetal intestine have been inferred from thin sections of fixed tissues, providing snapshots of changes over developmental stages. Three-dimensional information from thin serial sections can be challenging to interpret because of the difficulty of reconstructing serial sections perfectly and maintaining proper orientation of the tissue over serial sections. Recent findings by Grosse et al., 2011 highlight the importance of three- dimensional information in understanding morphogenesis of the developing villi of the intestine(1). Three-dimensional reconstruction of singly labeled intestinal cells demonstrated that the majority of the intestinal epithelial cells contact both the apical and basal surfaces. Furthermore, three-dimensional reconstruction of the actin cytoskeleton at the apical surface of the epithelium demonstrated that the intestinal lumen is continuous and that secondary lumens are an artifact of sectioning. Those two points, along with the demonstration of interkinetic nuclear migration in the intestinal epithelium, defined the developing intestinal epithelium as a pseudostratified epithelium and not stratified as previously thought(1). The ability to observe the epithelium three-dimensionally was seminal to demonstrating this point and redefining epithelial morphogenesis in the fetal intestine. With the evolution of multi-photon imaging technology and three-dimensional reconstruction software, the ability to visualize intact, developing organs is rapidly improving. Two-photon excitation allows less damaging penetration deeper into tissues with high resolution. Two-photon imaging and 3D reconstruction of the whole fetal mouse intestines in Walton et al., 2012 helped to define the pattern of villus outgrowth(2). Here we describe a whole organ culture system that allows ex vivo development of villi and extensions of that culture system to allow the intestines to be three-dimensionally imaged during their development.
Macroporous Hydrogel Scaffolds for Three-Dimensional Cell Culture and Tissue Engineering.
Fan, Changjiang; Wang, Dong-An
2017-10-01
Hydrogels have been promising candidate scaffolds for cell delivery and tissue engineering due to their tissue-like physical properties and capability for homogeneous cell loading. However, the encapsulated cells are generally entrapped and constrained in the submicron- or nanosized gel networks, seriously limiting cell growth and tissue formation. Meanwhile, the spatially confined settlement inhibits attachment and spreading of anchorage-dependent cells, leading to their apoptosis. In recent years, macroporous hydrogels have attracted increasing attention in use as cell delivery vehicles and tissue engineering scaffolds. The introduction of macropores within gel scaffolds not only improves their permeability for better nutrient transport but also creates space/interface for cell adhesion, proliferation, and extracellular matrix deposition. Herein, we will first review the development of macroporous gel scaffolds and outline the impact of macropores on cell behaviors. In the first part, the advantages and challenges of hydrogels as three-dimensional (3D) cell culture scaffolds will be described. In the second part, the fabrication of various macroporous hydrogels will be presented. Third, the enhancement of cell activities within macroporous gel scaffolds will be discussed. Finally, several crucial factors that are envisaged to propel the improvement of macroporous gel scaffolds are proposed for 3D cell culture and tissue engineering.
Informing Stem Cell-Based Tendon Tissue Engineering Approaches with Embryonic Tendon Development.
Okech, William; Kuo, Catherine K
Adult tendons fail to regenerate normal tissue after injury, and instead form dysfunctional scar tissue with abnormal mechanical properties. Surgical repair with grafts is the current standard to treat injuries, but faces significant limitations including pain and high rates of re-injury. To address this, we aim to regenerate new, normal tendons to replace dysfunctional tendons. A common approach to tendon tissue engineering is to design scaffolds and bioreactors based on adult tendon properties that can direct adult stem cell tenogenesis. Despite significant progress, advances have been limited due, in part, to a need for markers and potent induction cues. Our goal is to develop novel tendon tissue engineering approaches informed by embryonic tendon development. We are characterizing structure-property relationships of embryonic tendon to identify design parameters for three-dimensional scaffolds and bioreactor mechanical loading systems to direct adult stem cell tenogenesis. We will review studies in which we quantified changes in the mechanical and biochemical properties of tendon during embryonic development and elucidated specific mechanisms of functional property elaboration. We then examined the effects of these mechanical and biochemical factors on embryonic tendon cell behavior. Using custom-designed bioreactors, we also examined the effects of dynamic mechanical loading and growth factor treatment on embryonic tendon cells. Our findings have established cues to induce tenogenesis as well as metrics to evaluate differentiation. We finish by discussing how we have evaluated the tenogenic differentiation potential of adult stem cells by comparing their responses to that of embryonic tendon cells in these culture systems.
Three-dimensional finite element analysis of the shear bond test.
DeHoff, P H; Anusavice, K J; Wang, Z
1995-03-01
The purpose of this study was to use finite element analyses to model the planar shear bond test and to evaluate the effects of modulus values, bonding agent thickness, and loading conditions on the stress distribution in the dentin adjacent to the bonding agent-dentin interface. All calculations were performed with the ANSYS finite element program. The planar shear bond test was modeled as a cylinder of resin-based composite bonded to a cylindrical dentin substrate. The effects of material, geometry and loading variables were determined primarily by use of a three-dimensional structural element. Several runs were also made using an axisymmetric element with harmonic loading and a plane strain element to determine whether two-dimensional analyses yield valid results. Stress calculations using three-dimensional finite element analyses confirmed the presence of large stress concentration effects for all stress components at the bonding agent-dentin interface near the application of the load. The maximum vertical shear stress generally occurs approximately 0.3 mm below the loading site and then decreases sharply in all directions. The stresses reach relatively uniform conditions within about 0.5 mm of the loading site and then increase again as the lower region of the interface is approached. Calculations using various loading conditions indicated that a wire-loop method of loading leads to smaller stress concentration effects, but a shear bond strength determined by dividing a failure load by the cross-sectional area grossly underestimates the true interfacial bond strength. Most dental researchers are using tensile and shear bond tests to predict the effects of process and material variables on the clinical performance of bonding systems but no evidence has yet shown that bond strength is relevant to clinical performance. A critical factor in assessing the usefulness of bond tests is a thorough understanding of the stress states that cause failure in the bond test and then to assess whether these stress states also exist in the clinical situation. Finite element analyses can help to answer this question but much additional work is needed to identify the failure modes in service and to relate these failures to particular loading conditions. The present study represents only a first step in understanding the stress states in the planar shear bond test.
Atom Probe Analysis of Ex Situ Gas-Charged Stable Hydrides.
Haley, Daniel; Bagot, Paul A J; Moody, Michael P
2017-04-01
In this work, we report on the atom probe tomography analysis of two metallic hydrides formed by pressurized charging using an ex situ hydrogen charging cell, in the pressure range of 200-500 kPa (2-5 bar). Specifically we report on the deuterium charging of Pd/Rh and V systems. Using this ex situ system, we demonstrate the successful loading and subsequent atom probe analysis of deuterium within a Pd/Rh alloy, and demonstrate that deuterium is likely present within the oxide-metal interface of a native oxide formed on vanadium. Through these experiments, we demonstrate the feasibility of ex situ hydrogen analysis for hydrides via atom probe tomography, and thus a practical route to three-dimensional imaging of hydrogen in hydrides at the atomic scale.
Xiang, Xu; Ding, Xiaochu; Moser, Trevor; Gao, Qi; Shokuhfar, Tolou; Heiden, Patricia A
2015-04-01
Peptide-functionalized polymeric nanoparticles were designed and self-assembled into continuous nanoparticle fibers and three-dimensional scaffolds via ionic complementary peptide interaction. Different nanoparticle compositions can be designed to be appropriate for each desired drug, so that the release of each drug is individually controlled and the simultaneous sustainable release of multiple drugs is achieved in a single scaffold. A self-assembled scaffold membrane was incubated with NIH3T3 fibroblast cells in a culture dish that demonstrated non-toxicity and non-inhibition on cell proliferation. This type of nanoparticle scaffold combines the advantages of peptide self-assembly and the versatility of polymeric nanoparticle controlled release systems for tissue engineering. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
NASA Technical Reports Server (NTRS)
Chen, Silvia S.; Revoltella, Roberto P.; Papini, Sandra; Michelini, Monica; Fitzgerald, Wendy; Zimmerberg, Joshua; Margolis, Leonid
2003-01-01
In the course of normal embryogenesis, embryonic stem (ES) cells differentiate along different lineages in the context of complex three-dimensional (3D) tissue structures. In order to study this phenomenon in vitro under controlled conditions, 3D culture systems are necessary. Here, we studied in vitro differentiation of rhesus monkey ES cells in 3D collagen matrixes (collagen gels and porous collagen sponges). Differentiation of ES cells in these 3D systems was different from that in monolayers. ES cells differentiated in collagen matrixes into neural, epithelial, and endothelial lineages. The abilities of ES cells to form various structures in two chemically similar but topologically different matrixes were different. In particular, in collagen gels ES cells formed gland-like circular structures, whereas in collagen sponges ES cells were scattered through the matrix or formed aggregates. Soluble factors produced by feeder cells or added to the culture medium facilitated ES cell differentiation into particular lineages. Coculture with fibroblasts in collagen gel facilitated ES cell differentiation into cells of a neural lineage expressing nestin, neural cell adhesion molecule, and class III beta-tubulin. In collagen sponges, keratinocytes facilitated ES cell differentiation into cells of an endothelial lineage expressing factor VIII. Exogenous granulocyte-macrophage colony-stimulating factor further enhanced endothelial differentiation. Thus, both soluble factors and the type of extracellular matrix seem to be critical in directing differentiation of ES cells and the formation of tissue-like structures. Three-dimensional culture systems are a valuable tool for studying the mechanisms of these phenomena.
NASA Technical Reports Server (NTRS)
Hooper, Steven J.
1989-01-01
Delamination is a common failure mode of laminated composite materials. This type of failure frequently occurs at the free edges of laminates where singular interlaminar stresses are developed due to the difference in Poisson's ratios between adjacent plies. Typically the delaminations develop between 90 degree plies and adjacent angle plies. Edge delamination has been studied by several investigators using a variety of techniques. Recently, Chan and Ochoa applied the quasi-three-dimensional finite element model to the analysis of a laminate subject to bending, extension, and torsion. This problem is of particular significance relative to the structural integrity of composite helicopter rotors. The task undertaken was to incorporate Chan and Ochoa's formulation into a Raju Q3DG program. The resulting program is capable of modeling extension, bending, and torsional mechanical loadings as well as thermal and hygroscopic loadings. The addition of the torsional and bending loading capability will provide the capability to perform a delamination analysis of a general unsymmetric laminate containing four cracks, each of a different length. The solutions obtained using this program are evaluated by comparing them with solutions from a full three-dimensional finite element solution. This comparison facilitates the assessment of three dimensional affects such as the warping constraint imposed by the load frame grips. It wlso facilitates the evaluation of the external load representation employed in the Q3D formulation. Finally, strain energy release rates computed from the three-dimensional results are compared with those predicted using the quasi-three-dimensional formulation.
Lawrence, Marc; Gross, Gion-Pitschen; Lang, Martin; Kuhn, Andreas; Keller, Thierry; Morari, Manfred
2008-08-01
New multichannel textile neuroprotheses were developed, which comprise multiple sets of transcutaneous electrode arrays and connecting wires embroidered into a fabric layer. The electrode arrays were placed on the forearm above the extrinsic finger flexors and extensors. Activation regions for selective finger flexion and wrist extension were configured by switching a subset of the array elements between cathode, anode, and off states. We present a new isometric measurement system for the assessment of finger forces and wrist torques generated using the new neuroprostheses. Finger forces (from the middle phalanxes) were recorded using five load cells mounted on a "grasp handle" that can be arbitrarily positioned in space. The hand and the grasp handle were rigidly mounted to a 6-degree of freedom load cell, and the forces and torques about the wrist were recorded. A vacuum cushion was used to comfortably fixate the forearm. The position and orientation of the forearm, wrist, fingers, and handle were recorded using a new three-dimensional position measurement system (accuracy <+/-1 mm). The measurement system was integrated into the real-time multichannel transcutaneous electrode environment, which is able to control the spatiotemporal position of multiple activation regions. Using the combined system and textile neuroprosthesis, we were able to optimize the activation regions to produce selective finger and wrist articulation, enabling improved functional grasp.
NASA Technical Reports Server (NTRS)
Goodwin, Thomas J.
2003-01-01
The present investigation details the development of model systems for growing two- and three-dimensional human neural progenitor cells within a culture medium facilitated by a time-varying electromagnetic field (TVEMF). The cells and culture medium are contained within a two- or three-dimensional culture vessel, and the electromagnetic field is emitted from an electrode or coil. These studies further provide methods to promote neural tissue regeneration by means of culturing the neural cells in either configuration. Grown in two dimensions, neuronal cells extended longitudinally, forming tissue strands extending axially along and within electrodes comprising electrically conductive channels or guides through which a time-varying electrical current was conducted. In the three-dimensional aspect, exposure to TVEMF resulted in the development of three-dimensional aggregates, which emulated organized neural tissues. In both experimental configurations, the proliferation rate of the TVEMF cells was 2.5 to 4.0 times the rate of the non-waveform cells. Each of the experimental embodiments resulted in similar molecular genetic changes regarding the growth potential of the tissues as measured by gene chip analyses, which measured more than 10,000 human genes simultaneously.
Rahaman, Mostafizur; Govindasami, Periyasami; Almoiqli, Mohammed; Altalhi, Tariq; Mezni, Amine
2017-01-01
This article discusses the deposition of different amount of microstuctured poly(3,4-ethylenedioxythiophene) (PEDOT) on reticulated vitreous carbon (RVC) by electrochemical method to prepare three-dimensional (3D) PEDOT/RVC electrodes aimed to be used in capacitive deionization (CDI) technology. A CDI unit cell has been constructed here in this study. The performance of CDI cell in the ion removal of NaCl onto the sites of PEDOT/RVC electrode has been systematically investigated in terms of flow-rate, applied electrical voltage, and increasing PEDOT loading on PEDOT/RVC electrodes. It is observed that the increase in flow-rate, electric voltage, and PEDOT loading up to a certain level improve the ion removal performance of electrode in the CDI cell. The result shows that these electrodes can be used effectively for desalination technology, as the electrosorption capacity/desalination performance of these electrodes is quite high compared to carbon materials. Moreover, the stability of the electrodes has been tested and it is reported that these electrodes are regenerative. The effect of increasing NaCl concentration on the electrosorption capacity has also been investigated for these electrodes. Finally, it has been shown that 1 m3 PEDOT-120 min/RVC electrodes from 75 mg/L NaCl feed solution produce 421, 978 L water per day of 20 mg/L NaCl final concentration. PMID:28773205
Aldalbahi, Ali; Rahaman, Mostafizur; Govindasami, Periyasami; Almoiqli, Mohammed; Altalhi, Tariq; Mezni, Amine
2017-07-24
This article discusses the deposition of different amount of microstuctured poly(3,4-ethylenedioxythiophene) (PEDOT) on reticulated vitreous carbon (RVC) by electrochemical method to prepare three-dimensional (3D) PEDOT/RVC electrodes aimed to be used in capacitive deionization (CDI) technology. A CDI unit cell has been constructed here in this study. The performance of CDI cell in the ion removal of NaCl onto the sites of PEDOT/RVC electrode has been systematically investigated in terms of flow-rate, applied electrical voltage, and increasing PEDOT loading on PEDOT/RVC electrodes. It is observed that the increase in flow-rate, electric voltage, and PEDOT loading up to a certain level improve the ion removal performance of electrode in the CDI cell. The result shows that these electrodes can be used effectively for desalination technology, as the electrosorption capacity/desalination performance of these electrodes is quite high compared to carbon materials. Moreover, the stability of the electrodes has been tested and it is reported that these electrodes are regenerative. The effect of increasing NaCl concentration on the electrosorption capacity has also been investigated for these electrodes. Finally, it has been shown that 1 m³ PEDOT-120 min/RVC electrodes from 75 mg/L NaCl feed solution produce 421, 978 L water per day of 20 mg/L NaCl final concentration.
Thomopoulos, Stavros; Das, Rosalina; Birman, Victor; Smith, Lester; Ku, Katherine; Elson, Elliott L; Pryse, Kenneth M; Marquez, Juan Pablo; Genin, Guy M
2011-04-01
Although much is known about the effects of uniaxial mechanical loading on fibrocartilage development, the stress fields to which fibrocartilaginous regions are subjected to during development are mutiaxial. That fibrocartilage develops at tendon-to-bone attachments and in compressive regions of tendons is well established. However, the three-dimensional (3D) nature of the stresses needed for the development of fibrocartilage is not known. Here, we developed and applied an in vitro system to determine whether fibrocartilage can develop under a state of periodic hydrostatic tension in which only a single principal component of stress is compressive. This question is vital to efforts to mechanically guide morphogenesis and matrix expression in engineered tissue replacements. Mesenchymal stromal cells in a 3D culture were exposed to compressive and tensile stresses as a result of an external tensile hydrostatic stress field. The stress field was characterized through mechanical modeling. Tensile cyclic stresses promoted spindle-shaped cells, upregulation of scleraxis and type one collagen, and cell alignment with the direction of tension. Cells experiencing a single compressive stress component exhibited rounded cell morphology and random cell orientation. No difference in mRNA expression of the genes Sox9 and aggrecan was observed when comparing tensile and compressive regions unless the medium was supplemented with the chondrogenic factor transforming growth factor beta3. In that case, Sox9 was upregulated under static loading conditions and aggrecan was upregulated under cyclic loading conditions. In conclusion, the fibrous component of fibrocartilage could be generated using only mechanical cues, but generation of the cartilaginous component of fibrocartilage required biologic factors in addition to mechanical cues. These studies support the hypothesis that the 3D stress environment influences cell activity and gene expression in fibrocartilage development.
Das, Rosalina; Birman, Victor; Smith, Lester; Ku, Katherine; Elson, Elliott L.; Pryse, Kenneth M.; Marquez, Juan Pablo; Genin, Guy M.
2011-01-01
Although much is known about the effects of uniaxial mechanical loading on fibrocartilage development, the stress fields to which fibrocartilaginous regions are subjected to during development are mutiaxial. That fibrocartilage develops at tendon-to-bone attachments and in compressive regions of tendons is well established. However, the three-dimensional (3D) nature of the stresses needed for the development of fibrocartilage is not known. Here, we developed and applied an in vitro system to determine whether fibrocartilage can develop under a state of periodic hydrostatic tension in which only a single principal component of stress is compressive. This question is vital to efforts to mechanically guide morphogenesis and matrix expression in engineered tissue replacements. Mesenchymal stromal cells in a 3D culture were exposed to compressive and tensile stresses as a result of an external tensile hydrostatic stress field. The stress field was characterized through mechanical modeling. Tensile cyclic stresses promoted spindle-shaped cells, upregulation of scleraxis and type one collagen, and cell alignment with the direction of tension. Cells experiencing a single compressive stress component exhibited rounded cell morphology and random cell orientation. No difference in mRNA expression of the genes Sox9 and aggrecan was observed when comparing tensile and compressive regions unless the medium was supplemented with the chondrogenic factor transforming growth factor beta3. In that case, Sox9 was upregulated under static loading conditions and aggrecan was upregulated under cyclic loading conditions. In conclusion, the fibrous component of fibrocartilage could be generated using only mechanical cues, but generation of the cartilaginous component of fibrocartilage required biologic factors in addition to mechanical cues. These studies support the hypothesis that the 3D stress environment influences cell activity and gene expression in fibrocartilage development. PMID:21091338
From Three-Dimensional Cell Culture to Organs-on-Chips
Huh, Dongeun; Hamilton, Geraldine A.; Ingber, Donald E.
2014-01-01
Three-dimensional (3D) cell culture models have recently garnered great attention because they often promote levels of cell differentiation and tissue organization not possible in conventional two-dimensional (2D) culture systems. Here, we review new advances in 3D culture that leverage microfabrication technologies from the microchip industry and microfluidics approaches to create cell culture microenvironments that both support tissue differentiation and recapitulate the tissue-tissue interfaces, spatiotemporal chemical gradients, and mechanical microenvironments of living organs. These ‘organs-on-chips’ permit study of human physiology in an organ-specific context, enable development of novel in vitro disease models, and could potentially serve as replacements for animals used in drug development and toxin testing. PMID:22033488
Weise, Frank; Fernekorn, Uta; Hampl, Jörg; Klett, Maren; Schober, Andreas
2013-09-01
By the use of a MatriGrid® we have established a three-dimensional high density cell culture. The MatriGrid® is a culture medium permeable, polymeric scaffold with 187 microcavities. In these cavities (300 μm diameter and 207 μm deep) the cells can growth three-dimensionally. For these experiments we measured the oxygen consumption of HepG2 cell cultures in order to optimize cultivation conditions. We measured and compared the oxygen consumption, growth rate and vitality under three different cultivation conditions: monolayer, three-dimensional static and three-dimensional actively perfused. The results show that the cells in a three-dimensional cell culture consume less oxygen as in a monolayer cell culture and that the actively perfused three-dimensional cell culture in the MatriGrid® has a similar growth rate and vitality as the monolayer culture. Copyright © 2013 Wiley Periodicals, Inc.
In situ calibration of neutron activation system on the large helical device
NASA Astrophysics Data System (ADS)
Pu, N.; Nishitani, T.; Isobe, M.; Ogawa, K.; Kawase, H.; Tanaka, T.; Li, S. Y.; Yoshihashi, S.; Uritani, A.
2017-11-01
In situ calibration of the neutron activation system on the Large Helical Device (LHD) was performed by using an intense 252Cf neutron source. To simulate a ring-shaped neutron source, we installed a railway inside the LHD vacuum vessel and made a train loaded with the 252Cf source run along a typical magnetic axis position. Three activation capsules loaded with thirty pieces of indium foils stacked with total mass of approximately 18 g were prepared. Each capsule was irradiated over 15 h while the train was circulating. The activation response coefficient (9.4 ± 1.2) × 10-8 of 115In(n, n')115mIn reaction obtained from the experiment is in good agreement with results from three-dimensional neutron transport calculations using the Monte Carlo neutron transport simulation code 6. The activation response coefficients of 2.45 MeV birth neutron and secondary 14.1 MeV neutron from deuterium plasma were evaluated from the activation response coefficient obtained in this calibration experiment with results from three-dimensional neutron calculations using the Monte Carlo neutron transport simulation code 6.
Hippocampal place-cell firing during movement in three-dimensional space
NASA Technical Reports Server (NTRS)
Knierim, J. J.; McNaughton, B. L.
2001-01-01
"Place" cells of the rat hippocampus are coupled to "head direction" cells of the thalamus and limbic cortex. Head direction cells are sensitive to head direction in the horizontal plane only, which leads to the question of whether place cells similarly encode locations in the horizontal plane only, ignoring the z axis, or whether they encode locations in three dimensions. This question was addressed by recording from ensembles of CA1 pyramidal cells while rats traversed a rectangular track that could be tilted and rotated to different three-dimensional orientations. Cells were analyzed to determine whether their firing was bound to the external, three-dimensional cues of the environment, to the two-dimensional rectangular surface, or to some combination of these cues. Tilting the track 45 degrees generally provoked a partial remapping of the rectangular surface in that some cells maintained their place fields, whereas other cells either gained new place fields, lost existing fields, or changed their firing locations arbitrarily. When the tilted track was rotated relative to the distal landmarks, most place fields remapped, but a number of cells maintained the same place field relative to the x-y coordinate frame of the laboratory, ignoring the z axis. No more cells were bound to the local reference frame of the recording apparatus than would be predicted by chance. The partial remapping demonstrated that the place cell system was sensitive to the three-dimensional manipulations of the recording apparatus. Nonetheless the results were not consistent with an explicit three-dimensional tuning of individual hippocampal neurons nor were they consistent with a model in which different sets of cells are tightly coupled to different sets of environmental cues. The results are most consistent with the statement that hippocampal neurons can change their "tuning functions" in arbitrary ways when features of the sensory input or behavioral context are altered. Understanding the rules that govern the remapping phenomenon holds promise for deciphering the neural circuitry underlying hippocampal function.
Zhao, Lingling; Niu, Lijing; Liang, Hongze; Tan, Hui; Liu, Chaozong; Zhu, Feiyan
2017-11-01
pH and glucose dual-responsive injectable hydrogels were prepared through the cross-linking of Schiff's base and phenylboronate ester using phenylboronic-modified chitosan, poly(vinyl alcohol) and benzaldehyde-capped poly(ethylene glycol). Protein drugs and live cells could be incorporated into the hydrogels during the in situ cross-linking, displaying sustained and pH/glucose-triggered drug release from the hydrogels and cell viability and proliferation in the three-dimensional hydrogel matrix as well. Hence, the hydrogels with insulin and fibroblasts were considered as bioactive dressings for diabetic wound healing. A streptozotocin-induced diabetic rat model was used to evaluate the efficacy of hydrogel dressings in wound repair. The results revealed that the incorporation of insulin and L929 in the hydrogels could promote neovascularization and collagen deposition and enhance the wound-healing process of diabetic wounds. Thus, the drug- and cell-loaded hydrogels have promising potential in wound healing as a medicated system for various therapeutic proteins and live cells.
NASA Astrophysics Data System (ADS)
Renaud, Olivier; Heintzmann, Rainer; Sáez-Cirión, Asier; Schnelle, Thomas; Mueller, Torsten; Shorte, Spencer
2007-02-01
Three dimensional imaging provides high-content information from living intact biology, and can serve as a visual screening cue. In the case of single cell imaging the current state of the art uses so-called "axial through-stacking". However, three-dimensional axial through-stacking requires that the object (i.e. a living cell) be adherently stabilized on an optically transparent surface, usually glass; evidently precluding use of cells in suspension. Aiming to overcome this limitation we present here the utility of dielectric field trapping of single cells in three-dimensional electrode cages. Our approach allows gentle and precise spatial orientation and vectored rotation of living, non-adherent cells in fluid suspension. Using various modes of widefield, and confocal microscope imaging we show how so-called "microrotation" can provide a unique and powerful method for multiple point-of-view (three-dimensional) interrogation of intact living biological micro-objects (e.g. single-cells, cell aggregates, and embryos). Further, we show how visual screening by micro-rotation imaging can be combined with micro-fluidic sorting, allowing selection of rare phenotype targets from small populations of cells in suspension, and subsequent one-step single cell cloning (with high-viability). Our methodology combining high-content 3D visual screening with one-step single cell cloning, will impact diverse paradigms, for example cytological and cytogenetic analysis on haematopoietic stem cells, blood cells including lymphocytes, and cancer cells.
Hirakawa, Takeshi; Matsunaga, Sachihiro
2016-01-01
In plants, chromatin dynamics spatiotemporally change in response to various environmental stimuli. However, little is known about chromatin dynamics in the nuclei of plants. Here, we introduce a three-dimensional, live-cell imaging method that can monitor chromatin dynamics in nuclei via a chromatin tagging system that can visualize specific genomic loci in living plant cells. The chromatin tagging system is based on a bacterial operator/repressor system in which the repressor is fused to fluorescent proteins. A recent refinement of promoters for the system solved the problem of gene silencing and abnormal pairing frequencies between operators. Using this system, we can detect the spatiotemporal dynamics of two homologous loci as two fluorescent signals within a nucleus and monitor the distance between homologous loci. These live-cell imaging methods will provide new insights into genome organization, development processes, and subnuclear responses to environmental stimuli in plants.
Crack surface roughness in three-dimensional random fuse networks
NASA Astrophysics Data System (ADS)
Nukala, Phani Kumar V. V.; Zapperi, Stefano; Šimunović, Srđan
2006-08-01
Using large system sizes with extensive statistical sampling, we analyze the scaling properties of crack roughness and damage profiles in the three-dimensional random fuse model. The analysis of damage profiles indicates that damage accumulates in a diffusive manner up to the peak load, and localization sets in abruptly at the peak load, starting from a uniform damage landscape. The global crack width scales as Wtilde L0.5 and is consistent with the scaling of localization length ξ˜L0.5 used in the data collapse of damage profiles in the postpeak regime. This consistency between the global crack roughness exponent and the postpeak damage profile localization length supports the idea that the postpeak damage profile is predominantly due to the localization produced by the catastrophic failure, which at the same time results in the formation of the final crack. Finally, the crack width distributions can be collapsed for different system sizes and follow a log-normal distribution.
Three-dimensional hysteresis compensation enhances accuracy of robotic artificial muscles
NASA Astrophysics Data System (ADS)
Zhang, Jun; Simeonov, Anthony; Yip, Michael C.
2018-03-01
Robotic artificial muscles are compliant and can generate straight contractions. They are increasingly popular as driving mechanisms for robotic systems. However, their strain and tension force often vary simultaneously under varying loads and inputs, resulting in three-dimensional hysteretic relationships. The three-dimensional hysteresis in robotic artificial muscles poses difficulties in estimating how they work and how to make them perform designed motions. This study proposes an approach to driving robotic artificial muscles to generate designed motions and forces by modeling and compensating for their three-dimensional hysteresis. The proposed scheme captures the nonlinearity by embedding two hysteresis models. The effectiveness of the model is confirmed by testing three popular robotic artificial muscles. Inverting the proposed model allows us to compensate for the hysteresis among temperature surrogate, contraction length, and tension force of a shape memory alloy (SMA) actuator. Feedforward control of an SMA-actuated robotic bicep is demonstrated. This study can be generalized to other robotic artificial muscles, thus enabling muscle-powered machines to generate desired motions.
Lifetime prediction for the subsurface crack propagation using three-dimensional dynamic FEA model
NASA Astrophysics Data System (ADS)
Yin, Yuan; Chen, Yun-Xia; Liu, Le
2017-03-01
The subsurface crack propagation is one of the major interests for gear system research. The subsurface crack propagation lifetime is the number of cycles remaining for a spall to appear, which can be obtained through either stress intensity factor or accumulated plastic strain analysis. In this paper, the heavy loads are applied to the gear system. When choosing stress intensity factor, the high compressive stress suppresses Mode I stress intensities and severely reduces Mode II stress intensities in the heavily loaded lubricated contacts. Such that, the accumulated plastic strain is selected to calculate the subsurface crack propagation lifetime from the three-dimensional FEA model through ANSYS Workbench transient analysis. The three-dimensional gear FEA dynamic model with the subsurface crack is built through dividing the gears into several small elements. The calculation of the total cycles of the elements is proposed based on the time-varying accumulated plastic strain, which then will be used to calculate the subsurface crack propagation lifetime. During this process, the demonstration from a subsurface crack to a spall can be uncovered. In addition, different sizes of the elements around the subsurface crack are compared in this paper. The influences of the frictional coefficient and external torque on the crack propagation lifetime are also discussed. The results show that the lifetime of crack propagation decreases significantly when the external load T increasing from 100 N m to 150 N m. Given from the distributions of the accumulated plastic strain, the lifetime shares no significant difference when the frictional coefficient f ranging in 0.04-0.06.
NASA Astrophysics Data System (ADS)
Quan, Wei-cai; Zhang, Zhu-ying; Zhang, Ai-qun; Zhang, Qi-feng; Tian, Yu
2015-04-01
This paper proposes a geometrically exact formulation for three-dimensional static and dynamic analyses of the umbilical cable in a deep-sea remotely operated vehicle (ROV) system. The presented formulation takes account of the geometric nonlinearities of large displacement, effects of axial load and bending stiffness for modeling of slack cables. The resulting nonlinear second-order governing equations are discretized spatially by the finite element method and solved temporally by the generalized- α implicit time integration algorithm, which is adapted to the case of varying coefficient matrices. The ability to consider three-dimensional union action of ocean current and ship heave motion upon the umbilical cable is the key feature of this analysis. The presented formulation is firstly validated, and then three numerical examples for the umbilical cable in a deep-sea ROV system are demonstrated and discussed, including the steady configurations only under the action of depth-dependent ocean current, the dynamic responses in the case of the only ship heave motion, and in the case of the combined action of the ship heave motion and ocean current.
Overview of Aerothermodynamic Loads Definition Study
NASA Technical Reports Server (NTRS)
Povinelli, L. A.
1985-01-01
The Aerothermodynamic Loads Definition were studied to develop methods to more accurately predict the operating environment in the space shuttle main engine (SSME) components. Development of steady and time-dependent, three-dimensional viscous computer codes and experimental verification and engine diagnostic testing are considered. The steady, nonsteady, and transient operating loads are defined to accurately predict powerhead life. Improvements in the structural durability of the SSME turbine drive systems depends on the knowledge of the aerothermodynamic behavior of the flow through the preburner, turbine, turnaround duct, gas manifold, and injector post regions.
Dynamics of an HIV-1 infection model with cell mediated immunity
NASA Astrophysics Data System (ADS)
Yu, Pei; Huang, Jianing; Jiang, Jiao
2014-10-01
In this paper, we study the dynamics of an improved mathematical model on HIV-1 virus with cell mediated immunity. This new 5-dimensional model is based on the combination of a basic 3-dimensional HIV-1 model and a 4-dimensional immunity response model, which more realistically describes dynamics between the uninfected cells, infected cells, virus, the CTL response cells and CTL effector cells. Our 5-dimensional model may be reduced to the 4-dimensional model by applying a quasi-steady state assumption on the variable of virus. However, it is shown in this paper that virus is necessary to be involved in the modeling, and that a quasi-steady state assumption should be applied carefully, which may miss some important dynamical behavior of the system. Detailed bifurcation analysis is given to show that the system has three equilibrium solutions, namely the infection-free equilibrium, the infectious equilibrium without CTL, and the infectious equilibrium with CTL, and a series of bifurcations including two transcritical bifurcations and one or two possible Hopf bifurcations occur from these three equilibria as the basic reproduction number is varied. The mathematical methods applied in this paper include characteristic equations, Routh-Hurwitz condition, fluctuation lemma, Lyapunov function and computation of normal forms. Numerical simulation is also presented to demonstrate the applicability of the theoretical predictions.
Half-Cell RF Gun Simulations with the Electromagnetic Particle-in-Cell Code VORPAL
NASA Astrophysics Data System (ADS)
Paul, K.; Dimitrov, D. A.; Busby, R.; Bruhwiler, D. L.; Smithe, D.; Cary, J. R.; Kewisch, J.; Kayran, D.; Calaga, R.; Ben-Zvi, I.
2009-01-01
We have simulated Brookhaven National Laboratory's half-cell superconducting RF gun design for a proposed high-current ERL using the three-dimensional, electromagnetic particle-in-cell code VORPAL. VORPAL computes the fully self-consistent electromagnetic fields produced by the electron bunches, meaning that it accurately models space-charge effects as well as bunch-to-bunch beam loading effects and the effects of higher-order cavity modes, though these are beyond the scope of this paper. We compare results from VORPAL to the well-established space-charge code PARMELA, using RF fields produced by SUPERFISH, as a benchmarking exercise in which the two codes should agree well.
Sambi, Manpreet; Chow, Theresa; Whiteley, Jennifer; Li, Mira; Chua, Shawn; Raileanu, Vanessa; Rogers, Ian M
2017-08-01
The development of strategies for tissue regeneration and bio-artificial organ development is based on our understanding of embryogenesis. Differentiation protocols attempt to recapitulate the signaling modalities of gastrulation and organogenesis, coupled with cell selection regimens to isolate the cells of choice. This strategy is impeded by the lack of optimal in vitro culture systems since traditional culture systems do not allow for the three-dimensional interaction between cells and the extracellular matrix. While artificial three-dimensional scaffolds are available, using the natural extracellular matrix scaffold is advantageous because it has a distinct architecture that is difficult to replicate. The adult extracellular matrix is predicted to mediate signaling related to tissue repair not embryogenesis but existing similarities between the two argues that the extracellular matrix will influence the differentiation of stem and progenitor cells. Previous studies using undifferentiated embryonic stem cells grown directly on acellular kidney ECM demonstrated that the acellular kidney supported cell growth but limited differentiation occurred. Using mouse kidney extracellular matrix and mouse embryonic stem cells we report that the extracellular matrix can support the development of kidney structures if the stem cells are first differentiated to kidney progenitor cells before being applied to the acellular organ.
Force system generated by elastic archwires with vertical V bends: a three-dimensional analysis.
Upadhyay, Madhur; Shah, Raja; Peterson, Donald; Asaki, Takafumi; Yadav, Sumit; Agarwal, Sachin
2017-04-01
Our previous understanding of V-bend mechanics is primarily from two-dimensional (2D) analysis of archwire bracket interactions in the second order. These analyses do not take into consideration the three-dimensional (3D) nature of orthodontic appliances involving the third order. To quantify the force system generated in a 3D two bracket set up involving the molar and incisors with vertical V-bends. Maxillary molar and incisor brackets were arranged in a dental arch form and attached to load cells capable of measuring forces and moments in all three planes (x, y, and z) of space. Symmetrical V-bends (right and left sides) were placed at 11 different locations along rectangular beta-titanium archwires of various sizes at an angle of 150degrees. Each wire was evaluated for the 11 bend positions. Specifically, the vertical forces (Fz) and anterio-posterior moments (Mx) were analysed. Descriptive statistics were used to interpret the results. With increasing archwire size, Fz and Mx increased at the two brackets (P < 0.05). The vertical forces were linear and symmetric in nature, increasing in magnitude as the bends moved closer to either bracket. The Mx curves were asymmetric and non-linear displaying higher magnitudes for molar bracket. As the bends were moved closer to either bracket a distinct flattening of the incisor Mx curve was noted, implying no change in its magnitude. This article provides critical information on V-bend mechanics involving second order and third order archwire-bracket interactions. A model for determining this force system is described that might allow for easier translation to actual clinical practice. © The Author 2016. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com
Chebabhi, Ali; Fellah, Mohammed Karim; Kessal, Abdelhalim; Benkhoris, Mohamed F
2016-07-01
In this paper is proposed a new balancing three-level three dimensional space vector modulation (B3L-3DSVM) strategy which uses a redundant voltage vectors to realize precise control and high-performance for a three phase three-level four-leg neutral point clamped (NPC) inverter based Shunt Active Power Filter (SAPF) for eliminate the source currents harmonics, reduce the magnitude of neutral wire current (eliminate the zero-sequence current produced by single-phase nonlinear loads), and to compensate the reactive power in the three-phase four-wire electrical networks. This strategy is proposed in order to gate switching pulses generation, dc bus voltage capacitors balancing (conserve equal voltage of the two dc bus capacitors), and to switching frequency reduced and fixed of inverter switches in same times. A Nonlinear Back Stepping Controllers (NBSC) are used for regulated the dc bus voltage capacitors and the SAPF injected currents to robustness, stabilizing the system and to improve the response and to eliminate the overshoot and undershoot of traditional PI (Proportional-Integral). Conventional three-level three dimensional space vector modulation (C3L-3DSVM) and B3L-3DSVM are calculated and compared in terms of error between the two dc bus voltage capacitors, SAPF output voltages and THDv, THDi of source currents, magnitude of source neutral wire current, and the reactive power compensation under unbalanced single phase nonlinear loads. The success, robustness, and the effectiveness of the proposed control strategies are demonstrated through simulation using Sim Power Systems and S-Function of MATLAB/SIMULINK. Copyright © 2016 ISA. Published by Elsevier Ltd. All rights reserved.
NASA Technical Reports Server (NTRS)
Kennedy, Ronald; Padovan, Joe
1987-01-01
In a three-part series of papers, a generalized finite element solution strategy is developed to handle traveling load problems in rolling, moving and rotating structure. The main thrust of this section consists of the development of three-dimensional and shell type moving elements. In conjunction with this work, a compatible three-dimensional contact strategy is also developed. Based on these modeling capabilities, extensive analytical and experimental benchmarking is presented. Such testing includes traveling loads in rotating structure as well as low- and high-speed rolling contact involving standing wave-type response behavior. These point to the excellent modeling capabilities of moving element strategies.
NASA Technical Reports Server (NTRS)
Goodwin, T. J.; Prewett, T. L.; Spaulding, G. F.; Becker, J. L.
1997-01-01
The Rotating-Wall Vessel (RWV) is a novel in vitro cell culture system used to successfully culture a cell line derived from a heterologous mixed mullerian tumor cell of the ovary. Although the original tumor was comprised of both epithelial and mesodermal components, long-term culture in conventional flasks established a cell line from this tumor with homogeneous epitheliallike growth characteristics (1). Cells from Passage 36 were seeded into a Rotating-Wall Vessel containing Cytodex-3 microcarrier beads. Scanning electron micrographs of tumor cells cultured for 32 d in the RWV showed the presence of heterogeneous cell populations organized into three-dimensional tissuelike architecture. Immunocytochemical analysis confirmed the cellular heterogeneity, as demonstrated by expression of both epithelial and mesenchymal antigens. Reverse transcription polymerase chain reaction amplification demonstrated the presence of mRNA for cellular oncogenes HER-2/neu, H-ras, K-ras, and tumor suppressor p53. Thus, there are two advantages to propagation of tissue in the RWV culture system:(a) tissue diversification representing populations present in the original tumor, and (b) the three-dimensional freedom to organize tissues morphologically akin to those observed in vivo. These data indicate that the RWV culture system is suitable for generating large quantities of ovarian tumor cells in vitro that are amenable to immunocytochemical, oncogenic, morphologic characteristics demonstrated in vivo.
Balasubramanian, Saravana K; Coger, Robin N
2005-01-01
Bioartificial liver devices (BALs) have proven to be an effective bridge to transplantation for cases of acute liver failure. Enabling the long-term storage of these devices using a method such as cryopreservation will ensure their easy off the shelf availability. To date, cryopreservation of liver cells has been attempted for both single cells and sandwich cultures. This study presents the potential of using computational modeling to help develop a cryopreservation protocol for storing the three dimensional BAL: Hepatassist. The focus is upon determining the thermal and concentration profiles as the BAL is cooled from 37 degrees C-100 degrees C, and is completed in two steps: a cryoprotectant loading step and a phase change step. The results indicate that, for the loading step, mass transfer controls the duration of the protocol, whereas for the phase change step, when mass transfer is assumed negligible, the latent heat released during freezing is the control factor. The cryoprotocol that is ultimately proposed considers time, cooling rate, and the temperature gradients that the cellular space is exposed to during cooling. To our knowledge, this study is the first reported effort toward designing an effective protocol for the cryopreservation of a three-dimensional BAL device.
Analysis of interlaminar stresses in symmetric and unsymmetric laminates under various loadings
NASA Astrophysics Data System (ADS)
Leger, C. A.; Chan, W. S.
1993-04-01
A quasi-three-dimensional finite-element model is developed to investigate the interlaminar stresses in a composite laminate under combined loadings. An isoparametric quadrilateral element with eight nodes and three degrees of freedom per node is the finite element used in this study. The element is used to model a composite laminate cross section loaded by tension, torsion, transverse shear, and both beam and chord bending which are representative of loading in a helicopter rotor system. Symmetric and unsymmetric laminates are examined with comparisons made between the interlaminar stress distributions and magnitudes for each laminate. Unsymmetric results are compared favorably to limited results found in literature. The unsymmetric interlaminar normal stress distribution in a symmetric laminate containing a free edge delamination is also examined.
Hodde, Dorothee; Gerardo-Nava, José; Wöhlk, Vanessa; Weinandy, Stefan; Jockenhövel, Stefan; Kriebel, Andreas; Altinova, Haktan; Steinbusch, Harry W M; Möller, Martin; Weis, Joachim; Mey, Jörg; Brook, Gary A
2016-02-01
The generation of complex three-dimensional bioengineered scaffolds that are capable of mimicking the molecular and topographical cues of the extracellular matrix found in native tissues is a field of expanding research. The systematic development of such scaffolds requires the characterisation of cell behaviour in response to the individual components of the scaffold. In the present investigation, we studied cell-substrate interactions between purified populations of Schwann cells and three-dimensional fibrin hydrogel scaffolds, in the presence or absence of multiple layers of highly orientated electrospun polycaprolactone nanofibres. Embedded Schwann cells remained viable within the fibrin hydrogel for up to 7 days (the longest time studied); however, cell behaviour in the hydrogel was somewhat different to that observed on the two-dimensional fibrin substrate: Schwann cells failed to proliferate in the fibrin hydrogel, whereas cell numbers increased steadily on the two-dimensional fibrin substrate. Schwann cells within the fibrin hydrogel developed complex process branching patterns, but, when presented with orientated nanofibres, showed a strong tendency to redistribute themselves onto the nanofibres, where they extended long processes that followed the longitudinal orientation of the nanofibres. The process length along nanofibre-containing fibrin hydrogel reached near-maximal levels (for the present experimental conditions) as early as 1 day after culturing. The ability of this three-dimensional, extracellular matrix-mimicking scaffold to support Schwann cell survival and provide topographical cues for rapid process extension suggest that it may be an appropriate device design for the bridging of experimental lesions of the peripheral nervous system. © 2015 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.
NASTRAN computer system level 12.1
NASA Technical Reports Server (NTRS)
Butler, T. G.
1971-01-01
Program uses finite element displacement method for solving linear response of large, three-dimensional structures subject to static, dynamic, thermal, and random loadings. Program adapts to computers of different manufacture, permits up-dating and extention, allows interchange of output and input information between users, and is extensively documented.
Rotary culture enhances pre-osteoblast aggregation and mineralization.
Facer, S R; Zaharias, R S; Andracki, M E; Lafoon, J; Hunter, S K; Schneider, G B
2005-06-01
Three-dimensional environments have been shown to enhance cell aggregation and osteoblast differentiation. Thus, we hypothesized that three-dimensional (3D) growth environments would enhance the mineralization rate of human embryonic palatal mesenchymal (HEPM) pre-osteoblasts. The objective of this study was to investigate the potential use of rotary cell culture systems (RCCS) as a means to enhance the osteogenic potential of pre-osteoblast cells. HEPM cells were cultured in a RCCS to create 3D enviroments. Tissue culture plastic (2D) cultures served as our control. 3D environments promoted three-dimensional aggregate formations. Increased calcium and phosphorus deposition was significantly enhanced three- to 18-fold (P < 0.001) in 3D cultures as compared with 2D environments. 3D cultures mineralized in 1 wk as compared with the 2D cultures, which took 4 wks, a decrease in time of nearly 75%. In conclusion, our studies demonstrated that 3D environments enhanced osteoblast cell aggregation and mineralization.
Load evaluation of the da Vinci surgical system for transoral robotic surgery.
Fujiwara, Kazunori; Fukuhara, Takahiro; Niimi, Koji; Sato, Takahiro; Kitano, Hiroya
2015-12-01
Transoral robotic surgery, performed with the da Vinci surgical system (da Vinci), is a surgical approach for benign and malignant lesions of the oral cavity and laryngopharynx. It provides several unique advantages, which include a 3-dimensional magnified view and ability to see and work around curves or angles. However, the current da Vinci surgical system does not provide haptic feedback. This is problematic because the potential risks specific to the transoral use of the da Vinci include tooth injury, mucosal laceration, ocular injury and mandibular fracture. To assess the potential for intraoperative injuries, we measured the load of the endoscope and the instrument of the da Vinci Si surgical system. We pressed the endoscope and instrument of the da Vinci Si against Load cell six times each and measured the dynamic load and the time-to-maximum load. We also struck the da Vinci Si endoscope and instrument against the Load cell six times each and measured the impact load. The maximum dynamic load was 7.27 ± 1.31 kg for the endoscope and 1.90 ± 0.72 for the instrument. The corresponding time-to-maximum loads were 1.72 ± 0.22 and 1.29 ± 0.34 s, but the impact loads were significantly lower than the dynamic load. It remains possible that a major load is exerted on adjacent structures by continuous contact with the endoscope and instrument of da Vinci Si. However, there is a minor delay in reaching the maximum load. Careful monitoring by an on-site assistant may, therefore, help prevent contiguous injury.
NASA Technical Reports Server (NTRS)
Stein, M.; Stein, P. A.
1978-01-01
Approximate solutions for three nonlinear orthotropic plate problems are presented: (1) a thick plate attached to a pad having nonlinear material properties which, in turn, is attached to a substructure which is then deformed; (2) a long plate loaded in inplane longitudinal compression beyond its buckling load; and (3) a long plate loaded in inplane shear beyond its buckling load. For all three problems, the two dimensional plate equations are reduced to one dimensional equations in the y-direction by using a one dimensional trigonometric approximation in the x-direction. Each problem uses different trigonometric terms. Solutions are obtained using an existing algorithm for simultaneous, first order, nonlinear, ordinary differential equations subject to two point boundary conditions. Ordinary differential equations are derived to determine the variable coefficients of the trigonometric terms.
Three-dimensional cell culture models for investigating human viruses.
He, Bing; Chen, Guomin; Zeng, Yi
2016-10-01
Three-dimensional (3D) culture models are physiologically relevant, as they provide reproducible results, experimental flexibility and can be adapted for high-throughput experiments. Moreover, these models bridge the gap between traditional two-dimensional (2D) monolayer cultures and animal models. 3D culture systems have significantly advanced basic cell science and tissue engineering, especially in the fields of cell biology and physiology, stem cell research, regenerative medicine, cancer research, drug discovery, and gene and protein expression studies. In addition, 3D models can provide unique insight into bacteriology, virology, parasitology and host-pathogen interactions. This review summarizes and analyzes recent progress in human virological research with 3D cell culture models. We discuss viral growth, replication, proliferation, infection, virus-host interactions and antiviral drugs in 3D culture models.
Complex structures from patterned cell sheets
Misra, M.; Audoly, B.; Shvartsman, S. Y.
2017-01-01
The formation of three-dimensional structures from patterned epithelial sheets plays a key role in tissue morphogenesis. An important class of morphogenetic mechanisms relies on the spatio-temporal control of apical cell contractility, which can result in the localized bending of cell sheets and in-plane cell rearrangements. We have recently proposed a modified vertex model that can be used to systematically explore the connection between the two-dimensional patterns of cell properties and the emerging three-dimensional structures. Here we review the proposed modelling framework and illustrate it through the computational analysis of the vertex model that captures the salient features of the formation of the dorsal appendages during Drosophila oogenesis. This article is part of the themed issue ‘Systems morphodynamics: understanding the development of tissue hardware’. PMID:28348251
Server-based Approach to Web Visualization of Integrated Three-dimensional Brain Imaging Data
Poliakov, Andrew V.; Albright, Evan; Hinshaw, Kevin P.; Corina, David P.; Ojemann, George; Martin, Richard F.; Brinkley, James F.
2005-01-01
The authors describe a client-server approach to three-dimensional (3-D) visualization of neuroimaging data, which enables researchers to visualize, manipulate, and analyze large brain imaging datasets over the Internet. All computationally intensive tasks are done by a graphics server that loads and processes image volumes and 3-D models, renders 3-D scenes, and sends the renderings back to the client. The authors discuss the system architecture and implementation and give several examples of client applications that allow visualization and analysis of integrated language map data from single and multiple patients. PMID:15561787
Three-dimensional elastic-plastic finite-element analysis of fatigue crack propagation
NASA Technical Reports Server (NTRS)
Goglia, G. L.; Chermahini, R. G.
1985-01-01
Fatigue cracks are a major problem in designing structures subjected to cyclic loading. Cracks frequently occur in structures such as aircraft and spacecraft. The inspection intervals of many aircraft structures are based on crack-propagation lives. Therefore, improved prediction of propagation lives under flight-load conditions (variable-amplitude loading) are needed to provide more realistic design criteria for these structures. The main thrust was to develop a three-dimensional, nonlinear, elastic-plastic, finite element program capable of extending a crack and changing boundary conditions for the model under consideration. The finite-element model is composed of 8-noded (linear-strain) isoparametric elements. In the analysis, the material is assumed to be elastic-perfectly plastic. The cycle stress-strain curve for the material is shown Zienkiewicz's initial-stress method, von Mises's yield criterion, and Drucker's normality condition under small-strain assumptions are used to account for plasticity. The three-dimensional analysis is capable of extending the crack and changing boundary conditions under cyclic loading.
NASA Astrophysics Data System (ADS)
Fatemi, Javad
2011-05-01
The thermal protection system of the EXPERT re-entry vehicle is subjected to accelerations, vibrations, acoustic and shock loads during launch and aero-heating loads and aerodynamic forces during re-entry. To fully understand the structural and thermomechanical performances of the TPS, heat transfer analysis, thermal stress analysis, and thermal buckling analysis must be performed. This requires complex three-dimensional thermal and structural models of the entire TPS including the insulation and sensors. Finite element (FE) methods are employed to assess the thermal and structural response of the TPS to the mechanical and aerothermal loads. The FE analyses results are used for the design verification and design improvement of the EXPERT thermal protection system.
Chon, Brian H; Lee, Esther J; Jing, Liufang; Setton, Lori A; Chen, Jun
2013-10-02
Cell supplementation to the herniated or degenerated intervertebral disc (IVD) is a potential strategy to promote tissue regeneration and slow disc pathology. Human umbilical cord mesenchymal stromal cells (HUCMSCs) - originating from the Wharton's jelly - remain an attractive candidate for such endeavors with their ability to differentiate into multiple lineages. Previously, mesenchymal stem cells (MSCs) have been studied as a potential source for disc tissue regeneration. However, no studies have demonstrated that MSCs can regenerate matrix with unique characteristics matching that of immature nucleus pulposus (NP) tissues of the IVD. In our prior work, immature NP cells were found to express specific laminin isoforms and laminin-binding receptors that may serve as phenotypic markers for evaluating MSC differentiation to NP-like cells. The goal of this study is to evaluate these markers and matrix synthesis for HUCMSCs cultured in a laminin-rich pseudo-three-dimensional culture system. HUCMSCs were seeded on top of Transwell inserts pre-coated with Matrigel™, which contained mainly laminin-111. Cells were cultured under hypoxia environment with three differentiation conditions: NP differentiation media (containing 2.5% Matrigel™ solution to provide for a pseudo-three-dimensional laminin culture system) with no serum, or the same media supplemented with either insulin-like growth factor-1 (IGF-1) or transforming growth factor-β1 (TGF-β1). Cell clustering behavior, matrix production and the expression of NP-specific laminin and laminin-receptors were evaluated at days 1, 7, 13 and 21 of culture. Data show that a pseudo-three-dimensional culture condition (laminin-1 rich) promoted HUCMSC differentiation under no serum conditions. Starting at day 1, HUCMSCs demonstrated a cell clustering morphology similar to that of immature NP cells in situ and that observed for primary immature NP cells within the similar laminin-rich culture system (prior study). Differentiated HUCMSCs under all conditions were found to contain glycosaminoglycan, expressed extracellular matrix proteins of collagen II and laminin α5, and laminin receptors (integrin α3 and β4 subunits). However, neither growth factor treatment generated distinct differences in NP-like phenotype for HUCMSC as compared with no-serum conditions. HUCMSCs have the potential to differentiate into cells sharing features with immature NP cells in a laminin-rich culture environment and may be useful for IVD cellular therapy.
Wei, Bo; Yao, Qingqiang; Guo, Yang; Mao, Fengyong; Liu, Shuai; Xu, Yan; Wang, Liming
2015-08-01
The goal of this study was to investigate the chondrogenic potential of three-dimensional polycaprolactone-hydroxyapatite (PCL-HA) scaffolds loaded with bone marrow cells in vitro and the effect of PCL-HA scaffolds on osteochondral repair in vivo. Here, bone marrow was added to the prepared PCL-HA scaffolds and cultured in chondrogenic medium for 10 weeks. Osteochondral defects were created in the trochlear groove of 29 knees in 17 New Zealand white rabbits, which were then divided into four groups that underwent: implantation of PCL-HA scaffolds (left knee, n = 17; Group 1), microfracture (right knee, n = 6; Group 2), autologous osteochondral transplantation (right knee, n = 6; Group 3), and no treatment (right knee, n = 5; Control). Extracellular matrix produced by bone marrow cells covered the surface and filled the pores of PCL-HA scaffolds after 10 weeks in culture. Moreover, many cell-laden cartilage lacunae were observed, and cartilage matrix was concentrated in the PCL-HA scaffolds. After a 12-week repair period, Group 1 showed excellent vertical and lateral integration with host bone, but incomplete cartilage regeneration and matrix accumulation. An uneven surface of regenerated cartilage and reduced distribution of cartilage matrix were observed in Group 2. In addition, abnormal bone growth and unstable integration between repaired and host tissues were detected. For Group 3, the integration between transplanted and host cartilage was interrupted. Our findings indicate that the PCL-HA scaffolds loaded with bone marrow cells improved chondrogenesis in vitro and implantation of PCL-HA scaffolds for osteochondral repairenhanced integration with host bone. However, cartilage regeneration remained unsatisfactory. The addition of trophic factors or the use of precultured cell-PCL-HA constructs for accelerated osteochondral repair requires further investigation. © The Author(s) 2015.
Applications to car bodies - Generalized layout design of three-dimensional shells
NASA Technical Reports Server (NTRS)
Fukushima, Junichi; Suzuki, Katsuyuki; Kikuchi, Noboru
1993-01-01
We shall describe applications of the homogenization method, formulated in Part 1, to design layout of car bodies represented by three-dimensional shell structures based on a multi-loading optimization.
Static stability of a three-dimensional space truss. M.S. Thesis - Case Western Reserve Univ., 1994
NASA Technical Reports Server (NTRS)
Shaker, John F.
1995-01-01
In order to deploy large flexible space structures it is necessary to develop support systems that are strong and lightweight. The most recent example of this aerospace design need is vividly evident in the space station solar array assembly. In order to accommodate both weight limitations and strength performance criteria, ABLE Engineering has developed the Folding Articulating Square Truss (FASTMast) support structure. The FASTMast is a space truss/mechanism hybrid that can provide system support while adhering to stringent packaging demands. However, due to its slender nature and anticipated loading, stability characterization is a critical part of the design process. Furthermore, the dire consequences surely to result from a catastrophic instability quickly provide the motivation for careful examination of this problem. The fundamental components of the space station solar array system are the (1) solar array blanket system, (2) FASTMast support structure, and (3) mast canister assembly. The FASTMast once fully deployed from the canister will provide support to the solar array blankets. A unique feature of this structure is that the system responds linearly within a certain range of operating loads and nonlinearly when that range is exceeded. The source of nonlinear behavior in this case is due to a changing stiffness state resulting from an inability of diagonal members to resist applied loads. The principal objective of this study was to establish the failure modes involving instability of the FASTMast structure. Also of great interest during this effort was to establish a reliable analytical approach capable of effectively predicting critical values at which the mast becomes unstable. Due to the dual nature of structural response inherent to this problem, both linear and nonlinear analyses are required to characterize the mast in terms of stability. The approach employed herein is one that can be considered systematic in nature. The analysis begins with one and two-dimensional failure models of the system and its important components. From knowledge gained through preliminary analyses a foundation is developed for three-dimensional analyses of the FASTMast structure. The three-dimensional finite element (FE) analysis presented here involves a FASTMast system one-tenth the size of the actual flight unit. Although this study does not yield failure analysis results that apply directly to the flight article, it does establish a method by which the full-scale mast can be evaluated.
Static stability of a three-dimensional space truss
NASA Astrophysics Data System (ADS)
Shaker, John F.
1995-05-01
In order to deploy large flexible space structures it is necessary to develop support systems that are strong and lightweight. The most recent example of this aerospace design need is vividly evident in the space station solar array assembly. In order to accommodate both weight limitations and strength performance criteria, ABLE Engineering has developed the Folding Articulating Square Truss (FASTMast) support structure. The FASTMast is a space truss/mechanism hybrid that can provide system support while adhering to stringent packaging demands. However, due to its slender nature and anticipated loading, stability characterization is a critical part of the design process. Furthermore, the dire consequences surely to result from a catastrophic instability quickly provide the motivation for careful examination of this problem. The fundamental components of the space station solar array system are the (1) solar array blanket system, (2) FASTMast support structure, and (3) mast canister assembly. The FASTMast once fully deployed from the canister will provide support to the solar array blankets. A unique feature of this structure is that the system responds linearly within a certain range of operating loads and nonlinearly when that range is exceeded. The source of nonlinear behavior in this case is due to a changing stiffness state resulting from an inability of diagonal members to resist applied loads. The principal objective of this study was to establish the failure modes involving instability of the FASTMast structure. Also of great interest during this effort was to establish a reliable analytical approach capable of effectively predicting critical values at which the mast becomes unstable. Due to the dual nature of structural response inherent to this problem, both linear and nonlinear analyses are required to characterize the mast in terms of stability. The approach employed herein is one that can be considered systematic in nature. The analysis begins with one and two-dimensional failure models of the system and its important components. From knowledge gained through preliminary analyses a foundation is developed for three-dimensional analyses of the FASTMast structure. The three-dimensional finite element (FE) analysis presented here involves a FASTMast system one-tenth the size of the actual flight unit. Although this study does not yield failure analysis results that apply directly to the flight article, it does establish a method by which the full-scale mast can be evaluated.
Faghihi, Faezeh; Mirzaei, Esmaeil; Ai, Jafar; Lotfi, Abolfazl; Sayahpour, Forough Azam; Barough, Somayeh Ebrahimi; Joghataei, Mohammad Taghi
2016-04-01
Many people worldwide suffer from motor neuron-related disorders such as amyotrophic lateral sclerosis and spinal cord injuries. Recently, several attempts have been made to recruit stem cells to modulate disease progression in ALS and also regenerate spinal cord injuries. Chorion-derived mesenchymal stem cells (C-MSCs), used to be discarded as postpartum medically waste product, currently represent a class of cells with self renewal property and immunomodulatory capacity. These cells are able to differentiate into mesodermal and nonmesodermal lineages such as neural cells. On the other hand, gelatin, as a simply denatured collagen, is a suitable substrate for cell adhesion and differentiation. It has been shown that electrospinning of scaffolds into fibrous structure better resembles the physiological microenvironment in comparison with two-dimensional (2D) culture system. Since there is no report on potential of human chorion-derived MSCs to differentiate into motor neuron cells in two- and three-dimensional (3D) culture systems, we set out to determine the effect of retinoic acid (RA) and sonic hedgehog (Shh) on differentiation of human C-MSCs into motor neuron-like cells cultured on tissue culture plates (2D) and electrospun nanofibrous gelatin scaffold (3D).
Three-Dimensional Temperature Field Simulation for the Rotor of an Asynchronous Motor
ERIC Educational Resources Information Center
Wang, Yanwu; Fan, Chunli; Yang, Li; Sun, Fengrui
2010-01-01
A three-dimensional heat transfer model is built according to the rotor structure of an asynchronous motor, and three-dimensional temperature fields of the rotor under different working conditions, such as the unloaded, rated loaded and that with broken rotor bars, are studied based on the finite element numerical method and experiments. The…
Continuum modeling of neuronal cell under blast loading
Jérusalem, Antoine; Dao, Ming
2012-01-01
Traumatic brain injuries have recently been put under the spotlight as one of the most important causes of accidental brain dysfunctions. Significant experimental and modeling efforts are thus ongoing to study the associated biological, mechanical and physical mechanisms. In the field of cell mechanics, progresses are also being made at the experimental and modeling levels to better characterize many of the cell functions such as differentiation, growth, migration and death, among others. The work presented here aims at bridging both efforts by proposing a continuum model of neuronal cell submitted to blast loading. In this approach, cytoplasm, nucleus and membrane (plus cortex) are differentiated in a representative cell geometry, and different material constitutive models are adequately chosen for each one. The material parameters are calibrated against published experimental work of cell nanoindentation at multiple rates. The final cell model is ultimately subjected to blast loading within a complete fluid-structure interaction computational framework. The results are compared to the nanoindentation simulation and the specific effects of the blast wave on the pressure and shear levels at the interfaces are identified. As a conclusion, the presented model successfully captures some of the intrinsic intracellular phenomena occurring during its deformation under blast loading and potentially leading to cell damage. It suggests more particularly the localization of damage at the nucleus membrane similarly to what has already been observed at the overall cell membrane. This degree of damage is additionally predicted to be worsened by a longer blast positive phase duration. As a conclusion, the proposed model ultimately provides a new three dimensional computational tool to evaluate intracellular damage during blast loading. PMID:22562014
Geometric charges in theories of elasticity and plasticity
NASA Astrophysics Data System (ADS)
Moshe, Michael
The mechanics of many natural systems is governed by localized sources of stresses. Examples include ''plastic events'' that occur in amorphous solids under external stress, defects formation in crystalline material, and force-dipoles applied by cells adhered to an elastic substrate. Recent developments in a geometric formulation of elasticity theory paved the way for a unifying mathematical description of such singular sources of stress, as ''elastic charges''. In this talk I will review basic results in this emerging field, focusing on the geometry and mechanics of elastic charges in two-dimensional solid bodies. I will demonstrate the applicability of this new approach in three different problems: failure of an amorphous solid under load, mechanics of Kirigami, and wrinkle patterns in geometrically-incompatible elastic sheets.
Methodology for dynamic biaxial tension testing of pregnant uterine tissue.
Manoogian, Sarah; Mcnally, Craig; Calloway, Britt; Duma, Stefan
2007-01-01
Placental abruption accounts for 50% to 70% of fetal losses in motor vehicle crashes. Since automobile crashes are the leading cause of traumatic fetal injury mortality in the United States, research of this injury mechanism is important. Before research can adequately evaluate current and future restraint designs, a detailed model of the pregnant uterine tissues is necessary. The purpose of this study is to develop a methodology for testing the pregnant uterus in biaxial tension at a rate normally seen in a motor vehicle crash. Since the majority of previous biaxial work has established methods for quasi-static testing, this paper combines previous research and new methods to develop a custom designed system to strain the tissue at a dynamic rate. Load cells and optical markers are used for calculating stress strain curves of the perpendicular loading axes. Results for this methodology show images of a tissue specimen loaded and a finite verification of the optical strain measurement. The biaxial test system dynamically pulls the tissue to failure with synchronous motion of four tissue grips that are rigidly coupled to the tissue specimen. The test device models in situ loading conditions of the pregnant uterus and overcomes previous limitations of biaxial testing. A non-contact method of measuring strains combined with data reduction to resolve the stresses in two directions provides the information necessary to develop a three dimensional constitutive model of the material. Moreover, future research can apply this method to other soft tissues with similar in situ loading conditions.
Transcription upregulation via force-induced direct stretching of chromatin
NASA Astrophysics Data System (ADS)
Tajik, Arash; Zhang, Yuejin; Wei, Fuxiang; Sun, Jian; Jia, Qiong; Zhou, Wenwen; Singh, Rishi; Khanna, Nimish; Belmont, Andrew S.; Wang, Ning
2016-12-01
Mechanical forces play critical roles in the function of living cells. However, the underlying mechanisms of how forces influence nuclear events remain elusive. Here, we show that chromatin deformation as well as force-induced transcription of a green fluorescent protein (GFP)-tagged bacterial-chromosome dihydrofolate reductase (DHFR) transgene can be visualized in a living cell by using three-dimensional magnetic twisting cytometry to apply local stresses on the cell surface via an Arg-Gly-Asp-coated magnetic bead. Chromatin stretching depended on loading direction. DHFR transcription upregulation was sensitive to load direction and proportional to the magnitude of chromatin stretching. Disrupting filamentous actin or inhibiting actomyosin contraction abrogated or attenuated force-induced DHFR transcription, whereas activating endogenous contraction upregulated force-induced DHFR transcription. Our findings suggest that local stresses applied to integrins propagate from the tensed actin cytoskeleton to the LINC complex and then through lamina-chromatin interactions to directly stretch chromatin and upregulate transcription.
Influence of Thickness and Contact Surface Geometry of Condylar Stem of TMJ Implant on Its Stability
NASA Astrophysics Data System (ADS)
Arabshahi, Zohreh; Kashani, Jamal; Kadir, Mohammed Rafiq Abdul; Azari, Abbas
The aim of this study is to examine the effect thickness and contact surface geometry of condylar stem of TMJ implant on its stability in total reconstruction system and evaluate the micro strain resulted in bone at fixation screw holes in jaw bone embedded with eight different designs of temporomandibular joint implants. A three dimensional model of a lower mandible of an adult were developed from a Computed Tomography scan images. Eight different TMJ implant designs and fixation screws were modeled. Three dimensional finite element models of eight implanted mandibles were analyzed. The forces assigned to the masticatory muscles for incisal clenching were applied consisting of nine important muscular loads. In chosen loading condition, The results indicated that the anatomical curvature contact surface design of TMJ implant can moderately improve the stability and the strain resulted in fixation screw holes in thinner TMJ implant was diminished in comparison with other thicknesses.
Fiber pushout test: A three-dimensional finite element computational simulation
NASA Technical Reports Server (NTRS)
Mital, Subodh K.; Chamis, Christos C.
1990-01-01
A fiber pushthrough process was computationally simulated using three-dimensional finite element method. The interface material is replaced by an anisotropic material with greatly reduced shear modulus in order to simulate the fiber pushthrough process using a linear analysis. Such a procedure is easily implemented and is computationally very effective. It can be used to predict fiber pushthrough load for a composite system at any temperature. The average interface shear strength obtained from pushthrough load can easily be separated into its two components: one that comes from frictional stresses and the other that comes from chemical adhesion between fiber and the matrix and mechanical interlocking that develops due to shrinkage of the composite because of phase change during the processing. Step-by-step procedures are described to perform the computational simulation, to establish bounds on interfacial bond strength and to interpret interfacial bond quality.
NASA Technical Reports Server (NTRS)
Brown, Richard; Collier, Gary; Heckenlaible, Richard; Dougherty, Edward; Dolenz, James; Ross, Iain
2012-01-01
The ASCENT program solves the three-dimensional motion and attendant structural loading on a flexible vehicle incorporating, optionally, an active analog thrust control system, aerodynamic effects, and staging of multiple bodies. ASCENT solves the technical problems of loads, accelerations, and displacements of a flexible vehicle; staging of the upper stage from the lower stage; effects of thrust oscillations on the vehicle; a payload's relative motion; the effect of fluid sloshing on vehicle; and the effect of winds and gusts on the vehicle (on the ground or aloft) in a continuous analysis. The ATTACH ASCENT Loads program reads output from the ASCENT flexible body loads program, and calculates the approximate load indicators for the time interval under consideration. It calculates the load indicator values from pre-launch to the end of the first stage.
Three-dimensional Myoblast Aggregates--Effects of Modeled Microgravity
NASA Technical Reports Server (NTRS)
Byerly, Diane; Sognier, M. A.; Marquette, M. L.
2006-01-01
The overall objective of these studies is to elucidate the molecular and cellular alterations that contribute to muscle atrophy in astronauts caused by exposure to microgravity conditions in space. To accomplish this, a three-dimensional model test system was developed using mouse myoblast cells (C2C12). Myoblast cells were grown as three-dimensional aggregates (without scaffolding or other solid support structures) in both modeled microgravity (Rotary Cell Culture System, Synthecon, Inc.) and at unit gravity in coated Petri dishes. Evaluation of H&E stained thin sections of the aggregates revealed the absence of any necrosis. Confocal microscopy evaluations of cells stained with the Live/Dead assay (Molecular Probes) confirmed that viable cells were present throughout the aggregates with an average of only three dead cells observed per aggregate. Preliminary results from gene array analysis (Affymetrix chip U74Av2) showed that approximately 14% of the genes were down regulated (decreased more than 3 fold) and 4% were upregulated in cells exposed to modeled microgravity for 12 hours compared to unit gravity controls. Additional studies using fluorescent phallacidin revealed a decrease in F-actin in the cells exposed to modeled microgravity compared to unit gravity. Myoblast cells grown as aggregates in modeled microgravity exhibited spontaneous differentiation into syncitia while no differentiation was seen in the unit gravity controls. These studies show that 1)the model test system developed is suitable for assessing cellular and molecular alterations in myoblasts; 2) gene expression alterations occur rapidly (within 12 hours) following exposure to modeled microgravity; and 3) modeled microgravity conditions stimulated myoblast cell differentiation. Achieving a greater understanding of the molecular alterations leading to muscle atrophy will eventually enable the development of cell-based countermeasures, which may be valuable for treatment of muscle diseases on Earth and future space explorations.
Finite state modeling of aeroelastic systems
NASA Technical Reports Server (NTRS)
Vepa, R.
1977-01-01
A general theory of finite state modeling of aerodynamic loads on thin airfoils and lifting surfaces performing completely arbitrary, small, time-dependent motions in an airstream is developed and presented. The nature of the behavior of the unsteady airloads in the frequency domain is explained, using as raw materials any of the unsteady linearized theories that have been mechanized for simple harmonic oscillations. Each desired aerodynamic transfer function is approximated by means of an appropriate Pade approximant, that is, a rational function of finite degree polynomials in the Laplace transform variable. The modeling technique is applied to several two dimensional and three dimensional airfoils. Circular, elliptic, rectangular and tapered planforms are considered as examples. Identical functions are also obtained for control surfaces for two and three dimensional airfoils.
Freefield vibrations due to dynamic loading on a tunnel embedded in a stratified medium
NASA Astrophysics Data System (ADS)
Clouteau, D.; Arnst, M.; Al-Hussaini, T. M.; Degrande, G.
2005-05-01
An efficient and modular numerical prediction model is developed to predict vibration and re-radiated noise in adjacent buildings from excitation due to metro trains in tunnels for both newly built and existing situations. The three-dimensional dynamic tunnel-soil interaction problem is solved with a subdomain formulation, using a finite element formulation for the tunnel and a boundary element method for the soil. The periodicity of the tunnel and the soil in the longitudinal direction is exploited using the Floquet transform, limiting the discretization effort to a single bounded reference cell. It is demonstrated in the paper how the boundary element method can efficiently be extended to deal with periodic media, reusing the available three-dimensional Green's tensors for layered media. The efficiency of the method is demonstrated with a numerical example, where the case of harmonic and transient point loading on the invert of a shallow cut-and-cover masonry tunnel in Paris is considered. The work described here was carried out under the auspices of the CONVURT project sponsored by the European Community.
Three-dimensional nano-biointerface as a new platform for guiding cell fate.
Liu, Xueli; Wang, Shutao
2014-04-21
Three-dimensional nano-biointerface has been emerging as an important topic for chemistry, nanotechnology, and life sciences in recent years. Understanding the exchanges of materials, signals, and energy at biological interfaces has inspired and helped the serial design of three-dimensional nano-biointerfaces. The intimate interactions between cells and nanostructures bring many novel properties, making three-dimensional nano-biointerfaces a powerful platform to guide cell fate in a controllable and accurate way. These advantages and capabilities endow three-dimensional nano-biointerfaces with an indispensable role in developing advanced biological science and technology. This tutorial review is mainly focused on the recent progress of three-dimensional nano-biointerfaces and highlights the new explorations and unique phenomena of three-dimensional nano-biointerfaces for cell-related fundamental studies and biomedical applications. Some basic bio-inspired principles for the design and creation of three-dimensional nano-biointerfaces are also delivered in this review. Current and further challenges of three-dimensional nano-biointerfaces are finally addressed and proposed.
Frequency response of pig intervertebral disc cells subjected to dynamic hydrostatic pressure.
Kasra, Mehran; Merryman, W David; Loveless, Kristen N; Goel, Vijay K; Martin, James D; Buckwalter, Joseph A
2006-10-01
The pathogenesis of vibration-induced disorders of intervertebral disc at the cellular level is largely unknown. Dynamic loads with frequencies close to that of the in vivo human spine resonant frequency (4-6 Hz) have a destructive effect, which may induce extracellular disc matrix (ECM) degradation. To investigate this issue, three-dimensional (3D) alginate cultures of normal pig intervertebral disc nucleus and inner annulus cells were tested under dynamic hydrostatic loading. Alginate cultures of each region were divided into six groups; five groups were exposed to cyclic hydrostatic pressures of frequencies 1, 3, 5, 8, and 10 Hz with the same amplitude (1 MPa), and group 6 was the control group (no loading). Cultures of different groups were loaded for 3 days (30 min daily) in a hydraulic chamber. Effects of loading frequency on disc collagen and protein metabolism were investigated by measuring 3H-proline-labeled proteins associated with the cells in the extracellular matrix and release of 3H-proline-labeled molecules into culture medium. The results indicated a poor synthesis rate and more degradation near the 5 Hz frequency. The repeatability of experiments was verified by performing two experiments with the same protocol. Both experiments indicated that a threshold frequency of around 5 Hz disrupted protein metabolism. Copyright (c) 2006 Orthopaedic Research Society.
Magnetic Tissue Engineering for Voice Rehabilitation - First Steps in a Promising Field.
Dürr, Stephan; Bohr, Christopher; Pöttler, Marina; Lyer, Stefan; Friedrich, Ralf Philipp; Tietze, Rainer; Döllinger, Michael; Alexiou, Christoph; Janko, Christina
2016-06-01
The voice is one of the most important instruments of communication between humans. It is the product of intact and well-working vocal folds. A defect of these structures causes dysphonia, associated with a clear reduction of quality of life. Tissue engineering of the vocal folds utilizing magnetic cell levitation after nanoparticle loading might be a technique to overcome this challenging problem. Vocal fold fibroblasts (VFFs) were isolated from rabbit larynges and cultured. For magnetization, cells were incubated with superparamagnetic iron oxide nanoparticles (SPION) and the loading efficiency was determined by Prussian blue staining. Biocompatibility was analyzed in flow cytometry by staining with annexin V-fluorescein isothiocyanate propidium iodide, 1,1',3,3,3',3'-hexamethylindodicarbo-cyanine iodide [DiIC1(5)] and propidium idodide-Triton X-100 to monitor phosphatidylserine exposure, plasma membrane integrity, mitochondrial membrane potential and DNA degradation. Isolated VFFs can be successfully loaded with SPION, and optimal iron loading associated with minimized cytotoxicity represents a balancing act in magnetic tissue engineering. Our data are a firm basis for the next steps of investigations. Magnetic tissue engineering using magnetic nanoparticle-loaded cells which form three-dimensional structures in a magnetic field will be a promising approach in the future. Copyright© 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.
A three-dimensional finite element model for biomechanical analysis of the hip.
Chen, Guang-Xing; Yang, Liu; Li, Kai; He, Rui; Yang, Bin; Zhan, Yan; Wang, Zhi-Jun; Yu, Bing-Nin; Jian, Zhe
2013-11-01
The objective of this study was to construct a three-dimensional (3D) finite element model of the hip. The images of the hip were obtained from Chinese visible human dataset. The hip model includes acetabular bone, cartilage, labrum, and bone. The cartilage of femoral head was constructed using the AutoCAD and Solidworks software. The hip model was imported into ABAQUS analysis system. The contact surface of the hip joint was meshed. To verify the model, the single leg peak force was loaded, and contact area of the cartilage and labrum of the hip and pressure distribution in these structures were observed. The constructed 3D hip model reflected the real hip anatomy. Further, this model reflected biomechanical behavior similar to previous studies. In conclusion, this 3D finite element hip model avoids the disadvantages of other construction methods, such as imprecision of cartilage construction and the absence of labrum. Further, it provides basic data critical for accurately modeling normal and abnormal loads, and the effects of abnormal loads on the hip.
Modelling the role of Tax expression in HTLV-I persistence in vivo.
Li, Michael Y; Lim, Aaron G
2011-12-01
Human T-lymphotropic virus type I (HTLV-I) is a persistent human retrovirus characterized by life-long infection and risk of developing HAM/TSP, a progressive neurological and inflammatory disease, and adult T-cell leukemia (ATL). Chronically infected individuals often harbor high proviral loads despite maintaining a persistently activated immune response. Based on a new hypothesis for the persistence of HTLV-I infection, a three-dimensional compartmental model is constructed that describes the dynamic interactions among latently infected target cells, target-cell activation, and immune responses to HTLV-I, with an emphasis on understanding the role of Tax expression in the persistence of HTLV-I.
A fuzzy-logic antiswing controller for three-dimensional overhead cranes.
Cho, Sung-Kun; Lee, Ho-Hoon
2002-04-01
In this paper, a new fuzzy antiswing control scheme is proposed for a three-dimensional overhead crane. The proposed control consists of a position servo control and a fuzzy-logic control. The position servo control is used to control crane position and rope length, and the fuzzy-logic control is used to suppress load swing. The proposed control guarantees not only prompt suppression of load swing but also accurate control of crane position and rope length for simultaneous travel, traverse, and hoisting motions of the crane. Furthermore, the proposed control provides practical gain tuning criteria for easy application. The effectiveness of the proposed control is shown by experiments with a three-dimensional prototype overhead crane.
Liu, An; Xue, Guang-huai; Sun, Miao; Shao, Hui-feng; Ma, Chi-yuan; Gao, Qing; Gou, Zhong-ru; Yan, Shi-gui; Liu, Yan-ming; He, Yong
2016-01-01
Desktop three-dimensional (3D) printers (D3DPs) have become a popular tool for fabricating personalized consumer products, favored for low cost, easy operation, and other advantageous qualities. This study focused on the potential for using D3DPs to successfully, rapidly, and economically print customized implants at medical clinics. An experiment was conducted on a D3DP-printed anterior cruciate ligament surgical implant using a rabbit model. A well-defined, orthogonal, porous PLA screw-like scaffold was printed, then coated with hydroxyapatite (HA) to improve its osteoconductivity. As an internal fixation as well as an ideal cell delivery system, the osteogenic scaffold loaded with mesenchymal stem cells (MSCs) were evaluated through both in vitro and in vivo tests to observe bone-ligament healing via cell therapy. The MSCs suspended in Pluronic F-127 hydrogel on PLA/HA screw-like scaffold showed the highest cell proliferation and osteogenesis in vitro. In vivo assessment of rabbit anterior cruciate ligament models for 4 and 12 weeks showed that the PLA/HA screw-like scaffold loaded with MSCs suspended in Pluronic F-127 hydrogel exhibited significant bone ingrowth and bone-graft interface formation within the bone tunnel. Overall, the results of this study demonstrate that fabricating surgical implants at the clinic (fab@clinic) with D3DPs can be feasible, effective, and economical. PMID:26875826
Santos, Jorge M; Camões, Sérgio P; Filipe, Elysse; Cipriano, Madalena; Barcia, Rita N; Filipe, Mariana; Teixeira, Mariana; Simões, Sandra; Gaspar, Manuela; Mosqueira, Diogo; Nascimento, Diana S; Pinto-do-Ó, Perpétua; Cruz, Pedro; Cruz, Helder; Castro, Matilde; Miranda, Joana P
2015-05-09
The secretion of trophic factors by mesenchymal stromal cells has gained increased interest given the benefits it may bring to the treatment of a variety of traumatic injuries such as skin wounds. Herein, we report on a three-dimensional culture-based method to improve the paracrine activity of a specific population of umbilical cord tissue-derived mesenchymal stromal cells (UCX®) towards the application of conditioned medium for the treatment of cutaneous wounds. A UCX® three-dimensional culture model was developed and characterized with respect to spheroid formation, cell phenotype and cell viability. The secretion by UCX® spheroids of extracellular matrix proteins and trophic factors involved in the wound-healing process was analysed. The skin regenerative potential of UCX® three-dimensional culture-derived conditioned medium (CM3D) was also assessed in vitro and in vivo against UCX® two-dimensional culture-derived conditioned medium (CM2D) using scratch and tubulogenesis assays and a rat wound splinting model, respectively. UCX® spheroids kept in our three-dimensional system remained viable and multipotent and secreted considerable amounts of vascular endothelial growth factor A, which was undetected in two-dimensional cultures, and higher amounts of matrix metalloproteinase-2, matrix metalloproteinase-9, hepatocyte growth factor, transforming growth factor β1, granulocyte-colony stimulating factor, fibroblast growth factor 2 and interleukin-6, when compared to CM2D. Furthermore, CM3D significantly enhanced elastin production and migration of keratinocytes and fibroblasts in vitro. In turn, tubulogenesis assays revealed increased capillary maturation in the presence of CM3D, as seen by a significant increase in capillary thickness and length when compared to CM2D, and increased branching points and capillary number when compared to basal medium. Finally, CM3D-treated wounds presented signs of faster and better resolution when compared to untreated and CM2D-treated wounds in vivo. Although CM2D proved to be beneficial, CM3D-treated wounds revealed a completely regenerated tissue by day 14 after excisions, with a more mature vascular system already showing glands and hair follicles. This work unravels an important alternative to the use of cells in the final formulation of advanced therapy medicinal products by providing a proof of concept that a reproducible system for the production of UCX®-conditioned medium can be used to prime a secretome for eventual clinical applications.
Rezende, Carlos Eduardo Edwards; Chase-Diaz, Melody; Costa, Max Doria; Albarracin, Max Laurent; Paschoeto, Gabriela; Sousa, Edson Antonio Capello; Rubo, José Henrique; Borges, Ana Flávia Sanches
2015-10-01
This study aimed to analyze the stress distribution in single implant system and to evaluate the compatibility of an in vitro model with finite element (FE) model. The in vitro model consisted of Brånemark implant; multiunit set abutment of 5 mm height; metal-ceramic screw-retained crown, and polyurethane simulating the bone. Deformations were recorded in the peri-implant region in the mesial and distal aspects, after an axial 300 N load application at the center of the occlusal aspect of the crown, using strain gauges. This in vitro model was scanned with micro CT to design a three-dimensional FE model and the strains in the peri-implant bone region were registered to check the compatibility between both models. The FE model was used to evaluate stress distribution in different parts of the system. The values obtained from the in vitro model (20-587 με) and the finite element analysis (81-588 με) showed agreement among them. The highest stresses because of axial and oblique load, respectively were 5.83 and 40 MPa for the cortical bone, 55 and 1200 MPa for the implant, and 80 and 470 MPa for the abutment screw. The FE method proved to be effective for evaluating the deformation around single implant. Oblique loads lead to higher stress concentrations.
NASA Technical Reports Server (NTRS)
Taylor, Shawn C.
2011-01-01
A noncontacting, two-dimensional (2-D) laser inspection system has been designed and implemented to dimensionally profile thermal barriers being developed for space vehicle applications. In a vehicle as-installed state, thermal barriers are commonly compressed between load sensitive thermal protection system (TPS) panels to prevent hot gas ingestion through the panel interface during flight. Loads required to compress the thermal barriers are functions of their construction, as well as their dimensional characteristics relative to the gaps in which they are installed. Excessive loads during a mission could damage surrounding TPS panels and have catastrophic consequences. As such, accurate dimensional profiling of thermal barriers prior to use is important. Due to the compliant nature of the thermal barriers, traditional contact measurement techniques (e.g., calipers and micrometers) are subjective and introduce significant error and variability into collected dimensional data. Implementation of a laser inspection system significantly enhanced the method by which thermal barriers are dimensionally profiled, and improved the accuracy and repeatability of collected data. A statistical design of experiments study comparing laser inspection and manual caliper measurement techniques verified these findings.
Folic acid-modified soy protein nanoparticles for enhanced targeting and inhibitory.
Cheng, Xu; Wang, Xin; Cao, Zhipeng; Yao, Weijing; Wang, Jun; Tang, Rupei
2017-02-01
Soy protein isolate (SPI) was hydrolyzed by compound enzymes to give water soluble low molecular soy protein (SP). SP and folic acid (FA) modified SP was polymerized with N-3- acrylamidophenylboronic acid (APBA) monomer in aqueous solution to give SP nanoparticles (SP NPs) and FA modified nanoparticles (FA-SP NPs), respectively. These NPs display excellent stability in different conditions, and have a uniform spherical shape with a diameter around of 200nm. Doxorubicin (DOX) was then successfully loaded into SP and FA-SP NPs with a desirable loading content of 13.33% and 16.01%, respectively. The cellular uptake and cytotoxicity of DOX-loaded SP NPs and FA-SP NPs were investigated using the two-dimensional (2D) monolayer cell model and three-dimensional (3D) multicellular spheroids (MCs). In vivo, tumor accumulation and growth inhibitory were then examined using H22 tumor-bearing mice. All these results demonstrated that conjugation of FA can efficiently enhance SP-based NPs' tumor accumulation and antitumor effect. Copyright © 2016 Elsevier B.V. All rights reserved.
Theoretical Analysis of Novel Quasi-3D Microscopy of Cell Deformation
Qiu, Jun; Baik, Andrew D.; Lu, X. Lucas; Hillman, Elizabeth M. C.; Zhuang, Zhuo; Guo, X. Edward
2012-01-01
A novel quasi-three-dimensional (quasi-3D) microscopy technique has been developed to enable visualization of a cell under dynamic loading in two orthogonal planes simultaneously. The three-dimensional (3D) dynamics of the mechanical behavior of a cell under fluid flow can be examined at a high temporal resolution. In this study, a numerical model of a fluorescently dyed cell was created in 3D space, and the cell was subjected to uniaxial deformation or unidirectional fluid shear flow via finite element analysis (FEA). Therefore, the intracellular deformation in the simulated cells was exactly prescribed. Two-dimensional fluorescent images simulating the quasi-3D technique were created from the cell and its deformed states in 3D space using a point-spread function (PSF) and a convolution operation. These simulated original and deformed images were processed by a digital image correlation technique to calculate quasi-3D-based intracellular strains. The calculated strains were compared to the prescribed strains, thus providing a theoretical basis for the measurement of the accuracy of quasi-3D and wide-field microscopy-based intracellular strain measurements against the true 3D strains. The signal-to-noise ratio (SNR) of the simulated quasi-3D images was also modulated using additive Gaussian noise, and a minimum SNR of 12 was needed to recover the prescribed strains using digital image correlation. Our computational study demonstrated that quasi-3D strain measurements closely recovered the true 3D strains in uniform and fluid flow cellular strain states to within 5% strain error. PMID:22707985
Microstructural Analysis and Transport Resistances of Low-Platinum-Loaded PEFC Electrodes
DOE Office of Scientific and Technical Information (OSTI.GOV)
Cetinbas, Firat C.; Wang, Xiaohua; Ahluwalia, Rajesh K.
In this study, we present microstructural characterization for polymer electrolyte fuel cell (PEFC) cathodes with low platinum loadings (low-PGM). The characterization results are used to quantify the contribution of mass transport resistances to cell voltage losses observed in polarization curve data. Three-dimensional pore morphology and ionomer distribution are resolved using nano-scale X-ray computed tomography (nano-CT). Electrode structural properties are reported along with analysis of the impact of microstructure on the effective charge and reactant transport properties. These characterizations are incorporated with a two-dimensional multi-physics model that accounts for energy, charge, and mass transport along with the effect of liquid watermore » flooding. Defining a total mass transport resistance for the whole polarization curve, contributions of transport mechanisms are identified. Analysis of the experimental polarization curves at different operating pressures and temperatures indicates that the mass transport resistance in the cathode is dominated by the transport processes in the electrode. It is shown that flooding in the electrode is a major contributor to transport losses especially at elevated operating pressures while the pressure-independent resistance at the catalyst surface due to transport through the ionomer film plays a significant role, especially at low temperatures and low catalyst loading. In addition, by performing a parametric study for varying catalyst loadings, the importance of electrode roughness (i.e, electrochemically-active surface area/geometric electrode area) in determining the mass transport losses is highlighted.« less
Microstructural Analysis and Transport Resistances of Low-Platinum-Loaded PEFC Electrodes
Cetinbas, Firat C.; Wang, Xiaohua; Ahluwalia, Rajesh K.; ...
2017-12-09
In this study, we present microstructural characterization for polymer electrolyte fuel cell (PEFC) cathodes with low platinum loadings (low-PGM). The characterization results are used to quantify the contribution of mass transport resistances to cell voltage losses observed in polarization curve data. Three-dimensional pore morphology and ionomer distribution are resolved using nano-scale X-ray computed tomography (nano-CT). Electrode structural properties are reported along with analysis of the impact of microstructure on the effective charge and reactant transport properties. These characterizations are incorporated with a two-dimensional multi-physics model that accounts for energy, charge, and mass transport along with the effect of liquid watermore » flooding. Defining a total mass transport resistance for the whole polarization curve, contributions of transport mechanisms are identified. Analysis of the experimental polarization curves at different operating pressures and temperatures indicates that the mass transport resistance in the cathode is dominated by the transport processes in the electrode. It is shown that flooding in the electrode is a major contributor to transport losses especially at elevated operating pressures while the pressure-independent resistance at the catalyst surface due to transport through the ionomer film plays a significant role, especially at low temperatures and low catalyst loading. In addition, by performing a parametric study for varying catalyst loadings, the importance of electrode roughness (i.e, electrochemically-active surface area/geometric electrode area) in determining the mass transport losses is highlighted.« less
NASA Technical Reports Server (NTRS)
Ko, William L.
1994-01-01
The combined load (mechanical or thermal load) buckling equations were established for orthotropic rectangular sandwich panels under four different edge conditions by using the Rayleigh-Ritz method of minimizing the total potential energy of a structural system. Two-dimensional buckling interaction curves and three-dimensional buckling interaction surfaces were constructed for high-temperature honeycomb-core sandwich panels supported under four different edge conditions. The interaction surfaces provide overall comparison of the panel buckling strengths and the domains of symmetrical and antisymmetrical buckling associated with the different edge conditions. In addition, thermal buckling curves of these sandwich panels are presented. The thermal buckling conditions for the cases with and without thermal moments were found to be identical for the small deformation theory.
Puetzer, Jennifer; Williams, John; Gillies, Allison; Bernacki, Susan
2013-01-01
This study investigates the effects of cyclic hydrostatic pressure (CHP) on chondrogenic differentiation of human adipose-derived stem cells (hASCs) in three-dimensional (3-D) agarose constructs maintained in a complete growth medium without soluble chondrogenic inducing factors. hASCs were seeded in 2% agarose hydrogels and exposed to 7.5 MPa CHP for 4 h per day at a frequency of 1 Hz for up to 21 days. On days 0, 7, 14, and 21, the expression levels of collagen II, Sox9, aggrecan, and cartilage oligomeric matrix protein (COMP) were examined by real-time reverse transcriptase–polymerase chain reaction analysis. Gene expression analysis found collagen II mRNA expression in only the CHP-loaded construct at day 14 and at no other time during the study. CHP-loaded hASCs exhibited upregulated mRNA expression of Sox9, aggrecan, and COMP at day 7 relative to unloaded controls, suggesting that CHP initiated chondrogenic differentiation of hASCs in a manner similar to human bone marrow-derived mesenchymal stem cells (hMSC). By day 14, however, loaded hASC constructs exhibited significantly lower mRNA expression of the chondrogenic markers than unloaded controls. Additionally, by day 21, the samples exhibited little measurable mRNA expression at all, suggesting a decreased viability. Histological analysis validated the lack of mRNA expression at day 21 for both the loaded and unloaded control samples with a visible decrease in the cell number and change in morphology. A comparative study with hASCs and hMSCs further examined long-term cell viability in 3-D agarose constructs of both cell types. Decreased cell metabolic activity was observed throughout the 21-day experimental period in both the CHP-loaded and control constructs of both hMSCs and hASCs, suggesting a decrease in cell metabolic activity, alluding to a decrease in cell viability. This suggests that a 2% agarose hydrogel may not optimally support hASC or hMSC viability in a complete growth medium in the absence of soluble chondrogenic inducing factors over long culture durations. This is the first study to examine the ability of mechanical stimuli alone, in the absence of chondrogenic factors transforming growth factor beta (TGF-β)3, TGF-β1 and/or bone morphogenetic protein 6 (BMP6) to induce hASC chondrogenic differentiation. The findings of this study suggest that CHP initiates hASC chondrogenic differentiation, even in the absence of soluble chondrogenic inductive factors, confirming the importance of considering both mechanical stimuli and appropriate 3-D culture for cartilage tissue engineering using hASCs. PMID:22871265
NASA Technical Reports Server (NTRS)
Lee, Jong-Hun
1993-01-01
The basic governing equations for the second-order three-dimensional hypersonic thermal and chemical nonequilibrium boundary layer are derived by means of an order-of-magnitude analysis. A two-temperature concept is implemented into the system of boundary-layer equations by simplifying the rather complicated general three-temperature thermal gas model. The equations are written in a surface-oriented non-orthogonal curvilinear coordinate system, where two curvilinear coordinates are non-orthogonial and a third coordinate is normal to the surface. The equations are described with minimum use of tensor expressions arising from the coordinate transformation, to avoid unnecessary confusion for readers. The set of equations obtained will be suitable for the development of a three-dimensional nonequilibrium boundary-layer code. Such a code could be used to determine economically the aerodynamic/aerothermodynamic loads to the surfaces of hypersonic vehicles with general configurations. In addition, the basic equations for three-dimensional stagnation flow, of which solution is required as an initial value for space-marching integration of the boundary-layer equations, are given along with the boundary conditions, the boundary-layer parameters, and the inner-outer layer matching procedure. Expressions for the chemical reaction rates and the thermodynamic and transport properties in the thermal nonequilibrium environment are explicitly given.
NASA Technical Reports Server (NTRS)
Farrokh, Babak; Rahim, Nur Aida Abul; Segal, Ken; Fan, Terry; Jones, Justin; Hodges, Ken; Mashni, Noah; Garg, Naman; Sang, Alex
2013-01-01
Three distinct strain measurement methods (i.e., foil resistance strain gages, fiber optic strain sensors, and a three-dimensional digital image photogrammetry that gives full field strain and displacement measurements) were implemented to measure strains on the back and front surfaces of a longitudinally jointed curved test article subjected to edge-wise compression testing, at NASA Goddard Space Flight Center, according to ASTM C364. The pre-test finite element analysis (FEA) was conducted to assess ultimate failure load and predict strain distribution pattern throughout the test coupon. The predicted strain pattern contours were then utilized as guidelines for installing the strain measurement instrumentations. The foil resistance strain gages and fiber optic strain sensors were bonded on the specimen at locations with nearly the same analytically predicted strain values, and as close as possible to each other, so that, comparisons between the measured strains by strain gages and fiber optic sensors, as well as the three-dimensional digital image photogrammetric system are relevant. The test article was loaded to failure (at 167 kN), at the compressive strain value of 10,000 micro epsilon. As a part of this study, the validity of the measured strains by fiber optic sensors is examined against the foil resistance strain gages and the three-dimensional digital image photogrammetric data, and comprehensive comparisons are made with FEA predictions.
Cardoso, T C; Sakamoto, S S; Stockmann, D; Souza, T F B; Ferreira, H L; Gameiro, R; Vieira, F V; Louzada, M J Q; Andrade, A L; Flores, E F
2017-06-01
In this study, derived complex carcinoma (CC) and simple carcinoma (SC) cell lines were established and cultured under two-dimensional (2D) and three-dimensional (3D) conditions. The 3D was performed in six-well AlgiMatrix™ (LifeTechnologies®, Carlsbad, CA, USA) scaffolds, resulting in spheroids sized 50-125 µm for CC and 175-200 µm for SC. Cell viability was demonstrated up to 14 days for both models. Epidermal growth factor receptor (EGFR) was expressed in CC and SC in both systems. However, higher mRNA and protein levels were observed in SC 2D and 3D systems when compared with CC (P < 0.005). The connective tissue modulators, metalloproteinases-1, -2, -9 and -13 (MMPs), relaxin receptors 1 and 2 (RXR1 and RXR2) and E-cadherin (CDH1) were quantitated. All were upregulated similarly when canine mammary tumour (CMT)-derived cell lines were cultured under 3D AlgiMatrix, except CDH1 that was downregulated (P < 0.005). These results are promising towards the used of 3D system to increase a high throughput in vitro canine tumour model. © 2016 John Wiley & Sons Ltd.
Structural weights analysis of advanced aerospace vehicles using finite element analysis
NASA Technical Reports Server (NTRS)
Bush, Lance B.; Lentz, Christopher A.; Rehder, John J.; Naftel, J. Chris; Cerro, Jeffrey A.
1989-01-01
A conceptual/preliminary level structural design system has been developed for structural integrity analysis and weight estimation of advanced space transportation vehicles. The system includes a three-dimensional interactive geometry modeler, a finite element pre- and post-processor, a finite element analyzer, and a structural sizing program. Inputs to the system include the geometry, surface temperature, material constants, construction methods, and aerodynamic and inertial loads. The results are a sized vehicle structure capable of withstanding the static loads incurred during assembly, transportation, operations, and missions, and a corresponding structural weight. An analysis of the Space Shuttle external tank is included in this paper as a validation and benchmark case of the system.
Microfluidic 3D cell culture: potential application for tissue-based bioassays
Li, XiuJun (James); Valadez, Alejandra V.; Zuo, Peng; Nie, Zhihong
2014-01-01
Current fundamental investigations of human biology and the development of therapeutic drugs, commonly rely on two-dimensional (2D) monolayer cell culture systems. However, 2D cell culture systems do not accurately recapitulate the structure, function, physiology of living tissues, as well as highly complex and dynamic three-dimensional (3D) environments in vivo. The microfluidic technology can provide micro-scale complex structures and well-controlled parameters to mimic the in vivo environment of cells. The combination of microfluidic technology with 3D cell culture offers great potential for in vivo-like tissue-based applications, such as the emerging organ-on-a-chip system. This article will review recent advances in microfluidic technology for 3D cell culture and their biological applications. PMID:22793034
Three-Dimensional Cell Cultures in Drug Discovery and Development
Fang, Ye; Eglen, Richard M.
2017-01-01
The past decades have witnessed significant efforts toward the development of three-dimensional (3D) cell cultures as systems that better mimic in vivo physiology. Today, 3D cell cultures are emerging, not only as a new tool in early drug discovery but also as potential therapeutics to treat disease. In this review, we assess leading 3D cell culture technologies and their impact on drug discovery, including spheroids, organoids, scaffolds, hydrogels, organs-on-chips, and 3D bioprinting. We also discuss the implementation of these technologies in compound identification, screening, and development, ranging from disease modeling to assessment of efficacy and safety profiles. PMID:28520521
Mechanical stress as a regulator of cell motility
NASA Astrophysics Data System (ADS)
Putelat, T.; Recho, P.; Truskinovsky, L.
2018-01-01
The motility of a cell can be triggered or inhibited not only by an applied force but also by a mechanically neutral force couple. This type of loading, represented by an applied stress and commonly interpreted as either squeezing or stretching, can originate from extrinsic interaction of a cell with its neighbors. To quantify the effect of applied stresses on cell motility we use an analytically transparent one-dimensional model accounting for active myosin contraction and induced actin turnover. We show that stretching can polarize static cells and initiate cell motility while squeezing can symmetrize and arrest moving cells. We show further that sufficiently strong squeezing can lead to the loss of cell integrity. The overall behavior of the system depends on the two dimensionless parameters characterizing internal driving (chemical activity) and external loading (applied stress). We construct a phase diagram in this parameter space distinguishing between static, motile, and collapsed states. The obtained results are relevant for the mechanical understanding of contact inhibition and the epithelial-to-mesenchymal transition.
Zhang, Yanzhuo; Wang, Hong; Li, Chuanjun; Sun, Baoxiang; Wang, Yu; Wang, Siling; Gao, Cunqiang
2014-04-01
A novel mesocellular carbon foam (MSU-FC) with a large pore size and a three-dimensional porous structure for the oral delivery of poorly water-soluble drugs was prepared. The goal of this study was to improve in vitro dissolution and in vivo absorption of celecoxib (CEB), a model drug, by means of novel carbon-based nanoparticles prepared from the MSU-FC matrix. The MSU-FC matrix was synthesized by an inverse replica templating method using mesocellular silica template. A solvent immersion/evaporation method was used to load the drug molecules. The drug-loaded nanoparticles were characterized for morphology, surface area, particle size, mesoporous structure, crystallinity, solubility and dissolution. The effect of MSU-FC on cell viability was measured using the MTT conversion assay. Furthermore, the oral bioavailability of CEB-loaded MSU-FC in fasted rats was compared with that of the marketed product. Our results demonstrate that CEB incorporation into the prepared MSU-FC resulted in an approximately 9-fold increase in aqueous solubility in comparison with crystalline CEB. MSU-FC produced accelerated immediate release of CEB in comparison with crystalline CEB (pure CEB powder or marketed formulation) and the drug-loaded conventional mesoporous carbon particles. The relative bioavailability of CEB for CEB-loaded MSU-FC was 172%. In addition, MSU-FC nanoparticles exhibited very low toxicity. The MSU-FC nanomatrix has been shown to be a promising drug delivery vehicle for improving the dissolution and biopharmaceutical characteristics of poorly water-soluble drugs.
ERIC Educational Resources Information Center
Lazarowitz, Reuven; Naim, Raphael
2014-01-01
The cell topic was taught to 9th-grade students in three modes of instruction: (a) students "hands-on," who constructed three-dimensional cell organelles and macromolecules during the learning process; (b) teacher demonstration of the three-dimensional model of the cell structures; and (c) teaching the cell topic with the regular…
Computational strategies for three-dimensional flow simulations on distributed computer systems
NASA Technical Reports Server (NTRS)
Sankar, Lakshmi N.; Weed, Richard A.
1995-01-01
This research effort is directed towards an examination of issues involved in porting large computational fluid dynamics codes in use within the industry to a distributed computing environment. This effort addresses strategies for implementing the distributed computing in a device independent fashion and load balancing. A flow solver called TEAM presently in use at Lockheed Aeronautical Systems Company was acquired to start this effort. The following tasks were completed: (1) The TEAM code was ported to a number of distributed computing platforms including a cluster of HP workstations located in the School of Aerospace Engineering at Georgia Tech; a cluster of DEC Alpha Workstations in the Graphics visualization lab located at Georgia Tech; a cluster of SGI workstations located at NASA Ames Research Center; and an IBM SP-2 system located at NASA ARC. (2) A number of communication strategies were implemented. Specifically, the manager-worker strategy and the worker-worker strategy were tested. (3) A variety of load balancing strategies were investigated. Specifically, the static load balancing, task queue balancing and the Crutchfield algorithm were coded and evaluated. (4) The classical explicit Runge-Kutta scheme in the TEAM solver was replaced with an LU implicit scheme. And (5) the implicit TEAM-PVM solver was extensively validated through studies of unsteady transonic flow over an F-5 wing, undergoing combined bending and torsional motion. These investigations are documented in extensive detail in the dissertation, 'Computational Strategies for Three-Dimensional Flow Simulations on Distributed Computing Systems', enclosed as an appendix.
Computational strategies for three-dimensional flow simulations on distributed computer systems
NASA Astrophysics Data System (ADS)
Sankar, Lakshmi N.; Weed, Richard A.
1995-08-01
This research effort is directed towards an examination of issues involved in porting large computational fluid dynamics codes in use within the industry to a distributed computing environment. This effort addresses strategies for implementing the distributed computing in a device independent fashion and load balancing. A flow solver called TEAM presently in use at Lockheed Aeronautical Systems Company was acquired to start this effort. The following tasks were completed: (1) The TEAM code was ported to a number of distributed computing platforms including a cluster of HP workstations located in the School of Aerospace Engineering at Georgia Tech; a cluster of DEC Alpha Workstations in the Graphics visualization lab located at Georgia Tech; a cluster of SGI workstations located at NASA Ames Research Center; and an IBM SP-2 system located at NASA ARC. (2) A number of communication strategies were implemented. Specifically, the manager-worker strategy and the worker-worker strategy were tested. (3) A variety of load balancing strategies were investigated. Specifically, the static load balancing, task queue balancing and the Crutchfield algorithm were coded and evaluated. (4) The classical explicit Runge-Kutta scheme in the TEAM solver was replaced with an LU implicit scheme. And (5) the implicit TEAM-PVM solver was extensively validated through studies of unsteady transonic flow over an F-5 wing, undergoing combined bending and torsional motion. These investigations are documented in extensive detail in the dissertation, 'Computational Strategies for Three-Dimensional Flow Simulations on Distributed Computing Systems', enclosed as an appendix.
Three-dimensional direct cell patterning in collagen hydrogels with near-infrared femtosecond laser
Hribar, Kolin C.; Meggs, Kyle; Liu, Justin; Zhu, Wei; Qu, Xin; Chen, Shaochen
2015-01-01
We report a methodology for three-dimensional (3D) cell patterning in a hydrogel in situ. Gold nanorods within a cell-encapsulating collagen hydrogel absorb a focused near-infrared femtosecond laser beam, locally denaturing the collagen and forming channels, into which cells migrate, proliferate, and align in 3D. Importantly, pattern resolution is tunable based on writing speed and laser power, and high cell viability (>90%) is achieved using higher writing speeds and lower laser intensities. Overall, this patterning technique presents a flexible direct-write method that is applicable in tissue engineering systems where 3D alignment is critical (such as vascular, neural, cardiac, and muscle tissue). PMID:26603915
On the impact of multi-axial stress states on trailing edge bondlines in wind turbine rotor blades
NASA Astrophysics Data System (ADS)
Noever Castelos, Pablo; Balzani, Claudio
2016-09-01
For a reliable design of wind turbine systems all of their components have to be designed to withstand the loads appearing in the turbine's lifetime. When performed in an integral manner this is called systems engineering, and is exceptionally important for components that have an impact on the entire wind turbine system, such as the rotor blade. Bondlines are crucial subcomponents of rotor blades, but they are not much recognized in the wind energy research community. However, a bondline failure can lead to the loss of a rotor blade, and potentially of the entire turbine, and is extraordinarily relevant to be treated with strong emphasis when designing a wind turbine. Modern wind turbine rotor blades with lengths of 80 m and more offer a degree of flexibility that has never been seen in wind energy technology before. Large deflections result in high strains in the adhesive connections, especially at the trailing edge. The latest edition of the DNV GL guideline from end of 2015 demands a three-dimensional stress analysis of bondlines, whereas before an isolated shear stress proof was sufficient. In order to quantify the lack of safety from older certification guidelines this paper studies the influence of multi-axial stress states on the ultimate and fatigue load resistance of trailing edge adhesive bonds. For this purpose, detailed finite element simulations of the IWES IWT-7.5-164 reference wind turbine blades are performed. Different yield criteria are evaluated for the prediction of failure and lifetime. The results show that the multi-axial stress state is governed by span-wise normal stresses. Those are evidently not captured in isolated shear stress proofs, yielding non-conservative estimates of lifetime and ultimate load resistance. This finding highlights the importance to include a three-dimensional stress state in the failure analysis of adhesive bonds in modern wind turbine rotor blades, and the necessity to perform a three-dimensional characterization of adhesive materials.
NASA Astrophysics Data System (ADS)
Muda, I.; Dharsuky, A.; Siregar, H. S.; Sadalia, I.
2017-03-01
This study examines the pattern of readiness dimensional accuracy of financial statements of local government in North Sumatra with a routine pattern of two (2) months after the fiscal year ends and patterns of at least 3 (three) months after the fiscal year ends. This type of research is explanatory survey with quantitative methods. The population and the sample used is of local government officials serving local government financial reports. Combined Analysis And Cross-Loadings Loadings are used with statistical tools WarpPLS. The results showed that there was a pattern that varies above dimensional accuracy of the financial statements of local government in North Sumatra.
Bioengineered humanized livers as better three-dimensional drug testing model system.
Vishwakarma, Sandeep Kumar; Bardia, Avinash; Lakkireddy, Chandrakala; Nagarapu, Raju; Habeeb, Md Aejaz; Khan, Aleem Ahmed
2018-01-27
To develop appropriate humanized three-dimensional ex-vivo model system for drug testing. Bioengineered humanized livers were developed in this study using human hepatic stem cells repopulation within the acellularized liver scaffolds which mimics with the natural organ anatomy and physiology. Six cytochrome P-450 probes were used to enable efficient identification of drug metabolism in bioengineered humanized livers. The drug metabolism study in bioengineered livers was evaluated to identify the absorption, distribution, metabolism, excretion and toxicity responses. The bioengineered humanized livers showed cellular and molecular characteristics of human livers. The bioengineered liver showed three-dimensional natural architecture with intact vasculature and extra-cellular matrix. Human hepatic cells were engrafted similar to the human liver. Drug metabolism studies provided a suitable platform alternative to available ex-vivo and in vivo models for identifying cellular and molecular dynamics of pharmacological drugs. The present study paves a way towards the development of suitable humanized preclinical model systems for pharmacological testing. This approach may reduce the cost and time duration of preclinical drug testing and further overcomes on the anatomical and physiological variations in xenogeneic systems.
Three-dimensional alginate spheroid culture system of murine osteosarcoma.
Akeda, Koji; Nishimura, Akinobu; Satonaka, Haruhiko; Shintani, Ken; Kusuzaki, Katsuyuki; Matsumine, Akihiko; Kasai, Yuichi; Masuda, Koichi; Uchida, Atsumasa
2009-11-01
Osteosarcoma (OS) is the most common primary malignant tumor of the bone and often forms pulmonary metastases, which are the most important prognostic factor. For further elucidation of the mechanism underlying the progression and metastasis of human OS, a culture system mimicking the microenvironment of the tumor in vivo is needed. We report a novel three-dimensional (3D) alginate spheroid culture system of murine osteosarcoma. Two different metastatic clones, the parental Dunn and its derivative line LM8, which has a higher metastatic potential to the lungs, were encapsulated in alginate beads to develop the 3D culture system. The beads containing murine OS cells were also transplanted into mice to determine their metastatic potential in vivo. In this culture system, murine OS cells encapsulated in alginate beads were able to grow in a 3D structure with cells detaching from the alginate environment. The number of detaching cells was higher in the LM8 cell line than the Dunn cell line. In the in vivo alginate bead transplantation model, the rate of pulmonary metastasis was higher with LM8 cells compared with that of Dunn cells. The cell characteristics and kinetics in this culture system closely reflect the original malignant potential of the cells in vivo.
Waiwijit, Uraiwan; Maturos, Thitima; Pakapongpan, Saithip; Phokharatkul, Ditsayut; Wisitsoraat, Anurat; Tuantranont, Adisorn
2016-08-01
Recently, three-dimensional graphene interconnected network has attracted great interest as a scaffold structure for tissue engineering due to its high biocompatibility, high electrical conductivity, high specific surface area and high porosity. However, free-standing three-dimensional graphene exhibits poor flexibility and stability due to ease of disintegration during processing. In this work, three-dimensional graphene is composited with polydimethylsiloxane to improve the structural flexibility and stability by a new simple two-step process comprising dip coating of polydimethylsiloxane on chemical vapor deposited graphene/Ni foam and wet etching of nickel foam. Structural characterizations confirmed an interconnected three-dimensional multi-layer graphene structure with thin polydimethylsiloxane scaffold. The composite was employed as a substrate for culture of L929 fibroblast cells and its cytocompatibility was evaluated by cell viability (Alamar blue assay), reactive oxygen species production and vinculin immunofluorescence imaging. The result revealed that cell viability on three-dimensional graphene/polydimethylsiloxane composite increased with increasing culture time and was slightly different from a polystyrene substrate (control). Moreover, cells cultured on three-dimensional graphene/polydimethylsiloxane composite generated less ROS than the control at culture times of 3-6 h. The results of immunofluorescence staining demonstrated that fibroblast cells expressed adhesion protein (vinculin) and adhered well on three-dimensional graphene/polydimethylsiloxane surface. Good cell adhesion could be attributed to suitable surface properties of three-dimensional graphene/polydimethylsiloxane with moderate contact angle and small negative zeta potential in culture solution. The results of electrochemical study by cyclic voltammetry showed that an oxidation current signal with no apparent peak was induced by fibroblast cells and the oxidation current at an oxidation potential of +0.9 V increased linearly with increasing cell number. Therefore, the three-dimensional graphene/polydimethylsiloxane composite exhibits high cytocompatibility and can potentially be used as a conductive substrate for cell-based electrochemical sensing. © The Author(s) 2016.
Hawkins, Rhoda J.; Poincloux, Renaud; Bénichou, Olivier; Piel, Matthieu; Chavrier, Philippe; Voituriez, Raphaël
2011-01-01
We present a model of cell motility generated by actomyosin contraction of the cell cortex. We identify, analytically, dynamical instabilities of the cortex and show that they yield steady-state cortical flows, which, in turn, can induce cell migration in three-dimensional environments. This mechanism relies on the regulation of contractility by myosin, whose transport is explicitly taken into account in the model. Theoretical predictions are compared to experimental data of tumor cells migrating in three-dimensional matrigel and suggest that this mechanism could be a general mode of cell migration in three-dimensional environments. PMID:21889440
Tajbakhsh, Jian; Gertych, Arkadiusz; Fagg, W. Samuel; Hatada, Seigo; Fair, Jeffrey H.
2011-01-01
The genome organization in pluripotent cells undergoing the first steps of differentiation is highly relevant to the reprogramming process in differentiation. Considering this fact, chromatin texture patterns that identify cells at the very early stage of lineage commitment could serve as valuable tools in the selection of optimal cell phenotypes for regenerative medicine applications. Here we report on the first-time use of high-resolution three-dimensional fluorescence imaging and comprehensive topological cell-by-cell analyses with a novel image-cytometrical approach towards the identification of in situ global nuclear DNA methylation patterns in early endodermal differentiation of mouse ES cells (up to day 6), and the correlations of these patterns with a set of putative markers for pluripotency and endodermal commitment, and the epithelial and mesenchymal character of cells. Utilizing this in vitro cell system as a model for assessing the relationship between differentiation and nuclear DNA methylation patterns, we found that differentiating cell populations display an increasing number of cells with a gain in DNA methylation load: first within their euchromatin, then extending into heterochromatic areas of the nucleus, which also results in significant changes of methylcytosine/global DNA codistribution patterns. We were also able to co-visualize and quantify the concomitant stochastic marker expression on a per-cell basis, for which we did not measure any correlation to methylcytosine loads or distribution patterns. We observe that the progression of global DNA methylation is not correlated with the standard transcription factors associated with endodermal development. Further studies are needed to determine whether the progression of global methylation could represent a useful signature of cellular differentiation. This concept of tracking epigenetic progression may prove useful in the selection of cell phenotypes for future regenerative medicine applications. PMID:21779341
DOE Office of Scientific and Technical Information (OSTI.GOV)
Chung, J.S.; Huttelmaier, H.P.; Cheng, B.R.
1995-12-31
For a heavy object falling on a free-span pipeline, this study assesses three-dimensional (3-D) pipe-span responses with the torsional ({theta}x-) coupling of a pipeline through the biaxial (y) bending responses. The static pipe-span equilibrium is achieved with its self-weight and buoyancy and the external torsional moment induced by the cross-flow (y-directional) current on the sagged pipe span. Load steps taken for 2 different sequences of applying static loads induced different pipe deformations, and the pipe twists in entirely different pattern. The two types of impact loads are applied in the vertical (z-) direction to excite the pipe span in itsmore » static equilibrium: (1) triangular impulse loading and (2) ramp loading. Boundary condition of the span supports is ``fixed-fixed`` at both ends in both displacement and rotation. 3-D coupled axial (x-), bending (y- and z-) and torsional ({theta}x-) responses, both state and dynamic, to the z-directional impact loadings, are modeled and analyzed by a nonlinear FEM method for a 16-in pipeline. The 3-D responses are compared with 2-D responses. The comparison shows significant torsional vibrations caused by the cross-flow current, especially for longer spans. The torsional ({theta}x-) coupling is very sensitive to the time-step size in achieving numerical stability and accuracy, particularly for the ramp loading and for a shorter span. For very large impact loads, the response frequencies differ from the fundamental frequencies of the span, exhibiting beatings and strong bending-to-axial and to-twist couplings. Also, the eigenvalues for the linear system are not necessarily the resonance frequencies for these nonlinear coupled responses.« less
Lee, H; Nguyen, T T; Kim, M; Jeong, J-H; Park, J-B
2018-05-31
Quercetin has been reported to exert many beneficial effects on the protection against various diseases, such as diabetes, cancer, and inflammation. The aim of this study is to evaluate the potential osteogenic differentiation ability of mesenchymal stem cells in the presence of quercetin. Quercetin-loaded poly(lactic-co-glycolic acid) microspheres were prepared using an electrospraying technique. Characterization of the microspheres was evaluated with a scanning electron microscope and release profile. Three-dimensional cell spheroids were fabricated using silicon elastomer-based concave microwells. Qualitative results of cellular viability were seen under a confocal microscope, and quantitative cellular viability was evaluated using the Cell Counting Kit-8 assay. The alkaline phosphatase activity and Alizarin Red S staining were performed. A quantitative real-time polymerase chain reaction and a western blot analysis were performed. Spheroids were well formed irrespective of quercetin concentration. Most of the cells in spheroids emitted green fluorescence, and the morphology was round without significant changes. The application of quercetin-loaded microspheres produced a significant increase in the alkaline phosphatase activity. The real-time polymerase chain reaction results showed a significant increase in Runx2, and western blot results showed higher expression of Runx2 protein expression. Biodegradable microspheres loaded with quercetin produced prolonged release profiles with increased mineralization. Microspheres loaded with quercetin can be used for the enhancement of osteoblastic differentiation in cell therapy. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Confinement and Structural Changes in Vertically Aligned Dust Structures
NASA Astrophysics Data System (ADS)
Hyde, Truell
2013-10-01
In physics, confinement is known to influence collective system behavior. Examples include coulomb crystal variants such as those formed from ions or dust particles (classical), electrons in quantum dots (quantum) and the structural changes observed in vertically aligned dust particle systems formed within a glass box placed on the lower electrode of a Gaseous Electronics Conference (GEC) rf reference cell. Recent experimental studies have expanded the above to include the biological domain by showing that the stability and dynamics of proteins confined through encapsulation and enzyme molecules placed in inorganic cavities such as those found in biosensors are also directly influenced by their confinement. In this paper, the self-assembly and subsequent collective behavior of structures formed from n, charged dust particles interacting with one another and located within a glass box placed on the lower, powered electrode of a GEC rf reference cell is discussed. Self-organized formation of vertically aligned one-dimensional chains, two-dimensional zigzag structures, and three-dimensional helical structures of triangular, quadrangular, pentagonal, hexagonal, and heptagonal symmetries are shown to occur. System evolution is shown to progress from one-dimensional chain structures, through a zigzag transition to a two-dimensional, spindle like structures, and then to various three-dimensional, helical structures exhibiting various symmetries. Stable configurations are shown to be strongly dependent upon system confinement. The critical conditions for structural transitions as well as the basic symmetry exhibited by the one-, two-, and three-dimensional structures that subsequently develop will be shown to be in good agreement with molecular dynamics simulations.
Lumped versus distributed thermoregulatory control: results from a three-dimensional dynamic model.
Werner, J; Buse, M; Foegen, A
1989-01-01
In this study we use a three-dimensional model of the human thermal system, the spatial grid of which is 0.5 ... 1.0 cm. The model is based on well-known physical heat-transfer equations, and all parameters of the passive system have definite physical values. According to the number of substantially different areas and organs, 54 spatially different values are attributed to each physical parameter. Compatibility of simulation and experiment was achieved solely on the basis of physical considerations and physiological basic data. The equations were solved using a modification of the alternating direction implicit method. On the basis of this complex description of the passive system close to reality, various lumped and distributed parameter control equations were tested for control of metabolic heat production, blood flow and sweat production. The simplest control equations delivering results on closed-loop control compatible with experimental evidence were determined. It was concluded that it is essential to take into account the spatial distribution of heat production, blood flow and sweat production, and that at least for control of shivering, distributed controller gains different from the pattern of distribution of muscle tissue are required. For sweat production this is not so obvious, so that for simulation of sweating control after homogeneous heat load a lumped parameter control may be justified. Based on these conclusions three-dimensional temperature profiles for cold and heat load and the dynamics for changes of the environmental conditions were computed. In view of the exact simulation of the passive system and the compatibility with experimentally attainable variables there is good evidence that those values extrapolated by the simulation are adequately determined. The model may be used both for further analysis of the real thermoregulatory mechanisms and for special applications in environmental and clinical health care.
Xu, Yue; Balooch, Guive; Chiou, Michael; Bekerman, Elena; Ritchie, Robert O.; Longaker, Michael T.
2009-01-01
Cartilage is an avascular tissue with only a limited potential to heal and chondrocytes in vitro have poor proliferative capacity. Recently, adipose-derived stromal cells (ASC) have demonstrated a great potential for application to tissue engineering due to their ability to differentiate into cartilage, bone, and fat. In this study, we have utilized a high density three-dimensional (3D) micromass model system of early chondrogenesis with ASC. The material properties of these micromasses showed a significant increase in dynamic and static elastic modulus during the early chondrogenic differentiation process. These data suggest that the 3D micromass culture system represents an in vitro model of early chondrogenesis with dynamic cell signaling interactions associated with the mechanical properties of chondrocyte differentiation. PMID:17543281
DOE Office of Scientific and Technical Information (OSTI.GOV)
Xu, Yue; Balooch, Guive; Chiou, Michael
2007-07-27
Cartilage is an avascular tissue with only a limited potential to heal and chondrocytes in vitro have poor proliferative capacity. Recently, adipose-derived stromal cells (ASC) have demonstrated a great potential for application to tissue engineering due to their ability to differentiate into cartilage, bone, and fat. In this study, we have utilized a high density three-dimensional (3D) micromass model system of early chondrogenesis with ASC. The material properties of these micromasses showed a significant increase in dynamic and static elastic modulus during the early chondrogenic differentiation process. These data suggest that the 3D micromass culture system represents an in vitromore » model of early chondrogenesis with dynamic cell signaling interactions associated with the mechanical properties of chondrocyte differentiation.« less
Systems Imaging of the Immune Synapse.
Ambler, Rachel; Ruan, Xiangtao; Murphy, Robert F; Wülfing, Christoph
2017-01-01
Three-dimensional live cell imaging of the interaction of T cells with antigen-presenting cells (APCs) visualizes the subcellular distributions of signaling intermediates during T cell activation at thousands of resolved positions within a cell. These information-rich maps of local protein concentrations are a valuable resource in understanding T cell signaling. Here, we describe a protocol for the efficient acquisition of such imaging data and their computational processing to create four-dimensional maps of local concentrations. This protocol allows quantitative analysis of T cell signaling as it occurs inside live cells with resolution in time and space across thousands of cells.
Skeletal muscle satellite cells cultured in simulated microgravity
NASA Technical Reports Server (NTRS)
Molnar, Greg; Hartzell, Charles R.; Schroedl, Nancy A.; Gonda, Steve R.
1993-01-01
Satellite cells are postnatal myoblasts responsible for providing additional nuclei to growing or regenerating muscle cells. Satellite cells retain the capacity to proliferate and differentiate in vitro and therefore provide a useful model to study postnatal muscle development. Most culture systems used to study postnatal muscle development are limited by the two-dimensional (2-D) confines of the culture dish. Limiting proliferation and differentiation of satellite cells in 2-D could potentially limit cell-cell contacts important for developing the level of organization in skeletal muscle obtained in vivo. Culturing satellite cells on microcarrier beads suspended in the High-Aspect-Ratio-Vessel (HARV) designed by NASA provides a low shear, three-dimensional (3-D) environment to study muscle development. Primary cultures established from anterior tibialis muscles of growing rats (approximately 200 gm) were used for all studies and were composed of greater than 75 % satellite cells. Different inoculation densities did not affect the proliferative potential of satellite cells in the HARV. Plating efficiency, proliferation, and glucose utilization were compared between 2-D flat culture and 3-D HARV culture. Plating efficiency (cells attached - cells plated x 100) was similar between the two culture systems. Proliferation was reduced in HARV cultures and this reduction was apparent for both satellite cells and non-satellite cells. Furthermore, reduction in proliferation within the HARV could not be attributed to reduced substrate availability since glucose levels in media from HARV and 2-D cell culture were similar. Morphologically, microcarrier beads within the HARVS were joined together by cells into three-dimensional aggregates composed of greater than 10 beads/aggregate. Aggregation of beads did not occur in the absence of cells. Myotubes were often seen on individual beads or spanning the surface of two beads. In summary, proliferation and differentiation of satellite cells on microcarrier beads within the HARV bioreactor results in a three dimensional level of organization that could provide a more suitable model to study postnatal muscle development.
Bueno, Ericka M.; Saeidi, Nima; Melotti, Suzanna
2009-01-01
The in vitro production of highly organized collagen fibrils by corneal keratocytes in a three-dimensional scaffold-free culture system presents a unique opportunity for the direct observation of organized matrix formation. The objective of this investigation was to develop such a culture system in a glass substrate (for optical accessibility) and to directly examine the effect of reducing serum and/or increasing insulin on the stratification and secretion of aligned matrix by fourth- to fifth-passage bovine corneal stromal keratocytes. Medium concentrations of 0%, 1%, or 10% fetal bovine serum and 0% or 1% insulin–transferrin–selenium were investigated. High-resolution differential interference contrast microscopy, quick-freeze/deep-etch, and conventional transmission electron microscopy were used to monitor the evolution, morphology, and ultrastructure of the cell–matrix constructs. In a medium containing 1% each of serum and insulin–transferrin–selenium, stromal cells stratified and secreted abundant and locally aligned matrix, generating the thickest cell–matrix constructs (allowing handling with forceps). The results of this study have the potential to significantly advance the field of developmental functional engineering of load-bearing tissues by (i) elucidating cues that modulate in vitro cell secretion of organized matrix and (ii) establishing an optically accessible cell culture system for investigating the mechanism of cell secretion of aligned collagen fibrils. PMID:19480568
Hosoda, Koh; Shimizu, Masahiro; Ikemoto, Shuhei; Nagura, Takeo; Seki, Hiroyuki; Kitashiro, Masateru; Imanishi, Nobuaki; Aiso, Sadakazu; Jinzaki, Masahiro; Ogihara, Naomichi
2017-01-01
The anatomical design of the human foot is considered to facilitate generation of bipedal walking. However, how the morphology and structure of the human foot actually contribute to generation of bipedal walking remains unclear. In the present study, we investigated the three-dimensional kinematics of the foot bones under a weight-bearing condition using cadaver specimens, to characterize the innate mobility of the human foot inherently prescribed in its morphology and structure. Five cadaver feet were axially loaded up to 588 N (60 kgf), and radiographic images were captured using a biplane X-ray fluoroscopy system. The present study demonstrated that the talus is medioinferiorly translated and internally rotated as the calcaneus is everted owing to axial loading, causing internal rotation of the tibia and flattening of the medial longitudinal arch in the foot. Furthermore, as the talus is internally rotated, the talar head moves medially with respect to the navicular, inducing external rotation of the navicular and metatarsals. Under axial loading, the cuboid is everted simultaneously with the calcaneus owing to the osseous locking mechanism in the calcaneocuboid joint. Such detailed descriptions about the innate mobility of the human foot will contribute to clarifying functional adaptation and pathogenic mechanisms of the human foot. PMID:29134100
A Relationship Between Constraint and the Critical Crack Tip Opening Angle
NASA Technical Reports Server (NTRS)
Johnston, William M.; James, Mark A.
2009-01-01
Of the various approaches used to model and predict fracture, the Crack Tip Opening Angle (CTOA) fracture criterion has been successfully used for a wide range of two-dimensional thin-sheet and thin plate applications. As thicker structure is considered, modeling the full three-dimensional fracture process will become essential. This paper investigates relationships between the local CTOA evaluated along a three-dimensional crack front and the corresponding local constraint. Previously reported tunneling crack front shapes were measured during fracture by pausing each test and fatigue cycling the specimens to mark the crack surface. Finite element analyses were run to model the tunneling shape during fracture, with the analysis loading conditions duplicating those tests. The results show an inverse relationship between the critical fracture value and constraint which is valid both before maximum load and after maximum load.
Atomic Force Microscopy Based Cell Shape Index
NASA Astrophysics Data System (ADS)
Adia-Nimuwa, Usienemfon; Mujdat Tiryaki, Volkan; Hartz, Steven; Xie, Kan; Ayres, Virginia
2013-03-01
Stellation is a measure of cell physiology and pathology for several cell groups including neural, liver and pancreatic cells. In the present work, we compare the results of a conventional two-dimensional shape index study of both atomic force microscopy (AFM) and fluorescent microscopy images with the results obtained using a new three-dimensional AFM-based shape index similar to sphericity index. The stellation of astrocytes is investigated on nanofibrillar scaffolds composed of electrospun polyamide nanofibers that has demonstrated promise for central nervous system (CNS) repair. Recent work by our group has given us the ability to clearly segment the cells from nanofibrillar scaffolds in AFM images. The clear-featured AFM images indicated that the astrocyte processes were longer than previously identified at 24h. It was furthermore shown that cell spreading could vary significantly as a function of environmental parameters, and that AFM images could record these variations. The new three-dimensional AFM-based shape index incorporates the new information: longer stellate processes and cell spreading. The support of NSF PHY-095776 is acknowledged.
A CFD analysis of blade row interactions within a high-speed axial compressor
NASA Astrophysics Data System (ADS)
Richman, Michael Scott
Aircraft engine design provides many technical and financial hurdles. In an effort to streamline the design process, save money, and improve reliability and performance, many manufacturers are relying on computational fluid dynamic simulations. An overarching goal of the design process for military aircraft engines is to reduce size and weight while maintaining (or improving) reliability. Designers often turn to the compression system to accomplish this goal. As pressure ratios increase and the number of compression stages decrease, many problems arise, for example stability and high cycle fatigue (HCF) become significant as individual stage loading is increased. CFD simulations have recently been employed to assist in the understanding of the aeroelastic problems. For accurate multistage blade row HCF prediction, it is imperative that advanced three-dimensional blade row unsteady aerodynamic interaction codes be validated with appropriate benchmark data. This research addresses this required validation process for TURBO, an advanced three-dimensional multi-blade row turbomachinery CFD code. The solution/prediction accuracy is characterized, identifying key flow field parameters driving the inlet guide vane (IGV) and stator response to the rotor generated forcing functions. The result is a quantified evaluation of the ability of TURBO to predict not only the fundamental flow field characteristics but the three dimensional blade loading.
NASA Astrophysics Data System (ADS)
Disotell, Kevin J.; Nikoueeyan, Pourya; Naughton, Jonathan W.; Gregory, James W.
2016-05-01
Recognizing the need for global surface measurement techniques to characterize the time-varying, three-dimensional loading encountered on rotating wind turbine blades, fast-responding pressure-sensitive paint (PSP) has been evaluated for resolving unsteady aerodynamic effects in incompressible flow. Results of a study aimed at demonstrating the laser-based, single-shot PSP technique on a low Reynolds number wind turbine airfoil in static and dynamic stall are reported. PSP was applied to the suction side of a Delft DU97-W-300 airfoil (maximum thickness-to-chord ratio of 30 %) at a chord Reynolds number of 225,000 in the University of Wyoming open-return wind tunnel. Static and dynamic stall behaviors are presented using instantaneous and phase-averaged global pressure maps. In particular, a three-dimensional pressure topology driven by a stall cell pattern is detected near the maximum lift condition on the steady airfoil. Trends in the PSP-measured pressure topology on the steady airfoil were confirmed using surface oil visualization. The dynamic stall case was characterized by a sinusoidal pitching motion with mean angle of 15.7°, amplitude of 11.2°, and reduced frequency of 0.106 based on semichord. PSP images were acquired at selected phase positions, capturing the breakdown of nominally two-dimensional flow near lift stall, development of post-stall suction near the trailing edge, and a highly three-dimensional topology as the flow reattaches. Structural patterns in the surface pressure topologies are considered from the analysis of the individual PSP snapshots, enabled by a laser-based excitation system that achieves sufficient signal-to-noise ratio in the single-shot images. The PSP results are found to be in general agreement with observations about the steady and unsteady stall characteristics expected for the airfoil.
Morabito, Caterina; Steimberg, Nathalie; Mazzoleni, Giovanna; Guarnieri, Simone; Fanò-Illic, Giorgio; Mariggiò, Maria A
2015-01-01
We propose a human-derived neuro-/glial cell three-dimensional in vitro model to investigate the effects of microgravity on cell-cell interactions. A rotary cell-culture system (RCCS) bioreactor was used to generate a modelled microgravity environment, and morphofunctional features of glial-like GL15 and neuronal-like SH-SY5Y cells in three-dimensional individual cultures (monotypic aggregates) and cocultures (heterotypic aggregates) were analysed. Cell survival was maintained within all cell aggregates over 2 weeks of culture. Moreover, compared to cells as traditional static monolayers, cell aggregates cultured under modelled microgravity showed increased expression of specific differentiation markers (e.g., GL15 cells: GFAP, S100B; SH-SY5Y cells: GAP43) and modulation of functional cell-cell interactions (e.g., N-CAM and Cx43 expression and localisation). In conclusion, this culture model opens a wide range of specific investigations at the molecular, biochemical, and morphological levels, and it represents an important tool for in vitro studies into dynamic interactions and responses of nervous system cell components to microgravity environmental conditions.
Mazzoleni, Giovanna; Fanò-Illic, Giorgio; Mariggiò, Maria A.
2015-01-01
We propose a human-derived neuro-/glial cell three-dimensional in vitro model to investigate the effects of microgravity on cell-cell interactions. A rotary cell-culture system (RCCS) bioreactor was used to generate a modelled microgravity environment, and morphofunctional features of glial-like GL15 and neuronal-like SH-SY5Y cells in three-dimensional individual cultures (monotypic aggregates) and cocultures (heterotypic aggregates) were analysed. Cell survival was maintained within all cell aggregates over 2 weeks of culture. Moreover, compared to cells as traditional static monolayers, cell aggregates cultured under modelled microgravity showed increased expression of specific differentiation markers (e.g., GL15 cells: GFAP, S100B; SH-SY5Y cells: GAP43) and modulation of functional cell-cell interactions (e.g., N-CAM and Cx43 expression and localisation). In conclusion, this culture model opens a wide range of specific investigations at the molecular, biochemical, and morphological levels, and it represents an important tool for in vitro studies into dynamic interactions and responses of nervous system cell components to microgravity environmental conditions. PMID:25654124
A versatile electrostatic trap with open optical access
NASA Astrophysics Data System (ADS)
Li, Sheng-Qiang; Yin, Jian-Ping
2018-04-01
A versatile electrostatic trap with open optical access for cold polar molecules in weak-field-seeking state is proposed in this paper. The trap is composed of a pair of disk electrodes and a hexapole. With the help of a finite element software, the spatial distribution of the electrostatic field is calculated. The results indicate that a three-dimensional closed electrostatic trap is formed. Taking ND3 molecules as an example, the dynamic process of loading and trapping is simulated. The results show that when the velocity of the molecular beam is 10 m/s and the loading time is 0.9964 ms, the maximum loading efficiency reaches 94.25% and the temperature of the trapped molecules reaches about 30.3 mK. A single well can be split into two wells, which is of significant importance to the precision measurement and interference of matter waves. This scheme, in addition, can be further miniaturized to construct one-dimensional, two-dimensional, and three-dimensional spatial electrostatic lattices.
An Elaborate Data Set Characterizing the Mechanical Response of the Foot
Erdemir, Ahmet; Sirimamilla, Pavana A.; Halloran, Jason P.; van den Bogert, Antonie J.
2010-01-01
Background Mechanical properties of the foot are responsible for its normal function and play a role in various clinical problems. Specifically, we are interested in quantification of foot mechanical properties to assist the development of computational models for movement analysis and detailed simulations of tissue deformation. Current available data are specific to a foot region and the loading scenarios are limited to a single direction. A data set that incorporates regional response, to quantify individual function of foot components, as well as overall response, to illustrate their combined operation, does not exist. Furthermore, combined three-dimensional loading scenarios while measuring the complete three-dimensional deformation response are lacking. When combined with an anatomical image data set, development of anatomically realistic and mechanically validated models becomes possible. Therefore, the goal of this study was to record and disseminate the mechanical response of a foot specimen, supported by imaging data. Method of Approach Robotic testing was conducted at the rear foot, forefoot, metatarsal heads, and the foot as a whole. Complex foot deformations were induced by single mode loading, e.g. compression, and combined loading, e.g. compression and shear. Small and large indenters were used for heel and metatarsal head loading; an elevated platform was utilized to isolate the rear foot and forefoot; and a full platform compressed the whole foot. Three-dimensional tool movements and reaction loads were recorded simultaneously. Computed tomography scans of the same specimen were collected for anatomical reconstruction a-priori. Results Three-dimensional mechanical response of the specimen was nonlinear and viscoelastic. A low stiffness region was observed starting with contact between the tool and foot regions, increasing with loading. Loading and unloading response portrayed hysteresis. Loading range ensured capturing the toe and linear regions of the load deformation curves for the dominant loading direction, with the rates approximating those of walking. Conclusion A large data set was successfully obtained to characterize the overall as well as regional mechanical response of an intact foot specimen under single and combined loads. Medical imaging complemented the mechanical testing data to establish the potential relationship between the anatomical architecture and mechanical response, and for further development of foot models that are mechanically realistic and anatomically consistent. This combined data set has been documented and disseminated in the public domain to promote future development in foot biomechanics. PMID:19725699
Effect of same-sided and cross-body load carriage on 3D back shape in young adults.
O'Shea, C; Bettany-Saltikov, J A; Warren, J G
2006-01-01
Regular carriage of heavy loads such as backpacks, satchels and mailbags results in a variety of acute medical problems and increased potential for back injury. There is a paucity of information about the specific changes in back posture that occur in response to asymmetrical loading. The purpose of this study was to examine the changes in back shape that occurred in response to asymmetrical load carriage, either on one shoulder (same-side) or across the body (cross-body), in healthy young adults. A convenience sample of 21 physiotherapy students randomly performed three trials (unloaded, same-side loaded, cross-body loaded) in standing with a 15% body load. The Microscribe 3DX digitiser (Immersion Group Ltd) recorded the three dimensional coordinates of 15 Key anatomical landmarks on the back in the three conditions. A one-way ANOVA with repeated measures and post-hoc tests was implemented to highlight statistical differences in the data collected (p<0.05). Significant differences were found in the x, y and z coordinates of the anatomical landmarks in the upper back between unloaded and loaded conditions. Results demonstrated significantly less impact on spinal posture from cross-body loading as compared to same-sided loading. This study confirms that there are significant three-dimensional changes in back shape in response to asymmetrical loading. Further work is needed to evaluate the optimal carriage type and maximal body load that results in the least spinal impact and injury potential in young adults.
Three dimensional modeling of rigid pavement : executive summary, February 1995.
DOT National Transportation Integrated Search
1995-02-17
A finite-element program has been developed to model the response of rigid pavement to both static loads and temperature changes. The program is fully three-dimensional and incorporates not only the common twenty-node brick element but also a thin in...
Three-dimensional modeling of rigid pavement : final report, September 1995.
DOT National Transportation Integrated Search
1995-02-17
A finite-element program has been developed to model the response of rigid pavement to both static loads and temperature changes. The program is fully three-dimensional and incorporates not only the common twenty-node brick element but also a thin in...
Manning, Phillip Lars; Lowe, Tristan; Withers, Philip J.
2017-01-01
This paper uses X-ray computed tomography to track the mechanical response of a vertebrate (Barnacle goose) long bone subjected to an axial compressive load, which is increased gradually until failure. A loading rig was mounted in an X-ray computed tomography system so that a time-lapse sequence of three-dimensional (3D) images of the bone’s internal (cancellous or trabecular) structure could be recorded during loading. Five distinct types of deformation mechanism were observed in the cancellous part of the bone. These were (i) cracking, (ii) thinning (iii) tearing of cell walls and struts, (iv) notch formation, (v) necking and (vi) buckling. The results highlight that bone experiences brittle (notch formation and cracking), ductile (thinning, tearing and necking) and elastic (buckling) modes of deformation. Progressive deformation, leading to cracking was studied in detail using digital image correlation. The resulting strain maps were consistent with mechanisms occurring at a finer-length scale. This paper is the first to capture time-lapse 3D images of a whole long bone subject to loading until failure. The results serve as a unique reference for researchers interested in how bone responds to loading. For those using computer modelling, the study not only provides qualitative information for verification and validation of their simulations but also highlights that constitutive models for bone need to take into account a number of different deformation mechanisms. PMID:28652932
Mustansar, Zartasha; McDonald, Samuel A; Sellers, William Irvin; Manning, Phillip Lars; Lowe, Tristan; Withers, Philip J; Margetts, Lee
2017-01-01
This paper uses X-ray computed tomography to track the mechanical response of a vertebrate (Barnacle goose) long bone subjected to an axial compressive load, which is increased gradually until failure. A loading rig was mounted in an X-ray computed tomography system so that a time-lapse sequence of three-dimensional (3D) images of the bone's internal (cancellous or trabecular) structure could be recorded during loading. Five distinct types of deformation mechanism were observed in the cancellous part of the bone. These were (i) cracking, (ii) thinning (iii) tearing of cell walls and struts, (iv) notch formation, (v) necking and (vi) buckling. The results highlight that bone experiences brittle (notch formation and cracking), ductile (thinning, tearing and necking) and elastic (buckling) modes of deformation. Progressive deformation, leading to cracking was studied in detail using digital image correlation. The resulting strain maps were consistent with mechanisms occurring at a finer-length scale. This paper is the first to capture time-lapse 3D images of a whole long bone subject to loading until failure. The results serve as a unique reference for researchers interested in how bone responds to loading. For those using computer modelling, the study not only provides qualitative information for verification and validation of their simulations but also highlights that constitutive models for bone need to take into account a number of different deformation mechanisms.
Dean, J C; Wilcox, C H; Daniels, A U; Goodwin, R R; Van Wagoner, E; Dunn, H K
1991-01-01
A new experimental technique for measuring generalized three-dimensional motion of vertebral bodies during cyclic loading in vitro is presented. The system consists of an orthogonal array of three lasers mounted rigidly to one vertebra, and a set of three mutually orthogonal charge-coupled devices mounted rigidly to an adjacent vertebra. Each laser strikes a corresponding charge-coupled device screen. The mathematical model of the system is reduced to a linear set of equations with consequent matrix algebra allowing fast real-time data reduction during cyclic movements of the spine. The range and accuracy of the system is well suited for studying thoracolumbar motion segments. Distinct advantages of the system include miniaturization of the components, the elimination of the need for mechanical linkages between the bodies, and a high degree of accuracy which is not dependent on viewing volume as found in photogrammetric systems. More generally, the spectrum of potential applications of systems of this type to the real-time measurement of the relative motion of two bodies is extremely broad.
Chip-based three-dimensional cell culture in perfused micro-bioreactors.
Gottwald, Eric; Lahni, Brigitte; Thiele, David; Giselbrecht, Stefan; Welle, Alexander; Weibezahn, Karl-Friedrich
2008-05-21
We have developed a chip-based cell culture system for the three-dimensional cultivation of cells. The chip is typically manufactured from non-biodegradable polymers, e.g., polycarbonate or polymethyl methacrylate by micro injection molding, micro hot embossing or micro thermo-forming. But, it can also be manufactured from bio-degradable polymers. Its overall dimensions are 0.7 1 x 20 x 20 x 0.7 1 mm (h x w x l). The main features of the chips used are either a grid of up to 1156 cubic micro-containers (cf-chip) each the size of 120-300 x 300 x 300 micron (h x w x l) or round recesses with diameters of 300 micron and a depth of 300 micron (r-chip). The scaffold can house 10 Mio. cells in a three-dimensional configuration. For an optimal nutrient and gas supply, the chip is inserted in a bioreactor housing. The bioreactor is part of a closed sterile circulation loop that, in the simplest configuration, is additionally comprised of a roller pump and a medium reservoir with a gas supply. The bioreactor can be run in perfusion, superfusion, or even a mixed operation mode. We have successfully cultivated cell lines as well as primary cells over periods of several weeks. For rat primary liver cells we could show a preservation of organotypic functions for more than 2 weeks. For hepatocellular carcinoma cell lines we could show the induction of liver specific genes not or only slightly expressed in standard monolayer culture. The system might also be useful as a stem cell cultivation system since first differentiation experiments with stem cell lines were promising.
Kido, Masamitsu; Ikoma, Kazuya; Hara, Yusuke; Imai, Kan; Maki, Masahiro; Ikeda, Takumi; Fujiwara, Hiroyoshi; Tokunaga, Daisaku; Inoue, Nozomu; Kubo, Toshikazu
2014-12-01
Insoles are frequently used in orthotic therapy as the standard conservative treatment for symptomatic flatfoot deformity to rebuild the arch and stabilize the foot. However, the effectiveness of therapeutic insoles remains unclear. In this study, we assessed the effectiveness of therapeutic insoles for flatfoot deformity using subject-based three-dimensional (3D) computed tomography (CT) models by evaluating the load responses of the bones in the medial longitudinal arch in vivo in 3D. We studied eight individuals (16 feet) with mild flatfoot deformity. CT scans were performed on both feet under non-loaded and full-body-loaded conditions, first with accessory insoles and then with therapeutic insoles under the same conditions. Three-dimensional CT models were constructed for the tibia and the tarsal and metatarsal bones of the medial longitudinal arch (i.e., first metatarsal bone, cuneiforms, navicular, talus, and calcaneus). The rotational angles between the tarsal bones were calculated under loading with accessory insoles or therapeutic insoles and compared. Compared with the accessory insoles, the therapeutic insoles significantly suppressed the eversion of the talocalcaneal joint. This is the first study to precisely verify the usefulness of therapeutic insoles (arch support and inner wedges) in vivo. Copyright © 2014 Elsevier Ltd. All rights reserved.
pH-triggered chitosan nanogels via an ortho ester-based linkage for efficient chemotherapy.
Yang, Guanqing; Wang, Xin; Fu, Shengxiang; Tang, Rupei; Wang, Jun
2017-09-15
We report on new types of chitosan-based nanogels via an ortho ester-based linkage, used as drug carriers for efficient chemotherapy. First, we synthesized a novel diacrylamide containing ortho ester (OEAM) as an acid-labile cross-linker. Subsequently, methacrylated succinyl-chitosan (MASCS) was prepared and polymerized with OEAM at different molar ratios to give a series of pH-triggered MASCS nanogels. Doxorubicin (DOX) as a model anticancer drug was loaded into MASCS nanogels with a loading content of 16.5%. As expected, with the incorporation of ortho ester linkages, these nanogels showed pH-triggered degradation and drug release at acidic pH values. In vitro cellular uptake shows that the DOX-loaded nanogels could be preferentially internalized by two-dimensional (2D) cells and three-dimensional (3D) multicellular spheroids (MCs), resulting in higher inhibition of the proliferation of tumor cells. In vivo biodistribution and anti-tumor effect were determined in H22 tumor-bearing mice, and the results demonstrate that the acid-labile MASCS nanogels can significantly prolong the blood circulation time of DOX and improve the accumulation in tumor areas, leading to higher therapeutic efficacy. We designed new pH-triggered chitosan nanogels via an ortho ester-based cross-linker for efficient drug-loading and chemotherapy. These drug-loaded nanogels exhibit excellent pH-triggered drug release behavior due to the degradation of ortho ester linkages in mildly acidic environments. In vitro and in vivo results demonstrate that the nanogels could be efficiently internalized by 2D cells and 3D-MCs, improve drug concentration in solid tumors, and lead to higher therapeutic efficacy. To the best of our knowledge, this is the first report on using an ortho ester-based cross-linker to prepare pH-triggered chitosan nanogels as tumor carriers, which may provide a potential route for improved safety and to increase the therapeutic efficacy of anticancer therapy. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
NASA Technical Reports Server (NTRS)
1993-01-01
Based on Johnson Space Flight Center's development of a rotating bioreactor cell culture apparatus for Space Shuttle medical research, Johnson Space Flight Center engineers who worked on the original project formed a company called Synthecon, with the intention of commercializing the bioreactor technology. Synthecon grows three dimensional tissues in the bioreactor. These are superior to previous two-dimensional tissue samples in the study of human cell growth. A refined version of the Johnson Space Center technology, Synthecon's Rotary Cell Culture System includes a cell culture chamber that rotates around a horizontal axis. The cells establish an orbit that approximates free fall through the liquid medium in the chamber. The technology has significant applications for cancer research and treatment as well as AIDS research.
Three-dimensional friction measurement during hip simulation
Braun, Steffen; Al-Salehi, Loay; Reinders, Joern; Mueller, Ulrike; Kretzer, J. Philippe
2017-01-01
Objectives Wear of total hip replacements has been the focus of many studies. However, frictional effects, such as high loading on intramodular connections or the interface to the bone, as well as friction associated squeaking have recently increased interest about the amount of friction that is generated during daily activities. The aim of this study was thus to establish and validate a three-dimensional friction setup under standardized conditions. Materials and methods A standard hip simulator was modified to allow for high precision measurements of small frictional effects in the hip during three-dimensional hip articulation. The setup was verified by an ideal hydrostatic bearing and validated with a static-load physical pendulum and an extension-flexion rotation with a dynamic load profile. Additionally, a pendulum model was proposed for screening measurement of frictional effects based on the damping behavior of the angular oscillation without the need for any force/moment transducer. Finally, three-dimensional friction measurements have been realized for ceramic-on-polyethylene bearings of three different sizes (28, 36 and 40 mm). Results A precision of less than 0.2 Nm during three-dimensional friction measurements was reported, while increased frictional torque (resultant as well as taper torque) was measured for larger head diameters. These effects have been confirmed by simple pendulum tests and the theoretical model. A comparison with current literature about friction measurements is presented. Conclusions This investigation of friction is able to provide more information about a field that has been dominated by the reduction of wear. It should be considered in future pre-clinical testing protocols given by international organizations of standardization. PMID:28886102
Three-dimensional friction measurement during hip simulation.
Sonntag, Robert; Braun, Steffen; Al-Salehi, Loay; Reinders, Joern; Mueller, Ulrike; Kretzer, J Philippe
2017-01-01
Wear of total hip replacements has been the focus of many studies. However, frictional effects, such as high loading on intramodular connections or the interface to the bone, as well as friction associated squeaking have recently increased interest about the amount of friction that is generated during daily activities. The aim of this study was thus to establish and validate a three-dimensional friction setup under standardized conditions. A standard hip simulator was modified to allow for high precision measurements of small frictional effects in the hip during three-dimensional hip articulation. The setup was verified by an ideal hydrostatic bearing and validated with a static-load physical pendulum and an extension-flexion rotation with a dynamic load profile. Additionally, a pendulum model was proposed for screening measurement of frictional effects based on the damping behavior of the angular oscillation without the need for any force/moment transducer. Finally, three-dimensional friction measurements have been realized for ceramic-on-polyethylene bearings of three different sizes (28, 36 and 40 mm). A precision of less than 0.2 Nm during three-dimensional friction measurements was reported, while increased frictional torque (resultant as well as taper torque) was measured for larger head diameters. These effects have been confirmed by simple pendulum tests and the theoretical model. A comparison with current literature about friction measurements is presented. This investigation of friction is able to provide more information about a field that has been dominated by the reduction of wear. It should be considered in future pre-clinical testing protocols given by international organizations of standardization.
Image matrix processor for fast multi-dimensional computations
Roberson, George P.; Skeate, Michael F.
1996-01-01
An apparatus for multi-dimensional computation which comprises a computation engine, including a plurality of processing modules. The processing modules are configured in parallel and compute respective contributions to a computed multi-dimensional image of respective two dimensional data sets. A high-speed, parallel access storage system is provided which stores the multi-dimensional data sets, and a switching circuit routes the data among the processing modules in the computation engine and the storage system. A data acquisition port receives the two dimensional data sets representing projections through an image, for reconstruction algorithms such as encountered in computerized tomography. The processing modules include a programmable local host, by which they may be configured to execute a plurality of different types of multi-dimensional algorithms. The processing modules thus include an image manipulation processor, which includes a source cache, a target cache, a coefficient table, and control software for executing image transformation routines using data in the source cache and the coefficient table and loading resulting data in the target cache. The local host processor operates to load the source cache with a two dimensional data set, loads the coefficient table, and transfers resulting data out of the target cache to the storage system, or to another destination.
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.
Three Dimensional Immobilization of Beta-Galactosidase on a Silicon Surface (Preprint)
2006-12-01
initial activity after 10 days at 24°C. The ability to generate three- dimensional structures with enhanced loading capacity for biosensing molecules...dimensional structures for biosensors (Charles et al. 2004). Silicon samples that had been washed but not activated with APTS did not retain any enzyme...preparation. The use of silica particles to build a 3-dimensional structure not only provides an increased capacity for the immobilization of β
Chen, Po-Yen; Dang, Xiangnan; Klug, Matthew T; Qi, Jifa; Dorval Courchesne, Noémie-Manuelle; Burpo, Fred J; Fang, Nicholas; Hammond, Paula T; Belcher, Angela M
2013-08-27
By genetically encoding affinity for inorganic materials into the capsid proteins of the M13 bacteriophage, the virus can act as a template for the synthesis of nanomaterial composites for use in various device applications. Herein, the M13 bacteriophage is employed to build a multifunctional and three-dimensional scaffold capable of improving both electron collection and light harvesting in dye-sensitized solar cells (DSSCs). This has been accomplished by binding gold nanoparticles (AuNPs) to the virus proteins and encapsulating the AuNP-virus complexes in TiO2 to produce a plasmon-enhanced and nanowire (NW)-based photoanode. The NW morphology exhibits an improved electron diffusion length compared to traditional nanoparticle-based DSSCs, and the AuNPs increase the light absorption of the dye-molecules through the phenomenon of localized surface plasmon resonance. Consequently, we report a virus-templated and plasmon-enhanced DSSC with an efficiency of 8.46%, which is achieved through optimizing both the NW morphology and the concentration of AuNPs loaded into the solar cells. In addition, we propose a theoretical model that predicts the experimentally observed trends of plasmon enhancement.
Álvarez-Murga, M; Perrillat, J P; Le Godec, Y; Bergame, F; Philippe, J; King, A; Guignot, N; Mezouar, M; Hodeau, J L
2017-01-01
X-ray tomography is a non-destructive three-dimensional imaging/microanalysis technique selective to a wide range of properties such as density, chemical composition, chemical states and crystallographic structure with extremely high sensitivity and spatial resolution. Here the development of in situ high-pressure high-temperature micro-tomography using a rotating module for the Paris-Edinburgh cell combined with synchrotron radiation is described. By rotating the sample chamber by 360°, the limited angular aperture of ordinary high-pressure cells is surmounted. Such a non-destructive high-resolution probe provides three-dimensional insight on the morphological and structural evolution of crystalline as well as amorphous phases during high pressure and temperature treatment. To demonstrate the potentials of this new experimental technique the compression behavior of a basalt glass is investigated by X-ray absorption tomography, and diffraction/scattering tomography imaging of the structural changes during the polymerization of C 60 molecules under pressure is performed. Small size and weight of the loading frame and rotating module means that this apparatus is portable, and can be readily installed on most synchrotron facilities to take advantage of the diversity of three-dimensional imaging techniques available at beamlines. This experimental breakthrough should open new ways for in situ imaging of materials under extreme pressure-temperature-stress conditions, impacting diverse areas in physics, chemistry, geology or materials sciences.
Genovese, Katia; Leeflang, Sander; Zadpoor, Amir A
2017-05-01
A custom-designed micro-digital image correlation system was used to track the evolution of the full-surface three-dimensional strain field of Ti6Al4V additively manufactured lattice samples under mechanical loading. The high-magnification capabilities of the method allowed to resolve the strain distribution down to the strut level and disclosed a highly heterogeneous mechanical response of the lattice structure with local strain concentrations well above the nominal global strain level. In particular, we quantified that strain heterogeneity appears at a very early stage of the deformation process and increases with load, showing a strain accumulation pattern with a clear correlation to the later onset of the fracture. The obtained results suggest that the unique opportunities offered by the proposed experimental method, in conjunction with analytical and computational models, could serve to provide novel important information for the rational design of additively manufactured porous biomaterials. Copyright © 2017 Elsevier Ltd. All rights reserved.
Guseva, Natalia V.; Dessus-Babus, Sophie; Moore, Cheryl G.; Whittimore, Judy D.; Wyrick, Priscilla B.
2007-01-01
In vitro studies of obligate intracellular chlamydia biology and pathogenesis are highly dependent on the use of experimental models and growth conditions that mimic the mucosal architecture and environment these pathogens encounter during natural infections. In this study, the growth of Chlamydia trachomatis genital serovar E was monitored in mouse fibroblast McCoy cells and compared to more relevant host human epithelial endometrium-derived HEC-1B and cervix-derived HeLa cells, seeded and polarized on collagen-coated microcarrier beads, using a three-dimensional culture system. Microscopy analysis of these cell lines prior to infection revealed morphological differences reminiscent of their in vivo architecture. Upon infection, early chlamydial inclusion distribution was uniform in McCoy cells but patchy in both epithelial cell lines. Although no difference in chlamydial attachment to or entry into the two genital epithelial cell lines was noted, active bacterial genome replication and transcription, as well as initial transformation of elementary bodies to reticulate bodies, were detected earlier in HEC-1B than in HeLa cells, suggesting a faster growth, which led to higher progeny counts and titers in HEC-1B cells upon completion of the developmental cycle. Chlamydial development in the less relevant McCoy cells was very similar to that in HeLa cells, although higher progeny counts were obtained. In conclusion, this three-dimensional bead culture system represents an improved model for harvesting large quantities of infectious chlamydia progeny from their more natural polarized epithelial host cells. PMID:17088348
Taghdisi, Seyed Mohammad; Danesh, Noor Mohammad; Ramezani, Mohammad; Yazdian-Robati, Rezvan; Abnous, Khalil
2018-05-07
Active targeting of nanostructures containing chemotherapeutic agents can improve cancer treatment. Here, a three-way junction pocket DNA nanostructure was developed for efficient doxorubicin (Dox) delivery into cancer cells. The three-way junction pocket DNA nanostructure is composed of three strands of AS1411 aptamer as both a therapeutic aptamer and nucleolin target, the potential biomarker of prostate (PC-3 cells) and breast (4T1 cells) cancers. The properties of the Dox-loaded three-way junction pocket DNA nanostructure were characterized and verified to have several advantages, including high serum stability and a pH-responsive property. Cellular uptake studies showed that the Dox-loaded DNA nanostructure was preferably internalized into target cancer cells (PC-3 and 4T1 cells). MTT cell viability assay demonstrated that the Dox-loaded DNA nanostructure had significantly higher cytotoxicity for PC-3 and 4T1 cells compared to that of nontarget cells (CHO cells, Chinese hamster ovary cell). The in vivo antitumor effect showed that the Dox-loaded DNA nanostructure was more effective in prohibition of the tumor growth compared to free Dox. These findings showed that the Dox-loaded three-way junction pocket DNA nanostructure could significantly reduce the cytotoxic effects of Dox against nontarget cells.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Cobb, Corie L.; Solberg, Scott E.
3-dimensional (3D) electrode architectures have been explored as a means to decouple power and energy trade-offs in thick battery electrodes. Limited work has been published which systematically examines the impact of these architectures at the pouch cell level. This paper conducts an analysis on the potential capacity gains that can be realized with thick co-extruded electrodes in a pouch cell. Moreover, our findings show that despite lower active material composition for each cathode layer, the effective gain in thickness and active material loading enables pouch cell capacity gains greater than 10% with a Lithium Nickel Manganese Cobalt Oxide (NMC) materialsmore » system.« less
[Study on sweat gland regeneration induced by microenvironment of three-dimensional bioprinting].
Yao, B; Xie, J F; Huang, S; Fu, X B
2017-01-20
Sweat glands are abundant in the body surface and essential for thermoregulation. Sweat glands fail to conduct self-repair in patients with large area of burn and trauma, and the body temperature of patients increases in hot climate, which may cause shock or even death. Now, co-culture system, reprogramming, and tissue engineering have made progresses in inducing sweat gland regeneration, but the inductive efficiency and duration need to be improved. Cellular microenvironment can regulate cell biological behavior, including cell migration and cell differentiation. This article reviews the studies of establishment of microenvironment in vitro by three-dimensional bioprinting technology to induce sweat gland regeneration.
Shijun, Xu; Junsheng, Mu; Jianqun, Zhang; Ping, Bo
2016-03-01
Identifying a suitable polymeric biomaterial for myocardial patch repair following myocardial infarction, cerebral infarction, and cartilage injury is essential. This study aimed to investigate the effect of the novel polymer material, poly3-hydroxybutyrate-co-3-hydroxyhexanoate, on the adhesion, proliferation, and differentiation of mouse-induced pluripotent stem cells in vitro. Mouse-induced pluripotent stem cells were isolated, expanded, and cultured on either two-dimensional or three-dimensional poly3-hydroxybutyrate-co-3-hydroxyhexanoate films (membranes were perforated to imitate three-dimensional space). Following attachment onto the films, mouse-induced pluripotent stem cell morphology was visualized using scanning electron microscopy. Cell vitality was detected using the Cell Counting Kit-8 assay and cell proliferation was observed using fluorescent 4',6-diamidino-2-phenylindole (DAPI) staining. Mouse-induced pluripotent stem cells were induced into cardiomyocytes by differentiation medium containing vitamin C. A control group in the absence of an inducer was included. Mouse-induced pluripotent stem cell survival and differentiation were observed using immunofluorescence and flow cytometry, respectively. Mouse-induced pluripotent stem cells growth, proliferation, and differentiation were observed on both two-dimensional and three-dimensional poly3-hydroxybutyrate-co-3-hydroxyhexanoate films. Vitamin C markedly improved the efficiency of mouse-induced pluripotent stem cells differentiation into cardiomyocytes on poly3-hydroxybutyrate-co-3-hydroxyhexanoate films. Three-dimensional culture was better at promoting mouse-induced pluripotent stem cell proliferation and differentiation compared with two-dimensional culture. © The Author(s) 2016.
ERIC Educational Resources Information Center
Zacharis, Georgios K.; Mikropoulos, Tassos Anastasios; Kalyvioti, Katerina
2016-01-01
Studies showed that two-dimensional (2D) and three-dimensional (3D) educational content contributes to learning. Although there were many studies with 3D stereoscopic learning environments, only a few studies reported on the differences between real, 2D, and 3D scenes, as far as cognitive load and attentional demands were concerned. We used…
[Establishment and application of mechanical strain loading system of multi-channel cells].
Li, Yongming; Wang, Hua; Zhang, Xiaodong; Tang, Lin
2012-02-01
Based on single-chip microcomputer, we have established a mechanical strain loading system with multi-channel to study the biological behavior of cultured cells in vitro under mechanical strain. We developed a multi-channel cell strain loading device controlled by single-chip microcomputer. We controlled the vacuum pump with vacuum chamber to make negative pressure changing periodically in the vacuum chamber. The tested cells were seeded on the surface of an elastic membrane mounted on the vacuum chamber, and could be strained or relaxed by cyclic pressure. Since the cells are attached to the surface of the membrane, they presumably experience the same deformation as that was applied to the membrane. The system was easy to carry and to operate, with deformation rate (1%-21%) and frequency (0-0. 5Hz) which could be adjusted correctly according to experimental requirement, and could compare different deformation rate of three channels at the same time. The system ran stably and completely achieved design aims, and provided a method to study the biological behavior of cultured cells attached to the surface of the elastic membrane under mechanical strain in vitro.
A unified theory for laminated plates
NASA Astrophysics Data System (ADS)
Guiamatsia Tafeuvoukeng, Irene
A literature survey on plate and beam theories show how the advent of the finite element method and the variational method circa 1940 have been a great stimulant for the research in this field. The initial thin plate formulation has been incrementally expanded to treat the isotropic thick plate, the anisotropic single layer, and then laminated plates. It appears however that current formulations still fall into one of two categories: (1) The formulation is tailored for a specific laminate and/or loading case; (2) or the formulation is too complicated to be of practical relevance. In this work a new unifying approach to laminated plate formulation is presented. All laminated plates, including sandwich panels, subjected to any surface load and with any boundary conditions are treated within a single model. In addition, the fundamental behavior of the plate as a two-dimensional structural element is explained. The novel idea is the introduction of fundamental state solutions, which are analytical far field stress and strain solutions of the laminated plate subjected to a set of hierarchical primary loads, the fundamental loads. These loads are carefully selected to form a basis of the load space, and corresponding solutions are superposed to obtain extremely accurate predictions of the three dimensional solution. six,y,z =aklx,y sikl z where i = 1,..., 6; 1=1,...,l max is a substate of the kth fundamental state k=1,2,3,... Typically, a fundamental state solution is expressed as a through-thickness function (z), while the amplitudes of each fundamental load are found from two dimensional finite element solution as a function of in-plane coordinates (x,y). Three major contributions are produced in this work: (1) A complete calibration of the plate as a two-dimensional structure is performed with pure bending and constant shear fundamental states. (2) There are four independent ways to apply a constant shear resultant on a plate, as opposed to one for a beam. This makes it impossible to define a unique 2 x 2 transverse shear stiffness matrix. Therefore the traditional problem of the shear correction factor loses all relevance. It is however shown that an explicit transverse constitutive relation can be obtained for isotropic-layered laminates or single-layers. (3) Higher accuracy, three-dimensional solutions are obtained using a two-dimensional finite element model with a complexity level (degrees of freedom) similar to the Reissner-Mindlin plate. The proof of concept is realized using Pagano solution for rectangular plates under sinusoidal load, for a sandwich panel. Additional comparisons are also performed for four and six-layer symmetric and antisymmetric laminates, between the new plate theory results and full three-dimensional finite element solutions.
Three-Dimensional, Transgenic Cell Models to Quantify Space Genotoxic Effects
NASA Technical Reports Server (NTRS)
Gonda, S. R.; Sognier, M. A.; Wu, H.; Pingerelli, P. L.; Glickman, B. W.; Dawson, David L. (Technical Monitor)
1999-01-01
The space environment contains radiation and chemical agents known to be mutagenic and carcinogenic to humans. Additionally, microgravity is a complicating factor that may modify or synergize induced genotoxic effects. Most in vitro models fail to use human cells (making risk extrapolation to humans more difficult), overlook the dynamic effect of tissue intercellular interactions on genotoxic damage, and lack the sensitivity required to measure low-dose effects. Currently a need exists for a model test system that simulates cellular interactions present in tissue, and can be used to quantify genotoxic damage induced by low levels of radiation and chemicals, and extrapolate assessed risk to humans. A state-of-the-art, three-dimensional, multicellular tissue equivalent cell culture model will be presented. It consists of mammalian cells genetically engineered to contain multiple copies of defined target genes for genotoxic assessment,. NASA-designed bioreactors were used to coculture mammalian cells into spheroids, The cells used were human mammary epithelial cells (H184135) and Stratagene's (Austin, Texas) Big Blue(TM) Rat 2 lambda fibroblasts. The fibroblasts were genetically engineered to contain -a high-density target gene for mutagenesis (60 copies of lacl/LacZ per cell). Tissue equivalent spheroids were routinely produced by inoculation of 2 to 7 X 10(exp 5) fibroblasts with Cytodex 3 beads (150 micrometers in diameter). at a 20:1 cell:bead ratio, into 50-ml HARV bioreactors (Synthecon, Inc.). Fibroblasts were cultured for 5 days, an equivalent number of epithelial cells added, and the fibroblast/epithelial cell coculture continued for 21 days. Three-dimensional spheroids with diameters ranging from 400 to 600 micrometers were obtained. Histological and immunohistochemical Characterization revealed i) both cell types present in the spheroids, with fibroblasts located primarily in the center, surrounded by epithelial cells; ii) synthesis of extracellular matrix; and iii,, mitotic cells located throughout the spheroids. Spheroidal integrity and cell viability were retained for the 30-day test period after removal of spheroids from the bioreactor. Potential utility of this three-dimensional, transgenic model for genotoxicity was initially assessed by exposure of spheroids to 0-2 Gy neon at dose rates of 0.3 to 1.5 Gy/min (National Institute of Radiological Sciences, Chiba, Japan). Quantification of mutation at the lacl gene revealed a linear dose response for mutation induction. Limited sequencing analysis of mutant clones revealed higher frequencies of deletions and multiple base sequence changes with increasing dose. These results suggest that our three-dimensional, transgenic model is applicable to a wide variety of studies involving the quantification, identification, and characterization of genotoxicity incurred in space and on Earth. This model uniquely allows investigation of the interaction of relevant factors, namely cell-to-cell interactions and the mechanistic interaction of microgravity with radiation insults and DNA repair. Using this three-dimensional model will allow us to obtain dual genotoxic information (i.e., mutation rate plus chromosome aberration data) from the same system so that one endpoint can be used to reference the other, thereby increasing the fidelity of the data set. Moreover, the tissue-equivalent nature of the three-dimensional model provides high confidence for relevance of risk assessment, i.e., the establishment of quality factors directly applicable to the microgravity environment.
Du, Wen-Cheng; Yin, Ya-Xia; Zeng, Xian-Xiang; Shi, Ji-Lei; Zhang, Shuai-Feng; Wan, Li-Jun; Guo, Yu-Guo
2016-02-17
An optimized nanocarbon-sulfur cathode material with ultrahigh sulfur loading of up to 90 wt % is realized in the form of sulfur nanolayer-coated three-dimensional (3D) conducting network. This 3D nanocarbon-sulfur network combines three different nanocarbons, as follows: zero-dimensional carbon nanoparticle, one-dimensional carbon nanotube, and two-dimensional graphene. This 3D nanocarbon-sulfur network is synthesized by using a method based on soluble chemistry of elemental sulfur and three types of nanocarbons in well-chosen solvents. The resultant sulfur-carbon material shows a high specific capacity of 1115 mA h g(-1) at 0.02C and good rate performance of 551 mA h g(-1) at 1C based on the mass of sulfur-carbon composite. Good battery performance can be attributed to the homogeneous compositing of sulfur with the 3D hierarchical hybrid nanocarbon networks at nanometer scale, which provides efficient multidimensional transport pathways for electrons and ions. Wet chemical method developed here provides an easy and cost-effective way to prepare sulfur-carbon cathode materials with high sulfur loading for application in high-energy Li-S batteries.
The Quantum Socket: Wiring for Superconducting Qubits - Part 1
NASA Astrophysics Data System (ADS)
McConkey, T. G.; Bejanin, J. H.; Rinehart, J. R.; Bateman, J. D.; Earnest, C. T.; McRae, C. H.; Rohanizadegan, Y.; Shiri, D.; Mariantoni, M.; Penava, B.; Breul, P.; Royak, S.; Zapatka, M.; Fowler, A. G.
Quantum systems with ten superconducting quantum bits (qubits) have been realized, making it possible to show basic quantum error correction (QEC) algorithms. However, a truly scalable architecture has not been developed yet. QEC requires a two-dimensional array of qubits, restricting any interconnection to external classical systems to the third axis. In this talk, we introduce an interconnect solution for solid-state qubits: The quantum socket. The quantum socket employs three-dimensional wires and makes it possible to connect classical electronics with quantum circuits more densely and accurately than methods based on wire bonding. The three-dimensional wires are based on spring-loaded pins engineered to insure compatibility with quantum computing applications. Extensive design work and machining was required, with focus on material quality to prevent magnetic impurities. Microwave simulations were undertaken to optimize the design, focusing on the interface between the micro-connector and an on-chip coplanar waveguide pad. Simulations revealed good performance from DC to 10 GHz and were later confirmed against experimental measurements.
Three-dimensional organization of dermal fibroblasts by macromass culture.
Deshpande, Manisha
2008-01-01
The three-dimensional organization of cells by high-cell-seeding-density culture, termed 'macromass culture', is described. By macromass culture, dermal fibroblasts can be made to organize themselves into a unified three-dimensional form without the aid of a scaffold, and macroscopic constructs, named macromasses, can be made wholly from cells. The sole factor causing three-dimensional organization is culture of cells at high cell seeding density per unit area. No scaffold or extraneous matrix is used for the generation of macromasses; they are of completely cellular origin. No other agents or external influences such as tissue-inducing chemicals, tissue-inducing growth factors, substratum with special properties, rotational culture, centrifugation etc. are employed for macromass formation, and all seeded cells become part of the cohesive construct. These three-dimensional constructs have the potential for use as in vitro tissue analogues, and a possible application for in vitro cytotoxicity testing is demonstrated.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Gatsonis, Nikolaos A.; Spirkin, Anton
2009-06-01
The mathematical formulation and computational implementation of a three-dimensional particle-in-cell methodology on unstructured Delaunay-Voronoi tetrahedral grids is presented. The method allows simulation of plasmas in complex domains and incorporates the duality of the Delaunay-Voronoi in all aspects of the particle-in-cell cycle. Charge assignment and field interpolation weighting schemes of zero- and first-order are formulated based on the theory of long-range constraints. Electric potential and fields are derived from a finite-volume formulation of Gauss' law using the Voronoi-Delaunay dual. Boundary conditions and the algorithms for injection, particle loading, particle motion, and particle tracking are implemented for unstructured Delaunay grids. Error andmore » sensitivity analysis examines the effects of particles/cell, grid scaling, and timestep on the numerical heating, the slowing-down time, and the deflection times. The problem of current collection by cylindrical Langmuir probes in collisionless plasmas is used for validation. Numerical results compare favorably with previous numerical and analytical solutions for a wide range of probe radius to Debye length ratios, probe potentials, and electron to ion temperature ratios. The versatility of the methodology is demonstrated with the simulation of a complex plasma microsensor, a directional micro-retarding potential analyzer that includes a low transparency micro-grid.« less
Apical polarity in three-dimensional culture systems: where to now?
DOE Office of Scientific and Technical Information (OSTI.GOV)
Inman, J.L.; Bissell, Mina
2010-01-21
Delineation of the mechanisms that establish and maintain the polarity of epithelial tissues is essential to understanding morphogenesis, tissue specificity and cancer. Three-dimensional culture assays provide a useful platform for dissecting these processes but, as discussed in a recent study in BMC Biology on the culture of mammary gland epithelial cells, multiple parameters that influence the model must be taken into account.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Jiguet Jiglaire, Carine, E-mail: carine.jiguet-jiglaire@univ-amu.fr; CRO2, UMR 911, Faculté de Médecine de la Timone, 27 boulevard Jean Moulin, 13284 Marseille Cedex; INSERM, U911, 13005 Marseille
Identification of new drugs and predicting drug response are major challenges in oncology, especially for brain tumors, because total surgical resection is difficult and radiation therapy or chemotherapy is often ineffective. With the aim of developing a culture system close to in vivo conditions for testing new drugs, we characterized an ex vivo three-dimensional culture system based on a hyaluronic acid-rich hydrogel and compared it with classical two-dimensional culture conditions. U87-MG glioblastoma cells and seven primary cell cultures of human glioblastomas were subjected to radiation therapy and chemotherapy drugs. It appears that 3D hydrogel preserves the original cancer growth behaviormore » and enables assessment of the sensitivity of malignant gliomas to radiation and drugs with regard to inter-tumoral heterogeneity of therapeutic response. It could be used for preclinical assessment of new therapies. - Highlights: • We have compared primary glioblastoma cell culture in a 2D versus 3D-matrix system. • In 3D morphology, organization and markers better recapitulate the original tumor. • 3D-matrix culture might represent a relevant system for more accurate drug screening.« less
An overview of cartilage tissue engineering.
Kim, H W; Han, C D
2000-12-01
Articular cartilage regeneration refers to the formation of new tissue that is indistinguishable from the native articular cartilage with respect to zonal organization, biochemical composition, and mechanical properties. Due to a limited capacity to repair cartilage, scar tissue frequently has a poorly organized structure and lacks the functional characteristics of normal cartilage. The degree of success to date achieved using a purely cell- or biological-based approach has been modest. Potentially the development of a hybrid strategy, whereby, chondrocytes or chondrogenic stem cells are combined with a matrix, making cartilage in vitro, which is then subsequently transplanted, offers a route towards a new successful treatment modality. The success of this approach depends upon the material being biocompatible, processable into a suitable three-dimensional structure and eventually biodegradable without harmful effects. In addition, the material should have a sufficient porosity to facilitate high cell loading and tissue ingrowth, and it should be able to support cell proliferation, differentiation, and function. The cell-polymer-bioreactor system provides a basis for studying the structural and functional properties of the cartilaginous matrix during its development, because tissue concentrations of glycosaminoglycan and collagen can be modulated by altering the conditions of tissue cultivation.
Regulation of gene expression in intervertebral disc cells by low and high hydrostatic pressure.
Neidlinger-Wilke, Cornelia; Würtz, Karin; Urban, Jill P G; Börm, Wolfgang; Arand, Markus; Ignatius, Anita; Wilke, Hans-Joachim; Claes, Lutz E
2006-08-01
Intervertebral disc structures are exposed to wide ranges of intradiscal hydrostatic pressure during different loading exercises and are at their minimum during lying or relaxed sitting and at maximum during lifting weights with a round back. We hypothesize that these different loading magnitudes influence the intervertebral disc (IVD) by alteration of disc matrix turnover depending on their magnitudes. Therefore the aim of this study was to assess changes in gene expression of human nucleus cells after the application of low hydrostatic pressure (0.25 MPa) and high hydrostatic pressure (2.5 MPa). IVD cells isolated from the nucleus of human (n = 18) and bovine (n = 24 from four animals) disc biopsies were seeded into three-dimensional collagen type-I matrices and exposed to the different loading magnitudes by specially developed pressure chambers. The lower pressure range (0.25 MPa, 30 min, 0.1 Hz) was applied with a recently published device by using an external compression cylinder. For the application of higher loads (2.5 MPa, 30 min, 0.1 Hz) the cell-loaded collagen gels were sealed into sterile bags with culture medium and stimulated in a newly developed water-filled compression cylinder by using a loading frame. These methods allowed the comparison of loading regimes in a wide physiological range under an equal three-dimensional culture conditions. Cells were harvested 24 h after the end of stimulation and changes in the expression of genes known to influence IVD matrix turnover (collagen-I, collagen-II, aggrecan, MMP1, MMP2, MMP3, MMP13) were analyzed by real-time RT-PCR. A Wilcoxon signed-rank test(1) and a Wilcoxon 2-sample test(2) were performed to detect differences between the stimulated and control samples(1) and differences between low and high hydrostatic pressure(2). Multiple testing was considered by adjusting the p value appropriately. Both regimes of hydrostatic pressure influenced gene expression in nucleus cells with opposite tendencies for the matrix forming proteins aggrecan and collagen type-I in response to the two different pressure magnitudes: Low hydrostatic-pressure (0.25 MPa) tended to increase collagen-I and aggrecan expression of human nucleus cells (P < 0.05) but only to a small degree. High hydrostatic pressure (2.5 MPa) tended to decrease gene expression of all anabolic proteins with significant effects on aggrecan expression of nucleus cells (P = 0.004). Low hydrostatic pressure had no influence on the expression of matrix metalloproteinases (MMP1, MMP2, MMP3 and MMP13). In contrast, high hydrostatic pressure tended to increase the expression of MMP1, MMP3 and MMP13 of human nucleus cells with high individual-individual variations. The decreased expression of aggrecan (P = 0.008) and collagen type II (P = 0.023) and the increased MMP3 expression (P = 0.008) in response to high hydrostatic pressure could be confirmed in additional experiments with bovine nucleus cells. These results suggest that hydrostatic pressure as one of the physiological stimuli of the IVD may influence matrix turnover in a magnitude dependent way. Low hydrostatic pressure (0.25 MPa) has quite small influences with a tendency to anabolic effects, whereas high hydrostatic pressure (2.5 MPa) tends to decrease the matrix protein expression with a tendency to increase some matrix-turnover enzymes. Therefore, hydrostatic pressure may regulate disc matrix turnover in a dose-dependent way.
Ma, Lin; Weisman, Catherine; Baltean-Carlès, Diana; Delbende, Ivan; Bauwens, Luc
2015-08-01
The influence of a resistive load on the starting performance of a standing-wave thermoacoustic engine is investigated numerically. The model used is based upon a low Mach number assumption; it couples the two-dimensional nonlinear flow and heat exchange within the thermoacoustic active cell with one-dimensional linear acoustics in the loaded resonator. For a given engine geometry, prescribed temperatures at the heat exchangers, prescribed mean pressure, and prescribed load, results from a simulation in the time domain include the evolution of the acoustic pressure in the active cell. That signal is then analyzed, extracting growth rate and frequency of the dominant modes. For a given load, the temperature difference between the two sides is then varied; the most unstable mode is identified and so is the corresponding critical temperature ratio between heater and cooler. Next, varying the load, a stability diagram is obtained, potentially with a predictive value. Results are compared with those derived from Rott's linear theory as well as with experimental results found in the literature.
NASA Technical Reports Server (NTRS)
Gnoffo, Peter A.
2009-01-01
The quality of simulated hypersonic stagnation region heating on tetrahedral meshes is investigated by using a three-dimensional, upwind reconstruction algorithm for the inviscid flux vector. Two test problems are investigated: hypersonic flow over a three-dimensional cylinder with special attention to the uniformity of the solution in the spanwise direction and hypersonic flow over a three-dimensional sphere. The tetrahedral cells used in the simulation are derived from a structured grid where cell faces are bisected across the diagonal resulting in a consistent pattern of diagonals running in a biased direction across the otherwise symmetric domain. This grid is known to accentuate problems in both shock capturing and stagnation region heating encountered with conventional, quasi-one-dimensional inviscid flux reconstruction algorithms. Therefore the test problem provides a sensitive test for algorithmic effects on heating. This investigation is believed to be unique in its focus on three-dimensional, rotated upwind schemes for the simulation of hypersonic heating on tetrahedral grids. This study attempts to fill the void left by the inability of conventional (quasi-one-dimensional) approaches to accurately simulate heating in a tetrahedral grid system. Results show significant improvement in spanwise uniformity of heating with some penalty of ringing at the captured shock. Issues with accuracy near the peak shear location are identified and require further study.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Que, Emily L.; Bleher, Reiner; Duncan, Francesca E.
2014-12-15
Fertilization of a mammalian egg initiates a series of 'zinc sparks' that are necessary to induce the egg-to-embryo transition. Despite the importance of these zinc-efflux events little is known about their origin. To understand the molecular mechanism of the zinc spark we combined four physical approaches that resolve zinc distributions in single cells: a chemical probe for dynamic live-cell fluorescence imaging and a combination of scanning transmission electron microscopy with energy-dispersive spectroscopy, X-ray fluorescence microscopy and three-dimensional elemental tomography for high-resolution elemental mapping. We show that the zinc spark arises from a system of thousands of zinc-loaded vesicles, each ofmore » which contains, on average, 10(6) zinc atoms. These vesicles undergo dynamic movement during oocyte maturation and exocytosis at the time of fertilization. The discovery of these vesicles and the demonstration that zinc sparks originate from them provides a quantitative framework for understanding how zinc fluxes regulate cellular processes« less
Que, Emily L; Bleher, Reiner; Duncan, Francesca E; Kong, Betty Y; Gleber, Sophie C; Vogt, Stefan; Chen, Si; Garwin, Seth A; Bayer, Amanda R; Dravid, Vinayak P; Woodruff, Teresa K; O'Halloran, Thomas V
2015-02-01
Fertilization of a mammalian egg initiates a series of 'zinc sparks' that are necessary to induce the egg-to-embryo transition. Despite the importance of these zinc-efflux events little is known about their origin. To understand the molecular mechanism of the zinc spark we combined four physical approaches that resolve zinc distributions in single cells: a chemical probe for dynamic live-cell fluorescence imaging and a combination of scanning transmission electron microscopy with energy-dispersive spectroscopy, X-ray fluorescence microscopy and three-dimensional elemental tomography for high-resolution elemental mapping. We show that the zinc spark arises from a system of thousands of zinc-loaded vesicles, each of which contains, on average, 10(6) zinc atoms. These vesicles undergo dynamic movement during oocyte maturation and exocytosis at the time of fertilization. The discovery of these vesicles and the demonstration that zinc sparks originate from them provides a quantitative framework for understanding how zinc fluxes regulate cellular processes.
Four-Dimensional Imaging of T Cells in Kidney Transplant Rejection.
Hughes, Andrew D; Lakkis, Fadi G; Oberbarnscheidt, Martin H
2018-06-01
Kidney transplantation is the treatment of choice for ESRD but is complicated by the response of the recipient's immune system to nonself histocompatibility antigens on the graft, resulting in rejection. Multiphoton intravital microscopy, referred to as four-dimensional imaging because it records dynamic events in three-dimensional tissue volumes, has emerged as a powerful tool to study immunologic processes in living animals. Here, we will review advances in understanding the complex mechanisms of T cell-mediated rejection made possible by four-dimensional imaging of mouse renal allografts. We will summarize recent data showing that activated (effector) T cell migration to the graft is driven by cognate antigen presented by dendritic cells that surround and penetrate peritubular capillaries, and that T cell-dendritic cell interactions persist in the graft over time, maintaining the immune response in the tissue. Copyright © 2018 by the American Society of Nephrology.
NASA Technical Reports Server (NTRS)
Campbell, C. W.
1984-01-01
A three dimensional model which combines measurements of wind shear in the real atmosphere with three dimensional Monte Carlo simulated turbulence was developed. The wind field over the body of an aircraft can be simulated and all aerodynamic loads and moments calculated.
[Stress analysis of the mandible by 3D FEA in normal human being under three loading conditions].
Sun, Jian; Zhang, Fu-qiang; Wang, Dong-wei; Yu, Jia; Wang, Cheng-tao
2004-02-01
The condition and character of stress distribution in the mandibular in normal human being during centric, protrusive, laterotrusive occlusion were analysed. The three-dimensional finite element model of the mandibular was developed by helica CT scanning and CAD/CAM software, and three-dimensional finite element stress analysis was done by ANSYS software. Three-dimensional finite element model of the mandibular was generated. Under these three occlusal conditions, the stress of various regions in the mandible were distributed unequally, and the stress feature was different;while the stress of corresponding region in bilateral mandibular was in symmetric distribution. The stress value of condyle neck, the posterior surface of coronoid process and mandibular angle were high. The material properties of mandible were closely correlated to the value of stress. Stress distribution were similar according to the three different loading patterns, but had different effects on TMJ joint. The concentrated areas of stress were in the condyle neck, the posterior surface of coronoid process and mandibular angle.
Kamalaldin, Nurulain 'Atikah; Jaafar, Mariatti; Zubairi, Saiful Irwan; Yahaya, Badrul Hisham
2018-01-04
The use of bioceramics, especially the combination of hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP), as a three-dimensional scaffold in bone engineering is essential because together these elements constitute 60% of the bone content. Different ratios of HA and β-TCP were previously tested for their ability to produce suitable bioceramic scaffolds, which must be able to withstand high mechanical load. In this study, two ratios of HA/TCP (20:80 and 70:30) were used to create pellets, which then were evaluated in vitro to identify any adverse effects of using the material in bone grafting. Diametral tensile strength (DTS) and density testing was conducted to assess the mechanical strength and porosity of the pellets. The pellets then were tested for their toxicity to normal human fibroblast cells. In the toxicity assay, cells were incubated with the pellets for 3 days. At the end of the experiment, cell morphological changes were assessed, and the absorbance was read using PrestoBlue Cell Viability Reagent™. An inversely proportional relationship between DTS and porosity percentage was detected. Fibroblasts showed normal cell morphology in both treatments, which suggests that the HA/TCP pellets were not toxic. In the osteoblast cell attachment assay, cells were able to attach to the surface of both ratios, but cells were also able to penetrate inside the scaffold of the 70:30 pellets. This finding suggests that the 70:30 ratio had better osteoconduction properties than the 20:80 ratio.
Mechanical and thermal buckling analysis of sandwich panels under different edge conditions
NASA Technical Reports Server (NTRS)
Ko, William L.
1993-01-01
By using the Rayleigh-Ritz method of minimizing the total potential energy of a structural system, combined load (mechanical or thermal load) buckling equations are established for orthotropic rectangular sandwich panels supported under four different edge conditions. Two-dimensional buckling interaction curves and three dimensional buckling interaction surfaces are constructed for high-temperature honeycomb-core sandwich panels supported under four different edge conditions. The interaction surfaces provide easy comparison of the panel buckling strengths and the domains of symmetrical and antisymmetrical buckling associated with the different edge conditions. Thermal buckling curves of the sandwich panels also are presented. The thermal buckling conditions for the cases with and without thermal moments were found to be identical for the small deformation theory. In sandwich panels, the effect of transverse shear is quite large, and by neglecting the transverse shear effect, the buckling loads could be overpredicted considerably. Clamping of the edges could greatly increase buckling strength more in compression than in shear.
Three-dimensional confocal microscopy of the living cornea and ocular lens
NASA Astrophysics Data System (ADS)
Masters, Barry R.
1991-07-01
The three-dimensional reconstruction of the optic zone of the cornea and the ocular crystalline lens has been accomplished using confocal microscopy and volume rendering computer techniques. A laser scanning confocal microscope was used in the reflected light mode to obtain the two-dimensional images from the cornea and the ocular lens of a freshly enucleated rabbit eye. The light source was an argon ion laser with a 488 nm wavelength. The microscope objective was a Leitz X25, NA 0.6 water immersion lens. The 400 micron thick cornea was optically sectioned into 133 three micron sections. The semi-transparent cornea and the in-situ ocular lens was visualized as high resolution, high contrast two-dimensional images. The structures observed in the cornea include: superficial epithelial cells and their nuclei, basal epithelial cells and their 'beaded' cell borders, basal lamina, nerve plexus, nerve fibers, nuclei of stromal keratocytes, and endothelial cells. The structures observed in the in- situ ocular lens include: lens capsule, lens epithelial cells, and individual lens fibers. The three-dimensional data sets of the cornea and the ocular lens were reconstructed in the computer using volume rendering techniques. Stereo pairs were also created of the two- dimensional ocular images for visualization. The stack of two-dimensional images was reconstructed into a three-dimensional object using volume rendering techniques. This demonstration of the three-dimensional visualization of the intact, enucleated eye provides an important step toward quantitative three-dimensional morphometry of the eye. The important aspects of three-dimensional reconstruction are discussed.
Assunção, Wirley Gonçalves; Gomes, Erica Alves; Rocha, Eduardo Passos; Delben, Juliana Aparecida
2011-01-01
Three-dimensional finite element analysis was used to evaluate the effect of vertical and angular misfit in three-piece implant-supported screw-retained fixed prostheses on the biomechanical response in the peri-implant bone, implants, and prosthetic components. Four three-dimensional models were fabricated to represent a right posterior mandibular section with one implant in the region of the second premolar (2PM) and another in the region of the second molar (2M). The implants were splinted by a three-piece implant-supported metal-ceramic prosthesis and differed according to the type of misfit, as represented by four different models: Control = prosthesis with complete fit to the implants; UAM (unilateral angular misfit) = prosthesis presenting unilateral angular misfit of 100 μm in the mesial region of the 2M; UVM (unilateral vertical misfit) = prosthesis presenting unilateral vertical misfit of 100 μm in the mesial region of the 2M; and TVM (total vertical misfit) = prosthesis presenting total vertical misfit of 100 μm in the platform of the framework in the 2M. A vertical load of 400 N was distributed and applied on 12 centric points by the software Ansys, ie, a vertical load of 150 N was applied to each molar in the prosthesis and a vertical load of 100 N was applied at the 2PM. The stress values and distribution in peri-implant bone tissue were similar for all groups. The models with misfit exhibited different distribution patterns and increased stress magnitude in comparison to the control. The highest stress values in group UAM were observed in the implant body and retention screw. The groups UVM and TVM exhibited high stress values in the platform of the framework and the implant hexagon, respectively. The three types of misfit influenced the magnitude and distribution of stresses. The influence of misfit on peri-implant bone tissue was modest. Each type of misfit increased the stress values in different regions of the system.
Cell culture in autologous fibrin scaffolds for applications in tissue engineering.
de la Puente, Pilar; Ludeña, Dolores
2014-03-10
In tissue engineering techniques, three-dimensional scaffolds are needed to adjust and guide cell growth and to allow tissue regeneration. The scaffold must be biocompatible, biodegradable and must benefit the interactions between cells and biomaterial. Some natural biomaterials such as fibrin provide a structure similar to the native extracellular matrix containing the cells. Fibrin was first used as a sealant based on pools of commercial fibrinogen. However, the high risk of viral transmission of these pools led to the development of techniques of viral inactivation and elimination and the use of autologous fibrins. In recent decades, fibrin has been used as a release system and three-dimensional scaffold for cell culture. Fibrin scaffolds have been widely used for the culture of different types of cells, and have found several applications in tissue engineering. The structure and development of scaffolds is a key point for cell culture because scaffolds of autologous fibrin offer an important alternative due to their low fibrinogen concentrations, which are more suitable for cell growth. With this review our aim is to follow methods of development, analyze the commercial and autologous fibrins available and assess the possible applications of cell culture in tissue engineering in these three-dimensional structures. Copyright © 2013 Elsevier Inc. All rights reserved.
NASA Astrophysics Data System (ADS)
Sebastian Mannoor, Manu
Direct multidimensional integration of functional electronics and mechanical elements with viable biological systems could allow for the creation of bionic systems and devices possessing unique and advanced capabilities. For example, the ability to three dimensionally integrate functional electronic and mechanical components with biological cells and tissue could enable the creation of bionic systems that can have tremendous impact in regenerative medicine, prosthetics, and human-machine interfaces. However, as a consequence of the inherent dichotomy in material properties and limitations of conventional fabrication methods, the attainment of truly seamless integration of electronic and/or mechanical components with biological systems has been challenging. Nanomaterials engineering offers a general route for overcoming these dichotomies, primarily due to the existence of a dimensional compatibility between fundamental biological functional units and abiotic nanomaterial building blocks. One area of compelling interest for bionic systems is in the field of biomedical sensing, where the direct interfacing of nanosensors onto biological tissue or the human body could stimulate exciting opportunities such as on-body health quality monitoring and adaptive threat detection. Further, interfacing of antimicrobial peptide based bioselective probes onto the bionic nanosensors could offer abilities to detect pathogenic bacteria with bio-inspired selectivity. Most compellingly, when paired with additive manufacturing techniques such as 3D printing, these characteristics enable three dimensional integration and merging of a variety of functional materials including electronic, structural and biomaterials with viable biological cells, in the precise anatomic geometries of human organs, to form three dimensionally integrated, multi-functional bionic hybrids and cyborg devices with unique capabilities. In this thesis, we illustrate these approaches using three representative bionic systems: 1) Bionic Nanosensors: featuring bio-integrated graphene nanosensors for ubiquitous sensing, 2) Bionic Organs: featuring 3D printed bionic ears with three dimensionally integrated electronics and 3) Bionic Leaves: describing ongoing work in the direction of the creation of a bionic leaf enabled by the integration of plant derived photosynthetic functional units with electronic materials and components into a leaf-shaped hierarchical structure for harvesting photosynthetic bioelectricity.
Horbens, Melanie; Branke, Dominik; Gärtner, Roland; Voigt, Axel; Stenger, Florian; Neinhuis, Christoph
2015-10-01
Sclereid formation in addition to or in gaps of fragmented fibre rings is common in dicotyledonous plant stems. Whether this sclereid formation is force-triggered remains open so far. In fruit peduncles of several Malus species as modified plant stems, for example, the persistent fibre ring is displaced to the centre by formation of cortex parenchyma during growth. Parenchyma cells subsequently differentiate into an additional layer of brachysclereids, previously interpreted as an adaptation to continuously rising fruit loads. The present study pursues a multi-scale numerical modelling approach, to verify the important effect for different cellular architectures in both sclerenchyma categories on the stiffness of these tissues and the entire peduncle. First, different material properties are simulated analogue to plant tissues on the basis of three cell types. A regular three-dimensional and a random Voronoi microstructure combined with various mechanical cell wall parameters are applied. Using homogenisation simulations based on HILL's principle, numerical calculations predict a lower effective homogenised tissue stiffness of isodiametric brachysclereids compared to those of fibres, confirming experimentally obtained data from Malus fruit peduncles. Furthermore, a curved peduncle model with a complex arrangement of different material layers is generated. Diverse material sets are tested under three representative loadings, using an adaptive diffuse domain approach (AMDiS). The model explains the function of sclereids as considerable contributors to the stiffness against bending and tensile deformations, as well as torsion, especially in consequence of superimposed load conditions in the case of a curved plant stem. Copyright © 2015 Elsevier Inc. All rights reserved.
A Novel Three-Dimensional Immune Oncology Model for High-Throughput Testing of Tumoricidal Activity.
Sherman, Hilary; Gitschier, Hannah J; Rossi, Ann E
2018-01-01
The latest advancements in oncology research are focused on autologous immune cell therapy. However, the effectiveness of this type of immunotherapy for cancer remediation is not equivalent for all patients or cancer types. This suggests the need for better preclinical screening models that more closely recapitulate in vivo tumor biology. The established method for investigating tumoricidal activity of immunotherapies has been study of two-dimensional (2D) monolayer cultures of immortalized cancer cell lines or primary tumor cells in standard tissue culture vessels. Indeed, a proven means to examine immune cell migration and invasion are 2D chemotaxis assays in permeabilized supports or Boyden chambers. Nevertheless, the more in vivo -like three-dimensional (3D) multicellular tumor spheroids are quickly becoming the favored model to examine immune cell invasion and tumor cell cytotoxicity. Accordingly, we have developed a 3D immune oncology model by combining 96-well permeable support systems and 96-well low-attachment microplates. The use of the permeable support system enables assessment of immune cell migration, which was tested in this study as chemotactic response of natural killer NK-92MI cells to human stromal-cell derived factor-1 (SDF-1α). Immune invasion was assessed by measuring NK-92MI infiltration into lung carcinoma A549 cell spheroids that were formed in low-attachment microplates. The novel pairing of the permeable support system with low-attachment microplates permitted simultaneous investigation of immune cell homing, immune invasion of tumor spheroids, and spheroid cytotoxicity. In effect, the system represents a more comprehensive and in vivo -like immune oncology model that can be utilized for high-throughput study of tumoricidal activity.
Upreti, Meenakshi; Jamshidi-Parsian, Azemat; Koonce, Nathan A; Webber, Jessica S; Sharma, Sunil K; Asea, Alexzander AA; Mader, Mathew J; Griffin, Robert J
2011-01-01
Classic cancer research for several decades has focused on understanding the biology of tumor cells in vitro. However, extending these findings to in vivo settings has been impeded owing to limited insights on the impact of microenvironment on tumor cells. We hypothesized that tumor cell biology and treatment response would be more informative when done in the presence of stromal components, like endothelial cells, which exist in the tumor microenvironment. To that end, we have developed a system to grow three-dimensional cultures of GFP-4T1 mouse mammary tumor and 2H11 murine endothelial cells in hanging drops of medium in vitro. The presence of 2H11 endothelial cells in these three-dimensional cocultures was found to sensitize 4T1-GFP tumor cells to chemotherapy (Taxol) and, at the same time, protect cells from ionizing radiation. These spheroidal cultures can also be implanted into the dorsal skinfold window chamber of mice for fluorescence imaging of vascularization and disease progression/treatment response. We observed rapid neovascularization of the tumor-endothelial spheroids in comparison to tumor spheroids grown in nude mice. Molecular analysis revealed pronounced up-regulation of several proangiogenic factors in the tumor tissue derived from the tumor-endothelial spheroids compared with tumor-only spheroids. Furthermore, the rate of tumor growth from tumor-endothelial spheroids in mice was faster than the tumor cell-only spheroids, resulting in greater metastasis to the lung. This three-dimensional coculture model presents an improved way to investigate more pertinent aspects of the therapeutic potential for radiation and/or chemotherapy alone and in combination with antiangiogenic agents. PMID:22191001
Upreti, Meenakshi; Jamshidi-Parsian, Azemat; Koonce, Nathan A; Webber, Jessica S; Sharma, Sunil K; Asea, Alexzander Aa; Mader, Mathew J; Griffin, Robert J
2011-12-01
Classic cancer research for several decades has focused on understanding the biology of tumor cells in vitro. However, extending these findings to in vivo settings has been impeded owing to limited insights on the impact of microenvironment on tumor cells. We hypothesized that tumor cell biology and treatment response would be more informative when done in the presence of stromal components, like endothelial cells, which exist in the tumor microenvironment. To that end, we have developed a system to grow three-dimensional cultures of GFP-4T1 mouse mammary tumor and 2H11 murine endothelial cells in hanging drops of medium in vitro. The presence of 2H11 endothelial cells in these three-dimensional cocultures was found to sensitize 4T1-GFP tumor cells to chemotherapy (Taxol) and, at the same time, protect cells from ionizing radiation. These spheroidal cultures can also be implanted into the dorsal skinfold window chamber of mice for fluorescence imaging of vascularization and disease progression/treatment response. We observed rapid neovascularization of the tumor-endothelial spheroids in comparison to tumor spheroids grown in nude mice. Molecular analysis revealed pronounced up-regulation of several proangiogenic factors in the tumor tissue derived from the tumor-endothelial spheroids compared with tumor-only spheroids. Furthermore, the rate of tumor growth from tumor-endothelial spheroids in mice was faster than the tumor cell-only spheroids, resulting in greater metastasis to the lung. This three-dimensional coculture model presents an improved way to investigate more pertinent aspects of the therapeutic potential for radiation and/or chemotherapy alone and in combination with antiangiogenic agents.
Osteochondral Interface Tissue Engineering Using Macroscopic Gradients of Bioactive Signals
Dormer, Nathan H.; Singh, Milind; Wang, Limin; Berkland, Cory J.; Detamore, Michael S.
2013-01-01
Continuous gradients exist at osteochondral interfaces, which may be engineered by applying spatially patterned gradients of biological cues. In the present study, a protein-loaded microsphere-based scaffold fabrication strategy was applied to achieve spatially and temporally controlled delivery of bioactive signals in three-dimensional (3D) tissue engineering scaffolds. Bone morphogenetic protein-2 and transforming growth factor-β1-loaded poly(d,llactic- co-glycolic acid) microspheres were utilized with a gradient scaffold fabrication technology to produce microsphere-based scaffolds containing opposing gradients of these signals. Constructs were then seeded with human bone marrow stromal cells (hBMSCs) or human umbilical cord mesenchymal stromal cells (hUCMSCs), and osteochondral tissue regeneration was assessed in gradient scaffolds and compared to multiple control groups. Following a 6-week cell culture, the gradient scaffolds produced regionalized extracellular matrix, and outperformed the blank control scaffolds in cell number, glycosaminoglycan production, collagen content, alkaline phosphatase activity, and in some instances, gene expression of major osteogenic and chondrogenic markers. These results suggest that engineered signal gradients may be beneficial for osteochondral tissue engineering. PMID:20379780
Coates, Emily E; Fisher, John P
2010-11-01
Articular cartilage defects have limited capacity to self-repair, and cost society up to 60 billion dollars annually in both medical treatments and loss of working days. Recent developments in cartilage tissue engineering have resulted in many new products coming to market or entering clinical trials. However, there is a distinct lack of treatments which aim to recreate the complex zonal organization of articular cartilage. Cartilage tissue withstands repetitive strains throughout an individual's lifetime and provides frictionless movement between joints. The structure and composition of its intricately organized extracellular matrix varies with tissue depth to provide optimal resistance to loading, ensure ease of movement, and integrate with the subchondral bone. Each tissue zone is specially designed to resist the load it experiences, and maximize the tissue properties needed for its location. It is unlikely that a homogenous solution to tissue repair will be able to optimally restore the function of such a heterogeneous tissue. For zonal engineering of articular cartilage to become practical, maintenance of phenotypically stable zonal cell populations must be achieved. The chondrocyte phenotype varies considerably by zone, and it is the activity of these cells that help achieve the structural organization of the tissue. This review provides an examination of literature which has studied variations in cellular phenotype between cartilage zones. By doing so, we have identified critical differences between cell populations and highlighted areas of research which show potential in the field. Current research has made the morphological and metabolic variations between these cell populations clear, but an ideal way of maintaining these differences in vitro culture is yet to be established. Combinations of delivered growth factors, mechanical loading, and layered three-dimensional culture systems all show potential for achieving this goal. Furthermore, differentiation of progenitor cell populations into chondrocyte subpopulations may also hold promise for achieving large numbers of zonal chondrocytes. Success of the field lies in establishing methods of retaining phenotypically stable cell populations for in vitro culture.
Wang, Ying; Zhao, Qinfu; Hu, Yanchen; Sun, Lizhang; Bai, Ling; Jiang, Tongying; Wang, Siling
2013-01-01
The goal of the present study was to compare the drug release properties and stability of the nanoporous silica with different pore architectures as a matrix for improved delivery of poorly soluble drugs. For this purpose, three dimensional ordered macroporous (3DOM) silica with 3D continuous and interconnected macropores of different sizes (200 nm and 500 nm) and classic mesoporous silica (ie, Mobil Composition of Matter [MCM]-41 and Santa Barbara Amorphous [SBA]-15) with well-ordered two dimensional (2D) cylindrical mesopores were successfully fabricated and then loaded with the model drug indomethacin (IMC) via the solvent deposition method. Scanning electron microscopy (SEM), N2 adsorption, differential scanning calorimetry (DSC), and X-ray diffraction (XRD) were applied to systematically characterize all IMC-loaded nanoporous silica formulations, evidencing the successful inclusion of IMC into nanopores, the reduced crystallinity, and finally accelerated dissolution of IMC. It was worth mentioning that, in comparison to 2D mesoporous silica, 3DOM silica displayed a more rapid release profile, which may be ascribed to the 3D interconnected pore networks and the highly accessible surface areas. The results obtained from the stability test indicated that the amorphous state of IMC entrapped in the 2D mesoporous silica (SBA-15 and MCM-41) has a better physical stability than in that of 3DOM silica. Moreover, the dissolution rate and stability of IMC loaded in 3DOM silica was closely related to the pore size of macroporous silica. The colorimetric 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Cell Counting Kit (CCK)-8 assays in combination with direct morphology observations demonstrated the good biocompatibility of nanoporous silica, especially for 3DOM silica and SBA-15. The present work encourages further study of the drug release properties and stability of drug entrapped in different pore architecture of silica in order to realize their potential in oral drug delivery. PMID:24174875
Shanbhag, Sachin; Wang, Shaopeng; Kotov, Nicholas A
2005-12-01
Limited ingrowth of stromal cells is observed when a three-dimensionally ordered scaffold possessing inverted-colloidal-crystal geometry is used to culture adherent cells. In this work, a computational model explaining, as well as predicting, experimental cell distributions is developed. It incorporates a modified Contois cell-growth model that includes the effects of nutrient saturation, competitive product inhibition, and cell-contact inhibition to describe the scaffold-cell system. Our results agree with the hypothesis that the rapid growth of cells on the surface of the scaffold depletes the nutrient supply to the core, resulting in the preferential growth on the exterior of the scaffold. When the cells are cultured in a scaffold subjected to a uniform velocity field, they penetrate to a greater extent into the scaffold core. Alternative seeding and culture strategies are suggested and evaluated.
Yan, Xu; Zhang, Xinwen; Chi, Weichao; Ai, Hongjun; Wu, Lin
2015-05-01
This study aimed to compare the influence of alveolar ridge cortical bone and sinus floor cortical bone in sinus areabi-cortical dental implantation by means of 3D finite element analysis. Three-dimensional finite element (FE) models in a posterior maxillary region with sinus membrane and the same height of alveolar ridge of 10 mm were generated according to the anatomical data of the sinus area. They were either with fixed thickness of crestal cortical bone and variable thickness of sinus floor cortical bone or vice versa. Ten models were assumed to be under immediate loading or conventional loading. The standard implant model based on the Nobel Biocare implant system was created via computer-aided design software. All materials were assumed to be isotropic and linearly elastic. An inclined force of 129 N was applied. Von Mises stress mainly concentrated on the surface of crestal cortical bone around the implant neck. For all the models, both the axial and buccolingual resonance frequencies of conventional loading were higher than those of immediate loading; however, the difference is less than 5%. The results showed that bi-cortical implant in sinus area increased the stability of the implant, especially for immediately loading implantation. The thickness of both crestal cortical bone and sinus floor cortical bone influenced implant micromotion and stress distribution; however, crestal cortical bone may be more important than sinus floor cortical bone.
Three-dimensional coherent X-ray diffractive imaging of whole frozen-hydrated cells
Rodriguez, Jose A.; Xu, Rui; Chen, Chien-Chun; Huang, Zhifeng; Jiang, Huaidong; Chen, Allan L.; Raines, Kevin S.; Pryor Jr, Alan; Nam, Daewoong; Wiegart, Lutz; Song, Changyong; Madsen, Anders; Chushkin, Yuriy; Zontone, Federico; Bradley, Peter J.; Miao, Jianwei
2015-01-01
A structural understanding of whole cells in three dimensions at high spatial resolution remains a significant challenge and, in the case of X-rays, has been limited by radiation damage. By alleviating this limitation, cryogenic coherent diffractive imaging (cryo-CDI) can in principle be used to bridge the important resolution gap between optical and electron microscopy in bio-imaging. Here, the first experimental demonstration of cryo-CDI for quantitative three-dimensional imaging of whole frozen-hydrated cells using 8 keV X-rays is reported. As a proof of principle, a tilt series of 72 diffraction patterns was collected from a frozen-hydrated Neospora caninum cell and the three-dimensional mass density of the cell was reconstructed and quantified based on its natural contrast. This three-dimensional reconstruction reveals the surface and internal morphology of the cell, including its complex polarized sub-cellular structure. It is believed that this work represents an experimental milestone towards routine quantitative three-dimensional imaging of whole cells in their natural state with spatial resolutions in the tens of nanometres. PMID:26306199
Three-dimensional coherent X-ray diffractive imaging of whole frozen-hydrated cells
Rodriguez, Jose A.; Xu, Rui; Chen, Chien -Chun; ...
2015-09-01
Here, a structural understanding of whole cells in three dimensions at high spatial resolution remains a significant challenge and, in the case of X-rays, has been limited by radiation damage. By alleviating this limitation, cryogenic coherent diffractive imaging (cryo-CDI) can in principle be used to bridge the important resolution gap between optical and electron microscopy in bio-imaging. Here, the first experimental demonstration of cryo-CDI for quantitative three-dimensional imaging of whole frozen-hydrated cells using 8 Kev X-rays is reported. As a proof of principle, a tilt series of 72 diffraction patterns was collected from a frozen-hydrated Neospora caninum cell and themore » three-dimensional mass density of the cell was reconstructed and quantified based on its natural contrast. This three-dimensional reconstruction reveals the surface and internal morphology of the cell, including its complex polarized sub-cellular structure. Finally, it is believed that this work represents an experimental milestone towards routine quantitative three-dimensional imaging of whole cells in their natural state with spatial resolutions in the tens of nanometres.« less
Three-dimensional coherent X-ray diffractive imaging of whole frozen-hydrated cells.
Rodriguez, Jose A; Xu, Rui; Chen, Chien-Chun; Huang, Zhifeng; Jiang, Huaidong; Chen, Allan L; Raines, Kevin S; Pryor, Alan; Nam, Daewoong; Wiegart, Lutz; Song, Changyong; Madsen, Anders; Chushkin, Yuriy; Zontone, Federico; Bradley, Peter J; Miao, Jianwei
2015-09-01
A structural understanding of whole cells in three dimensions at high spatial resolution remains a significant challenge and, in the case of X-rays, has been limited by radiation damage. By alleviating this limitation, cryogenic coherent diffractive imaging (cryo-CDI) can in principle be used to bridge the important resolution gap between optical and electron microscopy in bio-imaging. Here, the first experimental demonstration of cryo-CDI for quantitative three-dimensional imaging of whole frozen-hydrated cells using 8 keV X-rays is reported. As a proof of principle, a tilt series of 72 diffraction patterns was collected from a frozen-hydrated Neospora caninum cell and the three-dimensional mass density of the cell was reconstructed and quantified based on its natural contrast. This three-dimensional reconstruction reveals the surface and internal morphology of the cell, including its complex polarized sub-cellular structure. It is believed that this work represents an experimental milestone towards routine quantitative three-dimensional imaging of whole cells in their natural state with spatial resolutions in the tens of nanometres.
NASA Astrophysics Data System (ADS)
Xu, Yong; Dong, Wen-Cai
2013-08-01
A frequency domain analysis method based on the three-dimensional translating-pulsating (3DTP) source Green function is developed to investigate wave loads and free motions of two ships advancing on parallel course in waves. Two experiments are carried out respectively to measure the wave loads and the freemotions for a pair of side-byside arranged ship models advancing with an identical speed in head regular waves. For comparison, each model is also tested alone. Predictions obtained by the present solution are found in favorable agreement with the model tests and are more accurate than the traditional method based on the three dimensional pulsating (3DP) source Green function. Numerical resonances and peak shift can be found in the 3DP predictions, which result from the wave energy trapped in the gap between two ships and the extremely inhomogeneous wave load distribution on each hull. However, they can be eliminated by 3DTP, in which the speed affects the free surface and most of the wave energy can be escaped from the gap. Both the experiment and the present prediction show that hydrodynamic interaction effects on wave loads and free motions are significant. The present solver may serve as a validated tool to predict wave loads and motions of two vessels under replenishment at sea, and may help to evaluate the hydrodynamic interaction effects on the ships safety in replenishment operation.
Three-Dimensional Cell Printing of Large-Volume Tissues: Application to Ear Regeneration.
Lee, Jung-Seob; Kim, Byoung Soo; Seo, Donghwan; Park, Jeong Hun; Cho, Dong-Woo
2017-03-01
The three-dimensional (3D) printing of large-volume cells, printed in a clinically relevant size, is one of the most important challenges in the field of tissue engineering. However, few studies have reported the fabrication of large-volume cell-printed constructs (LCCs). To create LCCs, appropriate fabrication conditions should be established: Factors involved include fabrication time, residence time, and temperature control of the cell-laden hydrogel in the syringe to ensure high cell viability and functionality. The prolonged time required for 3D printing of LCCs can reduce cell viability and result in insufficient functionality of the construct, because the cells are exposed to a harsh environment during the printing process. In this regard, we present an advanced 3D cell-printing system composed of a clean air workstation, a humidifier, and a Peltier system, which provides a suitable printing environment for the production of LCCs with high cell viability. We confirmed that the advanced 3D cell-printing system was capable of providing enhanced printability of hydrogels and fabricating an ear-shaped LCC with high cell viability. In vivo results for the ear-shaped LCC also showed that printed chondrocytes proliferated sufficiently and differentiated into cartilage tissue. Thus, we conclude that the advanced 3D cell-printing system is a versatile tool to create cell-printed constructs for the generation of large-volume tissues.
Image matrix processor for fast multi-dimensional computations
Roberson, G.P.; Skeate, M.F.
1996-10-15
An apparatus for multi-dimensional computation is disclosed which comprises a computation engine, including a plurality of processing modules. The processing modules are configured in parallel and compute respective contributions to a computed multi-dimensional image of respective two dimensional data sets. A high-speed, parallel access storage system is provided which stores the multi-dimensional data sets, and a switching circuit routes the data among the processing modules in the computation engine and the storage system. A data acquisition port receives the two dimensional data sets representing projections through an image, for reconstruction algorithms such as encountered in computerized tomography. The processing modules include a programmable local host, by which they may be configured to execute a plurality of different types of multi-dimensional algorithms. The processing modules thus include an image manipulation processor, which includes a source cache, a target cache, a coefficient table, and control software for executing image transformation routines using data in the source cache and the coefficient table and loading resulting data in the target cache. The local host processor operates to load the source cache with a two dimensional data set, loads the coefficient table, and transfers resulting data out of the target cache to the storage system, or to another destination. 10 figs.
Zhang, Yi-Jun; Xu, Jian; Wang, Yue; Lin, Xiang-Jin; Ma, Xin
2015-02-01
The aim of this study was to explore the correlation between the kinematics of the hindfoot joint and the medial arch angle change in stage II posterior tibial tendon dysfunction flatfoot three-dimensionally under loading. Computed tomography (CT) scans of 12 healthy feet and 12 feet with stage II posterior tibial tendon dysfunction flatfoot were taken both in non- and full-body-weight-bearing condition. The CT images of the hindfoot bones were reconstructed into three-dimensional models with Mimics and Geomagic reverse engineering software. The three-dimensional changes of the hindfoot joint were calculated to determine their correlation to the medial longitudinal arch angle. The medial arch angle change was larger in stage II posterior tibial tendon dysfunction flatfoot compared to that in healthy foot under loading. The rotation and translation of the talocalcaneal joint, the talonavicular joint and the calcanocuboid joint had little influence on the change of the medial arch angle in healthy foot. However, the eversion of the talocalcaneal joint, the proximal translation of the calcaneus relative to the talus and the dorsiflexion of talonavicular joint could increase the medial arch angle in stage II posterior tibial tendon dysfunction flatfoot under loading. Joint instability occurred in patients with stage II posterior tibial tendon dysfunction flatfoot under loading. Limitation of over movement of the talocalcaneal joint and the talonavicular joint may help correct the medial longitudinal arch in stage II posterior tibial tendon dysfunction flatfoot. Copyright © 2014 Elsevier Ltd. All rights reserved.
Data Report on Post-Irradiation Dimensional Change of AGC-1 Samples
DOE Office of Scientific and Technical Information (OSTI.GOV)
William Windes
This report summarizes the initial dimensional changes for loaded and unloaded AGC-1 samples. The dimensional change for all samples is presented as a function of dose. The data is further presented by graphite type and applied load levels to illustrate the differences between graphite forming processes and stress levels within the graphite components. While the three different loads placed on the samples have been verified [ ref: Larry Hull’s report] verification of the AGC-1 sample temperatures and dose levels are expected in the summer of 2012. Only estimated dose and temperature values for the samples are presented in this reportmore » to allow a partial analysis of the results.« less
NASA Astrophysics Data System (ADS)
Yamauchi, Toyohiko; Iwai, Hidenao; Yamashita, Yutaka
2013-03-01
We succeeded in utilizing our low-coherent quantitative phase microscopy (LC-QPM) to achieve label-free and three-dimensional imaging of string-like structures bridging the free-space between live cells. In past studies, three dimensional morphology of the string-like structures between cells had been investigated by electron microscopies and fluorescence microscopies and these structures were called "membrane nanotubes" or "tunneling nanotubes." However, use of electron microscopy inevitably kills these cells and fluorescence microscopy is itself a potentially invasive method. To achieve noninvasive imaging of live cells, we applied our LC-QPM which is a reflection-type, phase resolved and full-field interference microscope employing a low-coherent light source. LC-QPM is able to visualize the three-dimensional morphology of live cells without labeling by means of low-coherence interferometry. The lateral (diffraction limit) and longitudinal (coherence-length) spatial resolution of LC-QPM were respectively 0.49 and 0.93 micrometers and the repeatability of the phase measurement was 0.02 radians (1.0 nm). We successfully obtained three-dimensional morphology of live cultured epithelial cells (cell type: HeLa, derived from cervix cancer) and were able to clearly observe the individual string-like structures interconnecting the cells. When we performed volumetric imaging, a 80 micrometer by 60 micrometer by 6.5 micrometer volume was scanned every 5.67 seconds and 70 frames of a three-dimensional movie were recorded for a duration of 397 seconds. Moreover, the optical phase images gave us detailed information about the three-dimensional morphology of the string-like structure at sub-wavelength resolution. We believe that our LC-QPM will be a useful tool for the study of three-dimensional morphology of live cells.
Three-Dimensional Stress Fields and Slip Systems for Single Crystal Superalloy Notched Specimens
NASA Technical Reports Server (NTRS)
Magnan, Shannon M.; Throckmorton, David (Technical Monitor)
2002-01-01
Single crystal superalloys have become increasingly popular for turbine blade and vane applications due to their high strength, and creep and fatigue resistance at elevated temperatures. The crystallographic orientation of a single crystal material greatly affects its material properties, including elastic modulus, shear modulus, and ductility. These directional properties, along with the type of loading and temperature, dictate an anisotropic response in the yield strength, creep resistance, creep rupture ductility, fatigue resistance, etc. A significant amount of research has been conducted to determine the material properties in the <001> orientation, yet the material properties deviating from the <001> orientation have not been assessed for all cases. Based on the desired application and design criteria, a crystal orientation is selected to yield the maximum properties. Currently, single crystal manufacturing is able to control the primary crystallographic orientation within 15 of the target orientation, which is an acceptable deviation to meet both performance and cost guidelines; the secondary orientation is rarely specified. A common experiment is the standard load-controlled tensile test, in which specimens with different orientations can be loaded to observe the material response. The deformation behavior of single-crystal materials under tension and compression is known to be a function of not only material orientation, but also of varying microdeformation (i.e. dislocation) mechanisms. The underlying dislocation motion causes deformation via slip, and affects the activation of specific slip systems based on load and orientation. The slip can be analyzed by observing the visible traces left on the surface of the specimen from the slip activity within the single crystal material. The goal of this thesis was to predict the slip systems activated in three-dimensional stress fields of a notched tensile specimen, as a function of crystal orientation, using finite element analysis without addressing microstructural deformation mechanisms that govern their activation. Out of three orientations tested, the specimen with a [110] load orientation and a [001] growth direction had the lowest maximum resolved shear stress; this specimen orientation appears to be the best design candidate for a tensile application.
Long-term immunologically competent human peripheral lymphoid tissue cultures in a 3D bioreactor
Kuzin, Igor; Sun, Hongliang; Moshkani, Safiekhatoon; Feng, Changyong; Mantalaris, Athanasios; Wu, JH David; Bottaro, Andrea
2011-01-01
Peripheral lymphoid organs (PLOs), the primary sites of development of adaptive immune responses, display a complex structural organization reflecting separation of cellular subsets (e.g. T and B lymphocytes) and functional compartments which is critical for immune function. The generation of in vitro culture systems capable of recapitulating salient features of PLOs for experimental, biotechnological and clinical applications would be highly desirable, but has been hampered so far by the complexity of these systems. We have previously developed a three-dimensional bioreactor system for long-term, functional culture of human bone marrow cells on macroporous microspheres in a packed-bed bioreactor with frequent medium change. Here we adapt the same system for culture of human primary cells from PLOs (tonsil) in the absence of specific exogenous growth factors or activators. Cells in this system displayed higher viability over several weeks, and maintain population diversity and cell surface markers largely comparable to primary cells. Light microscopy showed cells organizing in large diverse clusters within the scaffold pores and presence of B cell-enriched areas. Strikingly, these cultures generated a significant number of antibody-producing B cells when challenged with a panel of diverse antigens, as expected from a lymphoid tissue. Thus the three-dimensional tonsil bioreactor culture system may serve as a useful model of PLOs by recapitulating their structural organization and function ex vivo. PMID:21309085
Long-term immunologically competent human peripheral lymphoid tissue cultures in a 3D bioreactor.
Kuzin, Igor; Sun, Hongliang; Moshkani, Safiekhatoon; Feng, Changyong; Mantalaris, Athanasios; Wu, J H David; Bottaro, Andrea
2011-06-01
Peripheral lymphoid organs (PLOs), the primary sites of development of adaptive immune responses, display a complex structural organization reflecting separation of cellular subsets (e.g., T and B lymphocytes) and functional compartments which is critical for immune function. The generation of in vitro culture systems capable of recapitulating salient features of PLOs for experimental, biotechnological, and clinical applications would be highly desirable, but has been hampered so far by the complexity of these systems. We have previously developed a three-dimensional bioreactor system for long-term, functional culture of human bone marrow cells on macroporous microspheres in a packed-bed bioreactor with frequent medium change. Here we adapt the same system for culture of human primary cells from PLOs (tonsil) in the absence of specific exogenous growth factors or activators. Cells in this system displayed higher viability over several weeks, and maintain population diversity and cell surface markers largely comparable to primary cells. Light microscopy showed cells organizing in large diverse clusters within the scaffold pores and presence of B cell-enriched areas. Strikingly, these cultures generated a significant number of antibody-producing B cells when challenged with a panel of diverse antigens, as expected from a lymphoid tissue. Thus the three-dimensional tonsil bioreactor culture system may serve as a useful model of PLOs by recapitulating their structural organization and function ex vivo. Copyright © 2011 Wiley Periodicals, Inc.
A Fast Proceduere for Optimizing Thermal Protection Systems of Re-Entry Vehicles
NASA Astrophysics Data System (ADS)
Ferraiuolo, M.; Riccio, A.; Tescione, D.; Gigliotti, M.
The aim of the present work is to introduce a fast procedure to optimize thermal protection systems for re-entry vehicles subjected to high thermal loads. A simplified one-dimensional optimization process, performed in order to find the optimum design variables (lengths, sections etc.), is the first step of the proposed design procedure. Simultaneously, the most suitable materials able to sustain high temperatures and meeting the weight requirements are selected and positioned within the design layout. In this stage of the design procedure, simplified (generalized plane strain) FEM models are used when boundary and geometrical conditions allow the reduction of the degrees of freedom. Those simplified local FEM models can be useful because they are time-saving and very simple to build; they are essentially one dimensional and can be used for optimization processes in order to determine the optimum configuration with regard to weight, temperature and stresses. A triple-layer and a double-layer body, subjected to the same aero-thermal loads, have been optimized to minimize the overall weight. Full two and three-dimensional analyses are performed in order to validate those simplified models. Thermal-structural analyses and optimizations are executed by adopting the Ansys FEM code.
Basel, Matthew T; Balivada, Sivasai; Wang, Hongwang; Shrestha, Tej B; Seo, Gwi Moon; Pyle, Marla; Abayaweera, Gayani; Dani, Raj; Koper, Olga B; Tamura, Masaaki; Chikan, Viktor; Bossmann, Stefan H; Troyer, Deryl L
2012-01-01
Using magnetic nanoparticles to absorb alternating magnetic field energy as a method of generating localized hyperthermia has been shown to be a potential cancer treatment. This report demonstrates a system that uses tumor homing cells to actively carry iron/iron oxide nanoparticles into tumor tissue for alternating magnetic field treatment. Paramagnetic iron/ iron oxide nanoparticles were synthesized and loaded into RAW264.7 cells (mouse monocyte/ macrophage-like cells), which have been shown to be tumor homing cells. A murine model of disseminated peritoneal pancreatic cancer was then generated by intraperitoneal injection of Pan02 cells. After tumor development, monocyte/macrophage-like cells loaded with iron/ iron oxide nanoparticles were injected intraperitoneally and allowed to migrate into the tumor. Three days after injection, mice were exposed to an alternating magnetic field for 20 minutes to cause the cell-delivered nanoparticles to generate heat. This treatment regimen was repeated three times. A survival study demonstrated that this system can significantly increase survival in a murine pancreatic cancer model, with an average post-tumor insertion life expectancy increase of 31%. This system has the potential to become a useful method for specifically and actively delivering nanoparticles for local hyperthermia treatment of cancer.
Haraguchi, Yuji; Kagawa, Yuki; Sakaguchi, Katsuhisa; Matsuura, Katsuhisa; Shimizu, Tatsuya; Okano, Teruo
2017-01-31
In this paper, we report an in vitro co-culture system that combines mammalian cells and algae, Chlorococcum littorale, to create a three-dimensional (3-D) tissue. While the C2C12 mouse myoblasts and rat cardiac cells consumed oxygen actively, intense oxygen production was accounted for by the algae even in the co-culture system. Although cell metabolism within thicker cardiac cell-layered tissues showed anaerobic respiration, the introduction of innovative co-cultivation partially changed the metabolism to aerobic respiration. Moreover, the amount of glucose consumption and lactate production in the cardiac tissues and the amount of ammonia in the culture media decreased significantly when co-cultivated with algae. In the cardiac tissues devoid of algae, delamination was observed histologically, and the release of creatine kinase (CK) from the tissues showed severe cardiac cell damage. On the other hand, the layered cell tissues with algae were observed to be in a good histological condition, with less than one-fifth decline in CK release. The co-cultivation with algae improved the culture condition of the thicker tissues, resulting in the formation of 160 μm-thick cardiac tissues. Thus, the present study proposes the possibility of creating an in vitro "symbiotic recycling system" composed of mammalian cells and algae.
Red blood cell generation by three-dimensional aggregate cultivation of late erythroblasts.
Lee, EunMi; Han, So Yeon; Choi, Hye Sook; Chun, Bokhwan; Hwang, Byunghee; Baek, Eun Jung
2015-02-01
Stem cell-derived erythroid cells hold great potential for the treatment of blood-loss anemia and for erythropoiesis research; however, cultures using conventional flat plates or bioreactors have failed to show promising results. By mimicking the in vivo bone marrow (BM) environment in which most erythroid cells are physically aggregated, we show that a three-dimensional (3D) aggregate culture system facilitates erythroid cell maturation and red blood cell (RBC) production more effectively than two-dimensional high-density cell cultivation. Late erythroblasts (polychromatic or orthochromatic erythroblasts) were differentiated from cord blood CD34(+) cells over 15 days and then allowed to form tight aggregates at a minimum density of 1×10(7) cells/mL for 2-3 days. To scale up the cell culture and to make the media supply efficient throughout the cell aggregates, several macroporous microcarriers and porous scaffolds were applied to the 3D culture system. In comparison to control culture conditions, erythroid cells in 3D aggregates were significantly more differentiated toward RBCs with significantly reduced nuclear dysplasia. When 3D culture was performed inside macroporous microcarriers, the cell culture scale was increased and cells exhibited enhanced differentiation and enucleation. Microcarriers with a pore diameter of approximately 400 μm produced more mature cells than those with a smaller pore diameter. In addition, this aggregate culture method minimized the culture space and media volume required. In conclusion, a 3D aggregate culture system can be used to generate transfusable human erythrocytes at the terminal maturation stage, mimicking the in vivo BM microenvironment. Porous structures can efficiently maximize the culture scale, enabling large-scale production of RBCs. These results enhance our understanding of the importance of physical contact among late erythroblasts for their final maturation into RBCs.
Study of fuel cell on-site, integrated energy systems in residential/commercial applications
NASA Technical Reports Server (NTRS)
Wakefield, R. A.; Karamchetty, S.; Rand, R. H.; Ku, W. S.; Tekumalla, V.
1980-01-01
Three building applications were selected for a detailed study: a low rise apartment building; a retail store, and a hospital. Building design data were then specified for each application, based on the design and construction of typical, actual buildings. Finally, a computerized building loads analysis program was used to estimate hourly end use load profiles for each building. Conventional and fuel cell based energy systems were designed and simulated for each building in each location. Based on the results of a computer simulation of each energy system, levelized annual costs and annual energy consumptions were calculated for all systems.
Rezaei, Mahsa; Abbasi, Alireza; Dinarvand, Rassoul; Jeddi-Tehrani, Mahmood; Janczak, Jan
2018-05-17
Designable coordination polymers with suitable chemical diversities and biocompatible structures have been proposed as a promising class of vehicles for drug delivery systems. Here, we hydrothermally synthesized a novel one-dimensional (1D) coordination polymer, [Zn(H 2 O) 6 K 2 (H 2 BTC) 2 (H 2 O) 4 ](H 2 BTC) 2 ·2H 2 O, where H 3 BTC = benzene-1,3,5-tricarboxylic acid (trimesic acid), cp.1. As the hydrogen bonds stabilized 1D chains in three dimensions, the cp.1 could be a good candidate for delivering small-molecule chemotherapeutics such as 5-fluorouracil (5-Fu). The synthesized cp.1 showed a remarkable 5-Fu loading of 66% with encapsulation efficiency of 98% and almost complete release process. The 5-Fu-loaded cp.1 displayed a time-dependent cytotoxicity effect against breast cancer cell lines MCF-7 and 4T1. The cellular uptake of cp.1 particles was investigated via confocal laser scanning microscopy using fluorescein isothiocyanate and LysoTracker Red staining. Furthermore, the in vivo antitumor impact of 5-Fu-loaded cp.1 was studied on 4T1 breast cancer BALB/c mice model. The intratumor treatment of 5-Fu-loaded cp.1 demonstrated beneficial antitumor efficacy by postponing tumor growth. These results suggest that the 5-Fu-loaded cp.1 microparticles with a great locoregional delivery can be efficient anticancer drug carriers for further clinical treatments.
Equations of motion of slung load systems with results for dual lift
NASA Technical Reports Server (NTRS)
Cicolani, Luigi S.; Kanning, Gerd
1990-01-01
General simulation equations are derived for the rigid body motion of slung load systems. These systems are viewed as consisting of several rigid bodies connected by straight-line cables or links. The suspension can be assumed to be elastic or inelastic, both cases being of interest in simulation and control studies. Equations for the general system are obtained via D'Alembert's principle and the introduction of generalized velocity coordinates. Three forms are obtained. Two of these generalize previous case-specific results for single helicopter systems with elastic or inelastic suspensions. The third is a new formulation for inelastic suspensions. It is derived from the elastic suspension equations by choosing the generalized coordinates so as to separate motion due to cable stretching from motion with invariant cable lengths. The result is computationally more efficient than the conventional formulation, and is readily integrated with the elastic suspension formulation and readily applied to the complex dual lift and multilift systems. Equations are derived for dual lift systems. Three proposed suspension arrangements can be integrated in a single equation set. The equations are given in terms of the natural vectors and matrices of three-dimensional rigid body mechanics and are tractable for both analysis and programming.
ERIC Educational Resources Information Center
Chen, Chwen Jen; Fauzy Wan Ismail, Wan Mohd
2008-01-01
The real-time interactive nature of three-dimensional virtual environments (VEs) makes this technology very appropriate for exploratory learning purposes. However, many studies have shown that the exploration process may cause cognitive overload that affects the learning of domain knowledge. This article reports a quasi-experimental study that…
NASA Astrophysics Data System (ADS)
Kiani, Keivan
2014-06-01
Novel nonlocal discrete and continuous models are proposed for dynamic analysis of two- and three-dimensional ensembles of single-walled carbon nanotubes (SWCNTs). The generated extra van der Waals forces between adjacent SWCNTs due to their lateral motions are evaluated via Lennard-Jones potential function. Using a nonlocal Rayleigh beam model, the discrete and continuous models are developed for both two- and three-dimensional ensembles of SWCNTs acted upon by transverse dynamic loads. The capabilities of the proposed continuous models in capturing the vibration behavior of SWCNTs ensembles are then examined through various numerical simulations. A reasonably good agreement between the results of the continuous models and those of the discrete ones is also reported. The effects of the applied load frequency, intertube spaces, and small-scale parameter on the transverse dynamic responses of both two- and three-dimensional ensembles of SWCNTs are explained. The proposed continuous models would be very useful for dynamic analyses of large populated ensembles of SWCNTs whose discrete models suffer from both computational efforts and labor costs.
Shih, Wenting; Yamada, Soichiro
2011-12-22
Traditionally, cell migration has been studied on two-dimensional, stiff plastic surfaces. However, during important biological processes such as wound healing, tissue regeneration, and cancer metastasis, cells must navigate through complex, three-dimensional extracellular tissue. To better understand the mechanisms behind these biological processes, it is important to examine the roles of the proteins responsible for driving cell migration. Here, we outline a protocol to study the mechanisms of cell migration using the epithelial cell line (MDCK), and a three-dimensional, fibrous, self-polymerizing matrix as a model system. This optically clear extracellular matrix is easily amenable to live-cell imaging studies and better mimics the physiological, soft tissue environment. This report demonstrates a technique for directly visualizing protein localization and dynamics, and deformation of the surrounding three-dimensional matrix. Examination of protein localization and dynamics during cellular processes provides key insight into protein functions. Genetically encoded fluorescent tags provide a unique method for observing protein localization and dynamics. Using this technique, we can analyze the subcellular accumulation of key, force-generating cytoskeletal components in real-time as the cell maneuvers through the matrix. In addition, using multiple fluorescent tags with different wavelengths, we can examine the localization of multiple proteins simultaneously, thus allowing us to test, for example, whether different proteins have similar or divergent roles. Furthermore, the dynamics of fluorescently tagged proteins can be quantified using Fluorescent Recovery After Photobleaching (FRAP) analysis. This measurement assays the protein mobility and how stably bound the proteins are to the cytoskeletal network. By combining live-cell imaging with the treatment of protein function inhibitors, we can examine in real-time the changes in the distribution of proteins and morphology of migrating cells. Furthermore, we also combine live-cell imaging with the use of fluorescent tracer particles embedded within the matrix to visualize the matrix deformation during cell migration. Thus, we can visualize how a migrating cell distributes force-generating proteins, and where the traction forces are exerted to the surrounding matrix. Through these techniques, we can gain valuable insight into the roles of specific proteins and their contributions to the mechanisms of cell migration.
1993-08-01
measure the inherent fracture toughness of a material. A thor- ough understanding of the test specimen behavior is a prerequisite to the application of...measured material properties in structural applications . Three- dimensional dynamic analyses are performed for three different specimen configurations...derstanding of the test specimen behavior is a prerequisite to the application of measured ma- terial properties in structural applications . Three
Dynamic analysis of geometrically non-linear three-dimensional beams under moving mass
NASA Astrophysics Data System (ADS)
Zupan, E.; Zupan, D.
2018-01-01
In this paper, we present a coupled dynamic analysis of a moving particle on a deformable three-dimensional frame. The presented numerical model is capable of considering arbitrary curved and twisted initial geometry of the beam and takes into account geometric non-linearity of the structure. Coupled with dynamic equations of the structure, the equations of moving particle are solved. The moving particle represents the dynamic load and varies the mass distribution of the structure and at the same time its path is adapting due to deformability of the structure. A coupled geometrically non-linear behaviour of beam and particle is studied. The equation of motion of the particle is added to the system of the beam dynamic equations and an additional unknown representing the coordinate of the curvilinear path of the particle is introduced. The specially designed finite-element formulation of the three-dimensional beam based on the weak form of consistency conditions is employed where only the boundary conditions are affected by the contact forces.
NASA Technical Reports Server (NTRS)
Shivakumar, K. N.; Newman, J. C., Jr.
1992-01-01
A three dimensional stress concentration analysis was conducted on straight shank and countersunk (rivet) holes in a large plate subjected to various loading conditions. Three dimensional finite element analysis were performed with 20 node isoparametric elements. The plate material was assumed to be linear elastic and isotropic, with a Poisson ratio of 0.3. Stress concentration along the bore of the hole were computed for several ratios of hole radius to plate thickness (0.1 to 2.5) and ratios of countersink depth to plate thickness (0.25 to 1). The countersink angles were varied from 80 to 100 degrees in some typical cases, but the angle was held constant at 100 degrees for most cases. For straight shank holes, three types of loading were considered: remote tension, remote bending, and wedge loading in the hole. Results for remote tension and wedge loading were used to estimate stress concentration for simulated rivet in pin loading. For countersunk holes only remote tension and bending were considered. Based on the finite element results, stress concentration equations were developed. Whenever possible, the present results were compared with other numerical solutions and experimental results from the literature.
Rizvi, Mohd Suhail; Pal, Anupam
2014-09-01
The fibrous matrices are widely used as scaffolds for the regeneration of load-bearing tissues due to their structural and mechanical similarities with the fibrous components of the extracellular matrix. These scaffolds not only provide the appropriate microenvironment for the residing cells but also act as medium for the transmission of the mechanical stimuli, essential for the tissue regeneration, from macroscopic scale of the scaffolds to the microscopic scale of cells. The requirement of the mechanical loading for the tissue regeneration requires the fibrous scaffolds to be able to sustain the complex three-dimensional mechanical loading conditions. In order to gain insight into the mechanical behavior of the fibrous matrices under large amount of elongation as well as shear, a statistical model has been formulated to study the macroscopic mechanical behavior of the electrospun fibrous matrix and the transmission of the mechanical stimuli from scaffolds to the cells via the constituting fibers. The study establishes the load-deformation relationships for the fibrous matrices for different structural parameters. It also quantifies the changes in the fiber arrangement and tension generated in the fibers with the deformation of the matrix. The model reveals that the tension generated in the fibers on matrix deformation is not homogeneous and hence the cells located in different regions of the fibrous scaffold might experience different mechanical stimuli. The mechanical response of fibrous matrices was also found to be dependent on the aspect ratio of the matrix. Therefore, the model establishes a structure-mechanics interdependence of the fibrous matrices under large deformation, which can be utilized in identifying the appropriate structure and external mechanical loading conditions for the regeneration of load-bearing tissues. Copyright © 2014 Elsevier Ltd. All rights reserved.
Load measurement system with load cell lock-out mechanism
NASA Technical Reports Server (NTRS)
Le, Thang; Carroll, Monty; Liu, Jonathan
1995-01-01
In the frame work of the project Shuttle Plume Impingement Flight Experiment (SPIFEX), a Load Measurement System was developed and fabricated to measure the impingement force of Shuttle Reaction Control System (RCS) jets. The Load Measurement System is a force sensing system that measures any combination of normal and shear forces up to 40 N (9 lbf) in the normal direction and 22 N (5 lbf) in the shear direction with an accuracy of +/- 0.04 N (+/- 0.01 lbf) Since high resolution is required for the force measurement, the Load Measurement System is built with highly sensitive load cells. To protect these fragile load cells in the non-operational mode from being damaged due to flight loads such as launch and landing loads of the Shuttle vehicle, a motor driven device known as the Load Cell Lock-Out Mechanism was built. This Lock-Out Mechanism isolates the load cells from flight loads and re-engages the load cells for the force measurement experiment once in space. With this highly effective protection system, the SPIFEX load measurement experiment was successfully conducted on STS-44 in September 1994 with all load cells operating properly and reading impingement forces as expected.
NASA Technical Reports Server (NTRS)
Markey, Melvin F.
1959-01-01
A theory is derived for determining the loads and motions of a deeply immersed prismatic body. The method makes use of a two-dimensional water-mass variation and an aspect-ratio correction for three-dimensional flow. The equations of motion are generalized by using a mean value of the aspect-ratio correction and by assuming a variation of the two-dimensional water mass for the deeply immersed body. These equations lead to impact coefficients that depend on an approach parameter which, in turn, depends upon the initial trim and flight-path angles. Comparison of experiment with theory is shown at maximum load and maximum penetration for the flat-bottom (0 deg dead-rise angle) model with bean-loading coefficients from 36.5 to 133.7 over a wide range of initial conditions. A dead-rise angle correction is applied and maximum-load data are compared with theory for the case of a model with 300 dead-rise angle and beam-loading coefficients from 208 to 530.
Edge delamination of composite laminates subject to combined tension and torsional loading
NASA Technical Reports Server (NTRS)
Hooper, Steven J.
1990-01-01
Delamination is a common failure mode of laminated composite materials. Edge delamination is important since it results in reduced stiffness and strength of the laminate. The tension/torsion load condition is of particular significance to the structural integrity of composite helicopter rotor systems. Material coupons can easily be tested under this type of loading in servo-hydraulic tension/torsion test stands using techniques very similar to those used for the Edge Delamination Tensile Test (EDT) delamination specimen. Edge delamination of specimens loaded in tension was successfully analyzed by several investigators using both classical laminate theory and quasi-three dimensional (Q3D) finite element techniques. The former analysis technique can be used to predict the total strain energy release rate, while the latter technique enables the calculation of the mixed-mode strain energy release rates. The Q3D analysis is very efficient since it produces a three-dimensional solution to a two-dimensional domain. A computer program was developed which generates PATRAN commands to generate the finite element model. PATRAN is a pre- and post-processor which is commonly used with a variety of finite element programs such as MCS/NASTRAN. The program creates a sufficiently dense mesh at the delamination crack tips to support a mixed-mode fracture mechanics analysis. The program creates a coarse mesh in those regions where the gradients in the stress field are low (away from the delamination regions). A transition mesh is defined between these regions. This program is capable of generating a mesh for an arbitrarily oriented matrix crack. This program significantly reduces the modeling time required to generate these finite element meshes, thus providing a realistic tool with which to investigate the tension torsion problem.
Panadero, J A; Lanceros-Mendez, S; Ribelles, J L Gomez
2016-03-01
Chondrogenesis of dedifferentiated chondrocytes and mesenchymal stem cells is influenced not only by soluble molecules like growth factors, but also by the cell environment itself. The latter is achieved through both mechanical cues - which act as stimulation factor and influences nutrient transport - and adhesion to extracellular matrix cues - which determine cell shape. Although the effects of soluble molecules and cell environment have been intensively addressed, few observations and conclusions about the interaction between the two have been achieved. In this work, we review the state of the art on the single effects between mechanical and biochemical cues, as well as on the combination of the two. Furthermore, we provide a discussion on the techniques currently used to determine the mechanical properties of materials and tissues generated in vitro, their limitations and the future research needs to properly address the identified problems. The importance of biomechanical cues in chondrogenesis is well known. This paper reviews the existing literature on the effect of mechanical stimulation on chondrogenic differentiation of mesenchymal stem cells in order to regenerate hyaline cartilage. Contradictory results found with respect to the effect of different modes of external loading can be explained by the different properties of the scaffolding system that holds the cells, which determine cell adhesion and morphology and spatial distribution of cells, as well as the stress transmission to the cells. Thus, this review seeks to provide an insight into the interplay between external loading program and scaffold properties during chondrogenic differentiation. The review of the literature reveals an important gap in the knowledge in this field and encourages new experimental studies. The main issue is that in each of the few cases in which the interplay is investigated, just two groups of scaffolds are compared, leaving intermediate adhesion conditions out of study. The authors propose broader studies implementing new high-throughput techniques for mechanical characterization of tissue engineering constructs and the inclusion of fatigue analysis as support methodology to more exhaustive mechanical characterization. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
Multi-physics optimization of three-dimensional microvascular polymeric components
NASA Astrophysics Data System (ADS)
Aragón, Alejandro M.; Saksena, Rajat; Kozola, Brian D.; Geubelle, Philippe H.; Christensen, Kenneth T.; White, Scott R.
2013-01-01
This work discusses the computational design of microvascular polymeric materials, which aim at mimicking the behavior found in some living organisms that contain a vascular system. The optimization of the topology of the embedded three-dimensional microvascular network is carried out by coupling a multi-objective constrained genetic algorithm with a finite-element based physics solver, the latter validated through experiments. The optimization is carried out on multiple conflicting objective functions, namely the void volume fraction left by the network, the energy required to drive the fluid through the network and the maximum temperature when the material is subjected to thermal loads. The methodology presented in this work results in a viable alternative for the multi-physics optimization of these materials for active-cooling applications.
Quantitative characterization of 3D deformations of cell interactions with soft biomaterials
NASA Astrophysics Data System (ADS)
Franck, Christian
In recent years, the importance of mechanical forces in directing cellular function has been recognized as a significant factor in biological and physiological processes. In fact, these physical forces are now viewed equally as important as biochemical stimuli in controlling cellular response. Not only do these cellular forces, or cell tractions, play an important role in cell migration, they are also significant to many other physiological and pathological processes, both at the tissue and organ level, including wound healing, inflammation, angiogenesis, and embryogenesis. A complete quantification of cell tractions during cell-material interactions can lead to a deeper understanding of the fundamental role these forces play in cell biology. Thus, understanding the function and role of a cell from a mechanical framework can have important implications towards the development of new implant materials and drug treatments. Previous research has contributed significant descriptions of cell-tissue interactions by quantifying cell tractions in two-dimensional environments; however, most physiological processes are three-dimensional in nature. Recent studies have shown morphological differences in cells cultured on two-dimensional substrates versus three-dimensional matrices, and that the intrinsic extracellular matrix interactions and migration behavior are different in three dimensions versus two dimensions. Hence, measurement techniques are needed to investigate cellular behavior in all three dimensions. This thesis presents a full-field imaging technique capable of quantitatively measuring cell traction forces in all three spatial dimensions, and hence addresses the need of a three-dimensional quantitative imaging technique to gain insight into the fundamental role of physical forces in biological processes. The technique combines laser scanning confocal microscopy (LSCM) with digital volume correlation (DVC) to track the motion of fluorescent particles during cell-induced or externally applied deformations. This method is validated by comparing experimentally measured non-uniform deformation fields near hard and soft spherical inclusions under uniaxial compression with the corresponding analytical solution. Utilization of a newly developed computationally efficient stretch-correlation and deconvolution algorithm is shown to improve the overall measurement accuracy, in particular under large deformations. Using this technique, the full three-dimensional substrate displacement fields are experimentally determined during the migration of individual fibroblast cells on polyacrylamide gels. This is the first study to show the highly three-dimensional structure of cell-induced displacement and traction fields. These new findings suggest a three-dimensional push-pull cell motility, which differs from the traditional theories based on two-dimensional data. These results provide new insight into the dynamic cell-matrix force exchange or mechanotransduction of migrating cells, and will aid in the development of new three-dimensional cell motility and adhesion models. As this study reveals, the mechanical interactions of cells and their extracellular matrix appear to be highly three-dimensional. It also shows that the LSCM-DVC technique is well suited for investigating the mechanics of cell-matrix interactions while providing a platform to access detailed information of the intricate biomechanical coupling for many cellular responses. Thus, this method has the capability to provide direct quantitative experimental data showing how cells interact with their surroundings in three dimensions and might stimulate new avenues of scientific thought in understanding the fundamental role physical forces play in regulating cell behavior.
NASA Astrophysics Data System (ADS)
Braun, Robert Joseph
The advent of maturing fuel cell technologies presents an opportunity to achieve significant improvements in energy conversion efficiencies at many scales; thereby, simultaneously extending our finite resources and reducing "harmful" energy-related emissions to levels well below that of near-future regulatory standards. However, before realization of the advantages of fuel cells can take place, systems-level design issues regarding their application must be addressed. Using modeling and simulation, the present work offers optimal system design and operation strategies for stationary solid oxide fuel cell systems applied to single-family detached dwellings. A one-dimensional, steady-state finite-difference model of a solid oxide fuel cell (SOFC) is generated and verified against other mathematical SOFC models in the literature. Fuel cell system balance-of-plant components and costs are also modeled and used to provide an estimate of system capital and life cycle costs. The models are used to evaluate optimal cell-stack power output, the impact of cell operating and design parameters, fuel type, thermal energy recovery, system process design, and operating strategy on overall system energetic and economic performance. Optimal cell design voltage, fuel utilization, and operating temperature parameters are found using minimization of the life cycle costs. System design evaluations reveal that hydrogen-fueled SOFC systems demonstrate lower system efficiencies than methane-fueled systems. The use of recycled cell exhaust gases in process design in the stack periphery are found to produce the highest system electric and cogeneration efficiencies while achieving the lowest capital costs. Annual simulations reveal that efficiencies of 45% electric (LHV basis), 85% cogenerative, and simple economic paybacks of 5--8 years are feasible for 1--2 kW SOFC systems in residential-scale applications. Design guidelines that offer additional suggestions related to fuel cell-stack sizing and operating strategy (base-load or load-following and cogeneration or electric-only) are also presented.
NASA Astrophysics Data System (ADS)
Song, Huimin
In the aerospace and automotive industries, many finite element analyses use lower-dimensional finite elements such as beams, plates and shells, to simplify the modeling. These simplified models can greatly reduce the computation time and cost; however, reduced-dimensional models may introduce inaccuracies, particularly near boundaries and near portions of the structure where reduced-dimensional models may not apply. Another factor in creation of such models is that beam-like structures frequently have complex geometry, boundaries and loading conditions, which may make them unsuitable for modeling with single type of element. The goal of this dissertation is to develop a method that can accurately and efficiently capture the response of a structure by rigorous combination of a reduced-dimensional beam finite element model with a model based on full two-dimensional (2D) or three-dimensional (3D) finite elements. The first chapter of the thesis gives the background of the present work and some related previous work. The second chapter is focused on formulating a system of equations that govern the joining of a 2D model with a beam model for planar deformation. The essential aspect of this formulation is to find the transformation matrices to achieve deflection and load continuity on the interface. Three approaches are provided to obtain the transformation matrices. An example based on joining a beam to a 2D finite element model is examined, and the accuracy of the analysis is studied by comparing joint results with the full 2D analysis. The third chapter is focused on formulating the system of equations for joining a beam to a 3D finite element model for static and free-vibration problems. The transition between the 3D elements and beam elements is achieved by use of the stress recovery technique of the variational-asymptotic method as implemented in VABS (the Variational Asymptotic Beam Section analysis). The formulations for an interface transformation matrix and the generalized Timoshenko beam are discussed in this chapter. VABS is also used to obtain the beam constitutive properties and warping functions for stress recovery. Several 3D-beam joint examples are presented to show the convergence and accuracy of the analysis. Accuracy is accessed by comparing the joint results with the full 3D analysis. The fourth chapter provides conclusions from present studies and recommendations for future work.
Dagdeviren, Omur E
2018-08-03
The effect of surface disorder, load, and velocity on friction between a single asperity contact and a model surface is explored with one-dimensional and two-dimensional Prandtl-Tomlinson (PT) models. We show that there are fundamental physical differences between the predictions of one-dimensional and two-dimensional models. The one-dimensional model estimates a monotonic increase in friction and energy dissipation with load, velocity, and surface disorder. However, a two-dimensional PT model, which is expected to approximate a tip-sample system more realistically, reveals a non-monotonic trend, i.e. friction is inert to surface disorder and roughness in wearless friction regime. The two-dimensional model discloses that the surface disorder starts to dominate the friction and energy dissipation when the tip and the sample interact predominantly deep into the repulsive regime. Our numerical calculations address that tracking the minimum energy path and the slip-stick motion are two competing effects that determine the load, velocity, and surface disorder dependence of friction. In the two-dimensional model, the single asperity can follow the minimum energy path in wearless regime; however, with increasing load and sliding velocity, the slip-stick movement dominates the dynamic motion and results in an increase in friction by impeding tracing the minimum energy path. Contrary to the two-dimensional model, when the one-dimensional PT model is employed, the single asperity cannot escape to the minimum energy minimum due to constraint motion and reveals only a trivial dependence of friction on load, velocity, and surface disorder. Our computational analyses clarify the physical differences between the predictions of the one-dimensional and two-dimensional models and open new avenues for disordered surfaces for low energy dissipation applications in wearless friction regime.
Quantification of collagen contraction in three-dimensional cell culture.
Kopanska, Katarzyna S; Bussonnier, Matthias; Geraldo, Sara; Simon, Anthony; Vignjevic, Danijela; Betz, Timo
2015-01-01
Many different cell types including fibroblasts, smooth muscle cells, endothelial cells, and cancer cells exert traction forces on the fibrous components of the extracellular matrix. This can be observed as matrix contraction both macro- and microscopically in three-dimensional (3D) tissues models such as collagen type I gels. The quantification of local contraction at the micron scale, including its directionality and speed, in correlation with other parameters such as cell invasion, local protein or gene expression, can provide useful information to study wound healing, organism development, and cancer metastasis. In this article, we present a set of tools to quantify the flow dynamics of collagen contraction, induced by cells migrating out of a multicellular cancer spheroid into a three-dimensional (3D) collagen matrix. We adapted a pseudo-speckle technique that can be applied to bright-field and fluorescent microscopy time series. The image analysis presented here is based on an in-house written software developed in the Matlab (Mathworks) programming environment. The analysis program is freely available from GitHub following the link: http://dx.doi.org/10.5281/zenodo.10116. This tool provides an automatized technique to measure collagen contraction that can be utilized in different 3D cellular systems. Copyright © 2015 Elsevier Inc. All rights reserved.
NASA Technical Reports Server (NTRS)
Torgan, C. E.; Burge, S. S.; Collinsworth, A. M.; Truskey, G. A.; Kraus, W. E.
2000-01-01
The growth and repair of adult skeletal muscle are due in part to activation of muscle precursor cells, commonly known as satellite cells or myoblasts. These cells are responsive to a variety of environmental cues, including mechanical stimuli. The overall goal of the research is to examine the role of mechanical signalling mechanisms in muscle growth and plasticity through utilisation of cell culture systems where other potential signalling pathways (i.e. chemical and electrical stimuli) are controlled. To explore the effects of decreased mechanical loading on muscle differentiation, mammalian myoblasts are cultured in a bioreactor (rotating cell culture system), a model that has been utilised to simulate microgravity. C2C12 murine myoblasts are cultured on microcarrier beads in a bioreactor and followed throughout differentiation as they form a network of multinucleated myotubes. In comparison with three-dimensional control cultures that consist of myoblasts cultured on microcarrier beads in teflon bags, myoblasts cultured in the bioreactor exhibit an attenuation in differentiation. This is demonstrated by reduced immunohistochemical staining for myogenin and alpha-actinin. Western analysis shows a decrease, in bioreactor cultures compared with control cultures, in levels of the contractile proteins myosin (47% decrease, p < 0.01) and tropomyosin (63% decrease, p < 0.01). Hydrodynamic measurements indicate that the decrease in differentiation may be due, at least in part, to fluid stresses acting on the myotubes. In addition, constraints on aggregate size imposed by the action of fluid forces in the bioreactor affect differentiation. These results may have implications for muscle growth and repair during spaceflight.
Ardakani, Amir G; Cheema, Umber; Brown, Robert A; Shipley, Rebecca J
2014-09-06
A challenge in three-dimensional tissue culture remains the lack of quantitative information linking nutrient delivery and cellular distribution. Both in vivo and in vitro, oxygen is delivered by diffusion from its source (blood vessel or the construct margins). The oxygen level at a defined distance from its source depends critically on the balance of diffusion and cellular metabolism. Cells may respond to this oxygen environment through proliferation, death and chemotaxis, resulting in spatially resolved gradients in cellular density. This study extracts novel spatially resolved and simultaneous data on tissue oxygenation, cellular proliferation, viability and chemotaxis in three-dimensional spiralled, cellular collagen constructs. Oxygen concentration gradients drove preferential cellular proliferation rates and viability in the higher oxygen zones and induced chemotaxis along the spiral of the collagen construct; an oxygen gradient of 1.03 mmHg mm(-1) in the spiral direction induced a mean migratory speed of 1015 μm day(-1). Although this movement was modest, it was effective in balancing the system to a stable cell density distribution, and provided insights into the natural cell mechanism for adapting cell number and activity to a prevailing oxygen regime.
Analysis of No-load Iron Losses of Turbine Generators by 3D Magnetic Field Analysis
NASA Astrophysics Data System (ADS)
Nakahara, Akihito; Mogi, Hisashi; Takahashi, Kazuhiko; Ide, Kazumasa; Kaneda, Junya; Hattori, Ken'Ichi; Watanabe, Takashi; Kaido, Chikara; Minematsu, Eisuke; Hanzawa, Kazufumi
This paper focuses on no-load iron losses of turbine generators. To calculate iron losses of turbine generators a program was developed. In the program, core loss curves of materials used for stator core were reproduced precisely by using tables of loss coefficients. Accuracy of calculation by this method was confirmed by comparing calculated values with measured in a model stator core. The iron loss of a turbine generator estimated with considering three-dimensional distribution of magnetic fluxes. And additional losses included in measured iron loss was evaluated with three-dimensional magnetic field analysis.
Cultured High-Fidelity Three-Dimensional Human Urogenital Tract Carcinomas and Process
NASA Technical Reports Server (NTRS)
Goodwin, Thomas J. (Inventor); Prewett, Tacey L. (Inventor); Spaulding, Glenn F. (Inventor); Wolf, David A. (Inventor)
1998-01-01
Artificial high-fidelity three-dimensional human urogenital tract carcinomas are propagated under in vitro-microgravity conditions from carcinoma cells. Artificial high-fidelity three-dimensional human urogenital tract carcinomas are also propagated from a coculture of normal urogenital tract cells inoculated with carcinoma cells. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel.
Landarani-Isfahani, Amir; Moghadam, Majid; Mohammadi, Shima; Royvaran, Maryam; Moshtael-Arani, Naimeh; Rezaei, Saghar; Tangestaninejad, Shahram; Mirkhani, Valiollah; Mohammadpoor-Baltork, Iraj
2017-08-29
Owing to properties of magnetic nanoparticles and elegant three-dimensional macromolecule architectural features, dendrimeric structures have been investigated as nanoscale drug delivery systems. In this work, a novel magnetic nanocarrier, generation two (G2) triazine dendrimer modified Fe 3 O 4 @SiO 2 magnetic nanoparticles (MNP-G2), was designed, fabricated, and characterized by Fourier transform infrared (FT-IR), thermal gravimetric analysis (TGA), vibrating sample magnetometer (VSM), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), and dynamic light scattering (DLS). The prepared MNP-G2 nanosystem offers a new formulation that combines the unique properties of MNPs and triazine dendrimer as a biocompatible material for biomedical applications. To demonstrate the potential of MNP-G2, the nanoparticles were loaded with methotrexate (MTX), a proven chemotherapy drug. The MTX-loaded MNP-G2 (MNP-G2/MTX) exhibited a high drug-loading capacity of MTX and the excellent ability for controlled drug release. The cytotoxicity of MNP-G2/MTX using an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide based assay and MCF-7, HeLa, and Caov-4 cell lines revealed that MNP-G2/MTX was more active against the tumor cells than the free drug in a mildly acidic environment. The results of hemolysis, hemagglutination, and coagulation assays confirmed the good blood safety of MNP-G2/MTX. Moreover, the cell uptake and intracellular distribution of MNP-G2/MTX were studied by flow cytometry analysis and confocal laser scanning microscopy (CLSM). This research suggests that MNP-G2/MTX with good biocompatibility and degradability can be selected as an ideal and effective drug carrier in targeted biomedicine studies especially anticancer applications.
NASA Technical Reports Server (NTRS)
Norton, J. M.; Tari, U.; Weber, R. M.
1979-01-01
A quasi three dimensional design system and multiple-circular-arc airfoil sections were used to design a fan rotor. An axisymmetric intrablade flow field calculation modeled the shroud of an isolated splitter and radial distribution. The structural analysis indicates that the design is satisfactory for evaluation of aerodynamic performance of the fan stage in a test facility.
Yang, Xu; Vezeridis, Peter S; Nicholas, Brian; Crisco, Joseph J; Moore, Douglas C; Chen, Qian
2006-01-01
Objective Mechanical loading of cartilage influences chondrocyte metabolism and gene expression. The gene encoding type X collagen is expressed specifically by hypertrophic chondrocytes and up regulated during osteoarthritis. In this study we tested the hypothesis that the mechanical microenvironment resulting from higher levels of local strain in a three dimensional cell culture construct would lead to an increase in the expression of type X collagen mRNA by chondrocytes in those areas. Methods Hypertrophic chondrocytes were isolated from embryonic chick sterna and seeded onto rectangular Gelfoam sponges. Seeded sponges were subjected to various levels of cyclic uniaxial tensile strains at 1 Hz with the computer-controlled Bio-Stretch system. Strain distribution across the sponge was quantified by digital image analysis. After mechanical loading, sponges were cut and the end and center regions were separated according to construct strain distribution. Total RNA was extracted from the cells harvested from these regions, and real-time quantitative RT-PCR was performed to quantify mRNA levels for type X collagen and a housing-keeping gene 18S RNA. Results Chondrocytes distributed in high (9%) local strain areas produced more than two times type X collagen mRNA compared to the those under no load conditions, while chondrocytes located in low (2.5%) local strain areas had no appreciable difference in type X collagen mRNA production in comparison to non-loaded samples. Increasing local strains above 2.5%, either in the center or end regions of the sponge, resulted in increased expression of Col X mRNA by chondrocytes in that region. Conclusion These findings suggest that the threshold of chondrocyte sensitivity to inducing type X collagen mRNA production is more than 2.5% local strain, and that increased local strains above the threshold results in an increase of Col X mRNA expression. Such quantitative analysis has important implications for our understanding of mechanosensitivity of cartilage and mechanical regulation of chondrocyte gene expression. PMID:17150098
Performance of mid infrared spectroscopy in skin cancer cell type identification
NASA Astrophysics Data System (ADS)
Kastl, Lena; Kemper, Björn; Lloyd, Gavin R.; Nallala, Jayakrupakar; Stone, Nick; Naranjo, Valery; Penaranda, Francisco; Schnekenburger, Jürgen
2017-02-01
Marker free optical spectroscopy is a powerful tool for the rapid inspection of pathologically suspicious skin lesions and the non-invasive detection of early skin tumors. This goal can be reached by the combination of signal localization and the spectroscopical detection of chemical cell signatures. We here present the development and application of mid infrared spectroscopy (midIR) for the analysis of skin tumor cell types and three dimensional tissue phantoms towards the application of midIR spectroscopy for fast and reliable skin diagnostics. We developed standardized in vitro skin systems with increasing complexity, from single skin cell types as fibroblasts, keratinocytes and melanoma cells, to mixtures of these and finally three dimensional skin cancer phantoms. The cell systems were characterized with different systems in the midIR range up to 12 μm. The analysis of the spectra by novel data processing algorithms demonstrated the clear separation of all cell types, especially melanoma cells. Special attention and algorithm training was required for closely related mesenchymal cell types as dedifferentiated melanoma cells and fibroblasts. Proof of concept experiments with mixtures of in vivo fluorescence labelled skin cell types allowed the test of the new algorithms performance for the identification of specific cell types. The intense training of the software systems with various samples resulted in a increased sensitivity and specificity of the combined midIR and software system. These data highlight the potential of midIR spectroscopy as sensitive and specific future optical biopsy technology.
NASA Technical Reports Server (NTRS)
Schweikhard, W. G.; Singnoi, W. N.
1985-01-01
A two axis thrust measuring system was analyzed by using a finite a element computer program to determine the sensitivities of the thrust vectoring nozzle system to misalignment of the load cells and applied loads, and the stiffness of the structural members. Three models were evaluated: (1) the basic measuring element and its internal calibration load cells; (2) the basic measuring element and its external load calibration equipment; and (3) the basic measuring element, external calibration load frame and the altitude facility support structure. Alignment of calibration loads was the greatest source of error for multiaxis thrust measuring systems. Uniform increases or decreases in stiffness of the members, which might be caused by the selection of the materials, have little effect on the accuracy of the measurements. It is found that the POLO-FINITE program is a viable tool for designing and analyzing multiaxis thrust measurement systems. The response of the test stand to step inputs that might be encountered with thrust vectoring tests was determined. The dynamic analysis show a potential problem for measuring the dynamic response characteristics of thrust vectoring systems because of the inherently light damping of the test stand.
A 3D tension bioreactor platform to study the interplay between ECM stiffness and tumor phenotype.
Cassereau, Luke; Miroshnikova, Yekaterina A; Ou, Guanqing; Lakins, Johnathon; Weaver, Valerie M
2015-01-10
Extracellular matrix (ECM) structure, composition, and stiffness have profound effects on tissue development and pathologies such as cardiovascular disease and cancer. Accordingly, a variety of synthetic hydrogel systems have been designed to study the impact of ECM composition, density, mechanics, and topography on cell and tissue phenotype. However, these synthetic systems fail to accurately recapitulate the biological properties and structure of the native tissue ECM. Natural three dimensional (3D) ECM hydrogels, such as collagen or hyaluronic acid, feature many of the chemical and physical properties of tissue, yet, these systems have limitations including the inability to independently control biophysical properties such as stiffness and pore size. Here, we present a 3D tension bioreactor system that permits precise mechanical tuning of collagen hydrogel stiffness, while maintaining consistent composition and pore size. We achieve this by mechanically loading collagen hydrogels covalently-conjugated to a polydimethylsiloxane (PDMS) membrane to induce hydrogel stiffening. We validated the biological application of this system with oncogenically transformed mammary epithelial cell organoids embedded in a 3D collagen I hydrogel, either uniformly stiffened or calibrated to create a gradient of ECM stiffening, to visually demonstrate the impact of ECM stiffening on transformation and tumor cell invasion. As such, this bioreactor presents the first tunable 3D natural hydrogel system that is capable of independently assessing the role of ECM stiffness on tissue phenotype. Copyright © 2014 Elsevier B.V. All rights reserved.
Yang, Suixing; Feng, Jing; Zhang, Zuo; Qu, Aili; Gong, Miao; Tang, Jie; Fan, Junheng; Li, Songqing; Zhao, Yanling
2013-04-01
To construct a three-dimensional finite element model of the upper airway and adjacent structure of an obstructive sleep apnea hypopnea syndrome (OSAHS) patient for biomechanical analysis. And to study the influence of glossopharyngeum of an OSAHS patient with three-dimensional finite element model during titrated mandible advancement. DICOM format image information of an OSAHS patient's upper airway was obtained by thin-section CT scanning and digital image processing were utilized to construct a three-dimensional finite element model by Mimics 10.0, Imageware 10.0 and Ansys software. The changes and the law of glossopharyngeum were observed by biomechanics and morphology after loading with titrated mandible advancement. A three-dimensional finite element model of the adjacent upper airway structure of OSAHS was established successfully. After loading, the transverse diameter of epiglottis tip of glossopharyngeum increased significantly, although the sagittal diameter decreased correspondingly. The principal stress was mainly distributed in anterior wall of the upper airway. The location of principal stress concentration did not change significantly with the increasing of distance. The stress of glossopharyngeum increased during titrated mandible advancement. A more precise three-dimensional finite model of upper airway and adjacent structure of an OSAHS patient is established and improved efficiency by Mimics, Imageware and Ansys software. The glossopharyngeum of finite element model of OSAHS is analyzed by titrated mandible advancement and can effectively show the relationship between mandible advancement and the glossopharyngeum.
Volumetric Stress-Strain Analysis of Optohydrodynamically Suspended Biological Cells
Liang, Yu; Saha, Asit K.
2011-01-01
Ongoing investigations are exploring the biomechanical properties of isolated and suspended biological cells in pursuit of understanding single-cell mechanobiology. An optical tweezer with minimal applied laser power has positioned biologic cells at the geometric center of a microfluidic cross-junction, creating a novel optohydrodynamic trap. The resulting fluid flow environment facilitates unique multiaxial loading of single cells with site-specific normal and shear stresses resulting in a physical albeit extensional state. A recent two-dimensional analysis has explored the cytoskeletal strain response due to these fluid-induced stresses [Wilson and Kohles, 2010, “Two-Dimensional Modeling of Nanomechanical Stresses-Strains in Healthy and Diseased Single-Cells During Microfluidic Manipulation,” J Nanotechnol Eng Med, 1(2), p. 021005]. Results described a microfluidic environment having controlled nanometer and piconewton resolution. In this present study, computational fluid dynamics combined with multiphysics modeling has further characterized the applied fluid stress environment and the solid cellular strain response in three dimensions to accompany experimental cell stimulation. A volumetric stress-strain analysis was applied to representative living cell biomechanical data. The presented normal and shear stress surface maps will guide future microfluidic experiments as well as provide a framework for characterizing cytoskeletal structure influencing the stress to strain response. PMID:21186894
Comparative proteome analysis of monolayer and spheroid culture of canine osteosarcoma cells.
Gebhard, Christiane; Miller, Ingrid; Hummel, Karin; Neschi Née Ondrovics, Martina; Schlosser, Sarah; Walter, Ingrid
2018-04-15
Osteosarcoma is an aggressive bone tumor with high metastasis rate in the lungs and affects both humans and dogs in a similar way. Three-dimensional tumor cell cultures mimic the in vivo situation of micro-tumors and metastases and are therefore better experimental in vitro models than the often applied two-dimensional monolayer cultures. The aim of the present study was to perform comparative proteomics of standard monolayer cultures of canine osteosarcoma cells (D17) and three-dimensional spheroid cultures, to better characterize the 3D model before starting with experiments like migration assays. Using DIGE in combination with MALDI-TOF/TOF we found 27 unique canine proteins differently represented between these two culture systems, most of them being part of a functional network including mainly chaperones, structural proteins, stress-related proteins, proteins of the glycolysis/gluconeogenesis pathway and oxidoreductases. In monolayer cells, a noticeable shift to more acidic pI values was noticed for several proteins of medium to high abundance; two proteins (protein disulfide isomerase A3, stress-induced-phosphoprotein 1) showed an increase of phosphorylated protein species. Protein distribution within the cells, as detected by immunohistochemistry, displayed a switch of stress-induced-phosphoprotein 1 from the cytoplasm (in monolayer cultures) to the nucleus (in spheroid cultures). Additionally, Western blot testing revealed upregulated concentrations of metastasin (S100A4), triosephosphate isomerase 1 and septin 2 in spheroid cultures, in contrast to decreased concentrations of CCT2, a subunit of the T-complex. Results indicate regulation of stress proteins in the process of three-dimensional organization characterized by a hypoxic and nutrient-deficient environment comparable to tumor micro-metastases. Osteosarcoma is an aggressive bone tumor that early spreads to the lungs. Three-dimensional tumor cell cultures represent the avascular stage of micro-tumors and metastases, and should therefore represent a better experimental in vitro model compared to two-dimensional monolayer cultures. Significant differences have been reported in response to drug and radiation treatment between these two culture systems. A gel-based proteomic investigation was performed to compare protein patterns of a canine osteosarcoma cell line cultivated under those two conditions, to learn more about altered cell composition and its impact on cell behaviour. Due to the fact that the canine osteosarcoma is an accepted model for the human disease, results will be relevant for the human species as well. Copyright © 2018 Elsevier B.V. All rights reserved.
NASA Technical Reports Server (NTRS)
Wong, Kin C.
2003-01-01
This paper documents the derivation of the data reduction equations for the calibration of the six-component thrust stand located in the CE-22 Advanced Nozzle Test Facility. The purpose of the calibration is to determine the first-order interactions between the axial, lateral, and vertical load cells (second-order interactions are assumed to be negligible). In an ideal system, the measurements made by the thrust stand along the three coordinate axes should be independent. For example, when a test article applies an axial force on the thrust stand, the axial load cells should measure the full magnitude of the force, while the off-axis load cells (lateral and vertical) should read zero. Likewise, if a lateral force is applied, the lateral load cells should measure the entire force, while the axial and vertical load cells should read zero. However, in real-world systems, there may be interactions between the load cells. Through proper design of the thrust stand, these interactions can be minimized, but are hard to eliminate entirely. Therefore, the purpose of the thrust stand calibration is to account for these interactions, so that necessary corrections can be made during testing. These corrections can be expressed in the form of an interaction matrix, and this paper shows the derivation of the equations used to obtain the coefficients in this matrix.
Hartmann, Sofie Bruun; Mohanty, Soumyaranjan; Skovgaard, Kerstin; Brogaard, Louise; Flagstad, Frederikke Bjergvang; Emnéus, Jenny; Wolff, Anders; Summerfield, Artur; Jungersen, Gregers
2016-01-01
In vitro generation of dendritic-like cells through differentiation of peripheral blood monocytes is typically done using two-dimensional polystyrene culture plates. In the process of optimising cell culture techniques, engineers have developed fluidic micro-devises usually manufactured in materials other than polystyrene and applying three-dimensional structures more similar to the in vivo environment. Polydimethylsiloxane (PDMS) is an often used polymer for lab-on-a-chip devices but not much is known about the effect of changing the culture surface material from polystyrene to PDMS. In the present study the differentiation of porcine monocytes to monocyte-derived dendritic cells (moDCs) was investigated using CD172apos pig blood monocytes stimulated with GM-CSF and IL-4. Monocytes were cultured on surfaces made of two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS and carbonised three-dimensional PDMS. Cells cultured conventionally (on two-dimensional polystyrene) differentiated into moDCs as expected. Interestingly, gene expression of a wide range of cytokines, chemokines, and pattern recognition receptors was influenced by culture surface material and architecture. Distinct clustering of cells, based on similar expression patterns of 46 genes of interest, was seen for cells isolated from two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS. Changing the material from polystyrene to PDMS resulted in cells with expression patterns usually associated with macrophage expression (upregulation of CD163 and downregulation of CD1a, FLT3, LAMP3 and BATF3). However, this was purely based on gene expression level, and no functional assays were included in this study which would be necessary in order to classify the cells as being macrophages. When changing to three-dimensional culture the cells became increasingly activated in terms of IL6, IL8, IL10 and CCR5 gene expression. Further stimulation with LPS resulted in a slight increase in the expression of maturation markers (SLA-DRB1, CD86 and CD40) as well as cytokines (IL6, IL8, IL10 and IL23A) but the influence of the surfaces was unchanged. These findings highlights future challenges of combining and comparing data generated from microfluidic cell culture-devices made using alternative materials to data generated using conventional polystyrene plates used by most laboratories today. PMID:27362493
Hartmann, Sofie Bruun; Mohanty, Soumyaranjan; Skovgaard, Kerstin; Brogaard, Louise; Flagstad, Frederikke Bjergvang; Emnéus, Jenny; Wolff, Anders; Summerfield, Artur; Jungersen, Gregers
2016-01-01
In vitro generation of dendritic-like cells through differentiation of peripheral blood monocytes is typically done using two-dimensional polystyrene culture plates. In the process of optimising cell culture techniques, engineers have developed fluidic micro-devises usually manufactured in materials other than polystyrene and applying three-dimensional structures more similar to the in vivo environment. Polydimethylsiloxane (PDMS) is an often used polymer for lab-on-a-chip devices but not much is known about the effect of changing the culture surface material from polystyrene to PDMS. In the present study the differentiation of porcine monocytes to monocyte-derived dendritic cells (moDCs) was investigated using CD172apos pig blood monocytes stimulated with GM-CSF and IL-4. Monocytes were cultured on surfaces made of two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS and carbonised three-dimensional PDMS. Cells cultured conventionally (on two-dimensional polystyrene) differentiated into moDCs as expected. Interestingly, gene expression of a wide range of cytokines, chemokines, and pattern recognition receptors was influenced by culture surface material and architecture. Distinct clustering of cells, based on similar expression patterns of 46 genes of interest, was seen for cells isolated from two- and three-dimensional polystyrene as well as two- and three-dimensional PDMS. Changing the material from polystyrene to PDMS resulted in cells with expression patterns usually associated with macrophage expression (upregulation of CD163 and downregulation of CD1a, FLT3, LAMP3 and BATF3). However, this was purely based on gene expression level, and no functional assays were included in this study which would be necessary in order to classify the cells as being macrophages. When changing to three-dimensional culture the cells became increasingly activated in terms of IL6, IL8, IL10 and CCR5 gene expression. Further stimulation with LPS resulted in a slight increase in the expression of maturation markers (SLA-DRB1, CD86 and CD40) as well as cytokines (IL6, IL8, IL10 and IL23A) but the influence of the surfaces was unchanged. These findings highlights future challenges of combining and comparing data generated from microfluidic cell culture-devices made using alternative materials to data generated using conventional polystyrene plates used by most laboratories today.
Thimm, Benjamin W; Hofmann, Sandra; Schneider, Philipp; Carretta, Roberto; Müller, Ralph
2012-03-01
Computed tomography (CT) represents a truly three-dimensional (3D) imaging technique that can provide high-resolution images on the cellular level. Thus, one approach to detect single cells is X-ray absorption-based CT, where cells are labeled with a dense, opaque material providing the required contrast for CT imaging. Within the present work, a novel cell-labeling method has been developed showing the feasibility of labeling fixed cells with iron oxide (FeO) particles for subsequent CT imaging and quantitative morphometry. A biotin-streptavidin detection system was exploited to bind FeO particles to its target endothelial cells. The binding of the particles was predominantly close to the cell centers on 2D surfaces as shown by light microscopy, scanning electron microscopy, and CT. When cells were cultured on porous, 3D polyurethane surfaces, significantly more FeO particles were detected compared with surfaces without cells and FeO particle labeling using CT. Here, we report on the implementation and evaluation of a novel cell detection method based on high-resolution CT. This system has potential in cell tracking for 3D in vitro imaging in the future.
Jia, Shengyong; Han, Hongjun; Hou, Baolin; Zhuang, Haifeng
2015-12-01
Laboratorial scale experiments were conducted to investigate a novel system three-dimensional catalytic electro-Fenton (3DCEF, catalyst of sewage sludge based activated carbon which loaded Fe3O4) integrating with membrane bioreactor (3DCEF-MBR) on advanced treatment of biologically pretreated coal gasification wastewater. The results indicated that 3DCEF-MBR represented high efficiencies in eliminating COD and total organic carbon, giving the maximum removal efficiencies of 80% and 75%, respectively. The integrated 3DCEF-MBR system significantly reduced the transmembrane pressure, giving 35% lower than conventional MBR after 30 days operation. The enhanced hydroxyl radical oxidation and bacteria self repair function were the mechanisms for 3DCEF-MBR performance. Therefore, the integrated 3DCEF-MBR was expected to be the promising technology for advanced treatment in engineering applications. Copyright © 2015. Published by Elsevier Ltd.
Three-dimensional hydrogel cell culture systems for modeling neural tissue
NASA Astrophysics Data System (ADS)
Frampton, John
Two-dimensional (2-D) neural cell culture systems have served as physiological models for understanding the cellular and molecular events that underlie responses to physical and chemical stimuli, control sensory and motor function, and lead to the development of neurological diseases. However, the development of three-dimensional (3-D) cell culture systems will be essential for the advancement of experimental research in a variety of fields including tissue engineering, chemical transport and delivery, cell growth, and cell-cell communication. In 3-D cell culture, cells are provided with an environment similar to tissue, in which they are surrounded on all sides by other cells, structural molecules and adhesion ligands. Cells grown in 3-D culture systems display morphologies and functions more similar to those observed in vivo, and can be cultured in such a way as to recapitulate the structural organization and biological properties of tissue. This thesis describes a hydrogel-based culture system, capable of supporting the growth and function of several neural cell types in 3-D. Alginate hydrogels were characterized in terms of their biomechanical and biochemical properties and were functionalized by covalent attachment of whole proteins and peptide epitopes. Methods were developed for rapid cross-linking of alginate hydrogels, thus permitting the incorporation of cells into 3-D scaffolds without adversely affecting cell viability or function. A variety of neural cell types were tested including astrocytes, microglia, and neurons. Cells remained viable and functional for longer than two weeks in culture and displayed process outgrowth in 3-D. Cell constructs were created that varied in cell density, type and organization, providing experimental flexibility for studying cell interactions and behavior. In one set of experiments, 3-D glial-endothelial cell co-cultures were used to model blood-brain barrier (BBB) structure and function. This co-culture system was designed for use as a tool to predict the transport and processing that occurs prior to drug uptake in the central nervous system (CNS), and to predict BBB permeability. Electrochemical techniques and immunohistochemistry were used to validate this model and provide detailed information about cellular organization and function. Electrochemical impedance spectroscopy (EIS) provided evidence that endothelial cells cultured in the presence of astrocytes formed tight junctions capable of occluding the flow of electrical current. In a second series of experiments, a microglia-astrocyte co-culture system was developed to assess the effects of glial cells on electrode impedance recorded from neural prosthetic devices in vitro. Impedance measurements were compared with confocal images to determine the effects of glial cell density and cell type on electrode performance. The results indicate that EIS data can be used to model components of the reactive cell responses in brain tissue, and that impedance measurements recorded in vitro can be compared to measurements recorded in vivo. Taken together, these results demonstrate that alginate hydrogels can be used for the creation of 3-D neural cell scaffolds, and that such cell scaffolds can be used to model a variety of three-dimensional neural tissues in vitro, that cannot be studied in 2-D cultures.
Lin, Shu-Ping; Kyriakides, Themis R; Chen, Jia-Jin J
2009-06-01
Despite many successful applications of microelectrode arrays (MEAs), typical two-dimensional in-vitro cultures do not project the full scale of the cell growth environment in the three-dimensional (3D) in-vivo setting. This study aims to on-line monitor in-vitro cell growth in a 3D matrix on the surface-modified MEAs with a dynamic perfusion culture system. A 3D matrix consisting of poly(ethylene glycol) hydrogel supplemented with poly-D-lysine was subsequently synthesized in situ on the self-assembled monolayer modified MEAs. FTIR spectrum analysis revealed a peak at 2100 cm(-1) due to the degradation of the structure of the 3D matrix. After 2 wks, microscopic examination revealed that the non-degraded area was around 1500 microm(2) and provided enough space for cell growth. Fluorescence microscopy revealed that the degraded 3D matrix was non-cytotoxic allowing the growth of NIH3T3 fibroblasts and cortical neurons in vitro. Time-course changes of total impedance including resistance and reactance were recorded for 8 days to evaluate the cell growth in the 3D matrix on the MEA. A consistent trend reflecting changes of reactance and total impedance was observed. These in-vitro assays demonstrate that our 3D matrix can construct a biomimetic system for cell growth and analysis of cell surface interactions.
Yoshitomi, Munetake; Ohta, Keisuke; Kanazawa, Tomonoshin; Togo, Akinobu; Hirashima, Shingo; Uemura, Kei-Ichiro; Okayama, Satoko; Morioka, Motohiro; Nakamura, Kei-Ichiro
2016-10-31
Endocrine and endothelial cells of the anterior pituitary gland frequently make close appositions or contacts, and the secretory granules of each endocrine cell tend to accumulate at the perivascular regions, which is generally considered to facilitate secretory functions of these cells. However, three-dimensional relationships between the localization pattern of secretory granules and blood vessels are not fully understood. To define and characterize these spatial relationships, we used scanning electron microscopy (SEM) three-dimensional reconstruction method based on focused ion-beam slicing and scanning electron microscopy (FIB/SEM). Full three-dimensional cellular architectures of the anterior pituitary tissue at ultrastructural resolution revealed that about 70% of endocrine cells were in apposition to the endothelial cells, while almost 30% of endocrine cells were entirely isolated from perivascular space in the tissue. Our three-dimensional analyses also visualized the distribution pattern of secretory granules in individual endocrine cells, showing an accumulation of secretory granules in regions in close apposition to the blood vessels in many cases. However, secretory granules in cells isolated from the perivascular region tended to distribute uniformly in the cytoplasm of these cells. These data suggest that the cellular interactions between the endocrine and endothelial cells promote an uneven cytoplasmic distribution of the secretory granules.
Formation of three-dimensional fetal myocardial tissue cultures from rat for long-term cultivation.
Just, Lothar; Kürsten, Anne; Borth-Bruhns, Thomas; Lindenmaier, Werner; Rohde, Manfred; Dittmar, Kurt; Bader, Augustinus
2006-08-01
Three-dimensional cardiomyocyte cultures offer new possibilities for the analysis of cardiac cell differentiation, spatial cellular arrangement, and time-specific gene expression in a tissue-like environment. We present a new method for generating homogenous and robust cardiomyocyte tissue cultures with good long-term viability. Ventricular heart cells prepared from fetal rats at embryonic day 13 were cultured in a scaffold-free two-step process. To optimize the cell culture model, several digestion protocols and culture conditions were tested. After digestion of fetal cardiac ventricles, the resultant cell suspension of isolated cardiocytes was shaken to initialize cell aggregate formation. In the second step, these three-dimensional cell aggregates were transferred onto a microporous membrane to allow further microstructure formation. Autonomously beating cultures possessed more than 25 cell layers and a homogenous distribution of cardiomyocytes without central necrosis after 8 weeks in vitro. The cardiomyocytes showed contractile elements, desmosomes, and gap junctions analyzed by immunohistochemistry and electron microscopy. The beat frequency could be modulated by adrenergic agonist and antagonist. Adenoviral green fluorescent protein transfer into cardiomyocytes was possible and highly effective. This three-dimensional tissue model proved to be useful for studying cell-cell interactions and cell differentiation processes in a three-dimensional cell arrangement.
Son, JoonGon; Kim, GeunHyung
2009-01-01
Various mechanical techniques have been used to fabricate biomedical scaffolds, including rapid prototyping (RP) devices that operate from CAD files of the target feature information. The three-dimensional (3-D) bio-plotter is one RP system that can produce design-based scaffolds with good mechanical properties for mimicking cartilage and bones. However, the scaffolds fabricated by RP have very smooth surfaces, which tend to discourage initial cell attachment. Initial cell attachment, migration, differentiation and proliferation are strongly dependent on the chemical and physical characteristics of the scaffold surface. In this study, we propose a new 3-D plotting method supplemented with a piezoelectric system for fabricating surface-modified scaffolds. The effects of the physically-modified surface on the mechanical and hydrophilic properties were investigated, and the results of cell culturing of chondrocytes indicate that this technique is a feasible new method for fabricating high-quality 3-D polymeric scaffolds.
Current and efficiency optimization under oscillating forces in entropic barriers
NASA Astrophysics Data System (ADS)
Nutku, Ferhat; Aydıner, Ekrem
2016-09-01
The transport of externally overdriven particles confined in entropic barriers is investigated under various types of oscillating and temporal forces. Temperature, load, and amplitude dependence of the particle current and energy conversion efficiency are investigated in three dimensions. For oscillating forces, the optimized temperature-load, amplitude-temperature, and amplitude-load intervals are determined when fixing the amplitude, load, and temperature, respectively. By using three-dimensional plots rather than two-dimensional ones, it is clearly shown that oscillating forces provide more efficiency compared with a temporal one in specified optimized parameter regions. Furthermore, the dependency of efficiency to the angle between the unbiased driving force and a constant force is investigated and an asymmetric angular dependence is found for all types of forces. Finally, it is shown that oscillating forces with a high amplitude and under a moderate load lead to higher efficiencies than a temporal force at both low and high temperatures for the entire range of contact angle. Project supported by the Istanbul University, Turkey (Grant No. 55383).
Three-Dimensional Terahertz Coded-Aperture Imaging Based on Single Input Multiple Output Technology.
Chen, Shuo; Luo, Chenggao; Deng, Bin; Wang, Hongqiang; Cheng, Yongqiang; Zhuang, Zhaowen
2018-01-19
As a promising radar imaging technique, terahertz coded-aperture imaging (TCAI) can achieve high-resolution, forward-looking, and staring imaging by producing spatiotemporal independent signals with coded apertures. In this paper, we propose a three-dimensional (3D) TCAI architecture based on single input multiple output (SIMO) technology, which can reduce the coding and sampling times sharply. The coded aperture applied in the proposed TCAI architecture loads either purposive or random phase modulation factor. In the transmitting process, the purposive phase modulation factor drives the terahertz beam to scan the divided 3D imaging cells. In the receiving process, the random phase modulation factor is adopted to modulate the terahertz wave to be spatiotemporally independent for high resolution. Considering human-scale targets, images of each 3D imaging cell are reconstructed one by one to decompose the global computational complexity, and then are synthesized together to obtain the complete high-resolution image. As for each imaging cell, the multi-resolution imaging method helps to reduce the computational burden on a large-scale reference-signal matrix. The experimental results demonstrate that the proposed architecture can achieve high-resolution imaging with much less time for 3D targets and has great potential in applications such as security screening, nondestructive detection, medical diagnosis, etc.
Repair of diaphyseal bone defects with calcitriol-loaded PLGA scaffolds and marrow stromal cells.
Yoon, Sun Jung; Park, Ki Suk; Kim, Moon Suk; Rhee, John M; Khang, Gilson; Lee, Hai Bang
2007-05-01
Calcitriol (1,25(OH)2D3)-loaded porous poly(D,L-lactide-co-glycolide) (PLGA) scaffolds prepared by solvent casting/salt leaching method were used to repair a 1.5 cm diaphyseal segmental bone defect as a fully absorbable osteogenic biomaterial. The in vitro release of sulforhodamine B (SRB) from PLGA scaffold was measured using spectrophotometer, considering SRB as a model drug. The SRB released from SRB-incorporated PLGA scaffold during 3 months was with relatively low initial burst. The calcitriol-loaded PLGA scaffolds with or without marrow stromal cells (MSCs) were implanted in a critical-sized intercalated bone defect in rabbit femur. Defects were assessed by radiographs until 9 weeks. The bony union of the defect was observed only in the calcitriol-loaded groups. RT-PCR results indicated that MSCs, which were seeded into calcitriol-loaded scaffold, expressed an increased level of alkaline phosphatase, osteonectin, and type I collagen mRNA at day 10. After 2 and 4 weeks, the implanted scaffolds were evaluated by histology. New osteoid matrix and direct calcium deposits were more evident in calcitriol/PLGA/MSC group. Three-dimensional computed tomography and frontal tomographic images of repaired femur showed that normal femur anatomy had been restored with cortical bone with no implanted PLGA remnants at 20 weeks. It can be concluded that the porous calcitriol-loaded PLGA scaffold combined with MSCs may be a novel method for repairing the large loaded bone defect.
Consideration of Moving Tooth Load in Gear Crack Propagation Predictions
NASA Technical Reports Server (NTRS)
Lewicki, David G.; Handschuh, Robert F.; Spievak, Lisa E.; Wawrzynek, Paul A.; Ingraffea, Anthony R.
2001-01-01
Robust gear designs consider not only crack initiation, but crack propagation trajectories for a fail-safe design. In actual gear operation, the magnitude as well as the position of the force changes as the gear rotates through the mesh. A study to determine the effect of moving gear tooth load on crack propagation predictions was performed. Two-dimensional analysis of an involute spur gear and three-dimensional analysis of a spiral-bevel pinion gear using the finite element method and boundary element method were studied and compared to experiments. A modified theory for predicting gear crack propagation paths based on the criteria of Erdogan and Sih was investigated. Crack simulation based on calculated stress intensity factors and mixed mode crack angle prediction techniques using a simple static analysis in which the tooth load was located at the highest point of single tooth contact was validated. For three-dimensional analysis, however, the analysis was valid only as long as the crack did not approach the contact region on the tooth.
Dentin barrier test with transfected bovine pulp-derived cells.
Schmalz, G; Schuster, U; Thonemann, B; Barth, M; Esterbauer, S
2001-02-01
Growth kinetics of SV40 large T-antigen-transfected bovine pulp-derived cells on dentin were investigated. These cells were used in a dentin barrier test device, and the system was evaluated by testing a set of dental filling materials. Cells (120 cells/mm2) were seeded on dentin slices and incubated for up to 21 days. Cell proliferation was recorded using MTT assay. For cytotoxicity tests 3500 cells/mm2 were seeded on dentin discs, which were then incorporated into the dentin barrier test device. After 72 h preincubation test materials were applied. After a 24 h exposure with or without perfusion of the pulpal part of the test device, cell survival was evaluated using MTT assay. The cells revealed similar growth kinetics on dentin slices and on tissue culture plates. In cytotoxicity tests the cells were more sensitive toward the test materials than previously used three-dimensional cultures of human foreskin fibroblasts and as anticipated from clinical experience. Further improvement is expected by using three-dimensional cultures of pulp-derived cells.
NASA Astrophysics Data System (ADS)
Fu, Bao-rong; Shen, Chao; Ren, Jing; Chen, Jia-yi; Zhao, Lin
2018-03-01
In this study, ferric oxide loading graphite particle electrodes (FO/GPEs) were prepared as cathode of a three-dimensional electrode MFC-Fenton system. The properties of the composite cathode were examined with higher surface area and more mesopores. FO/GPEs could work as both cathode and Fenton iron reagents, contributing to high oxidation activity and better performance of electricity generation. The application of FO/GPEs MFC-Fenton system on degrading p-nitrophenol presented high catalytic efficiency in a wide range of pH value. The removal of p-nitrophenol and TOC attained to about 85 % within 8 and 64 h at neutral pH, respectively. A neutral FO/GPEs MFC-Fenton oxidation mechanism was also proposed. Specifically, both the surface iron sites and dissolved iron ions catalyzed the decomposition of H2O2. As results, the generated hydroxyl radicals were used for p-nitrophenol degradation and the iron oxide was recycled.
Jalili, Reza B.; Forouzandeh, Farshad; Rezakhanlou, Alireza Moeen; Hartwell, Ryan; Medina, Abelardo; Warnock, Garth L.; Larijani, Bagher; Ghahary, Aziz
2010-01-01
OBJECTIVE The requirement of systemic immunosuppression after islet transplantation is of significant concern and a major drawback to clinical islet transplantation. Here, we introduce a novel composite three-dimensional islet graft equipped with a local immunosuppressive system that prevents islet allograft rejection without systemic antirejection agents. In this composite graft, expression of indoleamine 2,3 dioxygenase (IDO), a tryptophan-degrading enzyme, in syngeneic fibroblasts provides a low-tryptophan microenvironment within which T-cells cannot proliferate and infiltrate islets. RESEARCH DESIGN AND METHODS Composite three-dimensional islet grafts were engineered by embedding allogeneic mouse islets and adenoviral-transduced IDO–expressing syngeneic fibroblasts within collagen gel matrix. These grafts were then transplanted into renal subcapsular space of streptozotocin diabetic immunocompetent mice. The viability, function, and criteria for graft take were then determined in the graft recipient mice. RESULTS IDO-expressing grafts survived significantly longer than controls (41.2 ± 1.64 vs. 12.9 ± 0.73 days; P < 0.001) without administration of systemic immunesuppressive agents. Local expression of IDO suppressed effector T-cells at the graft site, induced a Th2 immune response shift, generated an anti-inflammatory cytokine profile, delayed alloantibody production, and increased number of regulatory T-cells in draining lymph nodes, which resulted in antigen-specific impairment of T-cell priming. CONCLUSIONS Local IDO expression prevents cellular and humoral alloimmune responses against islets and significantly prolongs islet allograft survival without systemic antirejection treatments. This promising finding proves the potent local immunosuppressive activity of IDO in islet allografts and sets the stage for development of a long-lasting nonrejectable islet allograft using stable IDO induction in bystander fibroblasts. PMID:20522587
Harley, Brendan A; Freyman, Toby M; Wong, Matthew Q; Gibson, Lorna J
2007-10-15
Cell-mediated contraction plays a critical role in many physiological and pathological processes, notably organized contraction during wound healing. Implantation of an appropriately formulated (i.e., mean pore size, chemical composition, degradation rate) three-dimensional scaffold into an in vivo wound site effectively blocks the majority of organized wound contraction and results in induced regeneration rather than scar formation. Improved understanding of cell contraction within three-dimensional constructs therefore represents an important area of study in tissue engineering. Studies of cell contraction within three-dimensional constructs typically calculate an average contractile force from the gross deformation of a macroscopic substrate by a large cell population. In this study, cellular solids theory has been applied to conventional column buckling relationships to quantify the magnitude of individual cell contraction events within a three-dimensional, collagen-glycosaminoglycan scaffold. This new technique can be used for studying cell mechanics with a wide variety of porous scaffolds that resemble low-density, open-cell foams. It extends previous methods for analyzing cell buckling of two-dimensional substrates to three-dimensional constructs. From data available in the literature, the mean contractile force (Fc) generated by individual dermal fibroblasts within the collagen-glycosaminoglycan scaffold was calculated to range between 11 and 41 nN (Fc=26+/-13 nN, mean+/-SD), with an upper bound of cell contractility estimated at 450 nN.
Advanced Software for Analysis of High-Speed Rolling-Element Bearings
NASA Technical Reports Server (NTRS)
Poplawski, J. V.; Rumbarger, J. H.; Peters, S. M.; Galatis, H.; Flower, R.
2003-01-01
COBRA-AHS is a package of advanced software for analysis of rigid or flexible shaft systems supported by rolling-element bearings operating at high speeds under complex mechanical and thermal loads. These loads can include centrifugal and thermal loads generated by motions of bearing components. COBRA-AHS offers several improvements over prior commercial bearing-analysis programs: It includes innovative probabilistic fatigue-life-estimating software that provides for computation of three-dimensional stress fields and incorporates stress-based (in contradistinction to prior load-based) mathematical models of fatigue life. It interacts automatically with the ANSYS finite-element code to generate finite-element models for estimating distributions of temperature and temperature-induced changes in dimensions in iterative thermal/dimensional analyses: thus, for example, it can be used to predict changes in clearances and thermal lockup. COBRA-AHS provides an improved graphical user interface that facilitates the iterative cycle of analysis and design by providing analysis results quickly in graphical form, enabling the user to control interactive runs without leaving the program environment, and facilitating transfer of plots and printed results for inclusion in design reports. Additional features include roller-edge stress prediction and influence of shaft and housing distortion on bearing performance.
Sun, Liangliang; Zhu, Guijie; Dovichi, Norman J.
2013-01-01
A capillary zone electrophoresis (CZE) electrospray ionization (ESI) tandem mass spectrometry (MS/MS) system was integrated with an immobilized trypsin microreactor. The system was evaluated and then applied for online digestion and analysis of picogram loadings of RAW 264.7 cell lysate. Protein samples were dissolved in a buffer containing 50% (v/v) acetonitrile (ACN), and then directly loaded into the capillary for digestion, followed by CZE separation and MS/MS identification. The organic solvent (50% (v/v) ACN) assisted the immobilized trypsin digestion and simplified the protein sample preparation protocol. Neither protein reduction nor alkylation steps were employed, which minimized sample loss and contamination. The integrated CZE-ESI-MS/MS system generated confident identification of bovine serum albumin (BSA) with 19% sequence coverage and 14 peptide IDs when 20 fmole was loaded. When only 1 fmole BSA was injected, one BSA peptide was consistently detected. For the analysis of a standard protein mixture, the integrated system produced efficient protein digestion and confident identification for proteins with different molecular weights and isoelectric points when low fmole amount was loaded for each protein. We further applied the system for triplicate analysis of a RAW 264.7 cell lysate; 2 ± 1 and 7 ± 2 protein groups were confidently identified from only 300 pg and 3 ng loadings, respectively. The 300 pg sample loading corresponds to the protein content of three RAW 264.7 cells. In addition to high sensitivity analysis, the integrated CZE-ESI-MS/MS system produces good reproducibility in terms of peptide and protein IDs, peptide migration time, and peptide intensity. PMID:23510126
Magnetic nanoparticle-loaded electrospun polymeric nanofibers for tissue engineering.
Zhang, Heng; Xia, JiYi; Pang, XianLun; Zhao, Ming; Wang, BiQiong; Yang, LingLin; Wan, HaiSu; Wu, JingBo; Fu, ShaoZhi
2017-04-01
Magnetic nanoparticles have been one of the most attractive nanomaterials for various biomedical applications including magnetic resonance imaging (MRI), diagnostic contrast enhancement, magnetic cell separation, and targeted drug delivery. Three-dimensional (3-D) fibrous scaffolds have broad application prospects in the biomedical field, such as drug delivery and tissue engineering. In this work, a novel three-dimensional composite membrane composed of the tri-block copolymer poly(ε-caprolactone)-poly(ethylene glycol)-poly(ε-caprolactone) (PCL-PEG-PCL, PCEC) and magnetic iron oxide nanoparticles (Fe 3 O 4 NPs) were fabricated using electrospinning technology. The physico-chemical properties of the PCEC/Fe 3 O 4 membranes were investigated by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and differential scanning calorimetry (DSC). Morphological observation using scanning electron microscopy (SEM) showed that the composite fibers containing 5% Fe 3 O 4 nanoparticles had a diameter of 250nm. In vitro cell culture of NIH 3T3 cells on the PCEC/Fe 3 O 4 membranes showed that the PCEC/Fe 3 O 4 fibers might be a suitable scaffold for cell adhesion. Moreover, MTT analysis also demonstrated that the membranes possessed lower cytotoxicity. Therefore, this study revealed that the magnetic PCEC/Fe 3 O 4 fibers might have great potential for using in skin tissue engineering. Copyright © 2016 Elsevier B.V. All rights reserved.
Three-Dimensional Culture of Human Breast Epithelial Cells: The How and the Why
Vidi, Pierre-Alexandre; Bissell, Mina J.; Lelièvre, Sophie A.
2013-01-01
Organs are made of the organized assembly of different cell types that contribute to the architecture necessary for functional differentiation. In those with exocrine function, such as the breast, cell–cell and cell–extracellular matrix (ECM) interactions establish mechanistic constraints and a complex biochemical signaling network essential for differentiation and homeostasis of the glandular epithelium. Such knowledge has been elegantly acquired for the mammary gland by placing epithelial cells under three-dimensional (3D) culture conditions. Three-dimensional cell culture aims at recapitulating normal and pathological tissue architectures, hence providing physiologically relevant models to study normal development and disease. The specific architecture of the breast epithelium consists of glandular structures (acini) connected to a branched ductal system. A single layer of basoapically polarized luminal cells delineates ductal or acinar lumena at the apical pole. Luminal cells make contact with myoepithelial cells and, in certain areas at the basal pole, also with basement membrane (BM) components. In this chapter, we describe how this exquisite organization as well as stages of disorganization pertaining to cancer progression can be reproduced in 3D cultures. Advantages and limitations of different culture settings are discussed. Technical designs for induction of phenotypic modulations, biochemical analyses, and state-of-the-art imaging are presented. We also explain how signaling is regulated differently in 3D cultures compared to traditional two-dimensional (2D) cultures. We believe that using 3D cultures is an indispensable method to unravel the intricacies of human mammary functions and would best serve the fight against breast cancer. PMID:23097109
NASA Astrophysics Data System (ADS)
Lauzurica, Sara; Márquez, Andrés.; Molpeceres, Carlos; Notario, Laura; Gómez-Fontela, Miguel; Lauzurica, Pilar
2017-02-01
The immune system is a very complex system that comprises a network of genetic and signaling pathways subtending a network of interacting cells. The location of the cells in a network, along with the gene products they interact with, rules the behavior of the immune system. Therefore, there is a great interest in understanding properly the role of a cell in such networks to increase our knowledge of the immune system response. In order to acquire a better understanding of these processes, cell printing with high spatial resolution emerges as one of the promising approaches to organize cells in two and three-dimensional patterns to enable the study the geometry influence in these interactions. In particular, laser assisted bio-printing techniques using sub-nanosecond laser sources have better characteristics for application in this field, mainly due to its higher spatial resolution, cell viability percentage and process automation. This work presents laser assisted bio-printing of antigen-presenting cells (APCs) in two-dimensional geometries, placing cellular components on a matrix previously generated on demand, permitting to test the molecular interactions between APCs and lymphocytes; as well as the generation of two-dimensional structures designed ad hoc in order to study the mechanisms of mobilization of immune system cells. The use of laser assisted bio-printing, along with APCs and lymphocytes emulate the structure of different niches of the immune system so that we can analyse functional requirement of these interaction.
Targeted Doxorubicin-Loaded Bacterially Derived Nano-Cells for the Treatment of Neuroblastoma.
Sagnella, Sharon M; Trieu, Jennifer; Brahmbhatt, Himanshu; MacDiarmid, Jennifer A; MacMillan, Alex; Whan, Renee M; Fife, Christopher M; McCarroll, Joshua A; Gifford, Andrew J; Ziegler, David S; Kavallaris, Maria
2018-05-01
Advanced stage neuroblastoma is an aggressive disease with limited treatment options for patients with drug-resistant tumors. Targeted delivery of chemotherapy for pediatric cancers offers promise to improve treatment efficacy and reduce toxicity associated with systemic chemotherapy. The EnGeneIC Dream Vector (EDV TM ) is a nanocell, which can package chemotherapeutic drugs and target tumors via attachment of bispecific proteins to the surface of the nanocell. Phase I trials in adults with refractory tumors have shown an acceptable safety profile. Herein we investigated the activity of EGFR-targeted and doxorubicin-loaded EDV TM ( EGFR EDV TM Dox ) for the treatment of neuroblastoma. Two independent neuroblastoma cell lines with variable expression of EGFR protein [SK-N-BE(2), high; SH-SY-5Y, low] were used. EGFR EDV TM Dox induced apoptosis in these cells compared to control, doxorubicin, or non-doxorubicin loaded EGFR EDV TM In three-dimensional tumor spheroids, imaging and fluorescence life-time microscopy revealed that EGFR EDV TM Dox had a marked enhancement of doxorubicin penetration compared to doxorubicin alone, and improved penetration compared to non-EGFR-targeted EDV TM Dox , with enhanced spheroid penetration leading to increased apoptosis. In two independent orthotopic human neuroblastoma xenograft models, short-term studies (28 days) of tumor-bearing mice led to a significant decrease in tumor size in EGFR EDV TM Dox -treated animals compared to control, doxorubicin, or non-EGFR EDV TM Dox There was increased TUNEL staining of tumors at day 28 compared to control, doxorubicin, or non-EGFR EDV TM Dox Moreover, overall survival was increased in neuroblastoma mice treated with EGFR EDV TM Dox ( P < 0007) compared to control. Drug-loaded bispecific-antibody targeted EDVs TM offer a highly promising approach for the treatment of aggressive pediatric malignancies such as neuroblastoma. Mol Cancer Ther; 17(5); 1012-23. ©2018 AACR . ©2018 American Association for Cancer Research.
Non-dimensional physics of pulsatile cardiovascular networks and energy efficiency.
Yigit, Berk; Pekkan, Kerem
2016-01-01
In Nature, there exist a variety of cardiovascular circulation networks in which the energetic ventricular load has both steady and pulsatile components. Steady load is related to the mean cardiac output (CO) and the haemodynamic resistance of the peripheral vascular system. On the other hand, the pulsatile load is determined by the simultaneous pressure and flow waveforms at the ventricular outlet, which in turn are governed through arterial wave dynamics (transmission) and pulse decay characteristics (windkessel effect). Both the steady and pulsatile contributions of the haemodynamic power load are critical for characterizing/comparing disease states and for predicting the performance of cardiovascular devices. However, haemodynamic performance parameters vary significantly from subject to subject because of body size, heart rate and subject-specific CO. Therefore, a 'normalized' energy dissipation index, as a function of the 'non-dimensional' physical parameters that govern the circulation networks, is needed for comparative/integrative biological studies and clinical decision-making. In this paper, a complete network-independent non-dimensional formulation that incorporates pulsatile flow regimes is developed. Mechanical design variables of cardiovascular flow systems are identified and the Buckingham Pi theorem is formally applied to obtain the corresponding non-dimensional scaling parameter sets. Two scaling approaches are considered to address both the lumped parameter networks and the distributed circulation components. The validity of these non-dimensional number sets is tested extensively through the existing empirical allometric scaling laws of circulation systems. Additional validation studies are performed using a parametric numerical arterial model that represents the transmission and windkessel characteristics, which are adjusted to represent different body sizes and non-dimensional haemodynamic states. Simulations demonstrate that the proposed non-dimensional indices are independent of body size for healthy conditions, but are sensitive to deviations caused by off-design disease states that alter the energetic load. Sensitivity simulations are used to identify the relationship between pulsatile power loss and non-dimensional characteristics, and optimal operational states are computed. © 2016 The Author(s).
Performance Estimation for Two-Dimensional Brownian Rotary Ratchet Systems
NASA Astrophysics Data System (ADS)
Tutu, Hiroki; Horita, Takehiko; Ouchi, Katsuya
2015-04-01
Within the context of the Brownian ratchet model, a molecular rotary system that can perform unidirectional rotations induced by linearly polarized ac fields and produce positive work under loads was studied. The model is based on the Langevin equation for a particle in a two-dimensional (2D) three-tooth ratchet potential of threefold symmetry. The performance of the system is characterized by the coercive torque, i.e., the strength of the load competing with the torque induced by the ac driving field, and the energy efficiency in force conversion from the driving field to the torque. We propose a master equation for coarse-grained states, which takes into account the boundary motion between states, and develop a kinetic description to estimate the mean angular momentum (MAM) and powers relevant to the energy balance equation. The framework of analysis incorporates several 2D characteristics and is applicable to a wide class of models of smooth 2D ratchet potential. We confirm that the obtained expressions for MAM, power, and efficiency of the model can enable us to predict qualitative behaviors. We also discuss the usefulness of the torque/power relationship for experimental analyses, and propose a characteristic for 2D ratchet systems.
Fatigue loading history reconstruction based on the rain-flow technique
NASA Technical Reports Server (NTRS)
Khosrovaneh, A. K.; Dowling, N. E.
1989-01-01
Methods are considered for reducing a non-random fatigue loading history to a concise description and then for reconstructing a time history similar to the original. In particular, three methods of reconstruction based on a rain-flow cycle counting matrix are presented. A rain-flow matrix consists of the numbers of cycles at various peak and valley combinations. Two methods are based on a two dimensional rain-flow matrix, and the third on a three dimensional rain-flow matrix. Histories reconstructed by any of these methods produce a rain-flow matrix identical to that of the original history, and as a result the resulting time history is expected to produce a fatigue life similar to that for the original. The procedures described allow lengthy loading histories to be stored in compact form.
Three-Dimensional Magnetic Levitation Culture System Simulating White Adipose Tissue.
Tseng, Hubert; Daquinag, Alexes C; Souza, Glauco R; Kolonin, Mikhail G
2018-01-01
White adipose tissue (WAT) has attracted interest for tissue engineering and cell-based therapies as an abundant source of adipose stem/stromal cells (ASC). However, technical challenges in WAT cell culture have limited its applications in regenerative medicine. Traditional two-dimensional (2D) cell culture models, which are essentially monolayers of cells on glass or plastic substrates, inadequately represent tissue architecture, biochemical concentration gradients, substrate stiffness, and most importantly for WAT research, cell phenotypic heterogeneity. Physiological cell culture platforms for WAT modeling must recapitulate the native diversity of cell types and their coordination within the organ. For this purpose, we developed a three-dimensional (3D) model using magnetic levitation. Here, we describe our protocol that we successfully employed to build adipose tissue organoids (adipospheres) that preserve the heterogeneity of the constituent cell types in vitro. We demonstrate the capacity of assembling adipospheres from multiple cell types, including ASCs, endohtelial cells, and leukocytes that recreate tissue organization. These adipospheres mimicked WAT organogenesis in that they enabled the formation of vessel-like endothelial structures with lumens and differentiation of unilocular adipocytes. Altogether, magnetic levitation is a cell culture platform that recreates tissue structure, function, and heterogeneity in vitro, and serves as a foundation for high-throughput WAT tissue culture and analysis.
Gao, Xuechuan; Hai, Xiao; Baigude, Huricha; Guan, Weihua; Liu, Zhiliang
2016-01-01
An advanced multifunctional, hollow metal-organic framework (MOF) drug delivery system with a high drug loading level and targeted delivery was designed and fabricated for the first time and applied to inhibit tumour cell growth. This hollow MOF targeting drug delivery system was prepared via a simple post-synthetic surface modification procedure, starting from hollow ZIF-8 successfully obtained for the first time via a mild phase transformation under solvothermal conditions. As a result, the hollow ZIF-8 exhibits a higher loading capacity for the model anticancer drug 5-fluorouracil (5-FU). Subsequently, 5-FU-loaded ZIF-8 was encapsulated into polymer layers (FA-CHI-5-FAM) with three components: a chitosan (CHI) backbone, the imaging agent 5-carboxyfluorescein (5-FAM), and the targeting reagent folic acid (FA). Thus, an advanced drug delivery system, ZIF-8/5-FU@FA-CHI-5-FAM, was fabricated. A cell imaging assay demonstrated that ZIF-8/5-FU@FA-CHI-5-FAM could target and be taken up by MGC-803 cells. Furthermore, the as-prepared ZIF-8/5-FU@FA-CHI-5-FAM exhibited stronger cell growth inhibitory effects on MGC-803 cells because of the release of 5-FU, as confirmed by a cell viability assay. In addition, a drug release experiment in vitro indicated that ZIF-8/5-FU@FA-CHI-5-FAM exhibited high loading capacity (51%) and a sustained drug release behaviour. Therefore, ZIF-8/5-FU@FA-CHI-5-FAM could provide targeted drug transportation, imaging tracking and localized sustained release. PMID:27876876
Bastami, Farshid; Paknejad, Zahrasadat; Jafari, Maissa; Salehi, Majid; Rezai Rad, Maryam; Khojasteh, Arash
2017-03-01
Fabrication of an ideal scaffold having proper composition, physical structure and able to have sustained release of growth factors still is challenging for bone tissue engineering. Current study aimed to design an appropriate three-dimensional (3-D) scaffold with suitable physical characteristics, including proper compressive strength, degradation rate, porosity, and able to sustained release of bone morphogenetic protein-2 (BMP2), for bone tissue engineering. A highly porous 3-D β-tricalcium phosphate (β-TCP) scaffolds, inside of which two perpendicular canals were created, was fabricated using foam-casting technique. Then, scaffolds were coated with gelatin layer. Next, BMP2-loaded chitosan (CS) nanoparticles were dispersed into collagen hydrogel and filled into the scaffold canals. Physical characteristics of fabricated constructs were evaluated. Moreover, the capability of given construct for bone regeneration has been evaluated in vitro in interaction with human buccal fat pad-derived stem cells (hBFPSCs). The results showed that gelatin-coated TCP scaffold with rhBMP2 delivery system not only could act as a mechanically and biologically compatible framework, but also act as an osteoinductive graft by sustained delivering of rhBMP2 in a therapeutic window for differentiation of hBFPSCs towards the osteoblast lineage. The proposed scaffold model can be suggested for delivering of cells and other growth factors such as vascular endothelial growth factor (VEGF), alone or in combination, for future investigations. Copyright © 2016 Elsevier B.V. All rights reserved.
Ali, Omar A.; Verbeke, Catia; Johnson, Chris; Sands, Warren; Lewin, Sarah A.; White, Des; Doherty, Edward; Dranoff, Glenn; Mooney, David J.
2014-01-01
The innate cellular and molecular components required to mediate effective vaccination against weak tumor-associated antigens remain unclear. In this study we utilized polymeric cancer vaccines incorporating different classes of adjuvants to induce tumor protection, in order to identify dendritic cell subsets and cytokines critical to this efficacy. Three-dimensional, porous polymer matrices loaded with tumor lysates and presenting distinct combinations of GM-CSF and various TLR agonists effected 70–90% prophylactic tumor protection in B16-F10 melanoma models. In aggressive, therapeutic B16 models, the vaccine systems incorporating GM-CSF in combination with P(I:C) or CpG-ODN induced the complete regression of solid tumors (≤40mm2) resulting in 33% long-term survival. Regression analysis revealed that the numbers of vaccine-resident CD8(+) DCs and plasmacytoid DCs, along with local IL-12, and G-CSF concentrations correlated strongly to vaccine efficacy regardless of adjuvant type. Further, vaccine studies in Batf3−/− mice revealed that CD8(+) DCs are required to effect tumor protection, as vaccines in these mice were deficient in cytotoxic T cell priming, and IL-12 induction in comparison to wild-type. These studies broadly demonstrate that three-dimensional polymeric vaccines provide a potent platform for prophylactic and therapeutic protection, and can be used as a tool to identify critical components of a desired immune response. Specifically, these results suggest that CD8(+) DCs, plasmacytoid DCs, IL-12, and G-CSF play important roles in priming effective anti-tumor responses with these vaccines. PMID:24480625
Zhan, Kang; Lin, Miao; Liu, MingMei; Sui, YangNan; Babekir, Haitham Mohammed; Zhao, GuoQi
2017-05-01
Primary bovine mammary epithelial cells (BMECs) are not ideal models for long-term studies of lactation mechanisms because these cells in a monolayer culture system cannot be polarized to simulate the physiological functions in vitro. We investigate the effects of different culture models and karyotypes on casein expression in a three-dimensional (3D) culture system. The immortalized cells' karyotypes were analyzed at passages 10, 20, 30 and 40 to detect the effects of chromosome stability. Western blotting examined that whether or not the immortalized cells at passages 5, 10, 20, 30, 40 and 50 could induce expression of casein in a 3D culture system. The proper polarization of the acinar structures was monitored. BMECs were successfully immortalized. The cell karyotype at passage 30 remained at 60 chromosomes and the average value was 57.1 ± 0.40 after passage 40. The polarized protein's levels were up-regulated in 3D culture compared to 2D culture. Expression of αs1, β and κ-casein could be detectable in a passage range in 3D culture. Expression of αs2-casein was undetectable in all experimental groups. However, all casein expressions were barely detectable in traditional 2D culture system. Therefore, 3D culture system is an important tool for the long-term study of lactation mechanisms in vitro. © 2016 Japanese Society of Animal Science.
Yeatts, Andrew B.; Choquette, Daniel T.; Fisher, John P.
2012-01-01
Background Mesenchymal stem cells (MSCs) are a promising cell source for bone and cartilage tissue engineering as they can be easily isolated from the body and differentiated into osteoblasts and chondrocytes. A cell based tissue engineering strategy using MSCs often involves the culture of these cells on three-dimensional scaffolds; however the size of these scaffolds and the cell population they can support can be restricted in traditional static culture. Thus dynamic culture in bioreactor systems provides a promising means to culture and differentiate MSCs in vitro. Scope of Review This review seeks to characterize key MSC differentiation signaling pathways and provides evidence as to how dynamic culture is augmenting these pathways. Following an overview of dynamic culture systems, discussion will be provided on how these systems can effectively modify and maintain important culture parameters including oxygen content and shear stress. Literature is reviewed for both a highlight of key signaling pathways and evidence for regulation of these signaling pathways via dynamic culture systems. Major Conclusions The ability to understand how these culture systems are affecting MSC signaling pathways could lead to a shear or oxygen regime to direct stem cell differentiation. In this way the efficacy of in vitro culture and differentiation of MSCs on three-dimensional scaffolds could be greatly increased. General Significance Bioreactor systems have the ability to control many key differentiation stimuli including mechanical stress and oxygen content. The further integration of cell signaling investigations within dynamic culture systems will lead to a quicker realization of the promise of tissue engineering and regenerative medicine. PMID:22705676
NASA Technical Reports Server (NTRS)
Arnold, James O.; Peterson, Keith H.; Yount, Bryan C.; Schneider, Nigel; Chavez-Garcia, Jose
2013-01-01
Arcjet testing and analysis of a three-dimensional (3D) woven carbon fabric has shown that it can be used as a thermal protection system and as a load bearing structural component for a low ballistic coefficient hypersonic decelerator called ADEPT (Adaptive Deployable Entry and Placement Technology). Results of arcjet tests proved that the 3D woven carbon fabric can withstand flight-like heating while under flight-like biaxial mechanical loads representative of those encountered during shallow entry flight path angles into the atmosphere of Venus. Importantly, the arcjet test results have been used to extend a preliminary material thermal response model based on previous testing of the same 3D woven carbon fabric under uni-axial mechanical loading.
Sun, X; Kang, Y; Bao, J; Zhang, Y; Yang, Y; Zhou, X
2013-01-01
Osteogenetic microenvironment is a complex constitution in which extracellular matrix (ECM) molecules, stem cells and growth factors each interact to direct the coordinate regulation of bone tissue development. Importantly, angiogenesis improvement and revascularization are critical for osteogenesis during bone tissue regeneration processes. In this study, we developed a three-dimensional (3D) multi-scale system model to study cell response to growth factors released from a 3D biodegradable porous calcium phosphate (CaP) scaffold. Our model reconstructed the 3D bone regeneration system and examined the effects of pore size and porosity on bone formation and angiogenesis. The results suggested that scaffold porosity played a more dominant role in affecting bone formation and angiogenesis compared with pore size, while the pore size could be controlled to tailor the growth factor release rate and release fraction. Furthermore, a combination of gradient VEGF with BMP2 and Wnt released from the multi-layer scaffold promoted angiogenesis and bone formation more readily than single growth factors. These results demonstrated that the developed model can be potentially applied to predict vascularized bone regeneration with specific scaffold and growth factors. PMID:23566802
Three-dimensional micro electromechanical system piezoelectric ultrasound transducer
NASA Astrophysics Data System (ADS)
Hajati, Arman; Latev, Dimitre; Gardner, Deane; Hajati, Azadeh; Imai, Darren; Torrey, Marc; Schoeppler, Martin
2012-12-01
Here we present the design and experimental acoustic test data for an ultrasound transducer technology based on a combination of micromachined dome-shaped piezoelectric resonators arranged in a flexible architecture. Our high performance niobium-doped lead zirconate titanate film is implemented in three-dimensional dome-shaped structures, which form the basic resonating cells. Adjustable frequency response is realized by mixing these basic cells and modifying their dimensions by lithography. Improved characteristics such as high sensitivity, adjustable wide-bandwidth frequency response, low transmit voltage compatible with ordinary integrated circuitry, low electrical impedance well matched to coaxial cabling, and intrinsic acoustic impedance match to water are demonstrated.
Ma, Da; Tang, Liang; Pan, Yan-Huan
2007-12-01
Three-dimensional finite method was used to analyze stress and strain distributions of periodontal ligament of abutments under dynamic loads. Finite element analysis was performed on the model under dynamic loads with vertical and oblique directions. The stress and strain distributions and stress-time curves were analyzed to study the biomechanical behavior of periodontal ligament of abutments. The stress and strain distributions of periodontal ligament under dynamic load were same with the static load. But the maximum stress and strain decreased apparently. The rate of change was between 60%-75%. The periodontal ligament had time-dependent mechanical behaviors. Some level of residual stress in periodontal ligament was left after one mastication period. The stress-free time under oblique load was shorter than that of vertical load. The maximum stress and strain decrease apparently under dynamic loads. The periodontal ligament has time-dependent mechanical behaviors during one mastication. There is some level of residual stress left after one mastication period. The level of residual stress is related to the magnitude and the direction of loads. The direction of applied loads is one important factor that affected the stress distribution and accumulation and release of abutment periodontal ligament.
NASA Technical Reports Server (NTRS)
Reddy, T. S. R.; Srivastava, R.; Mehmed, Oral
2002-01-01
An aeroelastic analysis system for flutter and forced response analysis of turbomachines based on a two-dimensional linearized unsteady Euler solver has been developed. The ASTROP2 code, an aeroelastic stability analysis program for turbomachinery, was used as a basis for this development. The ASTROP2 code uses strip theory to couple a two dimensional aerodynamic model with a three dimensional structural model. The code was modified to include forced response capability. The formulation was also modified to include aeroelastic analysis with mistuning. A linearized unsteady Euler solver, LINFLX2D is added to model the unsteady aerodynamics in ASTROP2. By calculating the unsteady aerodynamic loads using LINFLX2D, it is possible to include the effects of transonic flow on flutter and forced response in the analysis. The stability is inferred from an eigenvalue analysis. The revised code, ASTROP2-LE for ASTROP2 code using Linearized Euler aerodynamics, is validated by comparing the predictions with those obtained using linear unsteady aerodynamic solutions.
Chinese-English Electronics and Telecommunications Dictionary. Volume 2
1976-11-01
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Programmable Control in Extracellular Matrix-mimicking Polymer Hydrogels.
Hof, Kevin S; Bastings, Maartje M C
2017-06-28
The extracellular matrix (ECM) and cells have a reciprocal relationship, one shapes the other and vice versa. One of the main challenges of synthetic material systems for developmental cell culturing, organoid and stem cell work includes the implementation of this reciprocal nature. The largest hurdle to achieve true cell-instructive materials in biomaterials engineering is a lack of spatial and temporal control over material properties and the display of bioactive signals compared to the natural cell environment. ECM-mimicking hydrogels have been developed using a wide range of polymers, assembly and cross-linking strategies. While our synthetic toolbox is larger than nature, often our systems underperform when compared to ECM systems with natural components like Matrigel. Material properties and three-dimensional structure ill-represent the three-dimensional ECM reciprocal nature and ligand presentation is an oversimplified version of the complexity found in nature. We hypothesize that the lack of programmable control in properties and ligand presentation forms the basis of this mismatch in performance and analyze the presence of control in current state of the art ECM-mimicking systems based on covalent, supramolecular and recombinant polymers. We conclude that through combining the dynamics of supramolecular materials, robustness from covalent systems and the programmable spatial control of bio-activation in recombinant ECM materials, the optimal synthetic artificial ECM could be assembled.
NASA Astrophysics Data System (ADS)
Perillo, Evan P.; Liu, Yen-Liang; Huynh, Khang; Liu, Cong; Chou, Chao-Kai; Hung, Mien-Chie; Yeh, Hsin-Chih; Dunn, Andrew K.
2015-07-01
Molecular trafficking within cells, tissues and engineered three-dimensional multicellular models is critical to the understanding of the development and treatment of various diseases including cancer. However, current tracking methods are either confined to two dimensions or limited to an interrogation depth of ~15 μm. Here we present a three-dimensional tracking method capable of quantifying rapid molecular transport dynamics in highly scattering environments at depths up to 200 μm. The system has a response time of 1 ms with a temporal resolution down to 50 μs in high signal-to-noise conditions, and a spatial localization precision as good as 35 nm. Built on spatiotemporally multiplexed two-photon excitation, this approach requires only one detector for three-dimensional particle tracking and allows for two-photon, multicolour imaging. Here we demonstrate three-dimensional tracking of epidermal growth factor receptor complexes at a depth of ~100 μm in tumour spheroids.
Program to develop a performance and heat load prediction system for multistage turbines
NASA Technical Reports Server (NTRS)
Sharma, OM
1994-01-01
Flows in low-aspect ratio turbines, such as the SSME fuel turbine, are three dimensional and highly unsteady due to the relative motion of adjacent airfoil rows and the circumferential and spanwise gradients in total pressure and temperature, The systems used to design these machines, however, are based on the assumption that the flow is steady. The codes utilized in these design systems are calibrated against turbine rig and engine data through the use of empirical correlations and experience factors. For high aspect ratio turbines, these codes yield reasonably accurate estimates of flow and temperature distributions. However, future design trends will see lower aspect ratio (reduced number of parts) and higher inlet temperature which will result in increased three dimensionality and flow unsteadiness in turbines. Analysis of recently acquired data indicate that temperature streaks and secondary flows generated in combustors and up-stream airfoils can have a large impact on the time-averaged temperature and angle distributions in downstream airfoil rows.
Chvátal, Alexandr; Anděrová, Miroslava; Kirchhoff, Frank
2007-01-01
Pathological states in the central nervous system lead to dramatic changes in the activity of neuroactive substances in the extracellular space, to changes in ionic homeostasis and often to cell swelling. To quantify changes in cell morphology over a certain period of time, we employed a new technique, three-dimensional confocal morphometry. In our experiments, performed on enhanced green fluorescent protein/glial fibrillary acidic protein astrocytes in brain slices in situ and thus preserving the extracellular microenvironment, confocal morphometry revealed that the application of hypotonic solution evoked two types of volume change. In one population of astrocytes, hypotonic stress evoked small cell volume changes followed by a regulatory volume decrease, while in the second population volume changes were significantly larger without subsequent volume regulation. Three-dimensional cell reconstruction revealed that even though the total astrocyte volume increased during hypotonic stress, the morphological changes in various cell compartments and processes were more complex than have been previously shown, including swelling, shrinking and structural rearrangement. Our data show that astrocytes in brain slices in situ during hypotonic stress display complex behaviour. One population of astrocytes is highly capable of cell volume regulation, while the second population is characterized by prominent cell swelling, accompanied by plastic changes in morphology. It is possible to speculate that these two astrocyte populations play different roles during physiological and pathological states. PMID:17488344
Direct writing of bio-functional coatings for cardiovascular applications.
Perkins, Jessica; Hong, Yi; Ye, Sang-Ho; Wagner, William R; Desai, Salil
2014-12-01
The surface modification of metallic biomaterials is of critical importance to enhance the biocompatibility of surgical implant materials and devices. This article investigates the use of a direct-write inkjet technique for multilayer coatings of a biodegradable polymer (polyester urethane urea (PEUU)) embedded with an anti-proliferation drug paclitaxel (Taxol). The direct-write inkjet technique provides selective patterning capability for depositing multimaterial coatings on three-dimensional implant devices such as pins, screws, and stents for orthopedic and vascular applications. Drug release profiles were studied to observe the influence of drug loading and coating thickness for obtaining tunable release kinetics. Platelet deposition studies were conducted following ovine blood contact and significant reduction in platelet deposition was observed on the Taxol loaded PEUU substrate compared with the unloaded control. Rat smooth muscle cells were used for cell proliferation studies. Significant reduction in cell growth was observed following the release of anti-proliferative drug from the biopolymer thin film. This research provides a basis for developing anti-proliferative biocompatible coatings for different biomedical device applications. © 2014 Wiley Periodicals, Inc.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Dueñas, Maria Emilia; Essner, Jeffrey J.; Lee, Young Jin
The zebrafish ( Danio rerio) has been widely used as a model vertebrate system to study lipid metabolism, the roles of lipids in diseases, and lipid dynamics in embryonic development. Here, we applied high-spatial resolution matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry imaging (MSI) to map and visualize the three-dimensional spatial distribution of phospholipid classes, phosphatidylcholine (PC), phosphatidylethanolamines (PE), and phosphatidylinositol (PI), in newly fertilized individual zebrafish embryos. This is the first time MALDI-MSI has been applied for three dimensional chemical imaging of a single cell. PC molecular species are present inside the yolk in addition to the blastodisc, while PE andmore » PI species are mostly absent in the yolk. Two-dimensional MSI was also studied for embryos at different cell stages (1-, 2-, 4-, 8-, and 16-cell stage) to investigate the localization changes of some lipids at various cell developmental stages. Lastly, four different normalization approaches were compared to find reliable relative quantification in 2D- and 3D- MALDI MSI data sets.« less
Dueñas, Maria Emilia; Essner, Jeffrey J.; Lee, Young Jin
2017-11-02
The zebrafish ( Danio rerio) has been widely used as a model vertebrate system to study lipid metabolism, the roles of lipids in diseases, and lipid dynamics in embryonic development. Here, we applied high-spatial resolution matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry imaging (MSI) to map and visualize the three-dimensional spatial distribution of phospholipid classes, phosphatidylcholine (PC), phosphatidylethanolamines (PE), and phosphatidylinositol (PI), in newly fertilized individual zebrafish embryos. This is the first time MALDI-MSI has been applied for three dimensional chemical imaging of a single cell. PC molecular species are present inside the yolk in addition to the blastodisc, while PE andmore » PI species are mostly absent in the yolk. Two-dimensional MSI was also studied for embryos at different cell stages (1-, 2-, 4-, 8-, and 16-cell stage) to investigate the localization changes of some lipids at various cell developmental stages. Lastly, four different normalization approaches were compared to find reliable relative quantification in 2D- and 3D- MALDI MSI data sets.« less
Spanwise visualization of the flow around a three-dimensional foil with leading edge protuberances
NASA Astrophysics Data System (ADS)
Stanway, M. J.; Techet, A. H.
2006-11-01
Studies of model humpback whale fins have shown that leading edge protuberances, or tubercles, can lead to delayed stall and increased lift at higher angles of attack, compared to foils with geometrically smooth leading edges. Such enhanced performance characteristics could prove highly useful in underwater vehicles such as gliders or long range AUVs (autonomous underwater vehicles). In this work, Particle Imaging Velocimetry (PIV) is performed on two static wings in a water tunnel over a range of angles of attack. These three- dimensional, finite-aspect ratio wings are modeled after a humpback whale flipper and are identical in shape, tapered from root to tip, except for the leading edge. In one of the foils the leading edge is smooth, whereas in the other, regularly spaced leading edge bumps are machined to simulate the whale’s fin tubercles. Results from these PIV tests reveal distinct cells where coherent flow structures are destroyed as a result of the leading edge perturbations. Tests are performed at Reynolds numbers Re ˜ O(10^5), based on chordlength, in a recirculating water tunnel. An inline six-axis load cell is mounted to measure the forces on the foil over a range of static pitch angles. It is hypothesized that this spanwise breakup of coherent vortical structures is responsible for the delayed angle of stall. These quantitative experiments complement exiting qualitative studies with two dimensional foils.
Hwa, Yoon; Seo, Hyeon Kook; Yuk, Jong-Min; Cairns, Elton J
2017-11-08
The ambient-temperature rechargeable lithium/sulfur (Li/S) cell is a strong candidate for the beyond lithium ion cell since significant progress on developing advanced sulfur electrodes with high sulfur loading has been made. Here we report on a new sulfur electrode active material consisting of a cetyltrimethylammonium bromide-modified sulfur-graphene oxide-carbon nanotube (S-GO-CTA-CNT) nanocomposite prepared by freeze-drying. We show the real-time formation of nanocrystalline lithium sulfide (Li 2 S) at the interface between the S-GO-CTA-CNT nanocomposite and the liquid electrolyte by in situ TEM observation of the reaction. The combination of GO and CNT helps to maintain the structural integrity of the S-GO-CTA-CNT nanocomposite during lithiation/delithiation. A high S loading (11.1 mgS/cm 2 , 75% S) S-GO-CTA-CNT electrode was successfully prepared using a three-dimensional structured Al foam as a substrate and showed good S utilization (1128 mAh/g S corresponding to 12.5 mAh/cm 2 ), even with a very low electrolyte to sulfur weight ratio of 4. Moreover, it was demonstrated that the ionic liquid in the electrolyte improves the Coulombic efficiency and stabilizes the morphology of the Li metal anode.
Morshed, R A; Gutova, M; Juliano, J; Barish, M E; Hawkins-Daarud, A; Oganesyan, D; Vazgen, K; Yang, T; Annala, A; Ahmed, A U; Aboody, K S; Swanson, K R; Moats, R A; Lesniak, M S
2015-01-01
In preclinical studies, neural stem cell (NSC)-based delivery of oncolytic virus has shown great promise in the treatment of malignant glioma. Ensuring the success of this therapy will require critical evaluation of the spatial distribution of virus after NSC transplantation. In this study, the patient-derived GBM43 human glioma line was established in the brain of athymic nude mice, followed by the administration of NSCs loaded with conditionally replicating oncolytic adenovirus (NSC-CRAd-S-pk7). We determined the tumor coverage potential of oncolytic adenovirus by examining NSC distribution using magnetic resonance (MR) imaging and by three-dimensional reconstruction from ex vivo tissue specimens. We demonstrate that unmodified NSCs and NSC-CRAd-S-pk7 exhibit a similar distribution pattern with most prominent localization occurring at the tumor margins. We were further able to visualize the accumulation of these cells at tumor sites via T2-weighted MR imaging as well as the spread of viral particles using immunofluorescence. Our analyses reveal that a single administration of oncolytic virus-loaded NSCs allows for up to 31% coverage of intracranial tumors. Such results provide valuable insights into the therapeutic potential of this novel viral delivery platform.
Ewald method for polytropic potentials in arbitrary dimensionality
NASA Astrophysics Data System (ADS)
Osychenko, O. N.; Astrakharchik, G. E.; Boronat, J.
2012-02-01
The Ewald summation technique is generalized to power-law 1/| r | k potentials in three-, two- and one-dimensional geometries with explicit formulae for all the components of the sums. The cases of short-range, long-range and 'marginal' interactions are treated separately. The jellium model, as a particular case of a charge-neutral system, is discussed and the explicit forms of the Ewald sums for such a system are presented. A generalized form of the Ewald sums for a non-cubic (non-square) simulation cell for three- (two-) dimensional geometry is obtained and its possible field of application is discussed. A procedure for the optimization of the involved parameters in actual simulations is developed and an example of its application is presented.
Ma, Liang; Barker, Jeremy; Zhou, Changchun; Li, Wei; Zhang, Jing; Lin, Biaoyang; Foltz, Gregory; Küblbeck, Jenni; Honkakoski, Paavo
2013-01-01
A three-dimensional micro-scale perfusion-based two-chamber (3D-μPTC) tissue model system was developed to test the cytotoxicity of anticancer drugs in conjunction with liver metabolism. Liver cells with different cytochrome P450 (CYP) subtypes and glioblastoma multiforme (GBM) brain cancer cells were cultured in two separate chambers connected in tandem. Both chambers contained a 3D tissue engineering scaffold fabricated with biodegradable poly(lactic acid) (PLA) using a solvent-free approach. We used this model system to test the cytotoxicity of anticancer drugs, including temozolomide (TMZ) and ifosfamide (IFO). With the liver cells, TMZ showed a much lower toxicity to GBM cells under both 2D and 3D cell culture conditions. Comparing 2D, GBM cells cultured in 3D had much high viability under TMZ treatment. IFO was used to test the CYP-related metabolic effects. Cells with different expression levels of CYP3A4 differed dramatically in their ability to activate IFO, which led to strong metabolism-dependent cytotoxicity to GBM cells. These results demonstrate that our 3D-μPTC system could provide a more physiologically realistic in vitro environment than the current 2D monolayers for testing metabolism-dependent toxicity of anticancer drugs. It could therefore be used as an important platform for better prediction of drug dosing and schedule towards personalized medicine. PMID:22429982
Ikeda, Hiroko; Souda, Hikaru; Puspitasari, Anggraeini; Held, Kathryn D; Hidema, Jun; Nikawa, Takeshi; Yoshida, Yukari; Kanai, Tatsuaki; Takahashi, Akihisa
2017-02-01
Outer space is an environment characterized by microgravity and space radiation, including high-energy charged particles. Astronauts are constantly exposed to both microgravity and radiation during long-term stays in space. However, many aspects of the biological effects of combined microgravity and space radiation remain unclear. We developed a new three-dimensional (3D) clinostat synchronized heavy-ion irradiation system for use in ground-based studies of the combined exposures. Our new system uses a particle accelerator and a respiratory gating system from heavy-ion radiotherapy to irradiate samples being rotated in the 3D clinostat with carbon-ion beams only when the samples are in the horizontal position. A Peltier module and special sample holder were loaded on a static stage (standing condition) and the 3D clinostat (rotation condition) to maintain a suitable temperature under atmospheric conditions. The performance of the new device was investigated with normal human fibroblasts 1BR-hTERT in a disposable closed cell culture chamber. Live imaging revealed that cellular adhesion and growth were almost the same for the standing control sample and rotation sample over 48h. Dose flatness and symmetry were judged according to the relative density of Gafchromic films along the X-axis and Y-axis of the positions of the irradiated sample to confirm irradiation accuracy. Doses calculated using the carbon-ion calibration curve were almost the same for standing and rotation conditions, with the difference being less than 5% at 1Gy carbon-ion irradiation. Our new device can accurately synchronize carbon-ion irradiation and simulated microgravity while maintaining the temperature under atmospheric conditions at ground level. Copyright © 2017 The Committee on Space Research (COSPAR). Published by Elsevier Ltd. All rights reserved.
Probabilistic structural analysis methods for space propulsion system components
NASA Technical Reports Server (NTRS)
Chamis, C. C.
1986-01-01
The development of a three-dimensional inelastic analysis methodology for the Space Shuttle main engine (SSME) structural components is described. The methodology is composed of: (1) composite load spectra, (2) probabilistic structural analysis methods, (3) the probabilistic finite element theory, and (4) probabilistic structural analysis. The methodology has led to significant technical progress in several important aspects of probabilistic structural analysis. The program and accomplishments to date are summarized.
Tamura, Masato; Yanagawa, Fumiki; Sugiura, Shinji; Takagi, Toshiyuki; Sumaru, Kimio; Matsui, Hirofumi; Kanamori, Toshiyuki
2014-05-07
Cell sorting is an essential and efficient experimental tool for the isolation and characterization of target cells. A three-dimensional environment is crucial in determining cell behavior and cell fate in biological analysis. Herein, we have applied photodegradable hydrogels to optical cell separation from a 3D environment using a computer-controlled light irradiation system. The hydrogel is composed of photocleavable tetra-arm polyethylene glycol and gelatin, which optimized cytocompatibility to adjust a composition of crosslinker and gelatin. Local light irradiation could degrade the hydrogel corresponding to the micropattern image designed on a laptop; minimum resolution of photodegradation was estimated at 20 µm. Light irradiation separated an encapsulated fluorescent microbead without any contamination of neighbor beads, even at multiple targets. Upon selective separation of target cells in the hydrogels, the separated cells have grown on another dish, resulting in pure culture. Cell encapsulation, light irradiation and degradation products exhibited negligible cytotoxicity in overall process.
Basel, Matthew T; Balivada, Sivasai; Wang, Hongwang; Shrestha, Tej B; Seo, Gwi Moon; Pyle, Marla; Abayaweera, Gayani; Dani, Raj; Koper, Olga B; Tamura, Masaaki; Chikan, Viktor; Bossmann, Stefan H; Troyer, Deryl L
2012-01-01
Using magnetic nanoparticles to absorb alternating magnetic field energy as a method of generating localized hyperthermia has been shown to be a potential cancer treatment. This report demonstrates a system that uses tumor homing cells to actively carry iron/iron oxide nanoparticles into tumor tissue for alternating magnetic field treatment. Paramagnetic iron/ iron oxide nanoparticles were synthesized and loaded into RAW264.7 cells (mouse monocyte/ macrophage-like cells), which have been shown to be tumor homing cells. A murine model of disseminated peritoneal pancreatic cancer was then generated by intraperitoneal injection of Pan02 cells. After tumor development, monocyte/macrophage-like cells loaded with iron/ iron oxide nanoparticles were injected intraperitoneally and allowed to migrate into the tumor. Three days after injection, mice were exposed to an alternating magnetic field for 20 minutes to cause the cell-delivered nanoparticles to generate heat. This treatment regimen was repeated three times. A survival study demonstrated that this system can significantly increase survival in a murine pancreatic cancer model, with an average post-tumor insertion life expectancy increase of 31%. This system has the potential to become a useful method for specifically and actively delivering nanoparticles for local hyperthermia treatment of cancer. PMID:22287840
Small-Bolt Torque-Tension Tester
NASA Technical Reports Server (NTRS)
Posey, Alan J.
2009-01-01
The device described here measures the torque-tension relationship for fasteners as small as #0. The small-bolt tester consists of a plate of high-strength steel into which three miniature load cells are recessed. The depth of the recess is sized so that the three load cells can be shimmed, the optimum height depending upon the test hardware. The three miniature load cells are arranged in an equilateral triangular configuration with the test bolt aligned with the centroid of the three. This is a kinematic arrangement.
Fang, Baizeng; Kim, Jung Ho; Kim, Minsik; Kim, Minwoo; Yu, Jong-Sung
2009-03-07
Hierarchical nanostructured spherical carbon with hollow macroporous core in combination with mesoporous shell has been explored to support Pt cathode catalyst with high metal loading in proton exchange membrane fuel cell (PEMFC). The hollow core-mesoporous shell carbon (HCMSC) has unique structural characteristics such as large specific surface area and mesoporous volume, ensuring uniform dispersion of the supported high loading (60 wt%) Pt nanoparticles with small particle size, and well-developed three-dimensionally interconnected hierarchical porosity network, facilitating fast mass transport. The HCMSC-supported Pt(60 wt%) cathode catalyst has demonstrated markedly enhanced catalytic activity toward oxygen reduction and greatly improved PEMFC polarization performance compared with carbon black Vulcan XC-72 (VC)-supported ones. Furthermore, the HCMSC-supported Pt(40 wt%) or Pt(60 wt%) outperforms the HCMSC-supported Pt(20 wt%) even at a low catalyst loading of 0.2 mg Pt cm(-2) in the cathode, which is completely different from the VC-supported Pt catalysts. The capability of supporting high loading Pt is supposed to accelerate the commercialization of PEMFC due to the anticipated significant reduction in the amount of catalyst support required, diffusion layer thickness and fabricating cost of the supported Pt catalyst electrode.
Design of Aspirated Compressor Blades Using Three-dimensional Inverse Method
NASA Technical Reports Server (NTRS)
Dang, T. Q.; Rooij, M. Van; Larosiliere, L. M.
2003-01-01
A three-dimensional viscous inverse method is extended to allow blading design with full interaction between the prescribed pressure-loading distribution and a specified transpiration scheme. Transpiration on blade surfaces and endwalls is implemented as inflow/outflow boundary conditions, and the basic modifications to the method are outlined. This paper focuses on a discussion concerning an application of the method to the design and analysis of a supersonic rotor with aspiration. Results show that an optimum combination of pressure-loading tailoring with surface aspiration can lead to a minimization of the amount of sucked flow required for a net performance improvement at design and off-design operations.
Begum, M. Sameena; Dinesh, M. R.; Tan, Kenneth F. H.; Jairaj, Vani; Md Khalid, K.; Singh, Varun Pratap
2015-01-01
The finite element method (FEM) is a powerful computational tool for solving stress-strain problems; its ability to handle material inhomogeneity and complex shapes makes the FEM, the most suitable method for the analysis of internal stress levels in the tooth, periodontium, and alveolar bone. This article intends to explain the steps involved in the generation of a three-dimensional finite element model of tooth, periodontal ligament (PDL) and alveolar bone, as the procedure of modeling is most important because the result is based on the nature of the modeling systems. Finite element analysis offers a means of determining strain-stress levels in the tooth, ligament, and bone structures for a broad range of orthodontic loading scenarios without producing tissue damage. PMID:26538895
Khajavi, Noushafarin; Akbari, Mohammad; Abolhassani, Farid; Dehpour, Ahmad Reza; Koruji, Morteza; Habibi Roudkenar, Mehryar
2014-01-01
Objective Spermatogonial stem cells (SSCs) are the only cell type that can restore fertility to an infertile recipient following transplantation. Much effort has been made to develop a protocol for differentiating isolated SSCs in vitro. Recently, three-dimensional (3D) culture system has been introduced as an appropriate microenvironment for clonal expansion and differentiation of SSCs. This system provides structural support and multiple options for several manipulation such as addition of different cells. Somatic cells have a critical role in stimulating spermatogenesis. They provide complex cell to cell interaction, transport proteins and produce enzymes and regulatory factors. This study aimed to optimize the culture condition by adding somatic testicular cells to the collagen gel culture system in order to induce spermatogenesis progression. Materials and Methods In this experimental study, the disassociation of SSCs was performed by using a two-step enzymatic digestion of type I collagenase, hyaluronidase and DNase. Somatic testicular cells including Sertoli cells and peritubular cells were obtained after the second digestion. SSCs were isolated by Magnetic Activated Cell Sorting (MACS) using GDNF family receptor alpha-1 (Gfrα-1) antibody. Two experimental designs were investigated. 1. Gfrα-1 positive SSCs were cultured in a collagen solution. 2. Somatic testicular cells were added to the Gfrα-1 positive SSCs in a collagen solution. Spermatogenesis progression was determined after three weeks by staining of synaptonemal complex protein 3 (SCP3)-positive cells. Semi-quantitative Reverse Transcription PCR was undertaken for SCP3 as a meiotic marker and, Crem and Thyroid transcription factor-1 (TTF1) as post meiotic markers. For statistical analysis student t test was performed. Results Testicular supporter cells increased the expression of meiotic and post meiotic markers and had a positive effect on extensive colony formation. Conclusion Collagen gel culture system supported by somatic testicular cells provides a microenvironment that mimics seminiferous epithelium and induces spermatogenesis in vitro. PMID:24518977
A novel bioprinting method and system for forming hybrid tissue engineering constructs.
Shanjani, Y; Pan, C C; Elomaa, L; Yang, Y
2015-12-18
Three dimensional (3D) bioprinting is a promising approach to form tissue engineering constructs (TECs) via positioning biomaterials, growth factors, and cells with controlled spatial distribution due to its layer-by-layer manufacturing nature. Hybrid TECs composed of relatively rigid porous scaffolds for structural and mechanical integrity and soft hydrogels for cell- and growth factor-loading have a tremendous potential to tissue regeneration under mechanical loading. However, despite excessive progress in the field, the current 3D bioprinting techniques and systems fall short in integration of such soft and rigid multifunctional components. Here we present a novel 3D hybrid bioprinting technology (Hybprinter) and its capability enabling integration of soft and rigid components for TECs. Hybprinter employs digital light processing-based stereolithography (DLP-SLA) and molten material extrusion techniques for soft and rigid materials, respectively. In this study, poly-ethylene glycol diacrylate (PEGDA) and poly-(ε-caprolactone) (PCL) were used as a model material for soft hydrogel and rigid scaffold, respectively. It was shown that geometrical accuracy, swelling ratio and mechanical properties of the hydrogel component can be tailored by DLP-SLA module. We have demonstrated the printability of variety of complex hybrid construct designs using Hybprinter technology and characterized the mechanical properties and functionality of such constructs. The compressive mechanical stiffness of a hybrid construct (90% hydrogel) was significantly higher than hydrogel itself (∼6 MPa versus 100 kPa). In addition, viability of cells incorporated within the bioprinted hybrid constructs was determined approximately 90%. Furthermore, a functionality of a hybrid construct composed of porous scaffold with an embedded hydrogel conduit was characterized for vascularized tissue engineering applications. High material diffusion and high cell viability in about 2.5 mm distance surrounding the conduit indicated that culture media effectively diffused through the conduit and fed the cells. The results suggest that the developed technology is potent to form functional TECs composed of rigid and soft biomaterials.
NASA Astrophysics Data System (ADS)
Dahlin, J. T.; Drake, J. F.; Swisdak, M.
2017-09-01
Magnetic reconnection is an important driver of energetic particles in many astrophysical phenomena. Using kinetic particle-in-cell simulations, we explore the impact of three-dimensional reconnection dynamics on the efficiency of particle acceleration. In two-dimensional systems, Alfvénic outflows expel energetic electrons into flux ropes where they become trapped and disconnected from acceleration regions. However, in three-dimensional systems these flux ropes develop an axial structure that enables particles to leak out and return to acceleration regions. This requires a finite guide field so that particles may move quickly along the flux rope axis. We show that greatest energetic electron production occurs when the guide field is of the same order as the reconnecting component: large enough to facilitate strong transport, but not so large as to throttle the dominant Fermi mechanism responsible for efficient electron acceleration. This suggests a natural explanation for the envelope of electron acceleration during the impulsive phase of eruptive flares.
Tiberi, Gianluigi; Fontana, Nunzia; Costagli, Mauro; Stara, Riccardo; Biagi, Laura; Symms, Mark Roger; Monorchio, Agostino; Retico, Alessandra; Cosottini, Mirco; Tosetti, Michela
2015-07-01
Local specific absorption rate (SAR) evaluation in ultra high field (UHF) magnetic resonance (MR) systems is a major concern. In fact, at UHF, radiofrequency (RF) field inhomogeneity generates hot-spots that could cause localized tissue heating. Unfortunately, local SAR measurements are not available in present MR systems; thus, electromagnetic simulations must be performed for RF fields and SAR analysis. In this study, we used three-dimensional full-wave numerical electromagnetic simulations to investigate the dependence of local SAR at 7.0 T with respect to subject size in two different scenarios: surface coil loaded by adult and child calves and quadrature volume coil loaded by adult and child heads. In the surface coil scenario, maximum local SAR decreased with decreasing load size, provided that the RF magnetic fields for the different load sizes were scaled to achieve the same slice average value. On the contrary, in the volume coil scenario, maximum local SAR was up to 15% higher in children than in adults. © 2015 Wiley Periodicals, Inc.
Fathallah, F A; Marras, W S; Parnianpour, M
1999-09-01
Most biomechanical assessments of spinal loading during industrial work have focused on estimating peak spinal compressive forces under static and sagittally symmetric conditions. The main objective of this study was to explore the potential of feasibly predicting three-dimensional (3D) spinal loading in industry from various combinations of trunk kinematics, kinetics, and subject-load characteristics. The study used spinal loading, predicted by a validated electromyography-assisted model, from 11 male participants who performed a series of symmetric and asymmetric lifts. Three classes of models were developed: (a) models using workplace, subject, and trunk motion parameters as independent variables (kinematic models); (b) models using workplace, subject, and measured moments variables (kinetic models); and (c) models incorporating workplace, subject, trunk motion, and measured moments variables (combined models). The results showed that peak 3D spinal loading during symmetric and asymmetric lifting were predicted equally well using all three types of regression models. Continuous 3D loading was predicted best using the combined models. When the use of such models is infeasible, the kinematic models can provide adequate predictions. Finally, lateral shear forces (peak and continuous) were consistently underestimated using all three types of models. The study demonstrated the feasibility of predicting 3D loads on the spine under specific symmetric and asymmetric lifting tasks without the need for collecting EMG information. However, further validation and development of the models should be conducted to assess and extend their applicability to lifting conditions other than those presented in this study. Actual or potential applications of this research include exposure assessment in epidemiological studies, ergonomic intervention, and laboratory task assessment.
Song, Juqing; Gao, Huichang; Zhu, Guanglin; Cao, Xiaodong; Shi, Xuetao; Wang, Yingjun
2016-08-26
The development of modern biomedical nanotechnology requires three-dimensional macrostructures with nanotextures to meet the requirements for practical applications in intricate biological systems. Additionally, the restoration and regeneration of some specific body tissues and organs rely on the function of conductive polymers, which can provide electrical cues for cells. In this study, we fabricated three-dimensional composite nanofibre macrostructures of polycaprolactone (PCL) with different concentrations of polyaniline (PANi) by employing an improved electrospinning technology with a specially designed collector. The 3D structures possessed cap-like macrostructures with centimetre-scale thickness and interconnected pore nanotextures with nanometre-scale nanofibres. To estimate the biocompatibility of the 3D PCL/PANi composite nanofibre macrostructures, mouse myoblasts (C2C12 cells) were cultured as model cells. The initial responses of C2C12 cells to the 3D PCL/PANi composite macrostructures were significantly superior to those to pure PCL, that is, the cells exhibited typical myoblast-like morphologies with obvious pseudopodia and the moderate incorporation (less than 2.0 wt%) of conductive PANi facilitated cell proliferation, which indicated that PANi has appreciable cell affinity. Moreover, the addition of conductive PANi to the 3D composite nanofibre macrostructures considerably enhanced myoblast differentiation and myotube maturation. These results suggest that electrospun 3D PCL/PANi composite nanofibre macrostructures would have promising applications in tissue engineering.
Cobb, Corie L.; Solberg, Scott E.
2017-04-29
3-dimensional (3D) electrode architectures have been explored as a means to decouple power and energy trade-offs in thick battery electrodes. Limited work has been published which systematically examines the impact of these architectures at the pouch cell level. This paper conducts an analysis on the potential capacity gains that can be realized with thick co-extruded electrodes in a pouch cell. Moreover, our findings show that despite lower active material composition for each cathode layer, the effective gain in thickness and active material loading enables pouch cell capacity gains greater than 10% with a Lithium Nickel Manganese Cobalt Oxide (NMC) materialsmore » system.« less
Ultrahigh resolution optical coherence elastography using a Bessel beam for extended depth of field
NASA Astrophysics Data System (ADS)
Curatolo, Andrea; Villiger, Martin; Lorenser, Dirk; Wijesinghe, Philip; Fritz, Alexander; Kennedy, Brendan F.; Sampson, David D.
2016-03-01
Visualizing stiffness within the local tissue environment at the cellular and sub-cellular level promises to provide insight into the genesis and progression of disease. In this paper, we propose ultrahigh-resolution optical coherence elastography, and demonstrate three-dimensional imaging of local axial strain of tissues undergoing compressive loading. The technique employs a dual-arm extended focus optical coherence microscope to measure tissue displacement under compression. The system uses a broad bandwidth supercontinuum source for ultrahigh axial resolution, Bessel beam illumination and Gaussian beam detection, maintaining sub-2 μm transverse resolution over nearly 100 μm depth of field, and spectral-domain detection allowing high displacement sensitivity. The system produces strain elastograms with a record resolution (x,y,z) of 2×2×15 μm. We benchmark the advances in terms of resolution and strain sensitivity by imaging a suitable inclusion phantom. We also demonstrate this performance on freshly excised mouse aorta and reveal the mechanical heterogeneity of vascular smooth muscle cells and elastin sheets, otherwise unresolved in a typical, lower resolution optical coherence elastography system.
Three-dimensional behavior of ice crystals and biological cells during freezing of cell suspensions.
Ishiguro, H; Koike, K
1998-09-11
Behavior of ice crystals and human red blood cells during extracellular-freezing was investigated in three-dimensions using a confocal laser scanning microscope(CLSM), which noninvasively produces tomograms of biological materials. Physiological saline and physiological saline with 2.4 M glycerol were used for suspension. Various cooling rates for directional solidification were used for distinctive morphology of the ice crystals. Addition of acridine orange as a fluorescent dye into the cell suspension enabled ice crystal, cells and unfrozen solution to be distinguished by different colors. The results indicate that the microscopic structure is three-dimensional for flat, cellular, and dendritic solid-liquid interfaces and that a CLSM is very effective in studying three-dimensional structure during the freezing of cell suspensions.
Three-dimensional bioprinting in tissue engineering and regenerative medicine.
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.
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.
Akça, Kıvanç; Eser, Atılım; Çavuşoğlu, Yeliz; Sağırkaya, Elçin; Çehreli, Murat Cavit
2015-05-01
The aim of this study was to investigate conventionally and early loaded titanium and titanium-zirconium alloy implants by three-dimensional finite element stress analysis. Three-dimensional model of a dental implant was created and a thread area was established as a region of interest in trabecular bone to study a localized part of the global model with a refined mesh. The peri-implant tissues around conventionally loaded (model 1) and early loaded (model 2) implants were implemented and were used to explore principal stresses, displacement values, and equivalent strains in the peri-implant region of titanium and titanium-zirconium implants under static load of 300 N with or without 30° inclination applied on top of the abutment surface. Under axial loading, principal stresses in both models were comparable for both implants and models. Under oblique loading, principal stresses around titanium-zirconium implants were slightly higher in both models. Comparable stress magnitudes were observed in both models. The displacement values and equivalent strain amplitudes around both implants and models were similar. Peri-implant bone around titanium and titanium-zirconium implants experiences similar stress magnitudes coupled with intraosseous implant displacement values under conventional loading and early loading simulations. Titanium-zirconium implants have biomechanical outcome comparable to conventional titanium implants under conventional loading and early loading.
NASA Technical Reports Server (NTRS)
Gloss, B. B.; Johnson, F. T.
1976-01-01
The Boeing Commercial Airplane Company developed an inviscid three-dimensional lifting surface method that shows promise in being able to accurately predict loads, subsonic and supersonic, on wings with leading-edge separation and reattachment.
Mahmoudzadeh, Aziz; Mohammadpour, Hemn
2016-07-01
Tumor cells naturally live in three-dimensional (3D) microenvironments, while common laboratory tests and evaluations are done in two-dimensional (2D) plates. This study examined the impact of cultured 4T1 cancer cells in a 3D collagen-chitosan scaffold compared with 2D plate cultures. Collagen-chitosan scaffolds were provided and passed confirmatory tests. 4T1 tumor cells were cultured on scaffolds and then tumor cells growth rate, resistance to X-ray radiation, and cyclophosphamide as a chemotherapy drug were analyzed. Furthermore, 4T1 cells were extracted from the scaffold model and were injected into the mice. Tumor growth rate, survival rate, and systemic immune responses were evaluated. Our results showed that 4T1 cells infiltrated the scaffolds pores and constructed a 3D microenvironment. Furthermore, 3D cultured tumor cells showed a slower proliferation rate, increased levels of survival to the X-ray irradiation, and enhanced resistance to chemotherapy drugs in comparison with 2D plate cultures. Transfer of extracted cells to the mice caused enhanced tumor volume and decreased life span. This study indicated that collagen-chitosan nanoscaffolds provide a suitable model of tumor that would be appropriate for tumor studies. Copyright © 2016. Published by Elsevier B.V.
Monodisperse alginate microgel formation in a three-dimensional microfluidic droplet generator.
Lian, Meng; Collier, C Patrick; Doktycz, Mitchel J; Retterer, Scott T
2012-01-01
Droplet based microfluidic systems provide an ideal platform for partitioning and manipulating aqueous samples for analysis. Identifying stable operating conditions under which droplets are generated is challenging yet crucial for real-world applications. A novel three-dimensional microfluidic platform that facilitates the consistent generation and gelation of alginate-calcium hydrogel microbeads for microbial encapsulation, over a broad range of input pressures, in the absence of surfactants is described. The unique three-dimensional design of the fluidic network utilizes a height difference at the junction between the aqueous sample injection and organic carrier channels to induce droplet formation via a surface tension enhanced self-shearing mechanism. Combined within a flow-focusing geometry, under constant pressure control, this arrangement facilitates predictable generation of droplets over a much broader range of operating conditions than that of conventional two-dimensional systems. The impact of operating pressures and geometry on droplet gelation, aqueous and organic material flow rates, microbead size, and bead generation frequency are described. The system presented provides a robust platform for encapsulating single microbes in complex mixtures into individual hydrogel beads, and provides the foundation for the development of a complete system for sorting and analyzing microbes at the single cell level.
Monodisperse alginate microgel formation in a three-dimensional microfluidic droplet generator
Lian, Meng; Collier, C. Patrick; Doktycz, Mitchel J.; Retterer, Scott T.
2012-01-01
Droplet based microfluidic systems provide an ideal platform for partitioning and manipulating aqueous samples for analysis. Identifying stable operating conditions under which droplets are generated is challenging yet crucial for real-world applications. A novel three-dimensional microfluidic platform that facilitates the consistent generation and gelation of alginate-calcium hydrogel microbeads for microbial encapsulation, over a broad range of input pressures, in the absence of surfactants is described. The unique three-dimensional design of the fluidic network utilizes a height difference at the junction between the aqueous sample injection and organic carrier channels to induce droplet formation via a surface tension enhanced self-shearing mechanism. Combined within a flow-focusing geometry, under constant pressure control, this arrangement facilitates predictable generation of droplets over a much broader range of operating conditions than that of conventional two-dimensional systems. The impact of operating pressures and geometry on droplet gelation, aqueous and organic material flow rates, microbead size, and bead generation frequency are described. The system presented provides a robust platform for encapsulating single microbes in complex mixtures into individual hydrogel beads, and provides the foundation for the development of a complete system for sorting and analyzing microbes at the single cell level. PMID:24198865
Polyphosphazene/Nano-Hydroxyapatite Composite Microsphere Scaffolds for Bone Tissue Engineering
Nukavarapu, Syam P.; Kumbar, Sangamesh G.; Brown, Justin L.; Krogman, Nicholas R.; Weikel, Arlin L.; Hindenlang, Mark D.; Nair, Lakshmi S.; Allcock, Harry R; Laurencin, Cato T.
2009-01-01
The non-toxic, neutral degradation products of amino acid ester polyphosphazenes make them ideal candidates for in vivo orthopaedic applications. The quest for new osteocompatible materials for load bearing tissue engineering applications has led us to investigate mechanically competent amino acid ester substituted polyphosphazenes. In this study, we have synthesized three biodegradable polyphosphazenes substituted with side groups namely leucine, valine and phenylalanine ethyl esters. Of these polymers, the phenylalanine ethyl ester substituted polyphosphazene showed the highest glass transition temperature (41.6 °C) and hence was chosen as a candidate material for forming composite microspheres with 100 nm sized hydroxyapatite (nHAp). The fabricated composite microspheres were sintered into a three-dimensional (3-D) porous scaffold by adopting a dynamic solvent sintering approach. The composite microsphere scaffolds showed compressive moduli of 46–81 MPa with mean pore diameters in the range of 86–145 µm. The three-dimensional polyphosphazene-nHAp composite microsphere scaffolds showed good osteoblast cell adhesion, proliferation and alkaline phosphatase expression, and are potential suitors for bone tissue engineering applications. PMID:18517248
Li, Angsheng; Yin, Xianchen; Pan, Yicheng
2016-01-01
In this study, we propose a method for constructing cell sample networks from gene expression profiles, and a structural entropy minimisation principle for detecting natural structure of networks and for identifying cancer cell subtypes. Our method establishes a three-dimensional gene map of cancer cell types and subtypes. The identified subtypes are defined by a unique gene expression pattern, and a three-dimensional gene map is established by defining the unique gene expression pattern for each identified subtype for cancers, including acute leukaemia, lymphoma, multi-tissue, lung cancer and healthy tissue. Our three-dimensional gene map demonstrates that a true tumour type may be divided into subtypes, each defined by a unique gene expression pattern. Clinical data analyses demonstrate that most cell samples of an identified subtype share similar survival times, survival indicators and International Prognostic Index (IPI) scores and indicate that distinct subtypes identified by our algorithms exhibit different overall survival times, survival ratios and IPI scores. Our three-dimensional gene map establishes a high-definition, one-to-one map between the biologically and medically meaningful tumour subtypes and the gene expression patterns, and identifies remarkable cells that form singleton submodules. PMID:26842724
NASA Astrophysics Data System (ADS)
Nguyen, H. T.; Le, M. V.; Nguyen, T. A.; Nguyen, T. A. N.
2017-06-01
The solid oxide fuel cell is one of the promising technologies for future energy demand. Solid oxide fuel cell operated in the single-chamber mode exhibits several advantages over conventional single oxide fuel cell due to the simplified, compact, sealing-free cell structure. There are some studies on simulating the behavior of this type of fuel cell but they mainly focus on the 2D model. In the present study, a three-dimensional numerical model of a single chamber solid oxide fuel cell (SOFC) is reported and solved using COMSOL Multiphysics software. Experiments of a planar button solid oxide fuel cell were used to verify the simulation results. The system is fed by methane and oxygen and operated at 700°C. The cathode is LSCF6482, the anode is GDC-Ni, the electrolyte is LDM and the operating pressure is 1 atm. There was a good agreement between the cell temperature and current voltage estimated from the model and measured from the experiment. The results indicate that the model is applicable for the single chamber solid oxide fuel cell and it can provide a basic for the design, scale up of single chamber solid oxide fuel cell system.
Rational Design of Iron Oxide Nanoparticles as Targeted Nanomedicines for Cancer Therapy
NASA Astrophysics Data System (ADS)
Kievit, Forrest M.
2011-07-01
Nanotechnology provides a flexible platform for the development of effective therapeutic nanomaterials that can interact specifically with a target in a biological system and provoke a desired biological response. Of the nanomaterials studied, superparamagnetic iron oxide nanoparticles (SPIONs) have emerged as one of top candidates for cancer therapy due to their intrinsic superparamagnetism that enables non-invasive magnetic resonance imaging (MRI) and biodegradability favorable for in vivo application. This dissertation is aimed at development of SPION-based nanomedicines to overcome the current limitations in cancer therapy. These limitations include non-specificity of therapy which can harm healthy tissue, the difficulty in delivering nucleic acids for gene therapy, the formation of drug resistance, and the inability to detect and treat micrometastases. First, a SPION-based non-viral gene delivery vehicle was developed through functionalization of the SPION core with a co-polymer designed to provide stable binding of DNA and low toxicity which showed excellent gene delivery in vitro and in vivo. This SPION-based non-viral gene delivery vehicle was then activated with a targeting agent to improve gene delivery throughout a xenograft tumor of brain cancer. It was found that targeting did not promote the accumulation of SPIONs at the tumor site, but rather improved the distribution of SPIONs throughout the tumor so a higher proportion of cells received treatment. Next, the high surface area of SPIONs was utilized for loading large amounts of drug which was shown to overcome the multidrug resistance acquired by many cancer cells. Drug bound to SPIONs showed significantly higher multidrug resistant cell uptake as compared to free drug which translated into improved cell kill. Also, an antibody activated SPION was developed and was shown to be able to target micrometastases in a transgenic animal model of metastatic breast cancer. These SPION-based nanomedicines provide a platform for the future development of therapies that are hoped to overcome the current limitations in cancer therapy. Finally, a three-dimensional in vitro tumor tissue culture model was developed for mimicking the tumor microenvironment in which cultured cells showed higher malignancy than traditional two-dimensional and three-dimensional models. This in vitro model should provided researches with a better tool for testing novel nanomedicines in vitro before moving to the more costly in vivo experiments.
A three-dimensional non-isothermal model for a membraneless direct methanol redox fuel cell
NASA Astrophysics Data System (ADS)
Wei, Lin; Yuan, Xianxia; Jiang, Fangming
2018-05-01
In the membraneless direct methanol redox fuel cell (DMRFC), three-dimensional electrodes contribute to the reduction of methanol crossover and the open separator design lowers the system cost and extends its service life. In order to better understand the mechanisms of this configuration and further optimize its performance, the development of a three-dimensional numerical model is reported in this work. The governing equations of the multi-physics field are solved based on computational fluid dynamics methodology, and the influence of the CO2 gas is taken into consideration through the effective diffusivities. The numerical results are in good agreement with experimental data, and the deviation observed for cases of large current density may be related to the single-phase assumption made. The three-dimensional electrode is found to be effective in controlling methanol crossover in its multi-layer structure, while it also increases the flow resistance for the discharging products. It is found that the current density distribution is affected by both the electronic conductivity and the concentration of reactants, and the temperature rise can be primarily attributed to the current density distribution. The sensitivity and reliability of the model are analyzed through the investigation of the effects of cell parameters, including porosity values of gas diffusion layers and catalyst layers, methanol concentration and CO2 volume fraction, on the polarization characteristics.
Economic competitiveness of fuel cell onsite integrated energy systems
NASA Technical Reports Server (NTRS)
Bollenbacher, G.
1983-01-01
The economic competitiveness of fuel cell onsite integrated energy systems (OS/IES) in residential and commercial buildings is examined. The analysis is carried out for three different buildings with each building assumed to be at three geographic locations spanning a range of climatic conditions. Numerous design options and operating strategies are evaluated and two economic criteria are used to measure economic performance. In general the results show that fuel cell OS/IES's are competitive in most regions of the country if the OS/IES is properly designed. The preferred design is grid connected, makes effective use of the fuel cell's thermal output, and has a fuel cell powerplant sized for the building's base electrical load.
Load partitioning in Ai{sub 2}0{sub 3-}Al composites with three- dimensional periodic architecture.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Young, M. L.; Rao, R.; Almer, J. D.
2009-05-01
Interpenetrating composites are created by infiltration of liquid aluminum into three-dimensional (3-D) periodic Al{sub 2}O{sub 3} preforms with simple tetragonal symmetry produced by direct-write assembly. Volume-averaged lattice strains in the Al{sub 2}O{sub 3} phase of the composite are measured by synchrotron X-ray diffraction for various uniaxial compression stresses up to -350MPa. Load transfer, found by diffraction to occur from the metal phase to the ceramic phase, is in general agreement with simple rule-of-mixture models and in better agreement with more complex, 3-D finite-element models that account for metal plasticity and details of the geometry of both phases. Spatially resolved diffractionmore » measurements show variations in load transfer at two different positions within the composite.« less
In situ three-dimensional reciprocal-space mapping during mechanical deformation.
Cornelius, T W; Davydok, A; Jacques, V L R; Grifone, R; Schülli, T; Richard, M I; Beutier, G; Verdier, M; Metzger, T H; Pietsch, U; Thomas, O
2012-09-01
Mechanical deformation of a SiGe island epitaxically grown on Si(001) was studied by a specially adapted atomic force microscope and nanofocused X-ray diffraction. The deformation was monitored during in situ mechanical loading by recording three-dimensional reciprocal-space maps around a selected Bragg peak. Scanning the energy of the incident beam instead of rocking the sample allowed the safe and reliable measurement of the reciprocal-space maps without removal of the mechanical load. The crystal truncation rods originating from the island side facets rotate to steeper angles with increasing mechanical load. Simulations of the displacement field and the intensity distribution, based on the finite-element method, reveal that the change in orientation of the side facets of about 25° corresponds to an applied pressure of 2-3 GPa on the island top plane.
Cationized pullulan 3D matrices as new materials for gene transfer.
San Juan, Aurélie; Hlawaty, Hanna; Chaubet, Frédéric; Letourneur, Didier; Feldman, Laurent J
2007-08-01
This study deals with the development of a novel biocompatible cationized pullulan three-dimensional matrix for gene delivery. A water-soluble cationic polysaccharide, diethylaminoethyl-pullulan (DEAE-pullulan), was first synthesized and characterized. Fluorescence quenching and gel retardation assays evidenced the complexation in solution of DNA with DEAE-pullulan, but not with neutral pullulan. On cultured smooth muscle cells (SMCs) incubated with DEAE-pullulan and a plasmid vector expressing a secreted form of alkaline phosphatase (pSEAP), SEAP activity was 150-fold higher than with pSEAP alone or pSEAP with neutral pullulan. DEAE-pullulan was then chemically crosslinked using phosphorus oxychloride. The resulting matrices were obtained in less than a minute and molded as discs of 12 mm diameter and 2 mm thickness. Such DEAE-pullulan 3D matrices were loaded with up to 50 microg of plasmid DNA, with a homogeneous plasmid loading observed with YOYO-1 fluorescence staining. Moreover, the DEAE-pullulan matrix was shown to protect pSEAP from DNase I degradation. Incubation of cultured SMCs with pSEAP-loaded DEAE-pullulan matrices resulted in significant gene transfer without cell toxicity. This study suggests that these cationized pullulan 3D matrices could be useful biomaterials for local gene transfer.
NASA Astrophysics Data System (ADS)
Licciardi, Mariano; Scialabba, Cinzia; Giammona, Gaetano; Paolino, Marco; Razzano, Vincenzo; Grisci, Giorgio; Giuliani, Germano; Makovec, Francesco; Cappelli, Andrea
2017-06-01
A tri-component polymer brush (TCPB ), composed of a polybenzofulvene copolymer bearing low molecular weight hyaluronic acid (HA) on the surface of its cylindrical brush-like backbone and oligo-PEG fractions, was employed in the preparation of 350 nm nanostructured drug delivery systems capable of delivering the anticancer drug doxorubicin. The obtained drug delivery systems were characterized on the basis of drug loading and release, dimensions and zeta potential, morphology and in vitro cell activity, and uptake on three different human cell lines, namely the bronchial epithelial 16HBE, the breast adenocarcinoma MCF-7, and the colon cancer HCT116 cells. Finally, the ability of doxorubicin-loaded TCPB nanoparticles (DOXO-TCPB) to be internalized into cancer cells by CD44 receptor mediated uptake was assessed by means of uptake studies in HCT cells. These data were supported by anti-CD44-FITC staining assay. The proposed TCPB nanostructured drug delivery systems have many potential applications in nanomedicine, including cancer targeted drug delivery.
Alonzo, Luis F; Moya, Monica L; Shirure, Venktesh S; George, Steven C
2015-09-07
Tissue engineering can potentially recreate in vivo cellular microenvironments in vitro for an array of applications such as biological inquiry and drug discovery. However, the majority of current in vitro systems still neglect many biological, chemical, and mechanical cues that are known to impact cellular functions such as proliferation, migration, and differentiation. To address this gap, we have developed a novel microfluidic device that precisely controls the spatial and temporal interactions between adjacent three-dimensional cellular environments. The device consists of four interconnected microtissue compartments (~0.1 mm(3)) arranged in a square. The top and bottom pairs of compartments can be sequentially loaded with discrete cellularized hydrogels creating the opportunity to investigate homotypic (left to right or x-direction) and heterotypic (top to bottom or y-direction) cell-cell communication. A controlled hydrostatic pressure difference across the tissue compartments in both x and y direction induces interstitial flow and modulates communication via soluble factors. To validate the biological significance of this novel platform, we examined the role of stromal cells in the process of vasculogenesis. Our device confirms previous observations that soluble mediators derived from normal human lung fibroblasts (NHLFs) are necessary to form a vascular network derived from endothelial colony forming cell-derived endothelial cells (ECFC-ECs). We conclude that this platform could be used to study important physiological and pathological processes that rely on homotypic and heterotypic cell-cell communication.
Role of cellular adhesions in tissue dynamics spectroscopy
NASA Astrophysics Data System (ADS)
Merrill, Daniel A.; An, Ran; Turek, John; Nolte, David
2014-02-01
Cellular adhesions play a critical role in cell behavior, and modified expression of cellular adhesion compounds has been linked to various cancers. We tested the role of cellular adhesions in drug response by studying three cellular culture models: three-dimensional tumor spheroids with well-developed cellular adhesions and extracellular matrix (ECM), dense three-dimensional cell pellets with moderate numbers of adhesions, and dilute three-dimensional cell suspensions in agarose having few adhesions. Our technique for measuring the drug response for the spheroids and cell pellets was biodynamic imaging (BDI), and for the suspensions was quasi-elastic light scattering (QELS). We tested several cytoskeletal chemotherapeutic drugs (nocodazole, cytochalasin-D, paclitaxel, and colchicine) on three cancer cell lines chosen from human colorectal adenocarcinoma (HT-29), human pancreatic carcinoma (MIA PaCa-2), and rat osteosarcoma (UMR-106) to exhibit differences in adhesion strength. Comparing tumor spheroid behavior to that of cell suspensions showed shifts in the spectral motion of the cancer tissues that match predictions based on different degrees of cell-cell contacts. The HT-29 cell line, which has the strongest adhesions in the spheroid model, exhibits anomalous behavior in some cases. These results highlight the importance of using three-dimensional tissue models in drug screening with cellular adhesions being a contributory factor in phenotypic differences between the drug responses of tissue and cells.
NASA Technical Reports Server (NTRS)
Yang, Ren; Feeback, Daniel L.; Wang, Wan-Jun
2005-01-01
This paper details a novel three-dimensional (3D) hydro-focusing micro cell sorter for micro flow cytometry applications. The unit was microfabricated by means of SU-8 3D lithography. The 3D microstructure for coaxial sheathing was designed, microfabricated, and tested. Three-dimensional hydrofocusing capability was demonstrated with an experiment to sort labeled tanned sheep erythrocytes (red blood cells). This polymer hydro-focusing microstructure is easily microfabricated and integrated with other polymer microfluidic structures. Keywords: SU-8, three-dimensional hydro-focusing, microfluidic, microchannel, cytometer
Characterization of fluid flow by digital correlation of scattered light
NASA Technical Reports Server (NTRS)
Gilbert, John A.; Matthys, Donald R.
1989-01-01
The objective is to produce a physical system suitable for a space environment that can measure fluid velocities in a three-dimensional volume by the development of a particle correlation velocimetry technique. Experimental studies were conducted on a field test cell to demonstrate the suitability and accuracy of digital correlation techniques for measuring two-dimensional fluid flows. This objective was satisfied by: (1) the design of an appropriate illumination and detection system for making velocity measurements within a test cell; (2) the design and construction of a test cell; (3) the preliminary evaluations on fluid and seeding requirements; and (4) the performance of controlled tests using a multiple exposure correlation technique. This presentation is represented by viewgraphs with very little text.
Investigation of Micro-Scale Architectural Effects on Damage of Composites
NASA Technical Reports Server (NTRS)
Stier, Bertram; Bednarcyk, Brett A.; Simon, Jaan W.; Reese, Stefanie
2015-01-01
This paper presents a three-dimensional, energy based, anisotropic, stiffness reduction, progressive damage model for composite materials and composite material constituents. The model has been implemented as a user-defined constitutive model within the Abaqus finite element software package and applied to simulate the nonlinear behavior of a damaging epoxy matrix within a unidirectional composite material. Three different composite microstructures were considered as finite element repeating unit cells, with appropriate periodicity conditions applied at the boundaries. Results representing predicted transverse tensile, longitudinal shear, and transverse shear stress-strain curves are presented, along with plots of the local fields indicating the damage progression within the microstructure. It is demonstrated that the damage model functions appropriately at the matrix scale, enabling localization of the damage to simulate failure of the composite material. The influence of the repeating unit cell geometry and the effect of the directionality of the applied loading are investigated and discussed.
Modeling extracellular fields for a three-dimensional network of cells using NEURON.
Appukuttan, Shailesh; Brain, Keith L; Manchanda, Rohit
2017-10-01
Computational modeling of biological cells usually ignores their extracellular fields, assuming them to be inconsequential. Though such an assumption might be justified in certain cases, it is debatable for networks of tightly packed cells, such as in the central nervous system and the syncytial tissues of cardiac and smooth muscle. In the present work, we demonstrate a technique to couple the extracellular fields of individual cells within the NEURON simulation environment. The existing features of the simulator are extended by explicitly defining current balance equations, resulting in the coupling of the extracellular fields of adjacent cells. With this technique, we achieved continuity of extracellular space for a network model, thereby allowing the exploration of extracellular interactions computationally. Using a three-dimensional network model, passive and active electrical properties were evaluated under varying levels of extracellular volumes. Simultaneous intracellular and extracellular recordings for synaptic and action potentials were analyzed, and the potential of ephaptic transmission towards functional coupling of cells was explored. We have implemented a true bi-domain representation of a network of cells, with the extracellular domain being continuous throughout the entire model. This has hitherto not been achieved using NEURON, or other compartmental modeling platforms. We have demonstrated the coupling of the extracellular field of every cell in a three-dimensional model to obtain a continuous uniform extracellular space. This technique provides a framework for the investigation of interactions in tightly packed networks of cells via their extracellular fields. Copyright © 2017 Elsevier B.V. All rights reserved.
Study on dynamic performance of SOFC
NASA Astrophysics Data System (ADS)
Zhan, Haiyang; Liang, Qianchao; Wen, Qiang; Zhu, Runkai
2017-05-01
In order to solve the problem of real-time matching of load and fuel cell power, it is urgent to study the dynamic response process of SOFC in the case of load mutation. The mathematical model of SOFC is constructed, and its performance is simulated. The model consider the influence factors such as polarization effect, ohmic loss. It also takes the diffusion effect, thermal effect, energy exchange, mass conservation, momentum conservation. One dimensional dynamic mathematical model of SOFC is constructed by using distributed lumped parameter method. The simulation results show that the I-V characteristic curves are in good agreement with the experimental data, and the accuracy of the model is verified. The voltage response curve, power response curve and the efficiency curve are obtained by this way. It lays a solid foundation for the research of dynamic performance and optimal control in power generation system of high power fuel cell stack.
Three-dimensional cell to tissue development process
NASA Technical Reports Server (NTRS)
Goodwin, Thomas J. (Inventor); Parker, Clayton R. (Inventor)
2008-01-01
An improved three-dimensional cell to tissue development process using a specific time varying electromagnetic force, pulsed, square wave, with minimum fluid shear stress, freedom for 3-dimensional spatial orientation of the suspended particles and localization of particles with differing or similar sedimentation properties in a similar spatial region.
Shi, Wei-Bin; Le, Van-Minh; Gu, Chun-Hua; Zheng, Yuan-Hong; Lang, Mei-Dong; Lu, Yan-Hua; Liu, Jian-Wen
2014-04-01
The principal limitations of chemotherapy are dose-limiting systemic toxicity and the development of multidrug-resistant phenotypes. The aim of this study was to investigate the efficiency of a new sustained drug delivery system based on chitosan and ε-caprolactone to overcome multidrug resistance in monolayer and drug resistance associated with the three-dimensional (3D) tumor microenvironment in our established 3D models. The 5-fluorouracil (5-FU)-loaded nanoparticles (NPs) were characterized by transmission electron microscope and dynamic light scattering, and its released property was determined at different pH values. 5-FU/NPs exhibited well-sustained release properties and markedly enhanced the cytotoxicity of 5-FU against HCT116/L-OHP or HCT8/VCR MDR cells in two-dimensional (2D) and its parental cells in 3D collagen gel culture with twofold to threefold decrease in the IC50 values, as demonstrated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Hoechst/propidium iodide staining and flow cytometry analysis. Furthermore, the possible mechanism was explored by high-performance liquid chromatography and rhodamine 123 accumulation experiment. Overall, the results demonstrated that 5-FU/NPs increase intracellular concentration of 5-FU and enhance its anticancer efficiency by inducing apoptosis. It was suggested that this novel NPs are a promising carrier to decrease toxic of 5-FU and has the potential to reverse the forms of both intrinsic and acquired drug resistance in 2D and 3D cultures. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.
NASA Astrophysics Data System (ADS)
Wong, Derek N.
The US Navy is actively developing all electric fleets, raising serious questions about what is required of onboard power supplies in order to properly power the ship's electrical systems. This is especially relevant when choosing a viable power source to drive high power propulsion and electric weapon systems in addition to the conventional loads deployed aboard these types of vessels. Especially when high pulsed power loads are supplied, the issue of maintaining power quality becomes important and increasingly complex. Conventionally, a vessel's electrical power is generated using gas turbine or diesel driven motor-generator sets that are very inefficient when they are used outside of their most efficient load condition. What this means is that if the generator is not being utilized continuously at its most efficient load capacity, the quality of the output power may also be effected and fall outside of the acceptable power quality limits imposed through military standards. As a solution to this potential problem, the Navy has proposed using electrochemical storage devices since they are able to buffer conventional generators when the load is operating below the generator's most efficient power level or able to efficiently augment a generator when the load is operating in excess of the generator's most efficient power rating. Specifically, the US Navy is interested in using commercial off-the-shelf (COTS) lithium-ion batteries within an intelligently controlled energy storage module that could act as either a prime power supply for on-board pulsed power systems or as a backup generator to other shipboard power systems. Due to the unique load profile of high-rate pulsed power systems, the implementation of lithium-ion batteries within these complex systems requires them to be operated at very high rates and the effects these things have on cell degradation has been an area of focus. There is very little published research into the effects that high power transient or pulsed loading has on the degradation mechanisms of secondary lithium-ion cells. Prior to performing this work, it was unclear if the implementation of lithium-ion batteries in highly transient load conditions at high rate would accelerate cell degradation mechanisms that have been previously considered as minor issues. This work has focused on answering these previously unanswered questions. In early experiments performed here, COTS lithium-iron-phosphate (LFP) cells were studied under high-rate, transient load conditions and it was found that their capacity fade deviated from the traditional linear behavior and exponentially declined until no charge could be accepted when recharge was attempted at high rate. These findings indicated that subjecting LFP chemistries to transient, high rate charge/discharge profiles induced rapid changes in the electrode/electrolyte interface that rendered the cells useless when high rate recharge was required. These findings suggested there was more phenomena to learn about how these cells degraded under high rate pulsed conditions before they are fielded in Naval applications. Therefore, the research presented here has been focused on understanding the degradation mechanisms that are unique to LFP cells when they are cycled under pulsed load profiles at high charge and discharge rates. In particular, the work has been focused on identifying major degradation reactions that occur by studying the surface chemistry of cycled electrode materials. Efforts have been performed to map the impedance evolution of both cathode and anode half cells, respectively, using a novel three electrode technique that was developed for this research. Using this technique, the progression of degradation has been mapped using analysis of differential capacitance spectrums. In both the three electrode EIS mapping and differential capacitance analysis that has been performed, electrical component models have been developed. The results presented will show that there are unique degradation mechanisms induced through high rate pulsed loading conditions that are not normally seen in low rate continuous cycling of LFP cells.
NASA Astrophysics Data System (ADS)
Lazarowitz, Reuven; Naim, Raphael
2013-08-01
The cell topic was taught to 9th-grade students in three modes of instruction: (a) students "hands-on," who constructed three-dimensional cell organelles and macromolecules during the learning process; (b) teacher demonstration of the three-dimensional model of the cell structures; and (c) teaching the cell topic with the regular learning material in an expository mode (which use one- or two-dimensional cell structures as are presented in charts, textbooks and microscopic slides). The sample included 669, 9th-grade students from 25 classes who were taught by 22 Biology teachers. Students were randomly assigned to the three modes of instruction, and two tests in content knowledge in Biology were used. Data were treated with multiple analyses of variance. The results indicate that entry behavior in Biology was equal for all the study groups and types of schools. The "hands-on" learning group who build three-dimensional models through the learning process achieved significantly higher on academic achievements and on the high and low cognitive questions' levels than the other two groups. The study indicates the advantages students may have being actively engaged in the learning process through the "hands-on" mode of instruction/learning.
NASA Astrophysics Data System (ADS)
Swisdak, M.; Dahlin, J. T.; Drake, J. F.
2017-12-01
Magnetic reconnection is an important driver of energetic particles in many space and astrophysical phenomena. Using kinetic particle-in-cell simulations, we explore the effects that the dynamics in three-dimensions has on reconnection and the efficiency of particle acceleration. In two-dimensional systems, Alfvenic outflows expel energetic electrons into flux ropes where they become trapped and disconnected from acceleration regions. However, in three-dimensional systems these flux ropes develop axial structure that enables particles to leak out and return to acceleration regions. This requires a finite guide field so that particles may move quickly along the flux rope axis. The greatest energetic electron production occurs when the guide field is of the same order as the reconnecting component: large enough to facilitate strong transport, but not so large as to throttle the dominant Fermi mechanism responsible for efficient electron acceleration.
Karimpoor, Mahroo; Yebra-Fernandez, Eva; Parhizkar, Maryam; Orlu, Mine; Craig, Duncan; Khorashad, Jamshid S; Edirisinghe, Mohan
2018-04-01
The development of assays for evaluating the sensitivity of leukaemia cells to anti-cancer agents is becoming an important aspect of personalized medicine. Conventional cell cultures lack the three-dimensional (3D) structure of the bone marrow (BM), the extracellular matrix and stromal components which are crucial for the growth and survival of leukaemia stem cells. To accurately predict the sensitivity of the leukaemia cells in an in vitro assay a culturing system containing the essential components of BM is required. In this study, we developed a porous calcium alginate foam-based scaffold to be used for 3D culture. The new 3D culture was shown to be cell compatible as it supported the proliferation of both normal haematopoietic and leukaemia cells. Our cell differential assay for myeloid markers showed that the porous foam-based 3D culture enhanced myeloid differentiation in both leukaemia and normal haematopoietic cells compared to two-dimensional culture. The foam-based scaffold reduced the sensitivity of the leukaemia cells to the tested antileukaemia agents in K562 and HL60 leukaemia cell line model and also primary myeloid leukaemia cells. This observation supports the application of calcium alginate foams as scaffold components of the 3D cultures for investigation of sensitivity to antileukaemia agents in primary myeloid cells. © 2018 The Author(s).
NASA Astrophysics Data System (ADS)
Su, Yanfeng; Cai, Zhijian; Liu, Quan; Lu, Yifan; Guo, Peiliang; Shi, Lingyan; Wu, Jianhong
2018-04-01
In this paper, an autostereoscopic three-dimensional (3D) display system based on synthetic hologram reconstruction is proposed and implemented. The system uses a single phase-only spatial light modulator to load the synthetic hologram of the left and right stereo images, and the parallax angle between two reconstructed stereo images is enlarged by a grating to meet the split angle requirement of normal stereoscopic vision. To realize the crosstalk-free autostereoscopic 3D display with high light utilization efficiency, the groove parameters of the grating are specifically designed by the rigorous coupled-wave theory for suppressing the zero-order diffraction, and then the zero-order nulled grating is fabricated by the holographic lithography and the ion beam etching. Furthermore, the diffraction efficiency of the fabricated grating is measured under the illumination of a laser beam with a wavelength of 532 nm. Finally, the experimental verification system for the proposed autostereoscopic 3D display is presented. The experimental results prove that the proposed system is able to generate stereoscopic 3D images with good performances.
NASA Technical Reports Server (NTRS)
Pierzga, M. J.; Wood, J. R.
1984-01-01
An experimental investigation of the three dimensional flow field through a low aspect ratio, transonic, axial flow fan rotor has been conducted using an advanced laser anemometer (LA) system. Laser velocimeter measurements of the rotor flow field at the design operating speed and over a range of through flow conditions are compared to analytical solutions. The numerical technique used herein yields the solution to the full, three dimensional, unsteady Euler equations using an explicit time marching, finite volume approach. The numerical analysis, when coupled with a simplified boundary layer calculation, generally yields good agreement with the experimental data. The test rotor has an aspect ratio of 1.56, a design total pressure ratio of 1.629 and a tip relative Mach number of 1.38. The high spatial resolution of the LA data matrix (9 radial by 30 axial by 50 blade to blade) permits details of the transonic flow field such as shock location, turning distribution and blade loading levels to be investigated and compared to analytical results.
NASA Astrophysics Data System (ADS)
Huntzinger, D. N.; McCray, J. E.; Siegrist, R.; Lowe, K.; VanCuyk, S.
2001-05-01
Sixteen, one-dimensional column lysimeters have been developed to evaluate the influence of loading regime and infiltrative surface character on hydraulic performance in wastewater soil absorption systems. A duplicate design was utilized to evaluate two infiltrative surface conditions (gravel-free vs. gravel-laden) under four hydraulic loading regimes representative of possible field conditions. By loading the columns at rates of 25 to 200 cm/day, the 17 weeks of column operation actually reflect up to approximately 13 yrs of field operation (at 5 cm/day). Therefore, the cumulative mass throughput and infiltrative rate loss for each loading regime can be examined to determine the viability of accelerated loading as a means to compress the time scale of observation, while still producing meaningfully results for the field scale. During operation, the columns were loaded with septic tank effluent at a prescribed rate and routinely monitoring for applied effluent composition, infiltration rate, time-dependant soil water content, water volume throughput, and percolate composition. Bromide tracer tests were completed prior to system startup and at weeks 2, 6, and 17 of system operation. Hydraulic characterization of the columns is based on measurements of the hydraulic loading rate, volumetric throughput, soil water content, and bromide breakthrough curves. Incipient ponding of wastewater developed during the 1st week of operation for columns loaded at the highest hydraulic rate (loading regimes 1 and 2), and during the 3rd and 6th week of operation for loading regimes 3 and 4, respectfully. The bromide breakthrough curves exhibit later breakthrough and tailing as system life increases, indicating the development of spatially variability in hydraulic conductivity within the column and the development of a clogging zone at the infiltrative surface. Throughput is assessed for each loading regime to determine the infiltration rate loss versus days of operation. Loading regimes 1 and 2 approach a comparable long-term throughput rate less than 20 cm/day, while loading regimes 3 and 4 reach a long-term throughput rate of less than 10 cm/day. These one-dimensional columns allow for the analysis of infiltrative rate loss and hydraulic behavior as a result of infiltrative surface character and loading regime.
Three-dimensional organotypic culture: experimental models of mammalian biology and disease.
Shamir, Eliah R; Ewald, Andrew J
2014-10-01
Mammalian organs are challenging to study as they are fairly inaccessible to experimental manipulation and optical observation. Recent advances in three-dimensional (3D) culture techniques, coupled with the ability to independently manipulate genetic and microenvironmental factors, have enabled the real-time study of mammalian tissues. These systems have been used to visualize the cellular basis of epithelial morphogenesis, to test the roles of specific genes in regulating cell behaviours within epithelial tissues and to elucidate the contribution of microenvironmental factors to normal and disease processes. Collectively, these novel models can be used to answer fundamental biological questions and generate replacement human tissues, and they enable testing of novel therapeutic approaches, often using patient-derived cells.
Identification of dynamic load for prosthetic structures.
Zhang, Dequan; Han, Xu; Zhang, Zhongpu; Liu, Jie; Jiang, Chao; Yoda, Nobuhiro; Meng, Xianghua; Li, Qing
2017-12-01
Dynamic load exists in numerous biomechanical systems, and its identification signifies a critical issue for characterizing dynamic behaviors and studying biomechanical consequence of the systems. This study aims to identify dynamic load in the dental prosthetic structures, namely, 3-unit implant-supported fixed partial denture (I-FPD) and teeth-supported fixed partial denture. The 3-dimensional finite element models were constructed through specific patient's computerized tomography images. A forward algorithm and regularization technique were developed for identifying dynamic load. To verify the effectiveness of the identification method proposed, the I-FPD and teeth-supported fixed partial denture structures were investigated to determine the dynamic loads. For validating the results of inverse identification, an experimental force-measuring system was developed by using a 3-dimensional piezoelectric transducer to measure the dynamic load in the I-FPD structure in vivo. The computationally identified loads were presented with different noise levels to determine their influence on the identification accuracy. The errors between the measured load and identified counterpart were calculated for evaluating the practical applicability of the proposed procedure in biomechanical engineering. This study is expected to serve as a demonstrative role in identifying dynamic loading in biomedical systems, where a direct in vivo measurement may be rather demanding in some areas of interest clinically. Copyright © 2017 John Wiley & Sons, Ltd.
Three-dimensional finite-element analysis of chevron-notched fracture specimens
NASA Technical Reports Server (NTRS)
Raju, I. S.; Newman, J. C., Jr.
1984-01-01
Stress-intensity factors and load-line displacements were calculated for chevron-notched bar and rod fracture specimens using a three-dimensional finite-element analysis. Both specimens were subjected to simulated wedge loading (either uniform applied displacement or uniform applied load). The chevron-notch sides and crack front were assumed to be straight. Crack-length-to-specimen width ratios (a/w) ranged from 0.4 to 0.7. The width-to-thickness ratio (w/B) was 1.45 or 2. The bar specimens had a height-to-width ratio of 0.435 or 0.5. Finite-element models were composed of singularity elements around the crack front and 8-noded isoparametric elements elsewhere. The models had about 11,000 degrees of freedom. Stress-intensity factors were calculated by using a nodal-force method for distribution along the crack front and by using a compliance method for average values. The stress intensity factors and load-line displacements are presented and compared with experimental solutions from the literature. The stress intensity factors and load-line displacements were about 2.5 and 5 percent lower than the reported experimental values, respectively.
Finite element analysis of provisional structures of implant-supported complete prostheses.
Carneiro, Bruno Albuquerque; de Brito, Rui Barbosa; França, Fabiana Mantovani Gomes
2014-04-01
The use of provisional resin implant-supported complete dentures is a fast and safe procedure to restore mastication and esthetics of patients soon after surgery and during the adaptation phase to the new denture. This study assessed stress distribution of provisional implant-supported fixed dentures and the all-on-4 concept using self-curing acrylic resin (Tempron) and bis-acrylic resin (Luxatemp) to simulate functional loads through the three-dimensional finite element method. Solidworks software was used to build three-dimensional models using acrylic resin (Tempron, model A) and bis-acrylic resin (Luxatemp, model B) for denture captions. Two loading patterns were applied on each model: (1) right unilateral axial loading of 150 N on the occlusal surfaces of posterior teeth and (2) oblique loading vector of 150 N at 45°. The results showed that higher stress was found on the bone crest below oblique load application with a maximum value of 187.57 MPa on model A and 167.45 MPa on model B. It was concluded that model B improved stress distribution on the denture compared with model A.
Ritz, Ulrike; Gerke, Rebekka; Götz, Hermann; Stein, Stefan; Rommens, Pol Maria
2017-11-29
Although a lot of research has been performed, large segmental bone defects caused by trauma, infection, bone tumors or revision surgeries still represent big challenges for trauma surgeons. New and innovative bone substitutes are needed. Three-dimensional (3D) printing is a novel procedure to create 3D porous scaffolds that can be used for bone tissue engineering. In the present study, solid discs as well as porous cage-like 3D prints made of polylactide (PLA) are coated or filled with collagen, respectively, and tested for biocompatibility and endotoxin contamination. Microscopic analyses as well as proliferation assays were performed using various cell types on PLA discs. Stromal-derived factor (SDF-1) release from cages filled with collagen was analyzed and the effect on endothelial cells tested. This study confirms the biocompatibility of PLA and demonstrates an endotoxin contamination clearly below the FDA (Food and Drug Administration) limit. Cells of various cell types (osteoblasts, osteoblast-like cells, fibroblasts and endothelial cells) grow, spread and proliferate on PLA-printed discs. PLA cages loaded with SDF-1 collagen display a steady SDF-1 release, support cell growth of endothelial cells and induce neo-vessel formation. These results demonstrate the potential for PLA scaffolds printed with an inexpensive desktop printer in medical applications, for example, in bone tissue engineering.
Gerke, Rebekka; Götz, Hermann; Rommens, Pol Maria
2017-01-01
Although a lot of research has been performed, large segmental bone defects caused by trauma, infection, bone tumors or revision surgeries still represent big challenges for trauma surgeons. New and innovative bone substitutes are needed. Three-dimensional (3D) printing is a novel procedure to create 3D porous scaffolds that can be used for bone tissue engineering. In the present study, solid discs as well as porous cage-like 3D prints made of polylactide (PLA) are coated or filled with collagen, respectively, and tested for biocompatibility and endotoxin contamination. Microscopic analyses as well as proliferation assays were performed using various cell types on PLA discs. Stromal-derived factor (SDF-1) release from cages filled with collagen was analyzed and the effect on endothelial cells tested. This study confirms the biocompatibility of PLA and demonstrates an endotoxin contamination clearly below the FDA (Food and Drug Administration) limit. Cells of various cell types (osteoblasts, osteoblast-like cells, fibroblasts and endothelial cells) grow, spread and proliferate on PLA-printed discs. PLA cages loaded with SDF-1 collagen display a steady SDF-1 release, support cell growth of endothelial cells and induce neo-vessel formation. These results demonstrate the potential for PLA scaffolds printed with an inexpensive desktop printer in medical applications, for example, in bone tissue engineering. PMID:29186036
On the calculation of dynamic and heat loads on a three-dimensional body in a hypersonic flow
NASA Astrophysics Data System (ADS)
Bocharov, A. N.; Bityurin, V. A.; Evstigneev, N. M.; Fortov, V. E.; Golovin, N. N.; Petrovskiy, V. P.; Ryabkov, O. I.; Teplyakov, I. O.; Shustov, A. A.; Solomonov, Yu S.
2018-01-01
We consider a three-dimensional body in a hypersonic flow at zero angle of attack. Our aim is to estimate heat and aerodynamic loads on specific body elements. We are considering a previously developed code to solve coupled heat- and mass-transfer problem. The change of the surface shape is taken into account by formation of the iterative process for the wall material ablation. The solution is conducted on the multi-graphics-processing-unit (multi-GPU) cluster. Five Mach number points are considered, namely for M = 20-28. For each point we estimate body shape after surface ablation, heat loads on the surface and aerodynamic loads on the whole body and its elements. The latter is done using Gauss-type quadrature on the surface of the body. The comparison of the results for different Mach numbers is performed. We also estimate the efficiency of the Navier-Stokes code on multi-GPU and central processing unit architecture for the coupled heat and mass transfer problem.
Simulating Fatigue Crack Growth in Spiral Bevel Pinion
NASA Technical Reports Server (NTRS)
Ural, Ani; Wawrzynek, Paul A.; Ingraffe, Anthony R.
2003-01-01
This project investigates computational modeling of fatigue crack growth in spiral bevel gears. Current work is a continuation of the previous efforts made to use the Boundary Element Method (BEM) to simulate tooth-bending fatigue failure in spiral bevel gears. This report summarizes new results predicting crack trajectory and fatigue life for a spiral bevel pinion using the Finite Element Method (FEM). Predicting crack trajectories is important in determining the failure mode of a gear. Cracks propagating through the rim may result in catastrophic failure, whereas the gear may remain intact if one tooth fails and this may allow for early detection of failure. Being able to predict crack trajectories is insightful for the designer. However, predicting growth of three-dimensional arbitrary cracks is complicated due to the difficulty of creating three-dimensional models, the computing power required, and absence of closed- form solutions of the problem. Another focus of this project was performing three-dimensional contact analysis of a spiral bevel gear set incorporating cracks. These analyses were significant in determining the influence of change of tooth flexibility due to crack growth on the magnitude and location of contact loads. This is an important concern since change in contact loads might lead to differences in SIFs and therefore result in alteration of the crack trajectory. Contact analyses performed in this report showed the expected trend of decreasing tooth loads carried by the cracked tooth with increasing crack length. Decrease in tooth loads lead to differences between SIFs extracted from finite element contact analysis and finite element analysis with Hertz contact loads. This effect became more pronounced as the crack grew.
Three-Dimensional Modeling of Fluid and Heat Transport in an Accretionary Complex
NASA Astrophysics Data System (ADS)
Paula, C. A.; Ge, S.; Screaton, E. J.
2001-12-01
As sediments are scraped off of the subducting oceanic crust and accreted to the overriding plate, the rapid loading causes pore pressures in the underthrust sediments to increase. The change in pore pressure drives fluid flow and heat transport within the accretionary complex. Fluid is channeled along higher permeability faults and fractures and expelled at the seafloor. In this investigation, we examined the effects of sediment loading on fluid flow and thermal transport in the decollement at the Barbados Ridge subduction zone. Both the width and thickness of the Barbados Ridge accretionary complex increase from north to south. The presence of mud diapers south of the Tiburon Rise and an observed southward decrease in heat flow measurements indicate that the increased thickness of the southern Barbados accretionary prism affects the transport of chemicals and heat by fluids. The three-dimensional geometry and physical properties of the accretionary complex were utilized to construct a three-dimensional fluid flow/heat transport model. We calculated the pore pressure change due to a period of sediment loading and added this to steady-state pressure conditions to generate initial conditions for transient simulations. We then examined the diffusion of pore pressure and possible perturbation of the thermal regime over time due to loading of the underthrust sediments. The model results show that the sediment-loading event was sufficient to create small temperature fluctuations in the decollement zone. The magnitude of temperature fluctuation in the decollement was greatest at the deformation front but did not vary significantly from north to south of the Tiburon Rise.
Optimization of a Boiling Water Reactor Loading Pattern Using an Improved Genetic Algorithm
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kobayashi, Yoko; Aiyoshi, Eitaro
2003-08-15
A search method based on genetic algorithms (GA) using deterministic operators has been developed to generate optimized boiling water reactor (BWR) loading patterns (LPs). The search method uses an Improved GA operator, that is, crossover, mutation, and selection. The handling of the encoding technique and constraint conditions is designed so that the GA reflects the peculiar characteristics of the BWR. In addition, some strategies such as elitism and self-reproduction are effectively used to improve the search speed. LP evaluations were performed with a three-dimensional diffusion code that coupled neutronic and thermal-hydraulic models. Strong axial heterogeneities and three-dimensional-dependent constraints have alwaysmore » necessitated the use of three-dimensional core simulators for BWRs, so that an optimization method is required for computational efficiency. The proposed algorithm is demonstrated by successfully generating LPs for an actual BWR plant applying the Haling technique. In test calculations, candidates that shuffled fresh and burned fuel assemblies within a reasonable computation time were obtained.« less
Growth and Development of Three-Dimensional Plant Form.
Whitewoods, Christopher D; Coen, Enrico
2017-09-11
Plants can generate a spectacular array of complex shapes, many of which exhibit elaborate curvature in three dimensions, illustrated for example by orchid flowers and pitcher-plant traps. All of these structures arise through differential growth. Recent findings provide fresh mechanistic insights into how regional cell behaviours may lead to tissue deformations, including anisotropies and curvatures, which shape growing volumes and sheets of cells. Here were review our current understanding of how genes, growth, mechanics, and evolution interact to generate diverse structures. We illustrate problems and approaches with the complex three-dimensional trap of the bladderwort, Utricularia gibba, to show how a multidisciplinary approach can be extended to new model systems to understand how diverse plant shapes can develop and evolve. Copyright © 2017 Elsevier Ltd. All rights reserved.
Three-dimensional epithelial tissues generated from human embryonic stem cells.
Hewitt, Kyle J; Shamis, Yulia; Carlson, Mark W; Aberdam, Edith; Aberdam, Daniel; Garlick, Jonathan A
2009-11-01
The use of pluripotent human embryonic stem (hES) cells for tissue engineering may provide advantages over traditional sources of progenitor cells because of their ability to give rise to multiple cell types and their unlimited expansion potential. We derived cell populations with properties of ectodermal and mesenchymal cells in two-dimensional culture and incorporated these divergent cell populations into three-dimensional (3D) epithelial tissues. When grown in specific media and substrate conditions, two-dimensional cultures were enriched in cells (EDK1) with mesenchymal morphology and surface markers. Cells with a distinct epithelial morphology (HDE1) that expressed cytokeratin 12 and beta-catenin at cell junctions became the predominant cell type when EDK1 were grown on surfaces enriched in keratinocyte-derived extracellular matrix proteins. When these cells were incorporated into the stromal and epithelial tissue compartments of 3D tissues, they generated multilayer epithelia similar to those generated with foreskin-derived epithelium and fibroblasts. Three-dimensional tissues demonstrated stromal cells with morphologic features of mature fibroblasts, type IV collagen deposition in the basement membrane, and a stratified epithelium that expressed cytokeratin 12. By deriving two distinct cell lineages from a common hES cell source to fabricate complex tissues, it is possible to explore environmental cues that will direct hES-derived cells toward optimal tissue form and function.
Self-monitoring fiber reinforced polymer strengthening system for civil engineering infrastructures
NASA Astrophysics Data System (ADS)
Jiang, Guoliang; Dawood, Mina; Peters, Kara; Rizkalla, Sami
2008-03-01
Fiber reinforced polymer (FRP) materials are currently used for strengthening civil engineering infrastructures. The strengthening system is dependant on the bond characteristics of the FRP to the external surface of the structure to be effective in resisting the applied loads. This paper presents an innovative self-monitoring FRP strengthening system. The system consists of two components which can be embedded in FRP materials to monitor the global and local behavior of the strengthened structure respectively. The first component of the system is designed to evaluate the applied load acting on a structure based on elongation of the FRP layer along the entire span of the structure. Success of the global system has been demonstrated using a full-scale prestressed concrete bridge girder which was loaded up to failure. The test results indicate that this type of sensor can be used to accurately determine the load prior to failure within 15 percent of the measured value. The second sensor component consists of fiber Bragg grating sensors. The sensors were used to monitor the behavior of steel double-lap shear splices tested under tensile loading up to failure. The measurements were used to identify abnormal structural behavior such as epoxy cracking and FRP debonding. Test results were also compared to numerical values obtained from a three dimensional shear-lag model which was developed to predict the sensor response.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Katano, Takahito; Ootani, Akifumi; Department of Internal Medicine, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga 849-8501
2013-03-22
Highlights: ► We established a 3D culture system to allow long-term culture of stomach cells. ► In this culture system, gastric epithelial cells grew for about 3 months. ► The cultured cells differentiated into multi-units of the stomach. ► This culture method should be useful for elucidating the cause of gastric diseases. -- Abstract: Compared to the small intestine and colon, little is known about stem cells in the stomach because of a lack of specific stem cell markers and an in vitro system that allows long-term culture. Here we describe a long-term three-dimensional (3D) primary gastric culture system withinmore » the stem cell niche. Glandular stomach cells from neonatal mice cultured in collagen gel yielded expanding sphere-like structures for 3 months. The wall of the gastrospheres consisted of a highly polarized epithelial monolayer with an outer lining of myofibroblasts. The epithelial cells showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm as well as expression of MUC5AC, indicating differentiation into gastric surface mucous cells. These cells demonstrated the features of fully differentiated gastric surface mucous cells such as microvilli, junctional complexes, and glycogen and secretory granules. Fewer than 1% of cultured epithelial cells differentiated into enteroendocrine cells. Active proliferation of the epithelial cells and many apoptotic cells in the inner lumen revealed the rapid cell turnover in gastrospheres in vitro. This method enables us to investigate the role of signaling between cell–cell and epithelial–mesenchymal interactions in an environment that is extremely similar to the in vivo environment.« less
NASA Astrophysics Data System (ADS)
Matveev, A. D.
2016-11-01
To calculate the three-dimensional elastic body of heterogeneous structure under static loading, a method of multigrid finite element is provided, when implemented on the basis of algorithms of finite element method (FEM), using homogeneous and composite threedimensional multigrid finite elements (MFE). Peculiarities and differences of MFE from the currently available finite elements (FE) are to develop composite MFE (without increasing their dimensions), arbitrarily small basic partition of composite solids consisting of single-grid homogeneous FE of the first order can be used, i.e. in fact, to use micro approach in finite element form. These small partitions allow one to take into account in MFE, i.e. in the basic discrete models of composite solids, complex heterogeneous and microscopically inhomogeneous structure, shape, the complex nature of the loading and fixation and describe arbitrarily closely the stress and stain state by the equations of three-dimensional elastic theory without any additional simplifying hypotheses. When building the m grid FE, m of nested grids is used. The fine grid is generated by a basic partition of MFE, the other m —1 large grids are applied to reduce MFE dimensionality, when m is increased, MFE dimensionality becomes smaller. The procedures of developing MFE of rectangular parallelepiped, irregular shape, plate and beam types are given. MFE generate the small dimensional discrete models and numerical solutions with a high accuracy. An example of calculating the laminated plate, using three-dimensional 3-grid FE and the reference discrete model is given, with that having 2.2 milliards of FEM nodal unknowns.
NASA Astrophysics Data System (ADS)
Chiroux, Robert Charles
The objective of this research was to produce a three dimensional, non-linear, dynamic simulation of the interaction between a hyperelastic wheel rolling over compactable soil. The finite element models developed to produce the simulation utilized the ABAQUS/Explicit computer code. Within the simulation two separate bodies were modeled, the hyperelastic wheel and a compactable soil-bed. Interaction between the bodies was achieved by allowing them to come in contact but not to penetrate the contact surface. The simulation included dynamic loading of a hyperelastic, rubber tire in contact with compactable soil with an applied constant angular velocity or torque, including a tow load, applied to the wheel hub. The constraints on the wheel model produced a straight and curved path. In addition the simulation included a shear limit between the tire and soil allowing for the introduction of slip. Soil properties were simulated using the Drucker-Prager, Cap Plasticity model available within the ABAQUS/Explicit program. Numerical results obtained from the three dimensional model were compared with related experimental data and showed good correlation for similar conditions. Numerical and experimental data compared well for both stress and wheel rut formation depth under a weight of 5.8 kN and a constant angular velocity applied to the wheel hub. The simulation results provided a demonstration of the benefit of three-dimensional simulation in comparison to previous two-dimensional, plane strain simulations.
Three-dimensional effects for radio frequency antenna modeling
DOE Office of Scientific and Technical Information (OSTI.GOV)
Carter, M.D.; Batchelor, D.B.; Stallings, D.C.
1994-10-15
Electromagnetic field calculations for radio frequency (rf) antennas in two dimensions (2-D) neglect finite antenna length effects as well as the feeders leading to the main current strap. The 2-D calculations predict that the return currents in the sidewalls of the antenna structure depend strongly on the plasma parameters, but this prediction is suspect because of experimental evidence. To study the validity of the 2-D approximation, the Multiple Antenna Implementation System (MAntIS) has been used to perform three-dimensional (3-D) modeling of the power spectrum, plasma loading, and inductance for a relevant loop antenna design. Effects on antenna performance caused bymore » feeders to the main current strap and conducting sidewalls are considered. The modeling shows that the feeders affect the launched power spectrum in an indirect way by forcing the driven rf current to return in the antenna structure rather than the plasma, as in the 2-D model. It has also been found that poloidal dependencies in the plasma impedance matrix can reduce the loading predicted from that predicted in the 2-D model. For some plasma parameters, the combined 3-D effects can lead to a reduction in the predicted loading by as much as a factor of 2 from that given by the 2-D model, even with end-effect corrections for the 2-D model.« less
Three-dimensional effects for radio frequency antenna modeling
DOE Office of Scientific and Technical Information (OSTI.GOV)
Carter, M.D.; Batchelor, D.B.; Stallings, D.C.
1993-12-31
Electromagnetic field calculations for radio frequency (rf) antennas in two dimensions (2-D) neglect finite antenna length effects as well as the feeders leading to the main current strap. The 2-D calculations predict that the return currents in the sidewalls of the antenna structure depend strongly on the plasma parameters, but this prediction is suspect because of experimental evidence. To study the validity of the 2-D approximation, the Multiple Antenna Implementation System (MAntIS) has been used to perform three-dimensional (3-D) modeling of the power spectrum, plasma loading, and inductance for a relevant loop antenna design. Effects on antenna performance caused bymore » feeders to the main current strap and conducting sidewalls are considered. The modeling shows that the feeders affect the launched power spectrum in an indirect way by forcing the driven rf current to return in the antenna structure rather than the plasma, as in the 2-D model. It has also been found that poloidal dependencies in the plasma impedance matrix can reduce the loading predicted from that predicted in the 2-D model. For some plasma parameters, the combined 3-D effects can lead to a reduction in the predicted loading by as much as a factor of 2 from that given by the 2-D model, even with end-effect corrections for the 2-D model.« less
Zhang, Yanxin; Ma, Ye; Liu, Guangyu
2016-01-01
The objective of the study was to evaluate two types of cricket bowling techniques by comparing the lumbar spinal loading using a musculoskeletal modelling approach. Three-dimensional kinematic data were recorded by a Vicon motion capture system under two cricket bowling conditions: (1) participants bowled at their absolute maximal speeds (max condition), and (2) participants bowled at their absolute maximal speeds while simultaneously forcing their navel down towards their thighs starting just prior to ball release (max-trunk condition). A three-dimensional musculoskeletal model comprised of the pelvis, sacrum, lumbar vertebrae and torso segments, which enabled the motion of the individual lumbar vertebrae in the sagittal, frontal and coronal planes to be actuated by 210 muscle-tendon units, was used to simulate spinal loading based on the recorded kinematic data. The maximal lumbar spine compressive force is 4.89 ± 0.88BW for the max condition and 4.58 ± 0.54BW for the max-trunk condition. Results showed that there was no significant difference between the two techniques in trunk moments and lumbar spine forces. This indicates that the max-trunk technique may not increase lower back injury risks. The method proposed in this study could be served as a tool to evaluate lower back injury risks for cricket bowling as well as other throwing activities.
Liu, Jingyin; Pan, Shaoxia; Dong, Jing; Mo, Zhongjun; Fan, Yubo; Feng, Hailan
2013-03-01
The aim of this study was to evaluate strain distribution in peri-implant bone, stress in the abutments and denture stability of mandibular overdentures anchored by different numbers of implants under different loading conditions, through three-dimensional finite element analysis (3D FEA). Four 3D finite element models of mandibular overdentures were established, using between one and four Straumann implants with Locator attachments. Three types of load were applied to the overdenture in each model: 100N vertical and inclined loads on the left first molar and a 100N vertical load on the lower incisors. The biomechanical behaviours of peri-implant bone, implants, abutments and overdentures were recorded. Under vertical load on the lower incisors, the single-implant overdenture rotated over the implant from side to side, and no obvious increase of strain was found in peri-implant bone. Under the same loading conditions, the two-implant-retained overdenture showed more apparent rotation around the fulcrum line passing through the two implants, and the maximum equivalent stress in the abutments was higher than in the other models. In the three-implant-supported overdenture, no strain concentration was found in cortical bone around the middle implant under three loading conditions. Single-implant-retained mandibular overdentures do not show damaging strain concentration in the bone around the only implant and may be a cost-effective treatment option for edentulous patients. A third implant can be placed between the original two when patients rehabilitated by two-implant overdentures report constant and obvious denture rotation around the fulcrum line. Copyright © 2012 Elsevier Ltd. All rights reserved.
Huang, G S; Tseng, C S; Linju Yen, B; Dai, L G; Hsieh, P S; Hsu, S-h
2013-10-13
Three-dimensional (3D) cellular spheroids have recently emerged as a new trend to replace suspended single cells in modern cell-based therapies because of their greater regeneration capacities in vitro. They may lose the 3D structure during a change of microenvironment, which poses challenges to their translation in vivo. Besides, the conventional microporous scaffolds may have difficulty in accommodating these relatively large spheroids. Here we revealed a novel design of microenvironment for delivering and sustaining the 3D spheroids. Biodegradable scaffolds with macroporosity to accommodate mesenchymal stem cell (MSC) spheroids were made by solid freeform fabrication (SFF) from the solution of poly(D,L-lactide-co-glycolide). Their internal surface was modified with chitosan following air plasma treatment in order to preserve the morphology of the spheroids. It was demonstrated that human MSC spheroids loaded in SFF scaffolds produced a significantly larger amount of cartilage-associated extracellular matrix in vitro and in NOD/SCID mice compared to single cells in the same scaffolds. Implantation of MSC spheroid-loaded scaffolds into the chondral defects of rabbit knees showed superior cartilage regeneration. This study establishes new perspectives in designing the spheroid-sustaining microenvironment within a tissue engineering scaffold for in vivo applications.
Dickinson, J.E.; James, S.C.; Mehl, S.; Hill, M.C.; Leake, S.A.; Zyvoloski, G.A.; Faunt, C.C.; Eddebbarh, A.-A.
2007-01-01
A flexible, robust method for linking parent (regional-scale) and child (local-scale) grids of locally refined models that use different numerical methods is developed based on a new, iterative ghost-node method. Tests are presented for two-dimensional and three-dimensional pumped systems that are homogeneous or that have simple heterogeneity. The parent and child grids are simulated using the block-centered finite-difference MODFLOW and control-volume finite-element FEHM models, respectively. The models are solved iteratively through head-dependent (child model) and specified-flow (parent model) boundary conditions. Boundary conditions for models with nonmatching grids or zones of different hydraulic conductivity are derived and tested against heads and flows from analytical or globally-refined models. Results indicate that for homogeneous two- and three-dimensional models with matched grids (integer number of child cells per parent cell), the new method is nearly as accurate as the coupling of two MODFLOW models using the shared-node method and, surprisingly, errors are slightly lower for nonmatching grids (noninteger number of child cells per parent cell). For heterogeneous three-dimensional systems, this paper compares two methods for each of the two sets of boundary conditions: external heads at head-dependent boundary conditions for the child model are calculated using bilinear interpolation or a Darcy-weighted interpolation; specified-flow boundary conditions for the parent model are calculated using model-grid or hydrogeologic-unit hydraulic conductivities. Results suggest that significantly more accurate heads and flows are produced when both Darcy-weighted interpolation and hydrogeologic-unit hydraulic conductivities are used, while the other methods produce larger errors at the boundary between the regional and local models. The tests suggest that, if posed correctly, the ghost-node method performs well. Additional testing is needed for highly heterogeneous systems. ?? 2007 Elsevier Ltd. All rights reserved.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Giunta, G.; Belouettar, S.
In this paper, the static response of three-dimensional beams made of functionally graded materials is investigated through a family of hierarchical one-dimensional finite elements. A wide variety of elements is proposed differing by the kinematic formulation and the number of nodes per elements along the beam axis. Elements’ stiffness matrix and load vector are derived in a unified nuclear form that does not depend upon the a priori expansion order over the cross-section nor the finite element approximation along the beam axis. Results are validated towards three-dimensional finite element models as well as equivalent Navier-type analytical solutions. The numerical investigationsmore » show that accurate and efficient solutions (when compared with full three-dimensional FEM solutions) can be obtained by the proposed family of hierarchical one-dimensional elements’ family.« less
NASA Astrophysics Data System (ADS)
Jeong, Eun Sook; Kim, Jin Woong
2015-03-01
Hydrogel particles, also known as microgels, consist of cross-linked three-dimensional water-soluble polymer networks. They play an essential role in loading and delivering active ingredients in medicine, cosmetics, and foods. Despite their excellent biocompatibility as well as structural diversity, much wider applications are limited due mainly to their intrinsically loose network nature. This study introduces a practical and straightforward method that enables fabrication of hydrogel microparticles layered with a mechanically robust hybrid thin shell. Basically highly monodisperse hydrogel microparticles were produced in microcapillary devices. Then, their surface was coated with alternate polyelectrolyte layers through the layer-by-layer deposition. Finally a thin silica layer was again formed by reduction of silicate on the amino-functionalized polyelectrolyte layer. We have figured out that these hybrid hydrogel microparticles showed controlled loading and releasing behaviors for water-soluble probe molecules. Moreover, we have demonstrated that they can be applied for immobilization of biomacromolecules, such as bacteria and living cells, and even for targeted releasing.
[Establishment and validation of normal human L1-L5 lumbar three-dimensional finite element model].
Zhu, Zhenqi; Liu, Chenjun; Wang, Jiefu; Wang, Kaifeng; Huang, Zhixin; Wang, Weida; Liu, Haiying
2014-10-14
To create and validate a L1-L5 lumbar three-dimensional finite element model. The L1-L5 lumbar spines of a male healthy volunteer were scanned with computed tomography (CT). And a L1-L5 lumbar three-dimensional finite element model was created with the aid of software packages of Mimics, Geomagic and Ansys. Then border conditions were set, unit type was determined, finite element mesh was divided and a model was established for loading and calculating. Average model stiffness under the conditions of flexion, extension, lateral bending and axial rotation was calculated and compared with the outcomes of former articles for validation. A normal human L1-L5 lumbar three-dimensional finite element model was established to include 459 340 elements and 661 938 nodes. After constraining the inferior endplate of L5 vertebral body, 500 kg × m × s⁻² compressive loading was imposed averagely on the superior endplate of L1 vertebral body. Then 10 kg × m² × s⁻² moment simulating flexion, extension, lateral bending and axial rotation were imposed on the superior endplate of L1 vertebral body. Eventually the average stiffness of all directions was calculated and it was similar to the outcomes of former articles. The L1-L5 lumbar three-dimensional finite element model is validated so that it may used with biomechanical simulation and analysis of normal or surgical models.
Wang, Q; Yang, Y; Fei, Q; Li, D; Li, J J; Meng, H; Su, N; Fan, Z H; Wang, B Q
2017-06-06
Objective: To build a three-dimensional finite element models of a modified posterior cervical single open-door laminoplasty with short-segmental lateral mass screws fusion. Methods: The C(2)-C(7) segmental data were obtained from computed tomography (CT) scans of a male patient with cervical spondylotic myelopathy and spinal stenosis.Three-dimensional finite element models of a modified cervical single open-door laminoplasty (before and after surgery) were constructed by the combination of software package MIMICS, Geomagic and ABAQUS.The models were composed of bony vertebrae, articulating facets, intervertebral disc and associated ligaments.The loads of moments 1.5Nm at different directions (flexion, extension, lateral bending and axial rotation)were applied at preoperative model to calculate intersegmental ranges of motion.The results were compared with the previous studies to verify the validation of the models. Results: Three-dimensional finite element models of the modified cervical single open- door laminoplasty had 102258 elements (preoperative model) and 161 892 elements (postoperative model) respectively, including C(2-7) six bony vertebraes, C(2-3)-C(6-7) five intervertebral disc, main ligaments and lateral mass screws.The intersegmental responses at the preoperative model under the loads of moments 1.5 Nm at different directions were similar to the previous published data. Conclusion: Three-dimensional finite element models of the modified cervical single open- door laminoplasty were successfully established and had a good biological fidelity, which can be used for further study.
NASA Astrophysics Data System (ADS)
Gao, Xin; Chen, Min; Snyder, G. Jeffrey; Andreasen, Søren Juhl; Kær, Søren Knudsen
2013-07-01
To better manage the magnitude and direction of the heat flux in an exchanger-based methanol evaporator of a fuel cell system, thermoelectric (TE) modules can be deployed as TE heat flux regulators (TERs). The performance of the TE-integrated evaporator is strongly influenced by its heat exchange structure. The structure transfers the fuel cell exhaust heat to the evaporation chamber to evaporate the methanol, where TE modules are installed in between to facilitate the heat regulation. In this work, firstly, a numerical study is conducted to determine the working currents and working modes of the TERs under the system working condition fluctuations and during the system cold start. A three-dimensional evaporator model is generated in ANSYS FLUENT® by combining a compact TE model with various heat exchange structure geometries. The compact TE model can dramatically improve the computational efficiency, and uses a different material property acquisition method based on module manufacturers' datasheets. Secondly, a simulation study is carried out on the novel evaporator to minimize its thermal resistance and to assess the evaporator pressure drop. The factors studied include the type of fins in the heat exchange structure, the thickness of the fins, the axial conduction penalty, etc. Results show that the TE-integrated evaporator can work more efficiently and smoothly during both load fluctuations and system cold start, offering superior performance.
Murakami, Tatsuya C; Mano, Tomoyuki; Saikawa, Shu; Horiguchi, Shuhei A; Shigeta, Daichi; Baba, Kousuke; Sekiya, Hiroshi; Shimizu, Yoshihiro; Tanaka, Kenji F; Kiyonari, Hiroshi; Iino, Masamitsu; Mochizuki, Hideki; Tainaka, Kazuki; Ueda, Hiroki R
2018-04-01
A three-dimensional single-cell-resolution mammalian brain atlas will accelerate systems-level identification and analysis of cellular circuits underlying various brain functions. However, its construction requires efficient subcellular-resolution imaging throughout the entire brain. To address this challenge, we developed a fluorescent-protein-compatible, whole-organ clearing and homogeneous expansion protocol based on an aqueous chemical solution (CUBIC-X). The expanded, well-cleared brain enabled us to construct a point-based mouse brain atlas with single-cell annotation (CUBIC-Atlas). CUBIC-Atlas reflects inhomogeneous whole-brain development, revealing a significant decrease in the cerebral visual and somatosensory cortical areas during postnatal development. Probabilistic activity mapping of pharmacologically stimulated Arc-dVenus reporter mouse brains onto CUBIC-Atlas revealed the existence of distinct functional structures in the hippocampal dentate gyrus. CUBIC-Atlas is shareable by an open-source web-based viewer, providing a new platform for whole-brain cell profiling.
A photoreversible protein-patterning approach for guiding stem cell fate in three-dimensional gels
NASA Astrophysics Data System (ADS)
Deforest, Cole A.; Tirrell, David A.
2015-05-01
Although biochemically patterned hydrogels are capable of recapitulating many critical aspects of the heterogeneous cellular niche, exercising spatial and temporal control of the presentation and removal of biomolecular signalling cues in such systems has proved difficult. Here, we demonstrate a synthetic strategy that exploits two bioorthogonal photochemistries to achieve reversible immobilization of bioactive full-length proteins with good spatial and temporal control within synthetic, cell-laden biomimetic scaffolds. A photodeprotection-oxime-ligation sequence permits user-defined quantities of proteins to be anchored within distinct subvolumes of a three-dimensional matrix, and an ortho-nitrobenzyl ester photoscission reaction facilitates subsequent protein removal. By using this approach to pattern the presentation of the extracellular matrix protein vitronectin, we accomplished reversible differentiation of human mesenchymal stem cells to osteoblasts in a spatially defined manner. Our protein-patterning approach should provide further avenues to probe and direct changes in cell physiology in response to dynamic biochemical signalling.
NASA Technical Reports Server (NTRS)
Goodwin, Thomas J. (Inventor); Wolf, David A. (Inventor); Spaulding, Glenn F. (Inventor); Prewett, Tracey L. (Inventor)
1996-01-01
Normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural, and blood tissue. The cells are grown in vitro under microgravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel.
Common source-multiple load vs. separate source-individual load photovoltaic system
NASA Technical Reports Server (NTRS)
Appelbaum, Joseph
1989-01-01
A comparison of system performance is made for two possible system setups: (1) individual loads powered by separate solar cell sources; and (2) multiple loads powered by a common solar cell source. A proof for resistive loads is given that shows the advantage of a common source over a separate source photovoltaic system for a large range of loads. For identical loads, both systems perform the same.
Hydrogel-based three-dimensional cell culture for organ-on-a-chip applications.
Lee, Seung Hwan; Shim, Kyu Young; Kim, Bumsang; Sung, Jong Hwan
2017-05-01
Recent studies have reported that three-dimensionally cultured cells have more physiologically relevant functions than two-dimensionally cultured cells. Cells are three-dimensionally surrounded by the extracellular matrix (ECM) in complex in vivo microenvironments and interact with the ECM and neighboring cells. Therefore, replicating the ECM environment is key to the successful cell culture models. Various natural and synthetic hydrogels have been used to mimic ECM environments based on their physical, chemical, and biological characteristics, such as biocompatibility, biodegradability, and biochemical functional groups. Because of these characteristics, hydrogels have been combined with microtechnologies and used in organ-on-a-chip applications to more closely recapitulate the in vivo microenvironment. Therefore, appropriate hydrogels should be selected depending on the cell types and applications. The porosity of the selected hydrogel should be controlled to facilitate the movement of nutrients and oxygen. In this review, we describe various types of hydrogels, external stimulation-based gelation of hydrogels, and control of their porosity. Then, we introduce applications of hydrogels for organ-on-a-chip. Last, we also discuss the challenges of hydrogel-based three-dimensional cell culture techniques and propose future directions. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:580-589, 2017. © 2017 American Institute of Chemical Engineers.
Zhao, Shuan; Liu, Zhen-Xing; Gao, Hui; Wu, Yi; Fang, Yuan; Wu, Shuai-Shuai; Li, Ming-Jie; Bai, Jia-Hua; Liu, Yan; Evans, Alexander; Zeng, Shen-Ming
2015-07-15
No successful method exists to maintain the three-dimensional architecture of hatched embryos in vitro. Alginate, a linear polysaccharide derived from brown algae, has characteristics that make it an ideal material as a three-dimensional (3D) extracellular matrix for in vitro cell, tissue, or embryo culture. In this study, alginate hydrogel was used for IVC of posthatched bovine embryos to observe their development under the 3D system. In vitro-fertilized and parthenogenetically activated posthatched bovine blastocysts were cultured in an alginate encapsulation culture system (AECS), an alginate overlay culture system (AOCS), or control culture system. After 18 days of culture, the survival rate of embryos cultured in AECS was higher than that in the control group (P < 0.05), and the embryos were expanded and elongated in AECS with the maximal length of 1.125 mm. When the AECS shrinking embryos were taken out of the alginate beads on Day 18 and cultured in the normal culture system, 9.09% of them attached to the bottoms of the plastic wells and grew rapidly, with the largest area of an attached embryo being 66.00 mm(2) on Day 32. The embryos cultured in AOCS developed monovesicular or multivesicular morphologies. Total cell number of the embryos cultured in AECS on Day 19 was significantly higher than that of embryos on Day 8. Additionally, AECS and AOCS supported differentiation of the embryonic cells. Binuclear cells were visible in Day-26 adherent embryos, and the messenger RNA expression patterns of Cdx2 and Oct4 in AOCS-cultured embryos were similar to those in vivo embryos, whereas IFNT and ISG15 messenger RNA were still expressed in Day-26 and Day-32 prolong-cultured embryos. In conclusion, AECS and AOCS did support cell proliferation, elongation, and differentiation of hatched bovine embryos during prolonged IVC. The culture system will be useful to further investigate the molecular mechanisms controlling ruminant embryo elongation and implantation. Copyright © 2015 Elsevier Inc. All rights reserved.
NASA Astrophysics Data System (ADS)
Chen, Rong; Li, Kang; Xia, Kaiwen; Lin, Yuliang; Yao, Wei; Lu, Fangyun
2016-10-01
A dynamic load superposed on a static pre-load is a key problem in deep underground rock engineering projects. Based on a modified split Hopkinson pressure bar test system, the notched semi-circular bend (NSCB) method is selected to investigate the fracture initiation toughness of rocks subjected to pre-load. In this study, a two-dimensional ANSYS finite element simulation model is developed to calculate the dimensionless stress intensity factor. Three groups of NSCB specimen are tested under a pre-load of 0, 37 and 74 % of the maximum static load and with the loading rate ranging from 0 to 60 GPa m1/2 s-1. The results show that under a given pre-load, the fracture initiation toughness of rock increases with the loading rate, resembling the typical rate dependence of materials. Furthermore, the dynamic rock fracture toughness decreases with the static pre-load at a given loading rate. The total fracture toughness, defined as the sum of the dynamic fracture toughness and initial stress intensity factor calculated from the pre-load, increases with the pre-load at a given loading rate. An empirical equation is used to represent the effect of loading rate and pre-load force, and the results show that this equation can depict the trend of the experimental data.
[Advances in the research of application of hydrogels in three-dimensional bioprinting].
Yang, J; Zhao, Y; Li, H H; Zhu, S H
2016-08-20
Hydrogels are three-dimensional networks made of hydrophilic polymer crosslinked through covalent bonds or physical intermolecular attractions, which can contain growth media and growth factors to support cell growth. In bioprinting, hydrogels are used to provide accurate control over cellular microenvironment and to dramatically reduce experimental repetition times, meanwhile we can obtain three-dimensional cell images of high quality. Hydrogels in three-dimensional bioprinting have received a considerable interest due to their structural similarities to the natural extracellular matrix and polyporous frameworks which can support the cellular proliferation and survival. Meanwhile, they are accompanied by many challenges.
Ferraz, Marcia A M M; Henning, Heiko H W; Stout, Tom A E; Vos, Peter L A M; Gadella, Bart M
2017-07-01
The oviduct was long considered a largely passive conduit for gametes and embryos. However, an increasing number of studies into oviduct physiology have demonstrated that it specifically and significantly influences gamete interaction, fertilization and early embryo development. While oviduct epithelial cell (OEC) function has been examined during maintenance in conventional tissue culture dishes, cells seeded into these two-dimensional (2-D) conditions suffer a rapid loss of differentiated OEC characteristics, such as ciliation and secretory activity. Recently, three-dimensional (3-D) cell culture systems have been developed that make use of cell inserts to create basolateral and apical medium compartments with a confluent epithelial cell layer at the interface. Using such 3-D culture systems, OECs can be triggered to redevelop typical differentiated cell properties and levels of tissue organization can be developed that are not possible in a 2-D culture. 3-D culture systems can be further refined using new micro-engineering techniques (including microfluidics and 3-D printing) which can be used to produce 'organs-on-chips', i.e. live 3-D cultures that bio-mimic the oviduct. In this review, concepts for designing bio-mimic 3-D oviduct cultures are presented. The increased possibilities and concomitant challenges when trying to more closely investigate oviduct physiology, gamete activation, fertilization and embryo production are discussed.
Schmelzer, Eva; Finoli, Anthony; Nettleship, Ian; Gerlach, Jörg C
2015-04-01
The construction and long-term maintenance of three-dimensional in vitro bone marrow models is of great interest but still quite challenging. Here we describe the use of a multi-compartment hollow-fiber membrane based three-dimensional perfusion bioreactor for long-term culture of whole human bone marrow mononuclear cells. We also investigated bioreactors with incorporated open-porous foamed hydroxyapatite scaffolds, mimicking the in vivo bone matrix. Cells in bioreactors with and without scaffolds were cultured to 6 weeks and compared to Petri dish controls. Cells were analyzed for gene expression, surface markers by flow cytometry, metabolic activity, hematopoietic potential, viability, and attachment by immunocytochemistry. Cells in bioreactors were metabolic active during long-term culture. The percentages of hematopoietic stem cell and mature endothelial cell fractions were maintained in bioreactors. The expression of most of the analyzed genes stabilized and increased after long-term culture of 6 weeks. Compared to Petri dish culture controls, bioreactor perfusion culture improved in both the short and long-term, the colony formation unit capacity of hematopoietic progenitors. Cells attached to the ample surface area provided by hydroxyapatite scaffolds. The implementation of a hydroxyapatite scaffold did not influence colony formation capacity, percentages of cell type specific fractions, gene expression, cell viability or metabolic turnover when compared to control cells cultured in bioreactors without scaffolds. In conclusion, three-dimensional perfusion bioreactor culture enables long-term maintenance of primary human bone marrow cells, with hydroxyapatite scaffolds providing an in vivo-like scaffold for three-dimensional culture. © 2015 Wiley Periodicals, Inc.
On the variation in crack-opening stresses at different locations in a three-dimensional body
NASA Technical Reports Server (NTRS)
Chermahini, R. G.; Blom, Anders F.
1990-01-01
Crack propagation and closure behavior of thin, and thick middle crack tension specimens under constant amplitude loading were investigated using a three dimensional elastic plastic finite element analysis of fatigue crack propagation and closure. In the thin specimens the crack front closed first on the exterior (free) surface and closed last in the interior during the unloading portion of cyclic loading; a load reduced displacement technique was used to determine crack opening stresses at specified locations in the plate from the displacements calculated after the seven cycle. All the locations were on the plate external surface and were located near the crack tip, behind the crack tip, at the centerline of the crack. With this technique, the opening stresses at the specified points were found to be 0.52, 0.42, and 0.39 times the maximum applied stress.
Engineering cellular fibers for musculoskeletal soft tissues using directed self-assembly.
Schiele, Nathan R; Koppes, Ryan A; Chrisey, Douglas B; Corr, David T
2013-05-01
Engineering strategies guided by developmental biology may enhance and accelerate in vitro tissue formation for tissue engineering and regenerative medicine applications. In this study, we looked toward embryonic tendon development as a model system to guide our soft tissue engineering approach. To direct cellular self-assembly, we utilized laser micromachined, differentially adherent growth channels lined with fibronectin. The micromachined growth channels directed human dermal fibroblast cells to form single cellular fibers, without the need for a provisional three-dimensional extracellular matrix or scaffold to establish a fiber structure. Therefore, the resulting tissue structure and mechanical characteristics were determined solely by the cells. Due to the self-assembly nature of this approach, the growing fibers exhibit some key aspects of embryonic tendon development, such as high cellularity, the rapid formation (within 24 h) of a highly organized and aligned cellular structure, and the expression of cadherin-11 (indicating direct cell-to-cell adhesions). To provide a dynamic mechanical environment, we have also developed and characterized a method to apply precise cyclic tensile strain to the cellular fibers as they develop. After an initial period of cellular fiber formation (24 h postseeding), cyclic strain was applied for 48 h, in 8-h intervals, with tensile strain increasing from 0.7% to 1.0%, and at a frequency of 0.5 Hz. Dynamic loading dramatically increased cellular fiber mechanical properties with a nearly twofold increase in both the linear region stiffness and maximum load at failure, thereby demonstrating a mechanism for enhancing cellular fiber formation and mechanical properties. Tissue engineering strategies, designed to capture key aspects of embryonic development, may provide unique insight into accelerated maturation of engineered replacement tissue, and offer significant advances for regenerative medicine applications in tendon, ligament, and other fibrous soft tissues.
NASA Astrophysics Data System (ADS)
Wirtz, Denis
2014-03-01
Two-dimensional (2D) in vitro culture systems have for a number of years provided a controlled and versatile environment for mechanistic studies of cell adhesion, polarization, and migration, three interrelated cell functions critical to cancer metastasis. However, the organization and functions of focal adhesion proteins, protrusion machinery, and microtubule-based polarization in cells embedded in physiologically more relevant 3D extracellular matrices is qualitatively different from their organization and functions on conventional 2D planar substrates. This talk will describe the implications of the dependence of focal adhesion protein-based cell migration on micro-environmental dimensionality (1D vs. 2D vs.. 3D), how cell micromechanics plays a critical role in promoting local cell invasion, and associated validation in mouse models. We will discuss the implications of this work in cancer metastasis.
Application of cell co-culture system to study fat and muscle cells.
Pandurangan, Muthuraman; Hwang, Inho
2014-09-01
Animal cell culture is a highly complex process, in which cells are grown under specific conditions. The growth and development of these cells is a highly unnatural process in vitro condition. Cells are removed from animal tissues and artificially cultured in various culture vessels. Vitamins, minerals, and serum growth factors are supplied to maintain cell viability. Obtaining result homogeneity of in vitro and in vivo experiments is rare, because their structure and function are different. Living tissues have highly ordered complex architecture and are three-dimensional (3D) in structure. The interaction between adjacent cell types is quite distinct from the in vitro cell culture, which is usually two-dimensional (2D). Co-culture systems are studied to analyze the interactions between the two different cell types. The muscle and fat co-culture system is useful in addressing several questions related to muscle modeling, muscle degeneration, apoptosis, and muscle regeneration. Co-culture of C2C12 and 3T3-L1 cells could be a useful diagnostic tool to understand the muscle and fat formation in animals. Even though, co-culture systems have certain limitations, they provide a more realistic 3D view and information than the individual cell culture system. It is suggested that co-culture systems are useful in evaluating the intercellular communication and composition of two different cell types.
NASA Technical Reports Server (NTRS)
Goodwin, Thomas J. (Inventor)
1998-01-01
Normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural and blood tissue. The cells are grown in vitro under microgravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel.
NASA Technical Reports Server (NTRS)
Goodwin, Thomas J. (Inventor)
1998-01-01
Normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural and blood tissue.The cells are grown in vitro under microgravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel.
Mahyari, Mojtaba; Laeini, Mohammad Sadegh; Shaabani, Ahmad
2014-07-25
Copper(ii) tetrasulfophthalocyanine supported on three-dimensional nitrogen-doped graphene-based frameworks was synthesized and introduced as a bifunctional catalyst for selective aerobic oxidation of alkyl arenes and alcohols to the corresponding carbonyl compounds. The ease of catalyst separation, high turnover, low catalyst loading and recyclability could potentially render it applicable in industrial setting.
Subsurface Treatment of Domestic Wastewater Using Single Domicile Constructed Wetlands
NASA Astrophysics Data System (ADS)
Aseltyne, T.; Steer, D.; Fraser, L.
2001-05-01
Analysis of one year of input versus output water quality monitoring data from nine household wastewater treatment wetlands in western Ohio indicates that these systems substantially reduce effluent loads delivered to the local watershed. Overall performance as measured by output water quality improvement varies widely between the nine systems despite their close proximity and identical design. These three-cell systems (septic tank with 2 subsurface wetland cells) are found to reduce biological oxygen demand (BOD) 70-98%, fecal coliform 60-99.9%, NH3 29-97%, Phosphorus 21-99.9% and total suspended solids (TSS) up to 97%. NO3/NO2 readings were only taken at the second wetland cell, but show that NO3/NO2 levels are at 0.005-5.01 mg/l and well below the USEPA standards for discharge from a wetland. On average, the pH of the wastewater increases from 6.6 at the septic tank to 8.7 at the wetland output. Nearly all the monitoring data indicate clear decreases in nutrient loads and bacteria though individual systems are found to non-systematically fail to meet EPA discharge guidelines for one or more of the monitored loads. Preliminary analysis of the data indicates a decrease in overall efficiency of the wetlands in April that may be related to seasonal factors. These systems will be monitored for the next three years in order to relate changing performance trends to seasonal variability.
Multidisciplinary Design Optimization of A Highly Flexible Aeroservoelastic Wing
NASA Astrophysics Data System (ADS)
Haghighat, Sohrab
A multidisciplinary design optimization framework is developed that integrates control system design with aerostructural design for a highly-deformable wing. The objective of this framework is to surpass the existing aircraft endurance limits through the use of an active load alleviation system designed concurrently with the rest of the aircraft. The novelty of this work is two fold. First, a unified dynamics framework is developed to represent the full six-degree-of-freedom rigid-body along with the structural dynamics. It allows for an integrated control design to account for both manoeuvrability (flying quality) and aeroelasticity criteria simultaneously. Secondly, by synthesizing the aircraft control system along with the structural sizing and aerodynamic shape design, the final design has the potential to exploit synergies among the three disciplines and yield higher performing aircraft. A co-rotational structural framework featuring Euler--Bernoulli beam elements is developed to capture the wing's nonlinear deformations under the effect of aerodynamic and inertial loadings. In this work, a three-dimensional aerodynamic panel code, capable of calculating both steady and unsteady loadings is used. Two different control methods, a model predictive controller (MPC) and a 2-DOF mixed-norm robust controller, are considered in this work to control a highly flexible aircraft. Both control techniques offer unique advantages that make them promising for controlling a highly flexible aircraft. The control system works towards executing time-dependent manoeuvres along with performing gust/manoeuvre load alleviation. The developed framework is investigated for demonstration in two design cases: one in which the control system simply worked towards achieving or maintaining a target altitude, and another where the control system is also performing load alleviation. The use of the active load alleviation system results in a significant improvement in the aircraft performance relative to the optimum result without load alleviation. The results show that the inclusion of control system discipline along with other disciplines at early stages of aircraft design improves aircraft performance. It is also shown that structural stresses due to gust excitations can be better controlled by the use of active structural control systems which can improve the fatigue life of the structure.
Haraguchi, Yuji; Kagawa, Yuki; Sakaguchi, Katsuhisa; Matsuura, Katsuhisa; Shimizu, Tatsuya; Okano, Teruo
2017-01-01
In this paper, we report an in vitro co-culture system that combines mammalian cells and algae, Chlorococcum littorale, to create a three-dimensional (3-D) tissue. While the C2C12 mouse myoblasts and rat cardiac cells consumed oxygen actively, intense oxygen production was accounted for by the algae even in the co-culture system. Although cell metabolism within thicker cardiac cell-layered tissues showed anaerobic respiration, the introduction of innovative co-cultivation partially changed the metabolism to aerobic respiration. Moreover, the amount of glucose consumption and lactate production in the cardiac tissues and the amount of ammonia in the culture media decreased significantly when co-cultivated with algae. In the cardiac tissues devoid of algae, delamination was observed histologically, and the release of creatine kinase (CK) from the tissues showed severe cardiac cell damage. On the other hand, the layered cell tissues with algae were observed to be in a good histological condition, with less than one-fifth decline in CK release. The co-cultivation with algae improved the culture condition of the thicker tissues, resulting in the formation of 160 μm-thick cardiac tissues. Thus, the present study proposes the possibility of creating an in vitro “symbiotic recycling system” composed of mammalian cells and algae. PMID:28139713
NASA Astrophysics Data System (ADS)
Scolan, Y.-M.; Korobkin, A. A.
2003-02-01
Hydrodynamic impact phenomena are three dimensional in nature and naval architects need more advanced tools than a simple strip theory to calculate impact loads at the preliminary design stage. Three-dimensional analytical solutions have been obtained with the help of the so-called inverse Wagner problem as discussed by Scolan and Korobkin in 2001. The approach by Wagner provides a consistent way to evaluate the flow caused by a blunt body entering liquid through its free surface. However, this approach does not account for the spray jets and gives no idea regarding the energy evacuated from the main flow by the jets. Clear insight into the jet formation is required. Wagner provided certain elements of the answer for two-dimensional configurations. On the basis of those results, the energy distribution pattern is analysed for three-dimensional configurations in the present paper.
Development of a 3D cell printed construct considering angiogenesis for liver tissue engineering.
Lee, Jin Woo; Choi, Yeong-Jin; Yong, Woon-Jae; Pati, Falguni; Shim, Jin-Hyung; Kang, Kyung Shin; Kang, In-Hye; Park, Jaesung; Cho, Dong-Woo
2016-01-12
Several studies have focused on the regeneration of liver tissue in a two-dimensional (2D) planar environment, whereas actual liver tissue is three-dimensional (3D). Cell printing technology has been successfully utilized for building 3D structures; however, the poor mechanical properties of cell-laden hydrogels are a major concern. Here, we demonstrate the printing of a 3D cell-laden construct and its application to liver tissue engineering using 3D cell printing technology through a multi-head tissue/organ building system. Polycaprolactone (PCL) was used as a framework material because of its excellent mechanical properties. Collagen bioink containing three different types of cells-hepatocytes (HCs), human umbilical vein endothelial cells , and human lung fibroblasts--was infused into the canals of a PCL framework to induce the formation of capillary--like networks and liver cell growth. A co-cultured 3D microenvironment of the three types of cells was successfully established and maintained. The vascular formation and functional abilities of HCs (i.e., albumin secretion and urea synthesis) demonstrated that the heterotypic interaction among HCs and nonparenchymal cells increased the survivability and functionality of HCs within the collagen gel. Therefore, our results demonstrate the prospect of using cell printing technology for the creation of heterotypic cellular interaction within a structure for liver tissue engineering.
Cascade Analysis of a Floating Wind Turbine Rotor
NASA Astrophysics Data System (ADS)
Eliassen, Lene; Jakobsen, Jasna B.; Knauer, Andreas; Nielsen, Finn Gunnar
2014-12-01
Mounting a wind turbine on a floating foundation introduces more complexity to the aerodynamic loading. The floater motion contains a wide range of frequencies. To study some of the basic dynamic load effect on the blades due to these motions, a two-dimensional cascade approach, combined with a potential vortex method, is used. This is an alternative method to study the aeroelastic behavior of wind turbines that is different from the traditional blade element momentum method. The analysis tool demands little computational power relative to a full three dimensional vortex method, and can handle unsteady flows. When using the cascade plane, a "cut" is made at a section of the wind turbine blade. The flow is viewed parallel to the blade axis at this cut. The cascade model is commonly used for analysis of turbo machineries. Due to the simplicity of the code it requires little computational resources, however it has limitations in its validity. It can only handle two-dimensional potential flow, i.e. including neither three-dimensional effects, such as the tip loss effect, nor boundary layers and stall effects are modeled. The computational tool can however be valuable in the overall analysis of floating wind turbines, and evaluation of the rotor control system. A check of the validity of the vortex panel code using an airfoil profile is performed, comparing the variation of the lift force, to the theoretically derived Wagner function. To analyse the floating wind turbine, a floating structure with hub height 90 m is chosen. An axial motion of the rotor is considered.
Yang, Meng-Yin; Chiao, Ming-Tsang; Lee, Hsu-Tung; Chen, Chien-Min; Yang, Yi-Chin; Shen, Chiung-Chyi; Ma, Hsin-I
2015-04-01
Three-dimensional (3-D) tissue engineered constructs provide a platform for examining how the local extracellular matrix contributes to the malignancy of various cancers, including human glioblastoma multiforme. Here, we describe a simple and innovative 3-D culture environment and assess its potential for use with glioblastoma stem cells (GSCs) to examine the diversification inside the cell mass in the 3-D culture system. The dissociated human GSCs were cultured using gelatin foam. These cells were subsequently identified by immunohistochemical staining, reverse transcriptase-polymerase chain reaction, and Western blot assay. We demonstrate that the gelatin foam provides a suitable microenvironment, as a 3-D culture system, for GSCs to maintain their stemness. The gelatin foam culture system contributes a simplified assessment of cell blocks for immunohistochemistry assay. We show that the significant transcription activity of hypoxia and the protein expression of inflammatory responses are detected at the inside of the cell mass in vitro, while robust expression of PROM1/CD133 and hypoxia-induced factor-1 alpha are detected at the xenografted tumor in vivo. We also examine the common clinical trials under this culture platform and characterized a significant difference of drug resistance. The 3-D gelatin foam culture system can provide a more realistic microenvironment through which to study the in vivo behavior of GSCs to evaluate the role that biophysical factors play in the hypoxia, inflammatory responses and subsequent drug resistance. © 2014 Wiley Periodicals, Inc.