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Sample records for osteochondral tissue grafts

  1. Engineering custom-designed osteochondral tissue grafts

    PubMed Central

    Grayson, Warren L.; Chao, Pen-Hsiu Grace; Marolt, Darja; Kaplan, David L.; Vunjak-Novakovic, Gordana

    2009-01-01

    Tissue engineering is expected to help us outlive the failure of our organs by enabling the creation of tissue substitutes capable of fully restoring the original tissue function. Degenerative joint disease, which affects one-fifth of the US population and is the country’s leading cause of disability, drives current research of actively growing, functional tissue grafts for joint repair. Toward this goal, living cells are used in conjunction with bio-material scaffolds (serving as instructive templates for tissue development) and bioreactors (providing environmental control and molecular and physical regulatory signals). In this review, we discuss the requirements for engineering customized, anatomically-shaped, stratified grafts for joint repair and the challenges of designing these grafts to provide immediate functionality (load bearing, structural support) and long-term regeneration (maturation, integration, remodeling). PMID:18299159

  2. Challenges in engineering osteochondral tissue grafts with hierarchical structures.

    PubMed

    Gadjanski, Ivana; Vunjak-Novakovic, Gordana

    2015-01-01

    A major hurdle in treating osteochondral (OC) defects is the different healing abilities of two types of tissues involved - articular cartilage and subchondral bone. Biomimetic approaches to OC-construct engineering, based on recapitulation of biological principles of tissue development and regeneration, have potential for providing new treatments and advancing fundamental studies of OC tissue repair. This review on state of the art in hierarchical OC tissue graft engineering is focused on tissue engineering approaches designed to recapitulate the native milieu of cartilage and bone development. These biomimetic systems are discussed with relevance to bioreactor cultivation of clinically sized, anatomically shaped human cartilage/bone constructs with physiologic stratification and mechanical properties. The utility of engineered OC tissue constructs is evaluated for their use as grafts in regenerative medicine, and as high-fidelity models in biological research. A major challenge in engineering OC tissues is to generate a functionally integrated stratified cartilage-bone structure starting from one single population of mesenchymal cells, while incorporating perfusable vasculature into the bone, and in bone-cartilage interface. To this end, new generations of advanced scaffolds and bioreactors, implementation of mechanical loading regimens and harnessing of inflammatory responses of the host will likely drive the further progress.

  3. Challenges in engineering osteochondral tissue grafts with hierarchical structures Ivana Gadjanski, Gordana Vunjak Novakovic

    PubMed Central

    Gadjanski, Ivana; Vunjak-Novakovic, Gordana

    2015-01-01

    Introduction A major hurdle in treating osteochondral (OC) defects are the different healing abilities of two types of tissues involved - articular cartilage and subchondral bone. Biomimetic approaches to OC-construct-engineering, based on recapitulation of biological principles of tissue development and regeneration, have potential for providing new treatments and advancing fundamental studies of OC tissue repair. Areas covered This review on state of the art in hierarchical OC tissue graft engineering is focused on tissue engineering approaches designed to recapitulate the native milieu of cartilage and bone development. These biomimetic systems are discussed with relevance to bioreactor cultivation of clinically sized, anatomically shaped human cartilage/bone constructs with physiologic stratification and mechanical properties. The utility of engineered OC tissue constructs is evaluated for their use as grafts in regenerative medicine, and as high-fidelity models in biological research. Expert opinion A major challenge in engineering OC tissues is to generate a functionally integrated stratified cartilage-bone structure starting from one single population of mesenchymal cells, while incorporating perfusable vasculature into the bone, and in bone-cartilage interface. To this end, new generations of advanced scaffolds and bioreactors, implementation of mechanical loading regimens, and harnessing of inflammatory responses of the host will likely drive the further progress. PMID:26195329

  4. Fabrication of tissue engineered osteochondral grafts for restoring the articular surface of diarthrodial joints

    PubMed Central

    Roach, Brendan L.; Hung, Clark T.; Cook, James L.; Ateshian, Gerard A.; Tan, Andrea R.

    2015-01-01

    Osteochondral allograft implantation is an effective cartilage restoration technique for large defects (>10 cm2), though the demand far exceeds the supply of available quality donor tissue. Large bilayered engineered cartilage tissue constructs with accurate anatomical features (i.e. contours, thickness, architecture) could be beneficial in replacing damaged tissue. When creating these osteochondral constructs, however, it is pertinent to maintain biofidelity to restore functionality. Here, we describe a step-by-step framework for the fabrication of a large osteochondral construct with correct anatomical architecture and topology through a combination of high-resolution imaging, rapid prototyping, impression molding, and injection molding. PMID:25794950

  5. Porous titanium bases for osteochondral tissue engineering

    PubMed Central

    Nover, Adam B.; Lee, Stephanie L.; Georgescu, Maria S.; Howard, Daniel R.; Saunders, Reuben A.; Yu, William T.; Klein, Robert W.; Napolitano, Anthony P.; Ateshian, Gerard A.

    2015-01-01

    Tissue engineering of osteochondral grafts may offer a cell-based alternative to native allografts, which are in short supply. Previous studies promote the fabrication of grafts consisting of a viable cell-seeded hydrogel integrated atop a porous, bone-like metal. Advantages of the manufacturing process have led to the evaluation of porous titanium as the bone-like base material. Here, porous titanium was shown to support the growth of cartilage to produce native levels of Young’s modulus, using a clinically relevant cell source. Mechanical and biochemical properties were similar or higher for the osteochondral constructs compared to chondral-only controls. Further investigation into the mechanical influence of the base on the composite material suggests that underlying pores may decrease interstitial fluid pressurization and applied strains, which may be overcome by alterations to the base structure. Future studies aim to optimize titanium-based tissue engineered osteochondral constructs to best match the structural architecture and strength of native grafts. Statement of Significance The studies described in this manuscript follow up on previous studies from our lab pertaining to the fabrication of osteochondral grafts that consist of a bone-like porous metal and a chondrocyte-seeded hydrogel. Here, tissue engineered osteochondral grafts were cultured to native stiffness using adult chondrocytes, a clinically relevant cell source, and a porous titanium base, a material currently used in clinical implants. This porous titanium is manufactured via selective laser melting, offering the advantages of precise control over shape, pore size, and orientation. Additionally, this manuscript describes the mechanical influence of the porous base, which may have applicability to porous bases derived from other materials. PMID:26320541

  6. Mesenchymal Stem Cells for Osteochondral Tissue Engineering

    PubMed Central

    Ng, Johnathan; Bernhard, Jonathan; Vunjak-Novakovic, Gordana

    2017-01-01

    Summary Mesenchymal stem cells (MSC) are of major interest to regenerative medicine, because of the ease of harvesting from a variety of sources (including bone marrow and fat aspirates) and ability to form a range of mesenchymal tissues, in vitro and in vivo. We focus here on the use of MSCs for engineering of cartilage, bone, and complex osteochondral tissue constructs, using protocols that replicate some aspects of the natural mesodermal development. For engineering of human bone, we discuss some of the current advances, and highlight the use of perfusion bioreactors for supporting anatomically exact human bone grafts. For engineering of human cartilage, we discuss limitations of current approaches, and highlight engineering of stratified, mechanically functional human cartilage interfaced with bone by mesenchymal condensation of MSCs. Taken together, the current advances enable engineering physiologically relevant bone, cartilage and osteochondral composites, and physiologically relevant studies of osteochondral development and disease. PMID:27236665

  7. Review of the biomechanics and biotribology of osteochondral grafts used for surgical interventions in the knee

    PubMed Central

    Bowland, Philippa; Ingham, E; Jennings, Louise; Fisher, John

    2015-01-01

    A review of research undertaken to evaluate the biomechanical stability and biotribological behaviour of osteochondral grafts in the knee joint and a brief discussion of areas requiring further improvement in future studies are presented. The review takes into consideration osteochondral autografts, allografts, tissue engineered constructs and synthetic and biological scaffolds. PMID:26614801

  8. Osteochondral tissue engineering: current strategies and challenges.

    PubMed

    Nukavarapu, Syam P; Dorcemus, Deborah L

    2013-01-01

    Osteochondral defect management and repair remain a significant challenge in orthopedic surgery. Osteochondral defects contain damage to both the articular cartilage as well as the underlying subchondral bone. In order to repair an osteochondral defect the needs of the bone, cartilage and the bone-cartilage interface must be taken into account. Current clinical treatments for the repair of osteochondral defects have only been palliative, not curative. Tissue engineering has emerged as a potential alternative as it can be effectively used to regenerate bone, cartilage and the bone-cartilage interface. Several scaffold strategies, such as single phase, layered, and recently graded structures have been developed and evaluated for osteochondral defect repair. Also, as a potential cell source, tissue specific cells and progenitor cells are widely studied in cell culture models, as well with the osteochondral scaffolds in vitro and in vivo. Novel factor strategies being developed, including single factor, multi-factor, or controlled factor release in a graded fashion, not only assist bone and cartilage regeneration, but also establish osteochondral interface formation. The field of tissue engineering has made great strides, however further research needs to be carried out to make this strategy a clinical reality. In this review, we summarize current tissue engineering strategies, including scaffold design, bioreactor use, as well as cell and factor based approaches and recent developments for osteochondral defect repair. In addition, we discuss various challenges that need to be addressed in years to come. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Stratified Scaffolds for Osteochondral Tissue Engineering.

    PubMed

    Nooeaid, Patcharakamon; Schulze-Tanzil, Gundula; Boccaccini, Aldo R

    2015-01-01

    Stratified scaffolds are promising devices finding application in the field of osteochondral tissue engineering. In this scaffold type, different biomaterials are chosen to fulfill specific features required to mimic the complex osteochondral tissue interface, including cartilage, interlayer tissue, and subchondral bone. Here, the biomaterials and fabrication methods currently used to manufacture stratified multilayered scaffolds as well as cell seeding techniques for their characterization are presented.

  10. Generation and characterization of osteochondral grafts with human nasal chondrocytes.

    PubMed

    Barandun, Marina; Iselin, Lukas Daniel; Santini, Francesco; Pansini, Michele; Scotti, Celeste; Baumhoer, Daniel; Bieri, Oliver; Studler, Ueli; Wirz, Dieter; Haug, Martin; Jakob, Marcel; Schaefer, Dirk Johannes; Martin, Ivan; Barbero, Andrea

    2015-08-01

    We investigated whether nasal chondrocytes (NC) can be used to generate composite constructs with properties necessary for the repair of osteochondral (OC) lesions, namely maturation, integration and capacity to recover from inflammatory burst. OC grafts were fabricated by combining engineered cartilage tissues (generated by culturing NC or articular chondrocytes - AC - onto Chondro-Gide® matrices) with devitalized spongiosa cylinders (Tutobone®). OC tissues were then exposed to IL-1β for three days and cultured for additional 2 weeks in the absence of IL-1β. Cartilage maturation extent was assessed (immune) histologically, biochemically and by delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) while cartilage/bone integration was assessed using a peel-off mechanical test. The use of NC as compared to AC allowed for more efficient cartilage matrix accumulation and superior integration of the cartilage/bone layers. dGEMRIC and biochemical analyzes of the OC constructs showed a reduced glycosaminoglycan (GAG) contents upon IL-1β administration. Cartilaginous matrix contents and integration forces returned to baseline up on withdrawal of IL-1β. By having a cartilage layer well developed and strongly integrated to the subchondral layer, OC tissues generated with NC may successfully engraft in an inflammatory post-surgery joint environment. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  11. Recent progress in interfacial tissue engineering approaches for osteochondral defects.

    PubMed

    Castro, Nathan J; Hacking, S Adam; Zhang, Lijie Grace

    2012-08-01

    This review provides a brief synopsis of the anatomy and physiology of the osteochondral interface, scaffold-based and non-scaffold based approaches for engineering both tissues independently as well as recent developments in the manufacture of gradient constructs. Novel manufacturing techniques and nanotechnology will be discussed with potential application in osteochondral interfacial tissue engineering.

  12. Technique: Osteochondral Grafting of Capitate Chondrosis in PRC

    PubMed Central

    Tang, Peter; Imbriglia, Joseph E.

    2013-01-01

    Background Proximal row carpectomy (PRC) is a useful treatment option for wrist arthritis, but the operation is contraindicated when there is arthritis of the capitate head. We describe a technique that involves resurfacing of a capitate that has focal chondrosis, using an osteochondral graft harvested from the resected carpal bones. Materials and Methods PRC patients who had a focal area of capitate chondrosis underwent osteochondral grafting of the capitate. Pre- and postoperative pain level, employment status, motion, grip strength, and Modified Mayo Wrist Scores (MMWS) were assessed. Postoperative Disability of the Arm, Shoulder, and Hand (DASH) scores were also calculated. Description of Technique The articular surface of the capitate is assessed for need for grafting. The proximal row is resected with the lunate removed intact. The arthritic area is prepared. The graft is taken from the lunate and placed in the prepared site of the capitate. Results Eight patients (average age of 53 years) were followed for 18 months. Pain: Preoperatively, moderate to severe in 7 patients; postoperatively, mild to no pain in 7 patients. Motion: Preoperative, 84° (74% of the contralateral side); postoperative 75° (66%). Grip Strength: Preoperative, 29 kg (62%); postoperative, 34 kg (71%). Mayo Wrist Score: Preoperative, 51 (poor); postoperative, 68 (fair). Average postoperative DASH score was 19.5. Follow-up radiographs showed that 75% of patients had mild to no degeneration. Conclusions Osteochondral grafting in PRC offers satisfactory results in terms of pain relief, return to work, motion, and grip strength. Level of Evidence Therapeutic IV, Case series PMID:24436818

  13. Coronoid reconstruction using osteochondral grafts: a biomechanical study.

    PubMed

    Bellato, Enrico; Kim, Youngbok; Fitzsimmons, James S; Berglund, Lawrence J; Hooke, Alexander W; Bachman, Daniel R; O'Driscoll, Shawn W

    2017-07-19

    The purposes of this study were to test the hypothesis that coronoid deficiency in the setting of posteromedial rotatory instability (PMRI) must be reconstructed to restore articular contact pressures to normal and to compare 3 different osteochondral grafts for this purpose. After creation of a anteromedial fracture, six cadaveric elbows were tested under gravity varus stress using a custom-made machine designed to simulate muscle loads and to passively flex the elbow. Mean articular surface contact pressure data were collected and processed using TekScan sensors and software. After testing of the intact specimen (intact condition), a PMRI injury was created (PMRI condition). Testing was repeated after reconstruction of the lateral collateral ligament (LCL) (LCL-only condition), followed by reconstruction of the coronoid with 3 different osteochondral graft techniques (reconstructed conditions). Contact pressure was consistently significantly higher in the PMRI elbow compared with the intact, LCL-only, and reconstructed conditions (P < .006). The LCL-only elbow contact pressure was significantly higher than that of the intact and reconstructed conditions from 5° to 55° of flexion (P = .018). The contact pressure of the intact elbow was never significantly different from that of the reconstructed elbow, except at 5° of flexion (P ≤ .008). No significant difference was detected between each of the reconstructed techniques (P ≥ .15). However, the annular surface of the radial head was the only graft that yielded contact pressures not significantly different from normal at any flexion angle. Isolated reconstruction of the LCL did not restore native articular surface contact pressure, and reconstruction of the coronoid using osteochondral graft was necessary. There was no difference in contact pressures among the 3 coronoid reconstruction techniques. Copyright © 2017 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier

  14. Effect of Impaction Sequence on Osteochondral Graft Damage: The Role of Repeated and Varying Loads

    PubMed Central

    Kang, Richard W.; Friel, Nicole A.; Williams, James M.; Cole, Brian J.; Wimmer, Markus A.

    2013-01-01

    Background Osteochondral autografts and allografts require mechanical force for proper graft placement into the defect site; however, impaction compromises the tissue. This study aimed to determine the effect of impaction force and number of hits to seat the graft on cartilage integrity. Hypothesis Under constant impulse conditions, higher impaction load magnitudes are more detrimental to cell viability, matrix integrity and collagen network organization and will result in proteoglycan loss and nitric oxide release. Study Design Controlled laboratory study Methods Osteochondral explants, harvested from fresh bovine trochleas, were exposed to a series of consistent impact loads delivered by a pneumatically driven device. Each plug received the same overall impulse of 7 Ns, reflecting the mean of 23 clinically inserted plugs. Impaction loads of 37.5N, 75N, 150N, and 300N were matched with 74, 37, 21, and 11 hits respectively. Following impaction, the plugs were harvested and cartilage was analyzed for cell viability, histology by safranin-o and picosirius red, and release of sulfated glycosaminoglycans and nitric oxide. Data were compared with non-impacted control. Results Impacted plugs had significantly lower cell viability than non-impacted plugs. A dose response relationship in loss of cell viability with respect to load magnitude was seen immediately and after 4 days but lost after 8 days. Histologic analysis revealed intact cartilage surface in all samples (loaded or control), with loaded samples showing alterations in birefringence. While the sulfated GAG release was similar across varying impaction loads, release of nitric oxide increased with increasing impaction magnitudes and time. Conclusions Impaction loading parameters have a direct effect on the time course of the viability of the cartilage in the graft tissue. Clinical Relevance Optimal loading parameters for surgical impaction of osteochondral grafts are those with lower load magnitudes and a greater

  15. Computer tomographic evaluation of talar edge configuration for osteochondral graft transplantation.

    PubMed

    Wiewiorski, Martin; Hoechel, Sebastian; Wishart, Katarina; Leumann, André; Müller-Gerbl, Magdalena; Valderrabano, Victor; Nowakowski, Andrej Maria

    2012-09-01

    To successfully surgically reconstruct osteochondral lesions of the talus, the exact three-dimensional (3D) configuration of the upper articular surface of the talus has to be respected. We assessed the talar geometry by measuring the coronal and sagittal talar edge radius and the frontal talar profile in multiplanar reconstructions of computer tomographic (CT) studies of 79 patients (83 feet) with a healthy ankle joint. An image visualization software designated for coordinate measurement was used to perform the measurement. In the coronal plane, the mean lateral talar edge radius was 4.0 mm and the medial 4.5 mm. In the sagittal planes the mean lateral talar edge radius was 20.3 mm, the radius of the sulcus 20.7 mm and the medial talar edge radius 20.4 mm. The talus showed a concave shape in coronal cuts. These results show a significant difference between medial and lateral talar edge configuration in coronal planes. The measurements of the lateral and medial sagittal radius and the mid-sagittal radius in the sulcus tali show no statistically significant difference. The depth of the talar sulcus shows no correlation to age or sex. Different sizes of custom-made tissue-engineered grafts according to the location of the osteochondral lesion at the talus are needed for exact surgical reconstruction of the anatomy. Osteochondral lesions are three dimensional; therefore, a 3D preoperative planning tool by CT scan or MRI is mandatory. Copyright © 2012 Wiley Periodicals, Inc.

  16. Salvage of a post-traumatic arthritic wrist using the scaphoid as an osteochondral graft.

    PubMed

    Marcuzzi, A; Ozben, H; Russomando, A

    2014-09-01

    The authors describe a case of post-traumatic wrist arthritis with an osteochondral defect in the scaphoid fossa of the radius. The patient was treated with proximal row carpectomy, radial styloidectomy and reconstruction of the defect using the proximal half of the scaphoid as an autologous osteochondral graft. Pain relief was achieved while wrist motion and strength were improved. The carpal bones are a source of osteochondral grafts and can be used to expand the indications of motion-preserving wrist salvage procedures.

  17. Osteochondral tissue engineering with biphasic scaffold: current strategies and techniques.

    PubMed

    Shimomura, Kazunori; Moriguchi, Yu; Murawski, Christopher D; Yoshikawa, Hideki; Nakamura, Norimasa

    2014-10-01

    The management of osteoarthritis (OA) remains challenging and controversial. Although several clinical options exist for the treatment of OA, regeneration of the damaged articular cartilage has proved difficult due to the limited healing capacity. With the advancements in tissue engineering and cell-based technologies over the past decade, new therapeutic options for patients with osteochondral lesions potentially exist. This review will focus on the feasibility of tissue-engineered biphasic scaffolds, which can mimic the native osteochondral complex, for osteochondral repair and highlight the recent development of these techniques toward tissue regeneration. Moreover, basic anatomy, strategy for osteochondral repair, the design and fabrication methods of scaffolds, as well as the choice of cells, growth factor, and materials will be discussed. Specifically, we focus on the latest preclinical animal studies using large animals and clinical trials with high clinical relevance. In turn, this will facilitate an understanding of the latest trends in osteochondral repair and contribute to the future application of such clinical therapies in patients with OA.

  18. Customized Fabrication of Osteochondral Tissue for Articular Joint Surface Repair

    DTIC Science & Technology

    2015-09-01

    it is a disease of the cartilage, bone and surrounding soft tissue that disables 9-10% of the US population. In the US military , combat and non...cartilage – it is a disease of the cartilage, bone and surrounding soft tissue that disables 9-10% of the US population. In the US military , combat and...1 AD______________ AWARD NUMBER: W81XWH-14-1-0217 TITLE: “Customized Fabrication of Osteochondral Tissue for Articular Joint Surface Repair

  19. Hydrogen Supplementation of Preservation Solution Improves Viability of Osteochondral Grafts

    PubMed Central

    Yamada, Takuya; Onuma, Kenji; Kuzuno, Jun; Ujihira, Masanobu; Kurokawa, Ryosuke; Sakai, Rina; Takaso, Masashi

    2014-01-01

    Allogenic osteochondral tissue (OCT) is used for the treatment of large cartilage defects. Typically, OCTs collected during the disease-screening period are preserved at 4°C; however, the gradual reduction in cell viability during cold preservation adversely affects transplantation outcomes. Therefore, improved storage methods that maintain the cell viability of OCTs are needed to increase the availability of high-quality OCTs and improve treatment outcomes. Here, we evaluated whether long-term hydrogen delivery to preservation solution improved the viability of rat OCTs during cold preservation. Hydrogen-supplemented Dulbecco's Modified Eagles Medium (DMEM) and University of Wisconsin (UW) solution both significantly improved the cell viability of OCTs during preservation at 4°C for 21 days compared to nonsupplemented media. However, the long-term cold preservation of OCTs in DMEM containing hydrogen was associated with the most optimal maintenance of chondrocytes with respect to viability and morphology. Our findings demonstrate that OCTs preserved in DMEM supplemented with hydrogen are a promising material for the repair of large cartilage defects in the clinical setting. PMID:25506061

  20. The effect of devitalized trabecular bone on the formation of osteochondral tissue-engineered constructs

    PubMed Central

    Lima, Eric G.; Chao, Pen-hsiu Grace; Ateshian, Gerard A.; Bal, B. Sonny; Cook, James L.; Vunjak-Novakovic, Gordana; Hung, Clark T.

    2008-01-01

    In the current study, evidence is presented demonstrating that devitalized trabecular bone has an inhibitory effect on in vitro chondral tissue development when used as a base material for the tissue-engineering of osteochondral constructs for cartilage repair. Chondrocyte-seeded agarose hydrogel constructs were cultured alone or attached to an underlying bony base in a chemically defined medium formulation that has been shown to yield engineered cartilaginous tissue with native Young's modulus (EY) and glycosaminoglycan (GAG) content. By day 42 in culture the incorporation of a bony base significantly reduced these properties (EY = 87 ± 12 kPa, GAG = 1.9 ± 0.8%ww) compared to the gel-alone group (EY = 642 ± 97 kPa, GAG = 4.6 ± 1.4%ww). Similarly, the mechanical and biochemical properties of chondrocyte-seeded agarose constructs were inhibited when co-cultured adjacent to bone (unattached), suggesting that soluble factors rather than direct cell–bone interactions mediate the chondro-inhibitory bone effects. Altering the method of bone preparation, including demineralization, or the timing of bone introduction in co-culture did not ameliorate the effects. In contrast, osteochondral constructs with native cartilage properties (EY = 730 ± 65 kPa, GAG = 5.2 ± 0.9%ww) were achieved when a porous tantalum metal base material was adopted instead of bone. This work suggests that devitalized bone may not be a suitable substrate for long-term cultivation of osteochondral grafts. PMID:18718655

  1. Osteochondral Interface Tissue Engineering Using Macroscopic Gradients of Bioactive Signals

    PubMed Central

    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

  2. A Hydrogel-Mineral Composite Scaffold for Osteochondral Interface Tissue Engineering

    PubMed Central

    Khanarian, Nora T.; Jiang, Jie; Wan, Leo Q.; Mow, Van C.

    2012-01-01

    Osteoarthritis is the leading cause of physical disability among Americans, and tissue engineered cartilage grafts have emerged as a promising treatment option for this debilitating condition. Currently, the formation of a stable interface between the cartilage graft and subchondral bone remains a significant challenge. This study evaluates the potential of a hybrid scaffold of hydroxyapatite (HA) and alginate hydrogel for the regeneration of the osteochondral interface. Specifically, the effects of HA on the response of chondrocytes were determined, focusing on changes in matrix production and mineralization, as well as scaffold mechanical properties over time. Additionally, the optimal chondrocyte population for interface tissue engineering was evaluated. It was observed that the HA phase of the composite scaffold promoted the formation of a proteoglycan- and type II collagen–rich matrix when seeded with deep zone chondrocytes. More importantly, the elevated biosynthesis translated into significant increases in both compressive and shear moduli relative to the mineral-free control. Presence of HA also promoted chondrocyte hypertrophy and type X collagen deposition. These results demonstrate that the hydrogel–calcium phosphate composite supported the formation of a calcified cartilage-like matrix and is a promising scaffold design for osteochondral interface tissue engineering. PMID:21919797

  3. Viral Inactivation of Human Osteochondral Grafts with Methylene Blue and Light

    PubMed Central

    Zhao, Zhixing; Call, Gazell M.; Gao, Jizong; Yao, Jian Q.

    2014-01-01

    Objective: Cartilage injury is one of the most common disorders of synovial joints. Fresh osteochondral allografts are becoming a standard treatment; however, they are supply constrained with a potential risk of disease transmission. There are no known virucidal processes available for osteochondral allografts and most methods presently available are detrimental to cartilage. Methylene blue light treatment has been shown to be successful in the literature for viral inactivation of fresh frozen plasma. The purpose of this study was to determine the capacity of methylene blue light treatment to inactivate a panel of clinically relevant viruses inoculated onto osteochondral allografts. Design: Osteochondral grafts recovered from human cadaveric knees were inoculated with one of the following viruses: bovine viral diarrhea virus (BVDV), hepatitis A virus (HAV), human immunodeficiency virus type 1 (HIV-1), porcine parvovirus (PPV), and pseudorabies virus (PrV). The samples were processed through a methylene blue light treatment, which consisted of an initial soak in nonilluminated circulating methylene blue at ambient temperature, followed by light exposure with circulating methylene blue at cool temperatures. The final titer was compared with the recovery control for the viral log reduction. Results: HIV-1, BVDV, and PrV were reduced to nondetectable levels while HAV and PPV were reduced by 3.1 and 5.6 logs, respectively. Conclusions: The methylene blue light treatment was effective in reducing (a) enveloped DNA and RNA viruses to nondetectable levels and (b) nonenveloped DNA and RNA viruses of inoculated human osteochondral grafts by 3.1 to 5.6 logs. This study demonstrates the first practical method for significantly reducing viral load in osteochondral implants. PMID:26069682

  4. Viral Inactivation of Human Osteochondral Grafts with Methylene Blue and Light.

    PubMed

    Squillace, Donna M; Zhao, Zhixing; Call, Gazell M; Gao, Jizong; Yao, Jian Q

    2014-01-01

    Cartilage injury is one of the most common disorders of synovial joints. Fresh osteochondral allografts are becoming a standard treatment; however, they are supply constrained with a potential risk of disease transmission. There are no known virucidal processes available for osteochondral allografts and most methods presently available are detrimental to cartilage. Methylene blue light treatment has been shown to be successful in the literature for viral inactivation of fresh frozen plasma. The purpose of this study was to determine the capacity of methylene blue light treatment to inactivate a panel of clinically relevant viruses inoculated onto osteochondral allografts. Osteochondral grafts recovered from human cadaveric knees were inoculated with one of the following viruses: bovine viral diarrhea virus (BVDV), hepatitis A virus (HAV), human immunodeficiency virus type 1 (HIV-1), porcine parvovirus (PPV), and pseudorabies virus (PrV). The samples were processed through a methylene blue light treatment, which consisted of an initial soak in nonilluminated circulating methylene blue at ambient temperature, followed by light exposure with circulating methylene blue at cool temperatures. The final titer was compared with the recovery control for the viral log reduction. HIV-1, BVDV, and PrV were reduced to nondetectable levels while HAV and PPV were reduced by 3.1 and 5.6 logs, respectively. The methylene blue light treatment was effective in reducing (a) enveloped DNA and RNA viruses to nondetectable levels and (b) nonenveloped DNA and RNA viruses of inoculated human osteochondral grafts by 3.1 to 5.6 logs. This study demonstrates the first practical method for significantly reducing viral load in osteochondral implants.

  5. Tissue engineering osteochondral implants for temporomandibular joint repair.

    PubMed

    Schek, R M; Taboas, J M; Hollister, S J; Krebsbach, P H

    2005-11-01

    Tissue engineering has provided an alternative to traditional strategies to repair and regenerate temporomandibular joints (TMJ). A successful strategy to engineer osteochondral tissue, such as that found in the TMJ, will produce tissue that is both biologically and mechanically functional. Image-based design (IBD) and solid free-form (SFF) fabrication can be used to generate scaffolds that are load bearing and match patient and defect site geometry. The objective of this study was to demonstrate how scaffold design, materials, and biological factors can be used in an integrated approach to regenerate a multi-tissue interface. IBD and SFF were first used to create biomimetic scaffolds with appropriate bulk geometry and microarchitecture. Biphasic composite scaffolds were then manufactured with the same techniques and used to simultaneously generate bone and cartilage in discrete regions and provide for the development of a stable interface between cartilage and subchondral bone. Poly-l-lactic acid/hydroxyapatite composite scaffolds were differentially seeded with fibroblasts transduced with an adenovirus expressing bone morphogenetic protein-7 in the ceramic phase and fully differentiated chondrocytes in the polymeric phase, and were subcutaneously implanted into mice. Following implantation in the ectopic site, the biphasic scaffolds promoted the simultaneous growth of bone, cartilage, and a mineralized interface tissue. Within the ceramic phase, the pockets of tissue generated included blood vessels, marrow stroma, and adipose tissue. This combination of IBD and SFF-fabricated biphasic scaffolds with gene and cell therapy is a promising approach to regenerate osteochondral defects and, ultimately, the TMJ.

  6. Osteochondral grafting for cartilage defects in the patellar grooves of bilateral knee joints.

    PubMed

    Nakagawa, Yasuaki; Matsusue, Yoshitaka; Suzuki, Takashi; Kuroki, Hiroshi; Nakamura, Takashi

    2004-07-01

    Autogenous osteochondral grafts have become popular recently for use in small, isolated, contained articular cartilage defects. We treated a 35-year-old man who had cartilage defects, which were the same shape and probably the result of overuse, in the patellar grooves of both knee joints. The left side was 30 x 25 mm, and the right side was 17 x 17 mm in his right patellar groove, and 15 x 7 mm in his right medial femoral condyle. Therefore, we performed multiple osteochondral grafting of the bilateral lesions. Thirty-two months after his right knee operation (37 months after his left one), he had no pain or symptoms in his left knee and occasional mild pain and catching in his right knee. At second-look arthroscopy, the joint surface of the articular cartilage in the bilateral patellar groove was almost completely smooth. However, the whole of the weight-bearing area around the grafted plugs in the medial femoral condyle showed cartilage degeneration. Approximately 3 years after implantation of osteochondral grafts into similarly shaped cartilage lesions in the bilateral patellar grooves, the operative results were good. However, careful follow up is needed.

  7. Osteochondral tissue engineering: scaffolds, stem cells and applications

    PubMed Central

    Nooeaid, Patcharakamon; Salih, Vehid; Beier, Justus P; Boccaccini, Aldo R

    2012-01-01

    Osteochondral tissue engineering has shown an increasing development to provide suitable strategies for the regeneration of damaged cartilage and underlying subchondral bone tissue. For reasons of the limitation in the capacity of articular cartilage to self-repair, it is essential to develop approaches based on suitable scaffolds made of appropriate engineered biomaterials. The combination of biodegradable polymers and bioactive ceramics in a variety of composite structures is promising in this area, whereby the fabrication methods, associated cells and signalling factors determine the success of the strategies. The objective of this review is to present and discuss approaches being proposed in osteochondral tissue engineering, which are focused on the application of various materials forming bilayered composite scaffolds, including polymers and ceramics, discussing the variety of scaffold designs and fabrication methods being developed. Additionally, cell sources and biological protein incorporation methods are discussed, addressing their interaction with scaffolds and highlighting the potential for creating a new generation of bilayered composite scaffolds that can mimic the native interfacial tissue properties, and are able to adapt to the biological environment. PMID:22452848

  8. Integrated Bi-Layered Scaffold for Osteochondral Tissue Engineering

    PubMed Central

    Galperin, Anna; Oldinski, Rachael A.; Florczyk, Stephen J.; Bryers, James D.; Zhang, Miqin

    2013-01-01

    Osteochondral tissue engineering poses the challenge of combining both cartilage and bone tissue engineering fundamentals. In this study, a sphere-templating technique was applied to fabricate an integrated bi-layered scaffold based on degradable poly(hydroxyethyl methacrylate) hydrogel. One layer of the integrated scaffold was designed with a single defined, monodispersed pore size of 38 μm and pore surfaces coated with hydroxyapatite particles to promote regrowth of subchondral bone while the second layer had 200 μm pores with surfaces decorated with hyaluronan for articular cartilage regeneration. Mechanical properties of the construct as well as cyto-compatibility of the scaffold and its degradation products were elucidated. To examine the potential of the biphasic scaffold for regeneration of osteochondral tissue the designated cartilage and bone layers of the integrated bi-layered scaffold were seeded with chondrocytes differentiated from human mesenchymal stem cells and primary human mesenchymal stem cells, respectively. Both types of cells were co-cultured within the scaffold in standard medium without soluble growth/differentiation factors over four weeks. The ability of the integrated bi-layered scaffold to support simultaneous matrix deposition and adequate cell growth of two distinct cell lineages in each layer during four weeks of co-culture in vitro in the absence of soluble growth factors was demonstrated. PMID:23225568

  9. Traumatic proximal interphalangeal joint reconstruction with an autologous hemi-toe osteochondral graft: case report.

    PubMed

    Pirani, Asif A; Rao, Ajit; Sharma, Sheel

    2013-07-01

    We report a case of a traumatic proximal interphalangeal joint injury with loss of the middle phalangeal base and articular surface, which was reconstructed with an autologous hemi-toe osteochondral graft. The patient had a 72° improvement in proximal interphalangeal joint motion and excellent functional improvements. Postoperative computed tomography imaging indicated bony union and articular congruence. Copyright © 2013 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.

  10. Optical Clearing in Collagen- and Proteoglycan-Rich Osteochondral Tissues

    PubMed Central

    Neu, Corey P.; Novak, Tyler; Gilliland, Kateri Fites; Marshall, Peter; Calve, Sarah

    2014-01-01

    Objective Recent developments in optical clearing and microscopy technology have enabled the imaging of intact tissues at the millimeter scale to characterize cells via fluorescence labeling. While these techniques have facilitated the three-dimensional cellular characterization within brain and heart, study of dense connective tissues of the musculoskeletal system have been largely unexplored. Here, we quantify how optical clearing impacted the cell and tissue morphology of collagen-, proteoglycan-, and mineral-rich cartilage and bone from the articulating knee joint. Methods Water-based fructose solutions were used for optical clearing of bovine osteochondral tissues, followed by imaging with transmission and confocal microscopy. To confirm preservation of tissue structure during the clearing process, samples were mechanically tested in unconfined compression and visualized by cryoSEM. Results Optical clearing enhanced light transmission through cartilage, but not subchondral bone regions. Fluorescent staining and immunolabeling was preserved through sample preparations, enabling imaging to cartilage depths 5 times deeper than previously reported, limited only by the working distance of the microscope objective. Chondrocyte volume remained unchanged in response to, and upon the reversal, of clearing. Equilibrium modulus increased in cleared samples, and was attributed to exchange of interstitial fluid with the more viscous fructose solution, but returned to control levels upon unclearing. In addition, cryoSEM-based analysis of cartilage showed no ultrastructural changes. Conclusion We anticipate large-scale microscopy of diverse connective tissues will enable the study of intact, three-dimensional interfaces (e.g. osteochondral) and cellular connectivity as a function of development, disease, and regeneration, which have been previously hindered by specimen opacity. PMID:25454370

  11. Cartilage storage at 4 °C with regular culture medium replacement benefits chondrocyte viability of osteochondral grafts in vitro.

    PubMed

    Qi, Jianhong; Hu, Zunjie; Song, Hongqiang; Chen, Bin; Xie, Di; Zhou, Lu; Zhang, Yanming

    2016-09-01

    Maintenance of articular cartilage allografts in culture media is a common method of tissue storage; however, the technical parameters of graft storage remain controversial. In this study, we examined the optimal temperature and culture medium exchange rate for the storage of osteochondral allografts in vitro. Cylindrical osteochondral grafts (n = 120), harvested from the talar joint surface of ten Boer goats, were randomly classified into four groups and stored under the following conditions: Group A1 was maintained at 4 °C in culture medium that was refreshed every 2 days; Group A2 was maintained at 4 °C in the same culture medium, without refreshing; Group B1, was maintained at 37 °C in culture medium that was refreshed every 2 days; Group B2, was maintained at 37 °C in the same culture medium, without refreshing. Chondrocyte viability in the grafts was determined by ethidium bromide/fluorescein diacetate staining on days 7, 21, and 35. Proteoglycan content was measured by Safranin-O staining. Group A1 exhibited the highest chondrocyte survival rates of 90.88 %, 88.31 % and 78.69 % on days 7, 21, and 35, respectively. Safranin O staining revealed no significant differences between groups on days 21 and 35. These results suggest that storage of osteochondral grafts at 4 °C with regular culture medium replacement should be highly suitable for clinical application.

  12. Intra-articular osteoid osteoma of the lateral tibial plateau treated with arthroscopically assisted removal and retrograde osteochondral grafting.

    PubMed

    Adachi, Nobuo; Shimose, Shoji; Nakamae, Atsuo; Okuhara, Atsushi; Kamei, Goki; Ochi, Mitsuo

    2014-01-01

    The treatment of an intra-articular osteoid osteoma is sometimes challenging, because of its location. We report a patient with an intra-articular osteoid osteoma of the lateral tibial plateau which was excised under an arthroscopically assisted procedure. After total resection of the intra-articular osteoid osteoma, the osteochondral defect of the lateral tibial plateau was reconstructed with a retrograde autogenous osteochondral graft which was harvested from the non-weightbearing area of the distal femur.

  13. 3D Bioprinting for Cartilage and Osteochondral Tissue Engineering.

    PubMed

    Daly, Andrew C; Freeman, Fiona E; Gonzalez-Fernandez, Tomas; Critchley, Susan E; Nulty, Jessica; Kelly, Daniel J

    2017-08-14

    Significant progress has been made in the field of cartilage and bone tissue engineering over the last two decades. As a result, there is real promise that strategies to regenerate rather than replace damaged or diseased bones and joints will one day reach the clinic however, a number of major challenges must still be addressed before this becomes a reality. These include vascularization in the context of large bone defect repair, engineering complex gradients for bone-soft tissue interface regeneration and recapitulating the stratified zonal architecture present in many adult tissues such as articular cartilage. Tissue engineered constructs typically lack such spatial complexity in cell types and tissue organization, which may explain their relatively limited success to date. This has led to increased interest in bioprinting technologies in the field of musculoskeletal tissue engineering. The additive, layer by layer nature of such biofabrication strategies makes it possible to generate zonal distributions of cells, matrix and bioactive cues in 3D. The adoption of biofabrication technology in musculoskeletal tissue engineering may therefore make it possible to produce the next generation of biological implants capable of treating a range of conditions. Here, advances in bioprinting for cartilage and osteochondral tissue engineering are reviewed. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Repair of articular cartilage defects with tissue-engineered osteochondral composites in pigs.

    PubMed

    Cui, Weiding; Wang, Qing; Chen, Gang; Zhou, Shixiang; Chang, Qing; Zuo, Qiang; Ren, Kewei; Fan, Weimin

    2011-04-01

    To compare the results of repair of knee cartilage defects with tissue-engineered osteochondral composites and tissue-engineered cartilage in pigs. Autologous chondrocytes and osteoblasts were seeded on scaffolds of polylactic-co-glycolic acid (PLGA) and tricalcium phosphate (TCP) to generate tissue-engineered cartilage and tissue-engineered bone, respectively. The tissue-engineered osteochondral composite was formed by a chondrocyte-PLGA construct sutured to an osteoblast-TCP construct with an absorbable suture. Cartilage defects were surgically created at the weightbearing surface of the bilateral femoral medial condyles of 12 mini-pigs. Thus, 24 defects in 12 pigs were randomly assigned to three treatment groups: tissue-engineered osteochondral composite group, tissue-engineered cartilage group, and blank control group. Six months after surgery, the regenerated cartilage was scored macroscopically and histologically. The compressive properties and glycosaminoglycan (GAG) content of the cartilage were also assessed. The gross grading scale indicated that the mean scores of the tissue-engineered osteochondral composite group were significantly higher than those of the tissue-engineered cartilage group. According to the International Cartilage Repair Society (ICRS) Visual Histological Assessment Scale, the scores of the osteochondral composite group were significantly better than those of the tissue-engineered cartilage group and blank control group. Assessment of compressive properties and GAG content showed better repair results in the osteochondral composite group than those of the tissue-engineered cartilage group. Using tissue-engineered osteochondral composites to repair cartilage defects was better than that of tissue-engineered cartilage.

  15. Scaphoid reconstruction by a free vascularized osteochondral graft from the rib: a case report.

    PubMed

    Lanzetta, Marco

    2009-01-01

    Nonunion and avascular necrosis of the proximal pole of the scaphoid remain one of the most difficult problems in wrist reconstructive surgery. A number of interpositional vascularized bone grafts have been proposed for scaphoid nonunion, in order to promote faster union at the contact site with both fragments. However, once the proximal pole has undergone avascular changes and is completely necrotic, there is no alternative other than to remove it. At present, more radical operations have been advocated in these cases, such as proximal row carpectomy or intercarpal arthrodesis. We present a case where the necrotic proximal pole of the scaphoid was removed and replaced with a remodeled osteochondral-free vascularized graft from the rib based on the inferior and superior intercostal arteries, branches of the internal mammary artery.

  16. A biphasic scaffold based on silk and bioactive ceramic with stratified properties for osteochondral tissue regeneration.

    PubMed

    Li, Jiao Jiao; Kim, Kyungsook; Roohani-Esfahani, Seyed-Iman; Guo, Jin; Kaplan, David L; Zreiqat, Hala

    2015-07-14

    Significant clinical challenges encountered in the effective long-term treatment of osteochondral defects have inspired advancements in scaffold-based tissue engineering techniques to aid repair and regeneration. This study reports the development of a biphasic scaffold produced via a rational combination of silk fibroin and bioactive ceramic with stratified properties to satisfy the complex and diverse regenerative requirements of osteochondral tissue. Structural examination showed that the biphasic scaffold contained two phases with different pore morphologies to match the cartilage and bone segments of osteochondral tissue, which were joined at a continuous interface. Mechanical assessment showed that the two phases of the biphasic scaffold imitated the load-bearing behaviour of native osteochondral tissue and matched its compressive properties. In vitro testing showed that different compositions in the two phases of the biphasic scaffold could direct the preferential differentiation of human mesenchymal stem cells towards the chondrogenic or osteogenic lineage. By featuring simple and reproducible fabrication and a well-integrated interface, the biphasic scaffold strategy established in this study circumvented the common problems experienced with integrated scaffold designs and could provide an effective approach for the regeneration of osteochondral tissue.

  17. Advancements in Orthopedic Intervention: Retrograde Drilling and Bone Grafting of Osteochondral Lesions of the Knee Using Magnetic Resonance Imaging Guidance

    SciTech Connect

    Seebauer, Christian J.; Bail, Hermann J.; Rump, Jens C. Walter, Thula Teichgraeber, Ulf K. M.

    2010-12-15

    Computer-assisted surgery is currently a novel challenge for surgeons and interventional radiologists. Magnetic resonance imaging (MRI)-guided procedures are still evolving. In this experimental study, we describe and assess an innovative passive-navigation method for MRI-guided treatment of osteochondritis dissecans of the knee. A navigation principle using a passive-navigation device was evaluated in six cadaveric knee joint specimens for potential applicability in retrograde drilling and bone grafting of osteochondral lesions using MRI guidance. Feasibility and accuracy were evaluated in an open MRI scanner (1.0 T Philips Panorama HFO MRI System). Interactive MRI navigation allowed precise drilling and bone grafting of osteochondral lesions of the knee. All lesions were hit with an accuracy of 1.86 mm in the coronal plane and 1.4 mm the sagittal plane. Targeting of all lesions was possible with a single drilling. MRI allowed excellent assessment of correct positioning of the cancellous bone cylinder during bone grafting. The navigation device and anatomic structures could be clearly identified and distinguished throughout the entire drilling procedure. MRI-assisted navigation method using a passive navigation device is feasible for the treatment of osteochondral lesions of the knee under MRI guidance and allows precise and safe drilling without exposure to ionizing radiation. This method may be a viable alternative to other navigation principles, especially for pediatric and adolescent patients. This MRI-navigated method is also potentially applicable in many other MRI-guided interventions.

  18. Engineered osteochondral grafts using biphasic composite solid free-form fabricated scaffolds.

    PubMed

    Schek, Rachel M; Taboas, Juan M; Segvich, Sharon J; Hollister, Scott J; Krebsbach, Paul H

    2004-01-01

    Tissue engineering has provided an alternative to traditional strategies to repair cartilage damaged by injury or degenerative disease. A successful strategy to engineer osteochondral tissue will mimic the natural contour of the articulating surface, achieve native mechanical properties and functional load-bearing ability, and lead to integration with host cartilage and underlying subchondral bone. Image-based design (IBD) and solid free-form (SFF) fabrication can be used to generate scaffolds that are load bearing and match articular geometry. The objective of this study was to utilize materials and biological factors in an integrated approach to regenerate a multitissue interface. Biphasic composite scaffolds manufactured by IBD and SFF fabrication were used to simultaneously generate bone and cartilage in discrete regions and provide for the development of a stable interface between cartilage and subchondral bone. Poly-L-lactic acid/hydroxyapatite composite scaffolds were differentially seeded with fibroblasts transduced with an adenovirus expressing bone morphogenetic protein 7 (BMP-7) in the ceramic phase and fully differentiated chondrocytes in the polymeric phase. After subcutaneous implantation into mice, the biphasic scaffolds promoted the simultaneous growth of bone, cartilage, and a mineralized interface tissue. Within the ceramic phase, the pockets of tissue generated included blood vessels, marrow stroma, and adipose tissue. This combination of IBD and SFF-fabricated biphasic scaffolds with gene and cell therapy is a promising approach to regenerate osteochondral defects.

  19. Hyaline cartilage degenerates after autologous osteochondral transplantation.

    PubMed

    Tibesku, C O; Szuwart, T; Kleffner, T O; Schlegel, P M; Jahn, U R; Van Aken, H; Fuchs, S

    2004-11-01

    Autologous osteochondral grafting is a well-established clinical procedure to treat focal cartilage defects in patients, although basic research on this topic remains sparse. The aim of the current study was to evaluate (1) histological changes of transplanted hyaline cartilage of osteochondral grafts and (2) the tissue that connects the transplanted cartilage with the adjacent cartilage in a sheep model. Both knee joints of four sheep were opened surgically and osteochondral grafts were harvested and simultaneously transplanted to the contralateral femoral condyle. The animals were sacrificed after three months and the received knee joints were evaluated histologically. Histological evaluation showed a complete ingrowth of the osseous part of the osteochondral grafts. A healing or ingrowth at the level of the cartilage could not be observed. Histological evaluation of the transplanted grafts according to Mankin revealed significantly more and more severe signs of degeneration than the adjacent cartilage, such as cloning of chondrocytes and irregularities of the articular surface. We found no connecting tissue between the transplanted and the adjacent cartilage and histological signs of degeneration of the transplanted hyaline cartilage. In the light of these findings, long-term results of autologous osteochondral grafts in human beings have to be followed critically.

  20. Post-traumatic malunion of the distal radial intra-articular fractures treated with autologous costal osteochondral grafts and bioabsorbable plates.

    PubMed

    Furukawa, Kayoko; Sakai, Akinori; Menuki, Kunitaka; Oshige, Toshihisa; Zenke, Yukichi; Nakamura, Toshitaka

    2014-03-01

    Intra-articular distal radial fractures with partial bone loss at the wrist were reconstructed using osteochondral grafts in 2 patients who were followed up for at least 18 months. Both patients experienced posttraumatic arthrosis of the wrist joint. The materials of the intra-articular fixation were bioabsorbable plates and screws. Reconstruction of a partially destroyed articular surface using a costal osteochondral graft is reliable and allows filling and resurfacing an articular cartilage void.

  1. Osteochondral allograft transplantation in cartilage repair: Graft storage paradigm, translational models, and clinical applications.

    PubMed

    Bugbee, William D; Pallante-Kichura, Andrea L; Görtz, Simon; Amiel, David; Sah, Robert

    2016-01-01

    The treatment of articular cartilage injury and disease has become an increasingly relevant part of orthopaedic care. Articular cartilage transplantation, in the form of osteochondral allografting, is one of the most established techniques for restoration of articular cartilage. Our research efforts over the last two decades have supported the transformation of this procedure from experimental "niche" status to a cornerstone of orthopaedic practice. In this Kappa Delta paper, we describe our translational and clinical science contributions to this transformation: (1) to enhance the ability of tissue banks to process and deliver viable tissue to surgeons and patients, (2) to improve the biological understanding of in vivo cartilage and bone remodeling following osteochondral allograft (OCA) transplantation in an animal model system, (3) to define effective surgical techniques and pitfalls, and (4) to identify and clarify clinical indications and outcomes. The combination of coordinated basic and clinical studies is part of our continuing comprehensive academic OCA transplant program. Taken together, the results have led to the current standards for OCA processing and storage prior to implantation and also novel observations and mechanisms of the biological and clinical behavior of OCA transplants in vivo. Thus, OCA transplantation is now a successful and increasingly available treatment for patients with disabling osteoarticular cartilage pathology.

  2. Axial load-bearing capacity of an osteochondral autograft stabilized with a resorbable osteoconductive bone cement compared with a press-fit graft in a bovine model.

    PubMed

    Kiss, Marc-Olivier; Levasseur, Annie; Petit, Yvan; Lavigne, Patrick

    2012-05-01

    Osteochondral autografts in mosaicplasty are inserted in a press-fit fashion, and hence, patients are kept nonweightbearing for up to 2 months after surgery to allow bone healing and prevent complications. Very little has been published regarding alternative fixation techniques of those grafts. Osteochondral autografts stabilized with a resorbable osteoconductive bone cement would have a greater load-bearing capacity than standard press-fit grafts. Controlled laboratory study. Biomechanical testing was conducted on 8 pairs of cadaveric bovine distal femurs. For the first 4 pairs, 6 single osteochondral autografts were inserted in a press-fit fashion on one femur. On the contralateral femur, 6 grafts were stabilized with a calcium triglyceride osteoconductive bone cement. For the 4 remaining pairs of femurs, 4 groups of 3 adjacent press-fit grafts were inserted on one femur, whereas on the contralateral femur, grafts were cemented. After a maturation period of 48 hours, axial loading was applied on all single grafts and on the middle graft of each 3-in-a-row series. For the single-graft configuration, median loads required to sink the press-fit and cemented grafts by 2 and 3 mm were 281.87 N versus 345.56 N (P = .015) and 336.29 N versus 454.08 N (P = .018), respectively. For the 3-in-a-row configuration, median loads required to sink the press-fit and cemented grafts by 2 and 3 mm were 260.31 N versus 353.47 N (P = .035) and 384.83 N versus 455.68 N (P = .029), respectively. Fixation of osteochondral grafts using bone cement appears to improve immediate stability over the original mosaicplasty technique for both single- and multiple-graft configurations. Achieving greater primary stability of osteochondral grafts could potentially accelerate postoperative recovery, allowing early weightbearing and physical therapy.

  3. Results of Bone Peg Grafting for Capitellar Osteochondritis Dissecans in Adolescent Baseball Players.

    PubMed

    Oshiba, Hiroyuki; Itsubo, Toshiro; Ikegami, Shota; Nakamura, Koichi; Uchiyama, Shigeharu; Kato, Hiroyuki

    2016-12-01

    Bone peg grafting (BPG) has been advocated for early-stage humeral capitellar osteochondritis dissecans (COCD). However, the clinical and radiological results of BPG, along with its indications, have not been described in detail. COCD classified as International Cartilage Repair Society (ICRS) osteochondritis dissecans (OCD) I or II in adolescent baseball players can be treated successfully by BPG. Case series; Level of evidence, 4 METHODS: Eleven male baseball players (age range at surgery, 13-16 years) who underwent BPG for COCD were enrolled in this study. No improvement had been seen in any patient after 6 months of preoperative nonthrowing observation. During surgery, 2 to 5 bone pegs were inserted into the COCD lesion after confirmation of lesion stability to the bony floor. All patients were directly evaluated at 12 and 24 months after surgery by physical findings, radiological prognosis, and magnetic resonance imaging (MRI). Of the 11 patients, 10 could return to comparable baseball ability levels within 12 months. The Timmerman-Andrews score improved significantly from a mean ± SD of 171.8 ± 12.1 preoperatively to 192.3 ± 6.5 at the final observation. Radiological healing of the lesions was determined as complete in 8 patients and partial in 3. Patients possessing a centrally positioned lesion or a lesion <75% of the size of the capitellum tended most strongly to achieve complete radiological healing, while growth plate status appeared unrelated to outcome. The mean Henderson MRI score improved from 6.3 ± 1.5 to 4.8 ± 1.6 at 12 and 24 months after BPG, respectively. MRI findings also suggested that remodeling of COCD lesions had continued to up to 24 months postoperatively. BPG enabled 91% of COCD patients with ICRS OCD I or II to return to preoperative baseball abilities within 12 months. Integration of the grafted site may continue until at least 24 months postoperatively. An ICRS OCD I or II lesion with central positioning and/or occupying <75% of

  4. Stem cell-based microphysiological osteochondral system to model tissue response to interleukin-1β.

    PubMed

    Lin, Hang; Lozito, Thomas P; Alexander, Peter G; Gottardi, Riccardo; Tuan, Rocky S

    2014-07-07

    Osteoarthritis (OA) is a chronic degenerative disease of the articular joint that involves both bone and cartilage degenerative changes. An engineered osteochondral tissue within physiological conditions will be of significant utility in understanding the pathogenesis of OA and testing the efficacy of potential disease-modifying OA drugs (DMOADs). In this study, a multichamber bioreactor was fabricated and fitted into a microfluidic base. When the osteochondral construct is inserted, two chambers are formed on either side of the construct (top, chondral; bottom, osseous) that is supplied by different medium streams. These medium conduits are critical to create tissue-specific microenvironments in which chondral and osseous tissues will develop and mature. Human bone marrow stem cell (hBMSCs)-derived constructs were fabricated in situ and cultured within the bioreactor and induced to undergo spatially defined chondrogenic and osteogenic differentiation for 4 weeks in tissue-specific media. We observed tissue specific gene expression and matrix production as well as a basophilic interface suggesting a developing tidemark. Introduction of interleukin-1β (IL-1β) to either the chondral or osseous medium stream induced stronger degradative responses locally as well as in the opposing tissue type. For example, IL-1β treatment of the osseous compartment resulted in a strong catabolic response in the chondral layer as indicated by increased matrix metalloproteinase (MMP) expression and activity, and tissue-specific gene expression. This induction was greater than that seen with IL-1β application to the chondral component directly, indicative of active biochemical communication between the two tissue layers and supporting the osteochondral nature of OA. The microtissue culture system developed here offers novel capabilities for investigating the physiology of osteochondral tissue and pathogenic mechanisms of OA and serving as a high-throughput platform to test potential

  5. Dual growth factor delivery from bilayered, biodegradable hydrogel composites for spatially-guided osteochondral tissue repair

    PubMed Central

    Lu, Steven; Lam, Johnny; Trachtenberg, Jordan E.; Lee, Esther J.; Seyednejad, Hajar; van den Beucken, Jeroen J. J. P.; Tabata, Yasuhiko; Wong, Mark E.; Jansen, John A.; Mikos, Antonios G.; Kasper, F. Kurtis

    2014-01-01

    The present work investigated the use of biodegradable hydrogel composite scaffolds, based on the macromer oligo(poly(ethylene glycol) fumarate) (OPF), to deliver growth factors for the repair of osteochondral tissue in a rabbit model. In particular, bilayered OPF composites were used to mimic the structural layers of the osteochondral unit, and insulin-like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2) were loaded into gelatin microparticles and embedded within the OPF hydrogel matrix in a spatially controlled manner. Three different scaffold formations were implanted in a medial femoral condyle osteochondral defect: 1) IGF-1 in the chondral layer, 2) BMP-2 in the subchondral layer, and 3) IGF-1 and BMP-2 in their respective separate layers. The quantity and quality of osteochondral repair was evaluated at 6 and 12 weeks with histological scoring and micro-computed tomography (micro-CT). While histological scoring results at 6 weeks showed no differences between experimental groups, micro-CT analysis revealed that the delivery of BMP-2 alone increased the number of bony trabecular islets formed, an indication of early bone formation, over that of IGF-1 delivery alone. At 12 weeks post-implantation, minimal differences were detected between the three groups for cartilage repair. However, the dual delivery of IGF-1 and BMP-2 had a higher proportion of subchondral bone repair, greater bone growth at the defect margins, and lower bone specific surface than the single delivery of IGF-1. These results suggest that the delivery of BMP-2 enhances subchondral bone formation and that, while the dual delivery of IGF-1 and BMP-2 in separate layers does not improve cartilage repair under the conditions studied, they may synergistically enhance the degree of subchondral bone formation. Overall, bilayered OPF hydrogel composites demonstrate potential as spatially-guided, multiple growth factor release vehicles for osteochondral tissue repair. PMID:25047629

  6. Dual growth factor delivery from bilayered, biodegradable hydrogel composites for spatially-guided osteochondral tissue repair.

    PubMed

    Lu, Steven; Lam, Johnny; Trachtenberg, Jordan E; Lee, Esther J; Seyednejad, Hajar; van den Beucken, Jeroen J J P; Tabata, Yasuhiko; Wong, Mark E; Jansen, John A; Mikos, Antonios G; Kasper, F Kurtis

    2014-10-01

    The present work investigated the use of biodegradable hydrogel composite scaffolds, based on the macromer oligo(poly(ethylene glycol) fumarate) (OPF), to deliver growth factors for the repair of osteochondral tissue in a rabbit model. In particular, bilayered OPF composites were used to mimic the structural layers of the osteochondral unit, and insulin-like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2) were loaded into gelatin microparticles and embedded within the OPF hydrogel matrix in a spatially controlled manner. Three different scaffold formulations were implanted in a medial femoral condyle osteochondral defect: 1) IGF-1 in the chondral layer, 2) BMP-2 in the subchondral layer, and 3) IGF-1 and BMP-2 in their respective separate layers. The quantity and quality of osteochondral repair was evaluated at 6 and 12 weeks with histological scoring and micro-computed tomography (micro-CT). While histological scoring results at 6 weeks showed no differences between experimental groups, micro-CT analysis revealed that the delivery of BMP-2 alone increased the number of bony trabecular islets formed, an indication of early bone formation, over that of IGF-1 delivery alone. At 12 weeks post-implantation, minimal differences were detected between the three groups for cartilage repair. However, the dual delivery of IGF-1 and BMP-2 had a higher proportion of subchondral bone repair, greater bone growth at the defect margins, and lower bone specific surface than the single delivery of IGF-1. These results suggest that the delivery of BMP-2 enhances subchondral bone formation and that, while the dual delivery of IGF-1 and BMP-2 in separate layers does not improve cartilage repair under the conditions studied, they may synergistically enhance the degree of subchondral bone formation. Overall, bilayered OPF hydrogel composites demonstrate potential as spatially-guided, multiple growth factor release vehicles for osteochondral tissue repair.

  7. Spatial Engineering of Osteochondral Tissue Constructs Through Microfluidically Directed Differentiation of Mesenchymal Stem Cells.

    PubMed

    Goldman, Stephen M; Barabino, Gilda A

    2016-01-01

    The development of tissue engineered osteochondral units has been slowed by a number of technical hurdles associated with recapitulating their heterogeneous nature ex vivo. Subsequently, numerous approaches with respect to cell sourcing, scaffolding composition, and culture media formulation have been pursued, which have led to high variability in outcomes and ultimately the lack of a consensus bioprocessing strategy. As such, the objective of this study was to standardize the design process by focusing on differentially supporting formation of cartilaginous and bony matrix by a single cell source in a spatially controlled manner within a single material system. A cell-polymer solution of bovine mesenchymal stem cells and agarose was cast against micromolds of a serpentine network and stacked to produce tissue constructs containing two independent microfluidic networks. Constructs were fluidically connected to two controlled flow loops and supplied with independently tuned differentiation parameters for chondrogenic and osteogenic induction, respectively. Constructs receiving inductive media showed differential gene expression of both chondrogenic and osteogenic markers in opposite directions along the thickness of the construct that was recapitulated at the protein level with respect to collagens I, II, and X. A control group receiving noninductive media showed homogeneous expression of these biomarkers measured in lower concentrations at both the mRNA and protein level. This work represents an important step in the rational design of engineered osteochondral units through establishment of an enabling technology for further optimization of scaffolding formulations and bioprocessing conditions toward the production of commercially viable osteochondral tissue products.

  8. Ovine Mesenchymal Stromal Cells: Morphologic, Phenotypic and Functional Characterization for Osteochondral Tissue Engineering

    PubMed Central

    Sanjurjo-Rodríguez, Clara; Castro-Viñuelas, Rocío; Hermida-Gómez, Tamara; Fernández-Vázquez, Tania; Fuentes-Boquete, Isaac Manuel; de Toro-Santos, Francisco Javier; Blanco-García, Francisco Javier

    2017-01-01

    Introduction Knowledge of ovine mesenchymal stromal cells (oMSCs) is currently expanding. Tissue engineering combining scaffolding with oMSCs provides promising therapies for the treatment of osteochondral diseases. Purpose The aim was to isolate and characterize oMSCs from bone marrow aspirates (oBMSCs) and to assess their usefulness for osteochondral repair using β-tricalcium phosphate (bTCP) and type I collagen (Col I) scaffolds. Methods Cells isolated from ovine bone marrow were characterized morphologically, phenotypically, and functionally. oBMSCs were cultured with osteogenic medium on bTCP and Col I scaffolds. The resulting constructs were evaluated by histology, immunohistochemistry and electron microscopy studies. Furthermore, oBMSCs were cultured on Col I scaffolds to develop an in vitro cartilage repair model that was assessed using a modified International Cartilage Research Society (ICRS) II scale. Results oBMSCs presented morphology, surface marker pattern and multipotent capacities similar to those of human BMSCs. oBMSCs seeded on Col I gave rise to osteogenic neotissue. Assessment by the modified ICRS II scale revealed that fibrocartilage/hyaline cartilage was obtained in the in vitro repair model. Conclusions The isolated ovine cells were demonstrated to be oBMSCs. oBMSCs cultured on Col I sponges successfully synthesized osteochondral tissue. The data suggest that oBMSCs have potential for use in preclinical models prior to human clinical studies. PMID:28141815

  9. Mechanical loading regulates human MSC differentiation in a multi-layer hydrogel for osteochondral tissue engineering.

    PubMed

    Steinmetz, Neven J; Aisenbrey, Elizabeth A; Westbrook, Kristofer K; Qi, H Jerry; Bryant, Stephanie J

    2015-07-01

    A bioinspired multi-layer hydrogel was developed for the encapsulation of human mesenchymal stem cells (hMSCs) as a platform for osteochondral tissue engineering. The spatial presentation of biochemical cues, via incorporation of extracellular matrix analogs, and mechanical cues, via both hydrogel crosslink density and externally applied mechanical loads, were characterized in each layer. A simple sequential photopolymerization method was employed to form stable poly(ethylene glycol)-based hydrogels with a soft cartilage-like layer of chondroitin sulfate and low RGD concentrations, a stiff bone-like layer with high RGD concentrations, and an intermediate interfacial layer. Under a compressive load, the variation in hydrogel stiffness within each layer produced high strains in the soft cartilage-like layer, low strains in the stiff bone-like layer, and moderate strains in the interfacial layer. When hMSC-laden hydrogels were cultured statically in osteochondral differentiation media, the local biochemical and matrix stiffness cues were not sufficient to spatially guide hMSC differentiation after 21 days. However dynamic mechanical stimulation led to differentially high expression of collagens with collagen II in the cartilage-like layer, collagen X in the interfacial layer and collagen I in the bone-like layer and mineral deposits localized to the bone layer. Overall, these findings point to external mechanical stimulation as a potent regulator of hMSC differentiation toward osteochondral cellular phenotypes. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  10. A novel, visible light-induced, rapidly cross-linkable gelatin scaffold for osteochondral tissue engineering

    PubMed Central

    Mazaki, Tetsuro; Shiozaki, Yasuyuki; Yamane, Kentaro; Yoshida, Aki; Nakamura, Mariko; Yoshida, Yasuhiro; Zhou, Di; Kitajima, Takashi; Tanaka, Masato; Ito, Yoshihiro; Ozaki, Toshifumi; Matsukawa, Akihiro

    2014-01-01

    Osteochondral injuries remain difficult to repair. We developed a novel photo-cross-linkable furfurylamine-conjugated gelatin (gelatin-FA). Gelatin-FA was rapidly cross-linked by visible light with Rose Bengal, a light sensitizer, and was kept gelled for 3 weeks submerged in saline at 37°C. When bone marrow-derived stromal cells (BMSCs) were suspended in gelatin-FA with 0.05% Rose Bengal, approximately 87% of the cells were viable in the hydrogel at 24 h after photo-cross-linking, and the chondrogenic differentiation of BMSCs was maintained for up to 3 weeks. BMP4 fusion protein with a collagen binding domain (CBD) was retained in the hydrogels at higher levels than unmodified BMP4. Gelatin-FA was subsequently employed as a scaffold for BMSCs and CBD-BMP4 in a rabbit osteochondral defect model. In both cases, the defect was repaired with articular cartilage-like tissue and regenerated subchondral bone. This novel, photo-cross-linkable gelatin appears to be a promising scaffold for the treatment of osteochondral injury. PMID:24662725

  11. Biocompatibility of biomimetic multilayered alginate-chitosan/β-TCP scaffold for osteochondral tissue.

    PubMed

    Algul, Derya; Sipahi, Hande; Aydin, Ahmet; Kelleci, Feyza; Ozdatli, Sukran; Yener, Fatma Gulgun

    2015-08-01

    Biomimetic three-layered monolithic scaffold (TLS) intended for treatment of osteochondral defects was fabricated by using freeze drying method. The multilayered material was prepared with chitosan (C) and alginate (A) polyelectrolyte complex (CA/PEC) as a cartilaginous layer, a combination of CA/PEC (60 wt%) and β-tricalcium phosphate (β-TCP) (40 wt%) as an intermediate layer and a combination of CA/PEC (30 wt%) and β-TCP (70 wt%) as a subchondral layer in order to mimic the inherent gradient structure of healthy osteochondral tissue. Characterization of the scaffolds was performed using Fourier transform infrared (FT-IR) spectroscopy analysis, swelling and scanning electron microscopy (SEM) tests. In vitro cytotoxicity assay with L929 cells and EpiDerm skin irritation test (SIT) using the EpiDerm reconstructed human epidermal (RHE) model were performed to analyze biocompatibility of the scaffolds. Characterization results showed that there were strong ionic interactions among chitosan, alginate and β-TCP and the layers showed interconnected porous structure with different swelling ratios. The relative cell viability and SIT results were greater than 70% indicating that the scaffolds are considered nontoxic according to the International Organization for Standardization (ISO) standard. All results taken together, biomimetic TLS can be considered to be suitable for osteochondral applications. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    PubMed

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

    2015-09-01

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

  13. [Results of the experimental repair of osteochondral lesions in a pig model using tissue engineering].

    PubMed

    Villalobos Córdoba, F Enrique; Velasquillo Martínez, Cristina; Martínez López, Valentín; Lecona Butrón, Hugo; Reyes Marín, Baltasar; Estrada Villaseñor, Eréndira; Villegas, Castrejón Hilda; Solís Arrieta, Lilia; Espinosa Morales, Rolando; Ponce de León, Clemente Ibarra

    2007-01-01

    To repair experimental osteochondral knee lesions in pigs using tissue engineering. Eight 40-kg pigs underwent surgery. Cartilage and periosteal biopsies of their control knee were taken. Cartilage and periosteal cells were independently isolated, cultured and seeded in biodegradable PGA and PLA polymers that were fixed on the bottom of an osteochondral defect in the pig's experimental knee, with bioabsorbable Mitek implants. Four months later the pigs were sacrificed and the knees were analyzed with nuclear magnetic resonance imaging (NMRI), macroscopic assessment, histology, electron microscopy (EM), scanning electron microscopy (SEM) and SEM element analysis. All the defects were filled with cartilage-like tissue according to the NMRI evaluation and the visual examination. Hyaline-like cartilage was obtained in 3 defects and fibrocartilage in 5. The EM showed chondrocytes in the repair tissue. The SEM showed appropriate integration to the bone and the surrounding tissue. SEM element analysis showed sulphurized matrix attached to the bone with calcium and phosphates as predominant elements. Tissue engineering enabled the production of tissues similar to normal ones. The polymer fixation system was effective.

  14. Rabbit trochlear model of osteochondral allograft transplantation.

    PubMed

    To, Nhat; Curtiss, Shane; Neu, Corey P; Salgado, Christopher J; Jamali, Amir A

    2011-10-01

    Allografting and autografting of osteochondral tissues is a promising strategy to treat articular cartilage lesions in damaged joints. We developed a new model of fresh osteochondral allografting using the entire rabbit trochlea. The objective of the current study was to demonstrate that this model would achieve reproducible graft-host healing and maintain normal articular cartilage histologic, immunolocalization, and biochemical characteristics after transplantation under diverse storage and transplantation conditions. New Zealand white (n = 8) and Dutch belted (n = 8) rabbits underwent a 2-stage transplantation operation using osteochondral grafts that had been stored for 2 or 4 wk. Trochlear grafts harvested from the left knee were transplanted to the right knee as either autografts or allografts. Grafts were fixed with 22-gauge steel wire or 3-0 nylon suture. Rabbits were euthanized for evaluation at 1, 2, 4, 6, and 12 wk after transplantation. All grafts that remained in vivo for at least 4 wk demonstrated 100% interface healing by microCT. Trabecular bridging was present at the host-graft interface starting at 2 wk after transplantation, with no significant difference in cartilage histology between the various groups. The combined histology scores indicated minimal evidence of osteoarthritis. Immunostaining revealed that superficial zone protein was localized at the surface of all transplants. The rabbit trochlear model met our criteria for a successful model in regard to the ease of the procedure, low rate of surgical complications, relatively large articular cartilage surface area, and amount of host-graft bone interface available for analysis.

  15. Harnessing Cell–Biomaterial Interactions for Osteochondral Tissue Regeneration

    PubMed Central

    Kim, Kyobum; Yoon, Diana M.; Mikos, Antonios G.

    2013-01-01

    Articular cartilage that is damaged or diseased often requires surgical intervention to repair the tissue; therefore, tissue engineering strategies have been developed to aid in cartilage regeneration. Tissue engineering approaches often require the integration of cells, biomaterials, and growth factors to direct and support tissue formation. A variety of cell types have been isolated from adipose, bone marrow, muscle, and skin tissue to promote cartilage regeneration. The interaction of cells with each other and with their surrounding environment has been shown to play a key role in cartilage engineering. In tissue engineering approaches, biomaterials are commonly used to provide an initial framework for cell recruitment and proliferation and tissue formation. Modifications of the properties of biomaterials, such as creating sites for cell binding, altering their physicochemical characteristics, and regulating the delivery of growth factors, can have a significant influence on chondrogenesis. Overall, the goal is to completely restore healthy cartilage within an articular cartilage defect. This chapter aims to provide information about the importance of cell–biomaterial interactions for the chondrogenic differentiation of various cell populations that can eventually produce functional cartilage matrix that is indicative of healthy cartilage tissue. PMID:21975954

  16. Stem cell- and scaffold-based tissue engineering approaches to osteochondral regenerative medicine

    PubMed Central

    Sundelacruz, Sarah; Kaplan, David L.

    2009-01-01

    In osteochondral tissue engineering, cell recruitment, proliferation, differentiation, and patterning are critical for forming biologically and structurally viable constructs for repair of damaged or diseased tissue. However, since constructs prepared ex vivo lack the multitude of cues present in the in vivo microenvironment, cells often need to be supplied with external biological and physical stimuli to coax them towards targeted tissue functions. To determine which stimuli to present to cells, bioengineering strategies can benefit significantly from endogenous examples of skeletogenesis. As an example of developmental skeletogenesis, the developing limb bud serves as an excellent model system in which to study how an osteochondral structures form from undifferentiated precursor cells. Alongside skeletal formation during embryogenesis, bone also possesses innate regenerative capacity, displaying remarkable ability to heal after damage. Bone fracture healing shares many features with bone development, driving the hypothesis that the regenerative process generally recapitulates development. Similarities and differences between the two modes of bone formation may offer insight into the special requirements for healing damaged or diseased bone. Thus, endogenous fracture healing, as an example of regenerative skeletogenesis, may also inform bioengineering strategies. In this review, we summarize the key cellular events involving stem and progenitor cells in developmental and regenerative skeletogenesis, and discuss in parallel the corresponding cell- and scaffold-based strategies that tissue engineers employ to recapitulate these events in vitro. PMID:19508851

  17. All-arthroscopic AMIC(®) (AT-AMIC(®)) technique with autologous bone graft for talar osteochondral defects: clinical and radiological results.

    PubMed

    Usuelli, Federico Giuseppe; D'Ambrosi, Riccardo; Maccario, Camilla; Boga, Michele; de Girolamo, Laura

    2016-09-12

    Autologous Matrix-Induced Chondrogenesis (AMIC(®)) is known to provide satisfactory clinical results for the treatment of knee, hip, and ankle cartilage lesions. The purpose of this study was to evaluate clinical and radiological outcomes of patients treated with a new all-arthroscopic AMIC(®) (AT-AMIC(®)) technique with autologous bone graft for talar osteochondral defects at a follow-up of 24 months. Twenty patients underwent the AT-AMIC(®) procedure and autologous bone graft for type III and IV talar osteochondral lesions. Patients were evaluated pre-operatively and at 6, 12, and 24 months post-operatively using the American Orthopedic Foot and Ankle Society (AOFAS) score, the visual analog scale, and the SF-12 (Short Form-12). Radiological assessment included computed tomography (CT), magnetic resonance imaging (MRI), and magnetic resonance observation of cartilage repair tissue (MOCART). All scores significantly improved (p < 0.05) with respect to pre-operative values after 6 months. Further improvements were detected at 24 months (AOFAS, from 57.1 ± 14.9 before surgery to 86.6 ± 10.9 after 24 months; VAS, from 8.1 ± 1.4 to 2.5 ± 2.2; SF-12, from 29.9 ± 4.1 to 48.5 ± 6.9 and from 43.8 ± 2.9 to 53.1 ± 3.9, respectively, for Physical and Mental component score). Lesion area significantly reduced from 111.1 ± 43.2 mm(2) pre-operatively to 76.9 ± 38.1 mm(2) (p < 0.05) at final follow-up as assessed by CT, and from 154.1 ± 93.6 to 94.3 ± 61.3 mm(2) (p < 0.05) as assessed by MRI. The mean MOCART score was 42.8 ± 23.5 points and 50.9 ± 24.9 points, respectively, at 12 and 24 months after surgery (p < 0.05). AT-AMIC(®) with autologous bone grafting has proven to be a safe and effective minimal invasive technique, able to rapidly and significantly improve pain, function, and radiological healing of osteochondral talar lesions, with progressive further improvements up to 24 months. Orthopedic surgeons

  18. Construction of tissue-engineered osteochondral composites and repair of large joint defects in rabbit.

    PubMed

    Deng, Tianzheng; Lv, Jing; Pang, Jianliang; Liu, Bing; Ke, Jie

    2014-07-01

    In this study, a novel three-dimensional (3D) heterogeneous/bilayered scaffold was constructed to repair large defects in rabbit joints. The scaffold includes two distinct but integrated layers corresponding to the cartilage and bone components. The upper layer consists of gelatin, chondroitin sulphate and sodium hyaluronate (GCH), and the lower layer consists of gelatin and ceramic bovine bone (GCBB). The two form a 3D bilayered scaffold (GCH-GCBB), which mimics the natural osteochondral matrix for use as a scaffold for osteochondral tissue engineering. The purpose of this study was to evaluate the efficacy of this novel scaffold, combined with chondrocytes and bone marrow stem cells (BMSCs) to repair large defects in rabbit joints. Thirty-six large defects in rabbit femoral condyles were created; 12 defects were treated with the same scaffold combined with cells (group A); another 12 defects were treated with cell-free scaffolds (group B); the others were untreated (group C). At 6 and 12 weeks, in group A hyaline-like cartilage formation could be observed by histological examination; the newly formed cartilage, which stained for type II collagen, was detected by RT-PCR at high-level expression. Most of the GCBB was replaced by bone, while little remained in the underlying cartilage. At 36 weeks, GCBB was completely resorbed and a tidemark was observed in some areas. In contrast, groups B and C showed no cartilage formation but a great amount of fibrous tissue, with only a little bone formation. In summary, this study demonstrated that a novel scaffold, comprising a top layer of GCH, having mechanical properties comparable to native cartilage, and a bottom layer composed of GCBB, could be used to repair large osteochondral defects in joints.

  19. Injectable and 3D Bioprinted Polysaccharide Hydrogels: From Cartilage to Osteochondral Tissue Engineering.

    PubMed

    Radhakrishnan, Janani; Subramanian, Anuradha; Krishnan, Uma Maheswari; Sethuraman, Swaminathan

    2017-01-09

    Biomechanical performance of functional cartilage is executed by the exclusive anisotropic composition and spatially varying intricate architecture in articulating ends of diarthrodial joint. Osteochondral tissue constituting the articulating ends comprise superfical soft cartilage over hard subchondral bone sandwiching interfacial soft-hard tissue. The shock-absorbent, lubricating property of cartilage and mechanical stability of subchondral bone regions are rendered by extended chemical structure of glycosaminoglycans and mineral deposition, respectively. Extracellular matrix glycosaminoglycans analogous polysaccharides are major class of hydrogels investigated for restoration of functional cartilage. Recently, injectable hydrogels have gained momentum as it offers patient compliance, tunable mechanical properties, cell deliverability, and facile administration at physiological condition with long-term functionality and hyaline cartilage construction. Interestingly, facile modifiable functional groups in carbohydrate polymers impart tailorability of desired physicochemical properties and versatile injectable chemistry for the development of highly potent biomimetic in situ forming scaffold. The scaffold design strategies have also evolved from single component to bi- or multilayered and graded constructs with osteogenic properties for deep subchondral regeneration. This review highlights the significance of polysaccharide structure-based functions in engineering cartilage tissue, injectable chemistries, strategies for combining analogous matrices with cells/stem cells and biomolecules and multicomponent approaches for osteochondral mimetic constructs. Further, the rheology and precise spatiotemporal positioning of cells in hydrogel bioink for rapid prototyping of complex three-dimensional anisotropic cartilage have also been discussed.

  20. Effect of porosities of bilayered porous scaffolds on spontaneous osteochondral repair in cartilage tissue engineering

    PubMed Central

    Dong, Jian; Ding, Jiandong

    2015-01-01

    Poly(lactide-co-glycolide)-bilayered scaffolds with the same porosity or different ones on the two layers were fabricated, and the porosity effect on in vivo repairing of the osteochondral defect was examined in a comparative way for the first time. The constructs of scaffolds and bone marrow-derived mesenchymal stem cells were implanted into pre-created osteochondral defects in the femoral condyle of New Zealand white rabbits. After 12 weeks, all experimental groups exhibited good cartilage repairing according to macroscopic appearance, cross-section view, haematoxylin and eosin staining, toluidine blue staining, immunohistochemical staining and real-time polymerase chain reaction of characteristic genes. The group of 92% porosity in the cartilage layer and 77% porosity in the bone layer resulted in the best efficacy, which was understood by more biomechanical mimicking of the natural cartilage and subchondral bone. This study illustrates unambiguously that cartilage tissue engineering allows for a wide range of scaffold porosity, yet some porosity group is optimal. It is also revealed that the biomechanical matching with the natural composite tissue should be taken into consideration in the design of practical biomaterials, which is especially important for porosities of a multi-compartment scaffold concerning connected tissues. PMID:26813511

  1. Customized Fabrication of Osteochondral Tissue for Articular Joint Surface Repair

    DTIC Science & Technology

    2016-09-01

    Significant changes in use of biohazards and/or select agents 6. PRODUCTS:  Publications, conference papers , and presentations Report only the major...Scaffolds for Cartilage Tissue Engineering. Frontiers in Bioengineering and Biotechnology. Nothing to Report Other publications, conference papers ...prepped using chlorhexidine and betadiene scrubs, and the limb was draped using sterile towels . The initial incision was positioned on the medial

  2. Development of a Fresh Osteochondral Allograft Program Outside North America

    PubMed Central

    Tírico, Luís Eduardo Passarelli; Demange, Marco Kawamura; Santos, Luiz Augusto Ubirajara; de Rezende, Márcia Uchoa; Helito, Camilo Partezani; Gobbi, Riccardo Gomes; Pécora, José Ricardo; Croci, Alberto Tesconi; Bugbee, William Dick

    2015-01-01

    Objective To standardize and to develop a fresh osteochondral allograft protocol of procurement, processing and surgical utilization in Brazil. This study describes the steps recommended to make fresh osteochondral allografts a viable treatment option in a country without previous fresh allograft availability. Design The process involves regulatory process modification, developing and establishing procurement, and processing and surgical protocols. Results Legislation: Fresh osteochondral allografts were not feasible in Brazil until 2009 because the law prohibited preservation of fresh grafts at tissue banks. We approved an amendment that made it legal to preserve fresh grafts for 30 days from 2°C to 6°C in tissue banks. Procurement: We changed the protocol of procurement to decrease tissue contamination. All tissues were procured in an operating room. Processing: Processing of the grafts took place within 12 hours of tissue recovery. A serum-free culture media with antibiotics was developed to store the grafts. Surgeries: We have performed 8 fresh osteochondral allografts on 8 knees obtaining grafts from 5 donors. Mean preoperative International Knee Documentation Committee (IKDC) score was 31.99 ± 13.4, improving to 81.26 ± 14.7 at an average of 24 months’ follow-up. Preoperative Knee Injury and Oseoarthritis Outcome Score (KOOS) score was 46.8 ± 20.9 and rose to 85.24 ± 13.9 after 24 months. Mean preoperative Merle D’Aubigne-Postel score was 8.75 ± 2.25 rising to 16.1 ± 2.59 at 24 months’ follow-up. Conclusion To our knowledge, this is the first report of fresh osteochondral allograft transplantation in South America. We believe that this experience may be of value for physicians in countries that are trying to establish an osteochondral allograft transplant program. PMID:27375837

  3. Development of a Fresh Osteochondral Allograft Program Outside North America.

    PubMed

    Tírico, Luís Eduardo Passarelli; Demange, Marco Kawamura; Santos, Luiz Augusto Ubirajara; de Rezende, Márcia Uchoa; Helito, Camilo Partezani; Gobbi, Riccardo Gomes; Pécora, José Ricardo; Croci, Alberto Tesconi; Bugbee, William Dick

    2016-07-01

    To standardize and to develop a fresh osteochondral allograft protocol of procurement, processing and surgical utilization in Brazil. This study describes the steps recommended to make fresh osteochondral allografts a viable treatment option in a country without previous fresh allograft availability. The process involves regulatory process modification, developing and establishing procurement, and processing and surgical protocols. Legislation: Fresh osteochondral allografts were not feasible in Brazil until 2009 because the law prohibited preservation of fresh grafts at tissue banks. We approved an amendment that made it legal to preserve fresh grafts for 30 days from 2°C to 6°C in tissue banks. Procurement: We changed the protocol of procurement to decrease tissue contamination. All tissues were procured in an operating room. Processing: Processing of the grafts took place within 12 hours of tissue recovery. A serum-free culture media with antibiotics was developed to store the grafts. Surgeries: We have performed 8 fresh osteochondral allografts on 8 knees obtaining grafts from 5 donors. Mean preoperative International Knee Documentation Committee (IKDC) score was 31.99 ± 13.4, improving to 81.26 ± 14.7 at an average of 24 months' follow-up. Preoperative Knee Injury and Oseoarthritis Outcome Score (KOOS) score was 46.8 ± 20.9 and rose to 85.24 ± 13.9 after 24 months. Mean preoperative Merle D'Aubigne-Postel score was 8.75 ± 2.25 rising to 16.1 ± 2.59 at 24 months' follow-up. To our knowledge, this is the first report of fresh osteochondral allograft transplantation in South America. We believe that this experience may be of value for physicians in countries that are trying to establish an osteochondral allograft transplant program.

  4. Non-destructive monitoring of viability in an ex vivo organ culture model of osteochondral tissue.

    PubMed

    Elson, K M; Fox, N; Tipper, J L; Kirkham, J; Hall, R M; Fisher, J; Ingham, E

    2015-06-30

    Organ culture is an increasingly important tool in research, with advantages over monolayer cell culture due to the inherent natural environment of tissues. Successful organ cultures must retain cell viability. The aim of this study was to produce viable and non-viable osteochondral organ cultures, to assess the accumulation of soluble markers in the conditioned medium for predicting tissue viability. Porcine femoral osteochondral plugs were cultured for 20 days, with the addition of Triton X-100 on day 6 (to induce necrosis), camptothecin (to induce apoptosis) or no toxic additives. Tissue viability was assessed by the tissue destructive XTT (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxyanilide tetrazolium salt) assay method and LIVE/DEAD® staining of the cartilage at days 0, 6 and 20. Tissue structure was assessed by histological evaluation using haematoxylin & eosin and safranin O. Conditioned medium was assessed every 3-4 days for glucose depletion, and levels of lactate dehydrogenase (LDH), alkaline phosphatase (AP), glycosaminoglycans (GAGs), and matrix metalloproteinase (MMP)-2 and MMP-9. Necrotic cultures immediately showed a reduction in glucose consumption, and an immediate increase in LDH, GAG, MMP-2 and MMP-9 levels. Apoptotic cultures showed a delayed reduction in glucose consumption and delayed increase in LDH, a small rise in MMP-2 and MMP-9, but no significant effect on GAGs released into the conditioned medium. The data showed that tissue viability could be monitored by assessing the conditioned medium for the aforementioned markers, negating the need for tissue destructive assays. Physiologically relevant whole- or part-joint organ culture models, necessary for research and pre-clinical assessment of therapies, could be monitored this way, reducing the need to sacrifice tissues to determine viability, and hence reducing the sample numbers necessary.

  5. Evaluation of cartilage, synovium and adipose tissue as cellular sources for osteochondral repair.

    PubMed

    Innes, J F; Gordon, C; Vaughan-Thomas, A; Rhodes, N P; Clegg, P D

    2013-09-01

    Osteochondral lesions are a major cause of pain and disability in several species including dogs, horses and human beings. The objective of this study was to assess three potential sources of canine cells for their osteochondral regenerative potential. Cartilage, synovium and adipose tissue cells were grown in pellet culture in chondrogenic or osteogenic media. Cartilage-derived pellets displayed the best chondrogenic differentiation as indicated by significantly higher COL2A1 and SOX9 mRNA expression, greater glycosaminoglycan content, and higher retention of Safranin-O stain compared to the synovium and adipose-derived cells. Following application of the osteogenic media, all three cell sources exhibited small areas of positive alizarin red staining. Poor intracellular alkaline phosphatase activity was found in all three cell types when stimulated although osteocalcin and RUNX2 expression were significantly increased. Cells isolated and cultured from canine articular cartilage retained their specific chondrocytic phenotype. Furthermore, canine adipocytes and synovial cells did not undergo chondrogenic differentiation and did not exhibit evidence of multipotency. Although osteogenic differentiation was initiated at a genomic level, phenotypic osteoblastic differentiation was not observed. The findings of this study suggest that cells isolated from canine adipose tissue and synovium are sub-optimal substitutes for chondrocytes when engineering articular cartilage in vitro.

  6. Changes in condylar coefficient of friction after osteochondral graft transplantation and modulation with hyaluronan.

    PubMed

    Lane, John; Healey, Robert; Amiel, David

    2009-12-01

    To better understand the changes in the cartilage coefficient of friction (COF) after an osteochondral repair, an assessment of dynamic loads has been developed using a goat knee model. The application of hyaluronan (HA) was also assessed for its lubricative properties and the resulting COF of the knee after osteochondral repair. A total of 18 caprine knees were dissected and mounted into an Instron load frame (Instron, Norwood, MA) for testing. The COF was measured in 10 knees relative to the normal, unaltered joint and then calibrated to account for friction of the system. These experimental knees were tested in 5 modes: normal; empty 4.5-mm defect; and osteochondral repairs that were elevated, flush, or depressed relative to the cartilage surface. Saline solution lavage kept the knees moist during testing. The effect of HA was evaluated after mechanical testing. Eight knees were used to study the effect of lavage on the joints because of the significant increase in the COF that it produced. Whereas all modes increased the COF from normal levels, the most significant changes occurred when there was proud placement. Increases of 4 times the normal friction levels were measured. Increases in the COF were also associated with saline solution lavage (0.006 to 0.046). There was a significant reduction in friction after HA injection, which reduced the COF to near-normal levels. There is a significant increase in the COF associated with saline solution lavage and an osteochondral plug being left proud, which can be temporarily reduced with a lubricative material such as HA. Dramatic increases in the COF can potentially damage chondrocytes when the patient begins articulating the joint after surgery. Such injuries may affect the ability of the cartilage to heal fully. Reducing the elevated COF with lubricating materials, such as HA, is recommended based on the results of this study.

  7. Effect of lateral meniscectomy and osteochondral grafting of a lateral femoral condylar defect on contact mechanics: a cadaveric study in dogs

    PubMed Central

    2013-01-01

    Background Osteochondral autograft transfer (OAT) aims at restoring normal articular cartilage surface geometry and articular contact mechanics. To date, no studies have evaluated the contact mechanics of the canine stifle following OAT. Additionally, there are no studies that evaluated the role of the meniscus in contact mechanics following OAT in human or canine femorotibial joints. The objective of this study was to measure the changes in femorotibial contact areas (CA), mean contact pressure (MCP) and peak contact pressure (PCP) before and after osteochondral autograft transplantation (OAT) of a simulated lateral femoral condylar cartilage defect with an intact lateral meniscus and following lateral meniscectomy. Results With an intact lateral meniscus, creation of an osteochondral defect caused a decrease in MCP and PCP by 11% and 30%, respectively, compared to the intact stifle (p < 0.01). With an intact meniscus, implanting an osteochondral graft restored MCP and PCP to 96% (p = 0.56) and 92% (p = 0.41) of the control values. Lateral meniscectomy with grafting decreased CA by 54% and increased PCP by 79% compared to the intact stifle (p < 0.01). Conclusions OAT restored contact pressures in stifles with a simulated lateral condylar defect when the meniscus was intact. The lateral meniscus has a significant role in maintaining normal contact pressures in both stifles with a defect or following OAT. Meniscectomy should be avoided when a femoral condylar defect is present and when performing OAT. PMID:23522348

  8. The maturity of tissue-engineered cartilage in vitro affects the repairability for osteochondral defect.

    PubMed

    Jin, Cheng Zhe; Cho, Jae-Ho; Choi, Byung Hyune; Wang, Li Ming; Kim, Moon Suk; Park, So Ra; Yoon, Jeong Ho; Yun, Jung Ho; Oh, Hyun Ju; Min, Byoung-Hyun

    2011-12-01

    Cartilage tissue engineering using cells and biocompatible scaffolds has emerged as a promising approach to repair of cartilage damage. To date, however, no engineered cartilage has proven to be equivalent to native cartilage in terms of biochemical and compression properties, as well as histological features. An alternative strategy for cartilage engineering is to focus on the in vivo regeneration potential of immature engineered cartilage. Here, we used a rabbit model to evaluate the extent to which the maturity of engineered cartilage influenced the remodeling and integration of implanted extracellular matrix scaffolds containing allogenous chondrocytes. Full-thickness osteochondral defects were created in the trochlear groove of New Zealand white rabbits. Left knee defects were left untreated as a control (group 1), and right knee defects were implanted with tissue-engineered cartilage cultured in vitro for 2 days (group 2), 2 weeks (group 3), or 4 weeks (group 4). Histological, chemical, and compression assays of engineered cartilage in vitro showed that biochemical composition became more cartilagenous, and biomechanical property for compression gradually increased with culture time. In an in vivo study, gross imaging and histological observation at 1 and 3 months after implanting in vitro-cultured engineered cartilage showed that defects in groups 3 and 4 were repaired with hyaline cartilage-like tissue, whereas defects were only partially filled with fibrocartilage after 1 month in groups 1 and 2. At 3 months, group 4 showed striking features of hyaline cartilage tissue, with a mature matrix and a columnar arrangement of chondrocytes. Zonal distribution of type II collagen was most prominent, and the International Cartilage Repair Society score was also highest at this time. In addition, the subchondral bone was well ossified. In conclusion, in vivo engineered cartilage was remodeled when implanted; however, its extent to maturity varied with cultivation

  9. Growth Factor Gradients via Microsphere Delivery in Biopolymer Scaffolds for Osteochondral Tissue Engineering

    PubMed Central

    Wang, Xiaoqin; Wenk, Esther; Zhang, Xiaohui; Meinel, Lorenz; Vunjak-Novakovic, Gordana; Kaplan, David L.

    2009-01-01

    Temporally and spatially controlled delivery of growth factors in polymeric scaffolds is crucial for engineering composite tissue structures, such as osteochondral constructs. In the present study, microsphere-mediated growth factor delivery in polymer scaffolds and its impact on osteochondral differentiation of human bone marrow-derived mesenchymal stem cells (hMSCs) was evaluated. Two growth factors, bone morphogenetic protein 2 (rhBMP-2) and insulin-like growth factor I (rhIGF-I), were incorporated as a single concentration gradient or reverse gradient combining two factors in the scaffolds. To assess the gradient making system and the delivery efficiency of polylactic-co-glycolic acid (PLGA) and silk fibroin microspheres, initially an alginate gel was fabricated into a cylinder shape with microspheres incorporated as gradients. Compared to PLGA microspheres, silk microspheres were more efficient in delivering rhBMP-2, probably due to sustained release of the growth factor, while less efficient in delivering rhIGF-I, likely due to loading efficiency. The growth factor gradients formed were shallow, inducing non-gradient trends in hMSC osteochondral differentiation. Aqueous-derived silk porous scaffolds were used to incorporate silk microspheres using the same gradient process. Both growth factors formed deep and linear concentration gradients in the scaffold, as shown by enzyme-linked immunosorbent assay (ELISA). After seeding with hMSCs and culturing for 5 weeks in a medium containing osteogenic and chondrogenic components, hMSCs exhibited osteogenic and chondrogenic differentiation along the concentration gradients of rhBMP-2 in the single gradient of rhBMP-2 and reverse gradient of rhBMP-2/rhIGF-I, but not the rhIGF-I gradient system, confirming that silk microspheres were more efficient in delivering rhBMP-2 than rhIGF-I for hMSCs osteochondrogenesis. This novel silk microsphere/scaffold system offers a new option for the delivery of multiple growth factors

  10. The contribution of bone and cartilage to the near-infrared spectrum of osteochondral tissue

    PubMed Central

    McGoverin, Cushla M.; Lewis, Karl; Yang, Xu; Bostrom, Mathias P. G.; Pleshko, Nancy

    2014-01-01

    Near-infrared (NIR) spectroscopy has been utilized to assess hyaline cartilage quality in human and animal osteochondral tissues. However, due to the lack of NIR signal from bone phosphate, and the relatively deep penetration depth of the radiation, the separate contributions of cartilage and bone to the spectral signatures have not been well defined. The objectives of the current study were 1) to improve the understanding of the contributions of bone and cartilage to NIR spectra acquired from osteochondral tissue, and 2) to assess the ability of this non-destructive method to predict cartilage thickness and modified Mankin grade of human tibial plateau articular cartilage. NIR spectra were acquired from samples of bovine bone and cartilage with varying thicknesses, and from twenty-two tibial plateaus harvested from patients undergoing knee replacement surgery. Spectra were recorded from regions of the tibial plateaus with varying degrees of degradation, and the cartilage thickness and modified Mankin grade of these regions were assessed histologically. Spectra from bone and cartilage samples of known thicknesses were investigated to identify spectral regions that were distinct for these two tissues. Univariate and multivariate linear regression methods were used to correlate modified Mankin grade and cartilage thickness with NIR spectral changes. The ratio of the NIR absorbances associated with water at 5270 and 7085 cm−1 were the best differentiator of cartilage and bone spectra. NIR prediction models for thickness and Mankin grade calculated using partial least squares regression were more accurate than univariate-based prediction models, with root mean square errors of cross validation of 0.42 mm (thickness) and 1.3 (modified Mankin grade), respectively. We conclude that NIR spectroscopy may be used to simultaneously assess articular cartilage thickness and modified Mankin grade, based in part on differences in spectral contributions from bone and cartilage

  11. Osteochondral Tissue Engineering In Vivo: A Comparative Study Using Layered Silk Fibroin Scaffolds from Mulberry and Nonmulberry Silkworms

    PubMed Central

    Saha, Sushmita; Kundu, Banani; Kirkham, Jennifer; Wood, David; Kundu, Subhas C.; Yang, Xuebin B.

    2013-01-01

    The ability to treat osteochondral defects is a major clinical need. Existing polymer systems cannot address the simultaneous requirements of regenerating bone and cartilage tissues together. The challenge still lies on how to improve the integration of newly formed tissue with the surrounding tissues and the cartilage-bone interface. This study investigated the potential use of different silk fibroin scaffolds: mulberry (Bombyx mori) and non-mulberry (Antheraea mylitta) for osteochondral regeneration in vitro and in vivo. After 4 to 8 weeks of in vitro culture in chondro- or osteo-inductive media, non-mulberry constructs pre-seeded with human bone marrow stromal cells exhibited prominent areas of the neo tissue containing chondrocyte-like cells, whereas mulberry constructs pre-seeded with human bone marrow stromal cells formed bone-like nodules. In vivo investigation demonstrated neo-osteochondral tissue formed on cell-free multi-layer silk scaffolds absorbed with transforming growth factor beta 3 or recombinant human bone morphogenetic protein-2. Good bio-integration was observed between native and neo-tissue within the osteochondrol defect in patellar grooves of Wistar rats. The in vivo neo-matrix formed comprised of a mixture of collagen and glycosaminoglycans except in mulberry silk without growth factors, where a predominantly collagenous matrix was observed. Immunohistochemical assay showed stronger staining of type I and type II collagen in the constructs of mulberry and non-mulberry scaffolds with growth factors. The study opens up a new avenue of using inter-species silk fibroin blended or multi-layered scaffolds of a combination of mulberry and non-mulberry origin for the regeneration of osteochondral defects. PMID:24260335

  12. A new bi-layered scaffold for osteochondral tissue regeneration: In vitro and in vivo preclinical investigations.

    PubMed

    Sartori, M; Pagani, S; Ferrari, A; Costa, V; Carina, V; Figallo, E; Maltarello, M C; Martini, L; Fini, M; Giavaresi, G

    2017-01-01

    Current treatments for acute or degenerative chondral and osteochondral lesions are in need of improvement, as these types of injuries lead to disability and worsen the quality of life in a high percentage of patients. The aim of this study was to develop a new bi-layered scaffold for osteochondral tissue regeneration through a "biomimetic" and "bioinspired" approach. For chondral regeneration, the scaffold was realized with an organic compound (type I collagen), while for the regeneration of the subchondral layer, bioactive magnesium-doped hydroxyapatite (Mg/HA) crystals were co-precipitated with the organic component of the scaffold. The entire scaffold structure was stabilized with a cross-linking agent, highly reactive bis-epoxyde (1,4-butanediol diglycidyl ether - BDDGE 1wt%). The developed scaffold was then characterized for its physico-chemical characteristics. Its structure and adhesion strength between the integrated layers were investigated. At the same time, in vitro cell culture studies were carried out to examine the ability of chondral and bone scaffold layers to separately support adhesion, proliferation and differentiation of human mesenchymal stem cells (hMSCs) into chondrocytes and osteoblasts, respectively. Moreover, an in vivo study with nude mice, transplanted with osteochondral scaffolds plain or engineered with undifferentiated hMSCs, was also set up with 4 and 8-week time points. The results showed that chondral and bone scaffold layers represented biocompatible scaffolds able to sustain hMSCs attachment and proliferation. Moreover, the association of scaffold stimuli and differentiation medium, induced hMSCs chondrogenic and osteogenic differentiation and deposition of extracellular matrix (ECM). The ectopic implantation of the engineered osteochondral scaffolds indicated that hMSCs were able to colonize the osteochondral scaffold in depth. The scaffold appeared permissive to tissue growth and penetration, ensuring the diffusion of

  13. Novel nanostructured scaffold for osteochondral regeneration: pilot study in horses.

    PubMed

    Kon, E; Mutini, A; Arcangeli, E; Delcogliano, M; Filardo, G; Nicoli Aldini, N; Pressato, D; Quarto, R; Zaffagnini, S; Marcacci, M

    2010-06-01

    The present in vivo preliminary experiment is aimed at testing mechanical and biological behaviour of a new nano-structured composite multilayer biomimetic scaffold for the treatment of chondral and osteochondral defects. The three-dimensional biomimetic scaffold (Fin-Ceramica Faenza S.p.A., Faenza-Italy) was obtained by nucleating collagen fibrils with hydroxyapatite nanoparticles, in two configurations, bi- and tri-layered, to reproduce, respectively, chondral and osteochondral anatomy. Chondral defects (lateral condyle) and deep osteochondral defects (medial condyle) were made in the distal epiphysis of the third metacarpal bone of both forelimbs of two adult horses and treated respectively with the chondral and osteochondral grafts. Both animals were euthanised six months follow up. The images obtained at the second look arthroscopy evaluation, performed two months after surgery, demonstrated good filling of the chondral and osteo-chondral defects without any inflammatory reaction around and inside the lesions. At the histological analysis the growth of trabecular bone in the osteochondral lesion was evident. Only in one case, the whole thickness of the osteochondral lesion was filled by fibrocartilaginous tissue. The formation of a tidemark line was evident at the interface with the newly formed bone. Newly formed fibrocartilaginous tissue was present in the area of the chondral defect. Initial alignment of the collagen fibres was recognisable with polarised light in both groups. The results of the present pilot study showed that this novel osteochondral and chondral scaffold may act as a suitable matrix to facilitate orderly regeneration of bone and hyaline-like cartilage. Copyright 2010 John Wiley & Sons, Ltd.

  14. Combining Microfractures, Autologous Bone Graft, and Autologous Matrix-Induced Chondrogenesis for the Treatment of Juvenile Osteochondral Talar Lesions.

    PubMed

    D'Ambrosi, Riccardo; Maccario, Camilla; Ursino, Chiara; Serra, Nicola; Usuelli, Federico Giuseppe

    2017-05-01

    The purpose of this study was to evaluate the clinical and radiologic outcomes of patients younger than 20 years, treated with the arthroscopic-talus autologous matrix-induced chondrogenesis (AT-AMIC) technique and autologous bone graft for osteochondral lesion of the talus (OLT). Eleven patients under 20 years (range 13.3-20.0) underwent the AT-AMIC procedure and autologous bone graft for OLTs. Patients were evaluated preoperatively (T0) and at 6 (T1), 12 (T2), and 24 (T3) months postoperatively, using the American Orthopaedic Foot & Ankle Society Ankle and Hindfoot (AOFAS) score, the visual analog scale and the SF-12 respectively in its Mental and Physical Component Scores. Radiologic assessment included computed tomographic (CT) scan, magnetic resonance imaging (MRI) and intraoperative measurement of the lesion. A multivariate statistical analysis was performed. Mean lesion size measured during surgery was 1.1 cm(3) ± 0.5 cm(3). We found a significant difference in clinical and radiologic parameters with analysis of variance for repeated measures ( P < .001). All clinical scores significantly improved ( P < .05) from T0 to T3. Lesion area significantly reduced from 119.1 ± 29.1 mm(2) preoperatively to 77.9 ± 15.8 mm(2) ( P < .05) at final follow-up as assessed by CT, and from 132.2 ± 31.3 mm(2) to 85.3 ± 14.5 mm(2) ( P < .05) as assessed by MRI. Moreover, we noted an important correlation between intraoperative size of the lesion and body mass index (BMI) ( P = .011). The technique can be considered safe and effective with early good results in young patients. Moreover, we demonstrated a significant correlation between BMI and lesion size and a significant impact of OLTs on quality of life. Level IV, retrospective case series.

  15. Osteochondral repair using a scaffold-free tissue-engineered construct derived from synovial mesenchymal stem cells and a hydroxyapatite-based artificial bone.

    PubMed

    Shimomura, Kazunori; Moriguchi, Yu; Ando, Wataru; Nansai, Ryosuke; Fujie, Hiromichi; Hart, David A; Gobbi, Alberto; Kita, Keisuke; Horibe, Shuji; Shino, Konsei; Yoshikawa, Hideki; Nakamura, Norimasa

    2014-09-01

    For an ideal osteochondral repair, it is important to facilitate zonal restoration of the subchondral bone and the cartilage, layer by layer. Specifically, restoration of the osteochondral junction and secure integration with adjacent cartilage could be considered key factors. The purpose of the present study was to investigate the feasibility of a combined material comprising a scaffold-free tissue-engineered construct (TEC) derived from synovial mesenchymal stem cells (MSCs) and a hydroxyapatite (HA) artificial bone using a rabbit osteochondral defect model. Osteochondral defects were created on the femoral groove of skeletally mature rabbits. The TEC and HA artificial bone were hybridized to develop a combined implant just before use, which was then implanted into defects (N=23). In the control group, HA alone was implanted (N=18). Histological evaluation and micro-indentation testing was performed for the evaluation of repair tissue. Normal knees were used as an additional control group for biomechanical testing (N=5). At hybridization, the TEC rapidly attached onto the surface of HA artificial bone block, which was implantable to osteochondral defects. Osteochondral defects treated with the combined implants exhibited more rapid subchondral bone repair coupled with the development of cartilaginous tissue with good tissue integration to the adjacent host cartilage when assessed at 6 months post implantation. Conversely, the control group exhibited delayed subchondral bone repair. In addition, the repair cartilaginous tissue in this group had poor integration to adjacent cartilage and contained clustered chondrocytes, suggesting an early osteoarthritis (OA)-like degenerative change at 6 months post implantation. Biomechanically, the osteochondral repair tissue treated with the combined implants at 6 months restored tissue stiffness, similar to normal osteochondral tissue. The combined implants significantly accelerated and improved osteochondral repair

  16. Tissue engineering of small caliber vascular grafts.

    PubMed

    Hoerstrup, S P; Zünd, G; Sodian, R; Schnell, A M; Grünenfelder, J; Turina, M I

    2001-07-01

    Previous tissue engineering approaches to create small caliber vascular grafts have been limited by the structural and mechanical immaturity of the constructs. This study uses a novel in vitro pulse duplicator system providing a 'biomimetic' environment during tissue formation to yield more mature, implantable vascular grafts. Vascular grafts (I.D. 0.5 cm) were fabricated from novel bioabsorbable polymers (polyglycolic-acid/poly-4-hydroxybutyrate) and sequentially seeded with ovine vascular myofibroblasts and endothelial cells. After 4 days static culture, the grafts (n=24) were grown in vitro in a pulse duplicator system (bioreactor) for 4, 7, 14, 21, and 28 days. Controls (n=24) were grown in static culture conditions. Analysis of the neo-tissue included histology, scanning electron microscopy (SEM), and biochemical assays (DNA for cell content, 5-hydroxyproline for collagen). Mechanical testing was performed measuring the burst pressure and the suture retention strength. Histology showed viable, dense tissue in all samples. SEM demonstrated confluent smooth inner surfaces of the grafts exposed to pulsatile flow after 14 days. Biochemical analysis revealed a continuous increase of cell mass and collagen to 21 days compared to significantly lower values in the static controls. The mechanical properties of the pulsed vascular grafts comprised supra-physiological burst strength and suture retention strength appropriate for surgical implantation. This study demonstrates the feasibility of tissue engineering of viable, surgically implantable small caliber vascular grafts and the important effect of a 'biomimetic' in vitro environment on tissue maturation and extracellular matrix formation.

  17. Plant grafting: insights into tissue regeneration

    PubMed Central

    2016-01-01

    Abstract For millennia, people have cut and joined different plants together through a process known as grafting. The severed tissues adhere, the cells divide and the vasculature differentiates through a remarkable process of regeneration between two genetically distinct organisms as they become one. Grafting is becoming increasingly important in horticulture where it provides an efficient means for asexual propagation. Grafting also combines desirable roots and shoots to generate chimeras that are more vigorous, more pathogen resistant and more abiotic stress resistant. Thus, it presents an elegant and efficient way to improve plant productivity in vegetables and trees using traditional techniques. Despite this horticultural importance, we are only beginning to understand how plants regenerate tissues at the graft junction. By understanding grafting better, we can shed light on fundamental regeneration pathways and the basis for self/non‐self recognition. We can also better understand why many plants efficiently graft whereas others cannot, with the goal of improving grafting so as to broaden the range of grafted plants to create even more desirable chimeras. Here, I review the latest findings describing how plants graft and provide insight into future directions in this emerging field. PMID:28316790

  18. Plant grafting: insights into tissue regeneration.

    PubMed

    Melnyk, Charles W

    2017-02-01

    For millennia, people have cut and joined different plants together through a process known as grafting. The severed tissues adhere, the cells divide and the vasculature differentiates through a remarkable process of regeneration between two genetically distinct organisms as they become one. Grafting is becoming increasingly important in horticulture where it provides an efficient means for asexual propagation. Grafting also combines desirable roots and shoots to generate chimeras that are more vigorous, more pathogen resistant and more abiotic stress resistant. Thus, it presents an elegant and efficient way to improve plant productivity in vegetables and trees using traditional techniques. Despite this horticultural importance, we are only beginning to understand how plants regenerate tissues at the graft junction. By understanding grafting better, we can shed light on fundamental regeneration pathways and the basis for self/non-self recognition. We can also better understand why many plants efficiently graft whereas others cannot, with the goal of improving grafting so as to broaden the range of grafted plants to create even more desirable chimeras. Here, I review the latest findings describing how plants graft and provide insight into future directions in this emerging field.

  19. Hierarchical Structure of Articular Bone-Cartilage Interface and Its Potential Application for Osteochondral Tissue Engineering

    NASA Astrophysics Data System (ADS)

    Bian, Weiguo; Qin, Lian; Li, Dichen; Wang, Jin; Jin, Zhongmin

    2010-09-01

    The artificial biodegradable osteochondral construct is one of mostly promising lifetime substitute in the joint replacement. And the complex hierarchical structure of natural joint is important in developing the osteochondral construct. However, the architecture features of the interface between cartilage and bone, in particular those at the micro-and nano-structural level, remain poorly understood. This paper investigates these structural data of the cartilage-bone interface by micro computerized tomography (μCT) and Scanning Electron Microscope (SEM). The result of μCT shows that important bone parameters and the density of articular cartilage are all related to the position in the hierarchical structure. The conjunctions of bone and cartilage were defined by SEM. All of the study results would be useful for the design of osteochondral construct further manufactured by nano-tech. A three-dimensional model with gradient porous structure is constructed in the environment of Pro/ENGINEERING software.

  20. Bioprinting of a mechanically enhanced three-dimensional dual cell-laden construct for osteochondral tissue engineering using a multi-head tissue/organ building system

    NASA Astrophysics Data System (ADS)

    Shim, Jin-Hyung; Lee, Jung-Seob; Kim, Jong Young; Cho, Dong-Woo

    2012-08-01

    The aim of this study was to build a mechanically enhanced three-dimensional (3D) bioprinted construct containing two different cell types for osteochondral tissue regeneration. Recently, the production of 3D cell-laden structures using various scaffold-free cell printing technologies has opened up new possibilities. However, ideal 3D complex tissues or organs have not yet been printed because gel-state hydrogels have been used as the principal material and are unable to maintain the desired 3D structure due to their poor mechanical strength. In this study, thermoplastic biomaterial polycaprolactone (PCL), which shows relatively high mechanical properties as compared with hydrogel, was used as a framework for enhancing the mechanical stability of the bioprinted construct. Two different alginate solutions were then infused into the previously prepared framework consisting of PCL to create the 3D construct for osteochondral printing. For this work, a multi-head tissue/organ building system (MtoBS), which was particularly designed to dispense thermoplastic biomaterial and hydrogel having completely different rheology properties, was newly developed and used to bioprint osteochondral tissue. It was confirmed that the line width, position and volume control of PCL and alginate solutions were adjustable in the MtoBS. Most importantly, dual cell-laden 3D constructs consisting of osteoblasts and chondrocytes were successfully fabricated. Further, the separately dispensed osteoblasts and chondrocytes not only retained their initial position and viability, but also proliferated up to 7 days after being dispensed.

  1. Simultaneous Osteoperiosteal Autologous Iliac Crest Graft and Lateral Meniscus Allograft Transplantation for Osteochondral Lesion with Bony Defect and Lateral Discoid Meniscus Tear

    PubMed Central

    Lee, Dhong Won; Ha, Jeong Ku; Kim, Woo Jong

    2016-01-01

    The optimal treatment for combined osteochondritis dissecans (OCD) with considerable bony defect of the lateral femoral condyle (LFC) and torn discoid lateral meniscus is unclear. We present a case of a 15-year-old female who was a gymnast and had a large OCD lesion in the LFC combined with deficiency of the lateral meniscus. The patient underwent the "one-step" technique of osteoperiosteal autologous iliac crest graft and lateral meniscus allograft transplantation after a failure of meniscectomy with repair at another hospital. Twenty-four months postoperatively, clinical results were significantly improved. Follow-up imaging tests and second-look arthroscopy showed well incorporated structured bone graft and fibrous cartilage regeneration as well as stabilized lateral meniscus allograft. She could return to her sport without any pain or swelling. This "one-step" surgical technique is worth considering as a joint salvage procedure for massive OCD lesions with torn discoid lateral meniscus. PMID:27274475

  2. Sol gel-derived hydroxyapatite films over porous calcium polyphosphate substrates for improved tissue engineering of osteochondral-like constructs.

    PubMed

    Lee, Whitaik David; Gawri, Rahul; Pilliar, Robert M; Stanford, William L; Kandel, Rita A

    2017-08-14

    Integration of in vitro-formed cartilage on a suitable substrate to form tissue-engineered implants for osteochondral defect repair is a considerable challenge. In healthy cartilage, a zone of calcified cartilage (ZCC) act as an intermediary for mechanical force transfer from soft to hard tissue, as well as an effective interlocking structure to better resist interfacial shear forces. We have developed biphasic constructs that consist of scaffold-free cartilage tissue grown in vitro on, and interdigitated with, porous calcium polyphosphate (CPP) substrates. However, as CPP degrades, it releases inorganic polyphosphates (polyP) that can inhibit local mineralization, thereby preventing the formation of a ZCC at the interface. Thus, we hypothesize that coating CPP substrate with a layer of hydroxyapatite (HA) might prevent or limit this polyP release. To investigate this we tested both inorganic or organic sol-gel processing methods, as a barrier coating on CPP substrate to inhibit polyP release. Both types of coating supported the formation of ZCC in direct contact with the substrate, however the ZCC appeared more continuous in the tissue formed on the organic HA sol gel coated CPP. Tissues formed on coated substrates accumulated comparable quantities of extracellular matrix and mineral, but tissues formed on organic sol-gel (OSG)-coated substrates accumulated less polyP than tissues formed on inorganic sol-gel (ISG)-coated substrates. Constructs formed with OSG-coated CPP substrates had greater interfacial shear strength than those formed with ISG-coated and non-coated substrates. These results suggest that the OSG coating method can modify the location and distribution of ZCC and can be used to improve the mechanical integrity of tissue-engineered constructs formed on porous CPP substrates. Articular cartilage interfaces with bone through a zone of calcified cartilage. This study describes a method to generate an "osteochondral-like" implant that mimics this

  3. Osteochondritis Dissecans

    MedlinePlus

    ... symptom of osteochondritis dissecans might be triggered by physical activity — walking up stairs, climbing a hill or playing sports. Swelling and tenderness. The skin around your joint might be swollen and tender. ...

  4. Humeral Head Reconstruction With Osteochondral Allograft Transplantation.

    PubMed

    Saltzman, Bryan M; Riboh, Jonathan C; Cole, Brian J; Yanke, Adam B

    2015-09-01

    To synthesize, in a systematic review, the available clinical evidence of osteochondral allograft transplants for large osteochondral defects of the humeral head. The Medline, Embase, and Cochrane databases were searched for studies reporting clinical or radiographic outcomes of osteochondral allograft transplantation for humeral head defects. Descriptive statistics were provided for all outcomes. After checking for data normality, we compared postoperative and preoperative values using the Student t test. We included 12 studies (8 case reports and 4 case series) in this review. The study group consisted of 35 patients. The mean age was 35.4 ± 18.1 years; 77% of patients were male patients. Thirty-three patients had large Hill-Sachs lesions due to instability, 1 had an osteochondritis dissecans lesion, and 1 had an iatrogenic lesion after resection of synovial chondromatosis. The mean lesion size was 3 ± 1.4 cm (anteroposterior) by 2.25 ± 0.3 cm (medial-lateral), representing on average 40.5% ± 4.73% of the native articular surface. Of the 35 patients, 3 received a fresh graft, with all others receiving frozen grafts. Twenty-three femoral heads, 10 humeral heads, and 2 sets of osteochondral plugs were used. The mean length of follow-up was 57 months. Significant improvements were seen in forward flexion at 6 months (68° ± 18.1°, P < .001), forward flexion at 12 months (83.42° ± 18.3°, P < .001), and external rotation at 12 months (38.72° ± 18.8°, P < .001). American Shoulder and Elbow Surgeons scores improved by 14 points (P = .02). Radiographic studies at final follow-up showed allograft necrosis in 8.7% of cases, resorption in 36.2%, and glenohumeral arthritic changes in 35.7%. Complication rates were between 20% and 30%, and the reoperation rate was 26.67%. Although only 3 patients received fresh allografts, there were no reports of graft resorption, necrosis, or arthritic changes in these patients. Humeral head allograft-most commonly used in the

  5. Dynamic regulation of bone morphogenetic proteins in engineered osteochondral constructs by biomechanical stimulation.

    PubMed

    Nam, Jin; Perera, Priyangi; Rath, Bjoern; Agarwal, Sudha

    2013-03-01

    Osteochondral tissue-engineered grafts are proposed to hold greater potential to repair/regenerate damaged cartilage through enhanced biochemical and mechanical interactions with underlying subchondral bone as compared to simple engineered cartilage. Additionally, biomechanical stimulation of articular chondrocytes (ACs) or osteoblasts (OBs) was shown to induce greater morphogenesis of the engineered tissues composed of these cells. In this report, to define the advantages of biomechanical stimulation to osteochondral grafts for tissue engineering, we examined whether (1) ACs and OBs in three-dimensional (3D) osteochondral constructs support functional development of each other at the molecular level, and (2) biomechanical stimulation of osteochondral constructs further promotes the regenerative potential of such grafts. Various configurations of cell/scaffold assemblies, including chondral, osseous, and osteochondral constructs, were engineered with mechano-responsive electrospun poly(ɛ-caprolactone) scaffolds. These constructs were subjected to either static or dynamic (10% cyclic compressive strain at 1 Hz for 3 h/day) culture conditions for 2 weeks. The expression of bone morphogenetic proteins (BMPs) was examined to assess the regenerative potential of each treatment on the cells. Biomechanical stimulation augmented a marked upregulation of Bmp2, Bmp6, and Bmp7 as well as downregulation of BMP antagonist, Bmp3, in a time-specific manner in the ACs and OBs of 3D osteochondral constructs. More importantly, the presence of biomechanically stimulated OBs was especially crucial for the induction of Bmp6 in ACs, a BMP required for chondrocytic growth and differentiation. Biomechanical stimulation led to enhanced tissue morphogenesis possibly through this BMP regulation, evident by the improved effective compressive modulus of the osteochondral constructs (710 kPa of dynamic culture vs. 280 kPa of static culture). Similar BMP regulation was observed in the

  6. Microporous calcium phosphate ceramics as tissue engineering scaffolds for the repair of osteochondral defects: biomechanical results.

    PubMed

    Mayr, H O; Klehm, J; Schwan, S; Hube, R; Südkamp, N P; Niemeyer, P; Salzmann, G; von Eisenhardt-Rothe, R; Heilmann, A; Bohner, M; Bernstein, A

    2013-01-01

    This work investigated the suitability of microporous β-tricalcium phosphate (TCP) scaffolds pre-seeded with autologous chondrocytes for treatment of osteochondral defects in a large animal model. Microporous β-TCP cylinders (Ø 7 mm; length 25 mm) were seeded with autologous chondrocytes and cultured for 4 weeks in vitro. Only the upper end of the cylinder was seeded with chondrocytes. Chondrocytes formed a multilayer on the top. The implants were then implanted in defects (diameter 7 mm) created in the left medial femoral condyle of ovine knees. The implants were covered with synovial membrane from the superior recess of the same joint. For the right knees, an empty defect with the same dimensions served as control. Twenty-eight sheep were split into 6-, 12-, 26- and 52 week groups of seven animals. Indentation tests with a spherical (Ø 3mm) indenter were used to determine the biomechanical properties of regenerated tissue. A software-based limit switch was implemented to ensure a maximal penetration depth of 200 μm and maximal load of 1.5 N. The achieved load, the absorbed energy and the contact stiffness were measured. Newly formed cartilage was assessed with the International Cartilage Repair Society Visual Assessment Scale (ICRS score) and histomorphometric analysis. Results were analysed statistically using the t-test, Mann-Whitney U-test and Wilcoxon test. Statistical significance was set at p<0.05. After 6 weeks of implantation, the transplanted area tolerated an indentation load of 0.05±0.20 N. This value increased to 0.10±0.06 N after 12 weeks, to 0.27±0.18 N after 26 weeks, and 0.27±0.11 N after 52 weeks. The increase in the tolerated load was highly significant (p<0.0001), but the final value was not significantly different from that of intact cartilage (0.30±0.12 N). Similarly, the increase in contact stiffness from 0.87±0.29 N mm-(1) after 6 weeks to 3.14±0.86 N mm(-1) after 52 weeks was highly significant (p<0.0001). The absorbed energy

  7. Specific inductive potential of a novel nanocomposite biomimetic biomaterial for osteochondral tissue regeneration.

    PubMed

    Manferdini, C; Cavallo, C; Grigolo, B; Fiorini, M; Nicoletti, A; Gabusi, E; Zini, N; Pressato, D; Facchini, A; Lisignoli, G

    2016-05-01

    Osteochondral lesions require treatment to restore the biology and functionality of the joint. A novel nanostructured biomimetic gradient scaffold was developed to mimic the biochemical and biophysical properties of the different layers of native osteochondral structure. The present results show that the scaffold presents important physicochemical characteristics and can support the growth and differentiation of mesenchymal stromal cells (h-MSCs), which adhere and penetrate into the cartilaginous and bony layers. H-MSCs grown in chondrogenic or osteogenic medium decreased their proliferation during days 14-52 on both scaffold layers and in medium without inducing factors used as controls. Both chondrogenic and osteogenic differentiation of h-MSCs occurred from day 28 and were increased on day 52, but not in the control medium. Safranin O staining and collagen type II and proteoglycans immunostaining confirmed that chondrogenic differentiation was specifically induced only in the cartilaginous layer. Conversely, von Kossa staining, osteocalcin and osteopontin immunostaining confirmed that osteogenic differentiation occurred on both layers. This study shows the specific potential of each layer of the biomimetic scaffold to induce chondrogenic or osteogenic differentiation of h-MSCs. These processes depended mainly on the media used but not the biomaterial itself, suggesting that the local milieu is fundamental for guiding cell differentiation. Copyright © 2013 John Wiley & Sons, Ltd. Copyright © 2013 John Wiley & Sons, Ltd.

  8. Biomaterials/scaffolds. Design of bioactive, multiphasic PCL/collagen type I and type II-PCL-TCP/collagen composite scaffolds for functional tissue engineering of osteochondral repair tissue by using electrospinning and FDM techniques.

    PubMed

    Schumann, Detlef; Ekaputra, Andrew K; Lam, Christopher X F; Hutmacher, Dietmar W

    2007-01-01

    Current clinical therapies for traumatic or chronic injuries involving osteochondral tissue result in temporary pain reduction and filling of the defect but with biomechanically inferior repair tissue. Tissue engineering of osteochondral repair tissue using autologous cells and bioactive biomaterials has the potential to overcome the current limitations and results in native-like repair tissue with good integration capabilities. For this reason, we applied two modem biomaterial design techniques, namely, electrospinning and fused deposition modeling (FDM), to produce bioactive poly(epsilon-caprolactone)/collagen (PCL/Col) type I and type II-PCL-tri-calcium phosphate (TCP)/Col composites for precursor cell-based osteochondral repair. The application of these two design techniques (electrospinning and FDM) allowed us to specifically produce the a suitable three-dimensional (3D) environment for the cells to grow into a particular tissue (cartilage and bone) in vitro prior to in vivo implantation. We hypothesize that our new designed biomaterials, seeded with autologous bone marrow-derived precursor cells, in combination with bioreactor-stimulated cell-culture techniques can be used to produce clinically relevant osteochondral repair tissue.

  9. Soft Tissue Augmentation with Silk Composite Graft

    PubMed Central

    Park, Yong-Tae; Kweon, Hae Yong; Kim, Seong-Gon

    2014-01-01

    Purpose: The objective of this study was to evaluate the interaction between 4-hexylresorcinol (4HR) and antibody as that affects the performance of a silk-4HR combination graft for soft tissue augmentation in an animal model. Methods: The silk graft materials consisted of four types: silk+10% tricalcium phosphate (TCP) (ST0), silk+10% TCP+1% 4HR (ST1), silk+10% TCP+3% 4HR (ST3), and silk+10% TCP+6% 4-HR (ST6). The antibody binding assay tested the 4HR effect and scanning electron microscopic (SEM) exam was done for silk grafts. The animal experiment used a subcutaneous pocket mouse model. The graft – SH0 or SH1 or SH3 or SH6 – was placed in a subcutaneous pocket. The animals were killed at one, two, and four weeks, postoperatively. The specimens were subjected to histological analysis and lysozyme assay. Results: Groups with 4HR applied showed lower antibody binding affinity to antigen compared to groups without 4HR. In the SEM examination, there was no significant difference among groups. Histological examinations revealed many foreign body giant cells in ST0 and ST1 group at four weeks postoperatively. Both ST3 and ST6 groups developed significantly lower levels of giant cell values compared to ST0 and ST1 groups (P <0.001) at four weeks postoperatively. In the lysozyme assay, the ST1 and ST3 groups showed denser signals than the other groups. Conclusion: 4HR combined silk implants resulted in high levels of vascular and connective tissue regeneration. PMID:27489833

  10. UTE MRI of the Osteochondral Junction

    PubMed Central

    Biswas, Reni; Chen, Karen; Chang, Eric Y.; Chung, Christine B.

    2014-01-01

    The osteochondral junction is composed of numerous tissue components and serves important functions relating to structural stability and proper nutrition in joints such as the knee and spine. Conventional MR techniques have been inadequate at imaging the tissues of the osteochondral junction primarily because of the intrinsically short T2 nature of these tissues, rendering them “invisible” with the standard acquisitions. Ultrashort time to echo (UTE) MR techniques acquire sufficient MR signal of osteochondral tissues, thereby allowing direct evaluation. This article reviews the anatomy of the osteochondral junction of the knee and the spine, technical aspects of UTE MRI, and the application of UTE MRI for evaluation of the osteochondral junction. PMID:25061547

  11. A biomimetic multi-layered collagen-based scaffold for osteochondral repair.

    PubMed

    Levingstone, Tanya J; Matsiko, Amos; Dickson, Glenn R; O'Brien, Fergal J; Gleeson, John P

    2014-05-01

    Cartilage and osteochondral defects pose a significant challenge in orthopedics. Tissue engineering has shown promise as a potential method for the treatment of such defects; however, a long-lasting repair strategy has yet to be realized. This study focuses on the development of a layered construct for osteochondral repair, fabricated through a novel "iterative layering" freeze-drying technique. The process involved repeated steps of layer addition followed by freeze-drying, enabling control over material composition, pore size and substrate stiffness in each region of the construct, while also achieving a seamlessly integrated layer structure. The novel construct developed mimics the inherent gradient structure of healthy osteochondral tissue: a bone layer composed of type I collagen and hydroxyapatite (HA), an intermediate layer composed of type I collagen, type II collagen and HA and a cartilaginous region composed of type I collagen, type II collagen and hyaluronic acid. The material properties were designed to provide the biological cues required to encourage infiltration of host cells from the bone marrow while the biomechanical properties were designed to provide an environment optimized to promote differentiation of these cells towards the required lineage in each region. This novel osteochondral graft was shown to have a seamlessly integrated layer structure, high levels of porosity (>97%), a homogeneous pore structure and a high degree of pore interconnectivity. Moreover, homogeneous cellular distribution throughout the entire construct was evident following in vitro culture, demonstrating the potential of this multi-layered scaffold as an advanced strategy for osteochondral defect repair.

  12. Biomimetic biphasic scaffolds for osteochondral defect repair

    PubMed Central

    Li, Xuezhou; Ding, Jianxun; Wang, Jincheng; Zhuang, Xiuli; Chen, Xuesi

    2015-01-01

    The osteochondral defects caused by vigorous trauma or physical disease are difficult to be managed. Tissue engineering provides a possible option to regenerate the damaged osteochondral tissues. For osteochondral reconstruction, one intact scaffold should be considered to support the regeneration of both cartilage and subchondral bone. Therefore, the biphasic scaffolds with the mimic structures of osteochondral tissues have been developed to close this chasm. A variety of biomimetic bilayer scaffolds fabricated from natural or synthetic polymers, or the ones loading with growth factors, cells, or both of them make great progresses in osteochondral defect repair. In this review, the preparation and in vitro and/or in vivo verification of bioinspired biphasic scaffolds are summarized and discussed, as well as the prospect is predicted. PMID:26816644

  13. Studies in fat grafting: Part III. Fat grafting irradiated tissue--improved skin quality and decreased fat graft retention.

    PubMed

    Garza, Rebecca M; Paik, Kevin J; Chung, Michael T; Duscher, Dominik; Gurtner, Geoffrey C; Longaker, Michael T; Wan, Derrick C

    2014-08-01

    Following radiation therapy, skin becomes fibrotic and can present a difficult problem for reconstructive surgeons. There is an increasing belief that fat grafting under irradiated skin can reverse the damage caused by radiation. The present study evaluated the effect of fat grafting on irradiated skin, along with fat graft quality and retention rates in irradiated tissue. Nine adult Crl:NU-Foxn1 CD-1 mice underwent 30-Gy external beam irradiation of the scalp. Four weeks after irradiation, scalp skin from irradiated and nonirradiated mice was harvested and compared histologically for dermal thickness, collagen content, and vascular density. Human fat grafts were then injected in the subcutaneous plane of the scalp. Skin assessment was performed in the irradiated group at 2 and 8 weeks after grafting, and fat graft retention was measured at baseline and every 2 weeks up to 8 weeks after grafting using micro-computed tomography. Finally, fat graft samples were explanted at 8 weeks, and quality scoring was performed. Fat grafting resulted in decreased dermal thickness, decreased collagen content, and increased vascular density in irradiated skin. Computed tomographic analysis revealed significantly decreased fat graft survival in the irradiated group compared with the nonirradiated group. Histologic scoring of explanted fat grafts demonstrated no difference in quality between the irradiated and nonirradiated groups. Fat grafting attenuates dermal collagen deposition and vessel depletion characteristic of radiation fibrosis. Although fat graft retention rates are significantly lower in irradiated than in nonirradiated tissue, the quality of retained fat between the groups is similar.

  14. Osteochondral lesions of the talus: Current concept.

    PubMed

    Laffenêtre, O

    2010-09-01

    Osteochondral lesions of the talus (OTL) are among those injuries that we should not fail to recognize, especially following any type of hindfoot injury. They were thoroughly described 15 years ago in a round table session organized by Doré and Rosset for the Société orthopédique de l'Ouest. Their physiopathology has not yet been definitely determined, even though some of the pathogenic mechanisms are known. They are best characterized using the fractures, osteonecroses, geodes (FOG) radiological classification. Both their diagnosis and their surgical treatment remain a challenge to the orthopaedic surgeon: some basic surgical principles apply to all of the lesions, such as cartilage debridement and shaving of necrotic tissues, while others will be used depending on the location and size of the lesions as well as the surgeon's experience. Finally, no specific technique appears to be superior to the others. Arthroscopy appears to be the most effective procedure for lesions smaller than 1 cm(2), whereas larger lesions should be filled, either with cancellous bone or with an osteochondral graft or using autogenous chondrocyte implantation. The data available in the literature should also incite orthopaedists to consider the results of surgical management with some modesty, and conservative management should remain among the therapeutic options. Copyright © 2010 Elsevier Masson SAS. All rights reserved.

  15. Osteochondral Lesions of the Talar Dome.

    PubMed

    Stone

    1996-03-01

    Osteochondral lesions of the talar dome are relatively common causes of ankle pain and disability. Trauma is the most common cause, but ischemic necrosis, en-docrine disorders, and genetic factors may have etiologic significance. Medial lesions are usually located posteriorly on the dome of the talus, whereas lateral lesions are most frequently located anteriorly. Although the staging system described by Berndt and Harty remains popular, it may not accurately reflect the integrity of the articular cartilage. Small lesions of the talar dome may be present despite a normal appearance on plain radiography. Bone scintigraphy may show increased radionuclide uptake in the talar dome. Magnetic resonance imaging is also sensitive for identifying intraosseous abnormalities in the talus and has the added benefit of revealing other types of soft-tissue lesions not visible on routine radiographic studies. Computed tomography remains the imaging technique of choice when delineation of a bone fragment is desired. Nonoperative management of osteochondral lesions, including restricted weight-bearing and/or immobilization, is recommended unless a loose fragment is clearly present. Surgical options include drilling (usually reserved for intact lesions), debridement of the lesion with curettage or abrasion of the bone bed, internal fixation of the fragment, and bone grafting. Recent technical advances allow these procedures to be performed arthroscopically, with potential reduction of surgical trauma, length of hospital stay, and complication rates.

  16. Platelet-rich plasma increases transforming growth factor-beta1 expression at graft-host interface following autologous osteochondral transplantation in a rabbit model

    PubMed Central

    Boakye, Lorraine A; Ross, Keir A; Pinski, John M; Smyth, Niall A; Haleem, Amgad M; Hannon, Charles P; Fortier, Lisa A; Kennedy, John G

    2015-01-01

    AIM: To explore the effect of platelet-rich plasma on protein expression patterns of transforming growth factor-beta1 (TGF-β1) in cartilage following autologous osteochondral transplantation (AOT) in a rabbit knee cartilage defect model. METHODS: Twelve New Zealand white rabbits received bilateral AOT. In each rabbit, one knee was randomized to receive an autologous platelet rich plasma (PRP) injection and the contralateral knee received saline injection. Rabbits were euthanized at 3, 6 and 12 wk post-operatively. Articular cartilage sections were stained with TGF-β1 antibody. Histological regions of interest (ROI) (left, right and center of the autologous grafts interfaces) were evaluated using MetaMorph. Percentage of chondrocytes positive for TGF-β1 was then assessed. RESULTS: Percentage of chondrocytes positive for TGF-β1 was higher in PRP treated knees for selected ROIs (left; P = 0.03, center; P = 0.05) compared to control and was also higher in the PRP group at each post-operative time point (P = 6.6 × 10-4, 3.1 × 10-4 and 7.3 × 10-3 for 3, 6 and 12 wk, respectively). TGF-β1 expression was higher in chondrocytes of PRP-treated knees (36% ± 29% vs 15% ± 18%) (P = 1.8 × 10-6) overall for each post-operative time point and ROI. CONCLUSION: Articular cartilage of rabbits treated with AOT and PRP exhibit increased TGF-β1 expression compared to those treated with AOT and saline. Our findings suggest that adjunctive PRP may increase TGF-β1 expression, which may play a role in the chondrogenic effect of PRP in vivo. PMID:26716092

  17. Correction of infraorbital and malar deficiency using costal osteochondral graft along with orthognathic surgery in Crouzon syndrome.

    PubMed

    Song, Hyunsuk; Park, Myong Chul; Lee, Il Jae; Park, Dong Ha

    2014-09-01

    In syndromic craniosynostosis, such as Crouzon syndrome, midfacial hypoplasia can cause exophthalmos and concave facial profile. Though midfacial hypoplasia in Crouzon syndrome patients can be treated with midface advancement, known as a Le Fort II or Le Fort III osteotomy, such method can change nasal appearance and frequently fails to achieve class I occlusion after surgery. This report presents a case of an aesthetically and functionally successful midfacial augmentation using rib and cartilage graft along with orthognathic surgery (Le fort I and bilateral sagittal split ramus osteotomy) for patients with Crouzon syndrome. The patient was a 21-year-old male with Crouzon syndrome, who had undergone augmentation rhinoplasty 2 years ago. His main issues were midfacial retrusion and mild anterior open bite and cross bite and, furthermore, did not want any change in his nasal appearance. To augment midfacial volume, rib bone graft was inserted on the inferior orbital rim and costal cartilage graft was done on the zygomatic area. The costal osteocartilage was fixed with titanium screws. Additionally, Le Fort I osteotomy and bilateral sagittal split ramus osteotomy were done to treat the anterior open bite and cross bite. The maxillary segment was advanced 2 mm and posteriorly impacted 2.5 mm. Then, 5 mm of mandibular setback was done and the maxillomandibular segment was rotated clockwise. Finally, genioplasty with 5-mm advancement was done to compensate for the chin retrusion after performing the mandibular setback. The operation took 425 minutes and estimated blood loss was 500 mL. After 6 months since surgery, the patient had convex facial profile and class I occlusion. For the patient with mild midface hypoplasia, good nasal profile, and malocclusion, rib bone graft along with Le Fort I and bilateral sagittal ramus osteotomy can be a good surgical modality.

  18. [Tissue engineering applied to the trachea as a graft].

    PubMed

    Barrera-Ramírez, Elisa; Rico-Escobar, Edna; Garrido-Cardona, Rubén E

    2016-01-01

    Tissue engineering offers, through new technologies, an ex vivo generation of organs and functional tissues as grafts for transplants, for the improvement and substitution of biological functions, with an absence of immunological response. The treatment of extended tracheal lesions is a substitution of the affected segment; nevertheless, the allogeneic transplant has failed and the use of synthetic materials has not had good results. New tissue engineering technology is being developed to offer a tracheal graft for a posterior implantation. The purpose of this article is to review all the methods and components used by the engineering of tissue for tracheal grafts.

  19. Three-dimensional bioprinting of multilayered constructs containing human mesenchymal stromal cells for osteochondral tissue regeneration in the rabbit knee joint.

    PubMed

    Shim, Jin-Hyung; Jang, Ki-Mo; Hahn, Sei Kwang; Park, Ju Young; Jung, Hyuntae; Oh, Kyunghoon; Park, Kyeng Min; Yeom, Junseok; Park, Sun Hwa; Kim, Sung Won; Wang, Joon Ho; Kim, Kimoon; Cho, Dong-Woo

    2016-02-04

    The use of cell-rich hydrogels for three-dimensional (3D) cell culture has shown great potential for a variety of biomedical applications. However, the fabrication of appropriate constructs has been challenging. In this study, we describe a 3D printing process for the preparation of a multilayered 3D construct containing human mesenchymal stromal cells with a hydrogel comprised of atelocollagen and supramolecular hyaluronic acid (HA). This construct showed outstanding regenerative ability for the reconstruction of an osteochondral tissue in the knee joints of rabbits. We found that the use of a mechanically stable, host-guest chemistry-based hydrogel was essential and allowed two different types of extracellular matrix (ECM) hydrogels to be easily printed and stacked into one multilayered construct without requiring the use of potentially harmful chemical reagents or physical stimuli for post-crosslinking. To the best of our knowledge, this is the first study to validate the potential of a 3D printed multilayered construct consisting of two different ECM materials (atelocollagen and HA) for heterogeneous tissue regeneration using an in vivo animal model. We believe that this 3D printing-based platform technology can be effectively exploited for regeneration of various heterogeneous tissues as well as osteochondral tissue.

  20. Studies in Fat Grafting: Part III. Fat grafting irradiated tissue: Improved skin quality and decreased fat graft retention

    PubMed Central

    Garza, Rebecca M.; Paik, Kevin J.; Chung, Michael T.; Duscher, Dominik; Gurtner, Geoffrey C.; Longaker, Michael T.; Wan, Derrick C.

    2014-01-01

    Background Following radiation therapy, skin becomes fibrotic and can present a difficult problem for reconstructive surgeons. There is an increasing belief that fat grafting under radiated skin can reverse the damage caused by radiation. The present study evaluated the effect of fat grafting on irradiated skin, along with fat graft quality and retention rates in irradiated tissue. Methods Nine adult Crl:NU-Foxn1nu CD-1 mice received 30 Gy external beam radiation of the scalp. Four weeks following radiation, scalp skin from irradiated and non-irradiated mice was harvested and compared histologically for dermal thickness, collagen content, and vascular density. Human fat grafts were then injected in the subcutaneous plane of the scalp. Skin assessment was performed in the irradiated group at 2 and 8 weeks post-grafting, and fat graft retention was measured at baseline and every 2 weeks up to 8 weeks post-grafting using micro-CT. Finally, fat graft samples were explanted at 8 weeks, and quality scoring was performed. Results Fat grafting resulted in decreased dermal thickness, decreased collagen content, and increased vascular density in irradiated skin. CT analysis revealed significantly decreased fat graft survival in the irradiated group when compared to the non-irradiated group. Histological scoring of explanted fat grafts demonstrated no difference in quality between the irradiated and non-irradiated groups. Conclusions Fat grafting attenuates dermal collagen deposition and vessel depletion characteristic of radiation fibrosis. Although fat graft retention rates are significantly lower in irradiated than non-irradiated tissue, the quality of retained fat between the groups is similar. PMID:25068325

  1. Covalent Binding of Bone Morphogenetic Protein-2 and Transforming Growth Factor-β3 to 3D Plotted Scaffolds for Osteochondral Tissue Regeneration.

    PubMed

    Di Luca, Andrea; Klein-Gunnewiek, Michel; Vancso, Julius G; van Blitterswijk, Clemens A; Benetti, Edmondo M; Moroni, Lorenzo

    2017-09-02

    Engineering the osteochondral tissue presents some challenges mainly relying in its function of transition from the subchondral bone to articular cartilage and the gradual variation in several biological, mechanical, and structural features. A possible solution for osteochondral regeneration might be the design and fabrication of scaffolds presenting a gradient able to mimic this transition. Covalent binding of biological factors proved to enhance cell adhesion and differentiation in two-dimensional culture substrates. Here, we used polymer brushes as selective linkers of bone morphogenetic protein-2 (BMP-2) and transforming growth factor-β3 (TGF-β3) on the surface of 3D scaffolds fabricated via additive manufacturing (AM) and subsequent controlled radical polymerization. These growth factors (GFs) are known to stimulate the differentiation of human mesenchymal stromal cells (hMSCs) toward the osteogenic and chondrogenic lineages, respectively. BMP-2 and TGF-β3 were covalently bound both homogeneously within a poly(ethylene glycol) (PEG)-based brush-functionalized scaffolds, and following a gradient composition by varying their concentration along the axial section of the 3D constructs. Following an approach previously developed by our group and proved to be successful to generate fibronectin gradients, opposite brush-supported gradients of BMP-2 and TGF-β3 were finally generated and subsequently tested to differentiate cells in a gradient fashion. The brush-supported GFs significantly influenced hMSCs osteochondral differentiation when the scaffolds were homogenously modified, yet no effect was observed in the gradient scaffolds. Therefore, this technique seems promising to maintain the biological activity of growth factors covalently linked to 3D scaffolds, but needs to be further optimized in case biological gradients are desired. © 2017 The Authors. Biotechnology Journal published by Wiley-VCH GmbH & Co. KGaA, Weinheim.

  2. Cartilage repair with osteochondral autografts in sheep: effect of biophysical stimulation with pulsed electromagnetic fields.

    PubMed

    Benazzo, Franco; Cadossi, Matteo; Cavani, Francesco; Fini, Milena; Giavaresi, Gianluca; Setti, Stefania; Cadossi, Ruggero; Giardino, Roberto

    2008-05-01

    The effect of pulsed electromagnetic fields (PEMFs) on the integration of osteochondral autografts was evaluated in sheep. After osteochondral grafts were performed, the animals were treated with PEMFs for 6 h/day or sham-treated. Six animals were sacrificed at 1 month. Fourteen animals were treated for 2 months and sacrificed at 6 months. At 1 month, the osteogenic activity at the transplant-host subchondral bone interface was increased in PEMF-treated animals compared to controls. Articular cartilage was healthy in controls and stimulated animals. At 6 months, complete resorption was observed in four control grafts only. Cyst-like resorption areas were more frequent within the graft of sham-treated animals versus PEMF-treated. The average volume of the cysts was not significantly different between the two groups; nevertheless, analysis of the variance of the volumes demonstrated a significant difference. The histological score showed no significant differences between controls and stimulated animals, but the percentage of surface covered by fibrous tissue was higher in the control group than in the stimulated one. Interleukin-1 and tumor necrosis factor-alpha concentration in the synovial fluid was significantly lower, and transforming growth factor-beta1 was significantly higher, in PEMF-treated animals compared to controls. One month after osteochondral graft implantation, we observed larger bone formation in PEMF-treated grafts which favors early graft stabilization. In the long term, PEMF exposure limited the bone resorption in subchondral bone; furthermore, the cytokine profile in the synovial fluid was indicative of a more favorable articular environment for the graft.

  3. Tissue-engineered lymphatic graft for the treatment of lymphedema

    PubMed Central

    Kanapathy, Muholan; Patel, Nikhil M.; Kalaskar, Deepak M.; Mosahebi, Afshin; Mehrara, Babak J.; Seifalian, Alexander M.

    2015-01-01

    Background Lymphedema is a chronic debilitating condition and curative treatment is yet to be found. Tissue engineering approach, which combines cellular components, scaffold, and molecular signals hold great potential in the treatment of secondary lymphedema with the advent of lymphatic graft to reconstruct damaged collecting lymphatic vessel. This review highlights the ideal characteristics of lymphatic graft, the limitation and challenges faced, and the approaches in developing tissue-engineered lymphatic graft. Methods Literature on tissue engineering of lymphatic system and lymphatic tissue biology was reviewed. Results The prime challenge in the design and manufacturing of this graft is producing endothelialized conduit with intraluminal valves. Suitable scaffold material is needed to ensure stability and functionality of the construct. Endothelialization of the construct can be enhanced via biofunctionalization and nanotopography, which mimics extracellular matrix. Nanocomposite polymers with improved performance over existing biomaterials are likely to benefit the development of lymphatic graft. Conclusions With the in-depth understanding of tissue engineering, nanotechnology, and improved knowledge on the biology of lymphatic regeneration, the aspiration to develop successful lymphatic graft is well achievable. PMID:25248852

  4. Decellularized cartilage may be a chondroinductive material for osteochondral tissue engineering.

    PubMed

    Sutherland, Amanda J; Beck, Emily C; Dennis, S Connor; Converse, Gabriel L; Hopkins, Richard A; Berkland, Cory J; Detamore, Michael S

    2015-01-01

    Extracellular matrix (ECM)-based materials are attractive for regenerative medicine in their ability to potentially aid in stem cell recruitment, infiltration, and differentiation without added biological factors. In musculoskeletal tissue engineering, demineralized bone matrix is widely used, but recently cartilage matrix has been attracting attention as a potentially chondroinductive material. The aim of this study was thus to establish a chemical decellularization method for use with articular cartilage to quantify removal of cells and analyze the cartilage biochemical content at various stages during the decellularization process, which included a physically devitalization step. To study the cellular response to the cartilage matrix, rat bone marrow-derived mesenchymal stem cells (rBMSCs) were cultured in cell pellets containing cells only (control), chondrogenic differentiation medium (TGF-β), chemically decellularized cartilage particles (DCC), or physically devitalized cartilage particles (DVC). The chemical decellularization process removed the vast majority of DNA and about half of the glycosaminoglycans (GAG) within the matrix, but had no significant effect on the amount of hydroxyproline. Most notably, the DCC group significantly outperformed TGF-β in chondroinduction of rBMSCs, with collagen II gene expression an order of magnitude or more higher. While DVC did not exhibit a chondrogenic response to the extent that DCC did, DVC had a greater down regulation of collagen I, collagen X and Runx2. A new protocol has been introduced for cartilage devitalization and decellularization in the current study, with evidence of chondroinductivity. Such bioactivity along with providing the 'raw material' building blocks of regenerating cartilage may suggest a promising role for DCC in biomaterials that rely on recruiting endogenous cell recruitment and differentiation for cartilage regeneration.

  5. Decellularized Cartilage May Be a Chondroinductive Material for Osteochondral Tissue Engineering

    PubMed Central

    Sutherland, Amanda J.; Beck, Emily C.; Dennis, S. Connor; Converse, Gabriel L.; Hopkins, Richard A.; Berkland, Cory J.; Detamore, Michael S.

    2015-01-01

    Extracellular matrix (ECM)-based materials are attractive for regenerative medicine in their ability to potentially aid in stem cell recruitment, infiltration, and differentiation without added biological factors. In musculoskeletal tissue engineering, demineralized bone matrix is widely used, but recently cartilage matrix has been attracting attention as a potentially chondroinductive material. The aim of this study was thus to establish a chemical decellularization method for use with articular cartilage to quantify removal of cells and analyze the cartilage biochemical content at various stages during the decellularization process, which included a physically devitalization step. To study the cellular response to the cartilage matrix, rat bone marrow-derived mesenchymal stem cells (rBMSCs) were cultured in cell pellets containing cells only (control), chondrogenic differentiation medium (TGF-β), chemically decellularized cartilage particles (DCC), or physically devitalized cartilage particles (DVC). The chemical decellularization process removed the vast majority of DNA and about half of the glycosaminoglycans (GAG) within the matrix, but had no significant effect on the amount of hydroxyproline. Most notably, the DCC group significantly outperformed TGF-β in chondroinduction of rBMSCs, with collagen II gene expression an order of magnitude or more higher. While DVC did not exhibit a chondrogenic response to the extent that DCC did, DVC had a greater down regulation of collagen I, collagen X and Runx2. A new protocol has been introduced for cartilage devitalization and decellularization in the current study, with evidence of chondroinductivity. Such bioactivity along with providing the ‘raw material’ building blocks of regenerating cartilage may suggest a promising role for DCC in biomaterials that rely on recruiting endogenous cell recruitment and differentiation for cartilage regeneration. PMID:25965981

  6. Osteochondral Biopsy Analysis Demonstrates That BST-CarGel Treatment Improves Structural and Cellular Characteristics of Cartilage Repair Tissue Compared With Microfracture

    PubMed Central

    Méthot, Stéphane; Changoor, Adele; Tran-Khanh, Nicolas; Hoemann, Caroline D.; Stanish, William D.; Restrepo, Alberto; Shive, Matthew S.; Buschmann, Michael D.

    2016-01-01

    Objective The efficacy and safety of BST-CarGel, a chitosan-based medical device for cartilage repair, was compared with microfracture alone at 1 year during a multicenter randomized controlled trial (RCT) in the knee. The quality of repair tissue of osteochondral biopsies collected from a subset of patients was compared using blinded histological assessments. Methods The international RCT evaluated repair tissue quantity and quality by 3-dimensional quantitative magnetic resonance imaging as co-primary endpoints at 12 months. At an average of 13 months posttreatment, 21/41 BST-CarGel and 17/39 microfracture patients underwent elective second look arthroscopies as a tertiary endpoint, during which ICRS (International Cartilage Repair Society) macroscopic scoring was carried out, and osteochondral biopsies were collected. Stained histological sections were evaluated by blinded readers using ICRS I and II histological scoring systems. Collagen organization was evaluated using a polarized light microscopy score. Results BST-CarGel treatment resulted in significantly better ICRS macroscopic scores (P = 0.0002) compared with microfracture alone, indicating better filling, integration, and tissue appearance. Histologically, BST-CarGel resulted in a significant improvement of structural parameters—Surface Architecture (P = 0.007) and Surface/Superficial Assessment (P = 0.042)—as well as cellular parameters—Cell Viability (P = 0.006) and Cell Distribution (P = 0.032). No histological parameters were significantly better for the microfracture group. BST-CarGel treatment also resulted in a more organized repair tissue with collagen stratification more similar to native hyaline cartilage, as measured by polarized light microscopy scoring (P = 0.0003). Conclusion Multiple and independent analyses in this biopsy substudy demonstrated that BST-CarGel treatment results in improved structural and cellular characteristics of repair tissue at 1 year posttreatment compared with

  7. Controlled Release Strategies for Bone, Cartilage, and Osteochondral Engineering—Part I: Recapitulation of Native Tissue Healing and Variables for the Design of Delivery Systems

    PubMed Central

    Santo, Vítor E.; Mano, João F.; Reis, Rui L.

    2013-01-01

    The potential of growth factors to stimulate tissue healing through the enhancement of cell proliferation, migration, and differentiation is undeniable. However, critical parameters on the design of adequate carriers, such as uncontrolled spatiotemporal presence of bioactive factors, inadequate release profiles, and supraphysiological dosages of growth factors, have impaired the translation of these systems onto clinical practice. This review describes the healing cascades for bone, cartilage, and osteochondral interface, highlighting the role of specific growth factors for triggering the reactions leading to tissue regeneration. Critical criteria on the design of carriers for controlled release of bioactive factors are also reported, focusing on the need to provide a spatiotemporal control over the delivery and presentation of these molecules. PMID:23268651

  8. Vascular tissue engineering: towards the next generation vascular grafts.

    PubMed

    Naito, Yuji; Shinoka, Toshiharu; Duncan, Daniel; Hibino, Narutoshi; Solomon, Daniel; Cleary, Muriel; Rathore, Animesh; Fein, Corey; Church, Spencer; Breuer, Christopher

    2011-04-30

    The application of tissue engineering technology to cardiovascular surgery holds great promise for improving outcomes in patients with cardiovascular diseases. Currently used synthetic vascular grafts have several limitations including thrombogenicity, increased risk of infection, and lack of growth potential. We have completed the first clinical trial evaluating the feasibility of using tissue engineered vascular grafts (TEVG) created by seeding autologous bone marrow-derived mononuclear cells (BM-MNC) onto biodegradable tubular scaffolds. Despite an excellent safety profile, data from the clinical trial suggest that the primary graft related complication of the TEVG is stenosis, affecting approximately 16% of grafts within the first seven years after implantation. Continued investigation into the cellular and molecular mechanisms underlying vascular neotissue formation will improve our basic understanding and provide insights that will enable the rationale design of second generation TEVG. Published by Elsevier B.V.

  9. Strategies for osteochondral repair: Focus on scaffolds

    PubMed Central

    Seo, Seog-Jin; Mahapatra, Chinmaya; Singh, Rajendra K; Knowles, Jonathan C

    2014-01-01

    Interest in osteochondral repair has been increasing with the growing number of sports-related injuries, accident traumas, and congenital diseases and disorders. Although therapeutic interventions are entering an advanced stage, current surgical procedures are still in their infancy. Unlike other tissues, the osteochondral zone shows a high level of gradient and interfacial tissue organization between bone and cartilage, and thus has unique characteristics related to the ability to resist mechanical compression and restoration. Among the possible therapies, tissue engineering of osteochondral tissues has shown considerable promise where multiple approaches of utilizing cells, scaffolds, and signaling molecules have been pursued. This review focuses particularly on the importance of scaffold design and its role in the success of osteochondral tissue engineering. Biphasic and gradient composition with proper pore configurations are the basic design consideration for scaffolds. Surface modification is an essential technique to improve the scaffold function associated with cell regulation or delivery of signaling molecules. The use of functional scaffolds with a controllable delivery strategy of multiple signaling molecules is also considered a promising therapeutic approach. In this review, we updated the recent advances in scaffolding approaches for osteochondral tissue engineering. PMID:25343021

  10. Clinical feasibility of a novel biphasic osteochondral composite for matrix-associated autologous chondrocyte implantation.

    PubMed

    Chiang, H; Liao, C-J; Hsieh, C-H; Shen, C-Y; Huang, Y-Y; Jiang, C-C

    2013-04-01

    Matrix-associated autologous chondrocyte implantation has been used to treat cartilage defects. We developed a biphasic cylindrical osteochondral composite construct for such use, and conducted this study to determine its feasibility for treating osteochondral lesions in human knees. Ten patients with symptomatic osteochondral lesions at femoral condyles were treated by replacing pathological tissue with the construct of dl-poly-lactide-co-glycolide, whose lower body was impregnated with β-tricalcium phosphate and served as osseous phase. The construct had a chamber to load double-minced autologous cartilage, serving as source of chondrocytes. Osteochondral lesion was drill-fashioned a pit of identical dimension as the construct. Chondrocyte-laden construct was press-fit to fill the pit. Postoperative outcome was evaluated using Knee Injury and Osteoarthritis Outcome Score (KOOS) scale up to 24 months. Magnetic resonance image was taken, and sample tissue was collected with second-look arthroscopic needle biopsy at 12 months. Outcome parameters were primarily safety of surgery, and secondarily postoperative change in KOOS and regeneration of hyaline cartilage and cancellous bone. No patient experienced serious adverse events. Postoperative mean KOOS in "symptoms" subscale had not changed significantly from pre-operation until 24 months; whereas those in the other four subscales were significantly higher than pre-operation at 12 and 24 months. Second-look arthroscopy showed completely filled grafted sites, with regenerate cartilaginous surfaces flushed with surrounding native joint surface. Microscopically, regenerated cartilage appeared hyaline. This novel construct for chondrocyte implantation is safe for surgical application in knee. It repairs osteochondral lesions of femoral condyles by successful regeneration of hyaline cartilage. Copyright © 2013 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

  11. Osteochondritis Dissecans (OCD)

    MedlinePlus

    ... mice, motion, necrosis, osteochondritis, osteochondroses, pain, repetitive, sticking, stress, subchondral Family Health, Kids and Teens, Men September 2000 Copyright © American Academy of Family ...

  12. Extracellular Calcium Modulates Chondrogenic and Osteogenic Differentiation of Human Adipose-Derived Stem Cells: A Novel Approach for Osteochondral Tissue Engineering Using a Single Stem Cell Source.

    PubMed

    Mellor, Liliana F; Mohiti-Asli, Mahsa; Williams, John; Kannan, Arthi; Dent, Morgan R; Guilak, Farshid; Loboa, Elizabeth G

    2015-09-01

    We have previously shown that elevating extracellular calcium from a concentration of 1.8 to 8 mM accelerates and increases human adipose-derived stem cell (hASC) osteogenic differentiation and cell-mediated calcium accretion, even in the absence of any other soluble osteogenic factors in the culture medium. However, the effects of elevated calcium on hASC chondrogenic differentiation have not been reported. The goal of this study was to determine the effects of varied calcium concentrations on chondrogenic differentiation of hASC. We hypothesized that exposure to elevated extracellular calcium (8 mM concentration) in a chondrogenic differentiation medium (CDM) would inhibit chondrogenesis of hASC when compared to basal calcium (1.8 mM concentration) controls. We further hypothesized that a full osteochondral construct could be engineered by controlling local release of calcium to induce site-specific chondrogenesis and osteogenesis using only hASC as the cell source. Human ASC was cultured as micromass pellets in CDM containing transforming growth factor-β1 and bone morphogenetic protein 6 for 28 days at extracellular calcium concentrations of either 1.8 mM (basal) or 8 mM (elevated). Our findings indicated that elevated calcium induced osteogenesis and inhibited chondrogenesis in hASC. Based on these findings, stacked polylactic acid nanofibrous scaffolds containing either 0% or 20% tricalcium phosphate (TCP) nanoparticles were electrospun and tested for site-specific chondrogenesis and osteogenesis. Histological assays confirmed that human ASC differentiated locally to generate calcified tissue in layers containing 20% TCP, and cartilage in the layers with no TCP when cultured in CDM. This is the first study to report the effects of elevated calcium on chondrogenic differentiation of hASC, and to develop osteochondral nanofibrous scaffolds using a single cell source and controlled calcium release to induce site-specific differentiation. This approach

  13. Extracellular Calcium Modulates Chondrogenic and Osteogenic Differentiation of Human Adipose-Derived Stem Cells: A Novel Approach for Osteochondral Tissue Engineering Using a Single Stem Cell Source

    PubMed Central

    Mellor, Liliana F.; Mohiti-Asli, Mahsa; Williams, John; Kannan, Arthi; Dent, Morgan R.; Guilak, Farshid

    2015-01-01

    We have previously shown that elevating extracellular calcium from a concentration of 1.8 to 8 mM accelerates and increases human adipose-derived stem cell (hASC) osteogenic differentiation and cell-mediated calcium accretion, even in the absence of any other soluble osteogenic factors in the culture medium. However, the effects of elevated calcium on hASC chondrogenic differentiation have not been reported. The goal of this study was to determine the effects of varied calcium concentrations on chondrogenic differentiation of hASC. We hypothesized that exposure to elevated extracellular calcium (8 mM concentration) in a chondrogenic differentiation medium (CDM) would inhibit chondrogenesis of hASC when compared to basal calcium (1.8 mM concentration) controls. We further hypothesized that a full osteochondral construct could be engineered by controlling local release of calcium to induce site-specific chondrogenesis and osteogenesis using only hASC as the cell source. Human ASC was cultured as micromass pellets in CDM containing transforming growth factor-β1 and bone morphogenetic protein 6 for 28 days at extracellular calcium concentrations of either 1.8 mM (basal) or 8 mM (elevated). Our findings indicated that elevated calcium induced osteogenesis and inhibited chondrogenesis in hASC. Based on these findings, stacked polylactic acid nanofibrous scaffolds containing either 0% or 20% tricalcium phosphate (TCP) nanoparticles were electrospun and tested for site-specific chondrogenesis and osteogenesis. Histological assays confirmed that human ASC differentiated locally to generate calcified tissue in layers containing 20% TCP, and cartilage in the layers with no TCP when cultured in CDM. This is the first study to report the effects of elevated calcium on chondrogenic differentiation of hASC, and to develop osteochondral nanofibrous scaffolds using a single cell source and controlled calcium release to induce site-specific differentiation. This approach

  14. Tissue Engineered Bone Grafts: Biological Requirements, Tissue Culture and Clinical Relevance

    PubMed Central

    Fröhlich, Mirjam; Grayson, Warren L.; Wan, Leo Q.; Marolt, Darja; Drobnic, Matej; Vunjak-Novakovic, Gordana

    2009-01-01

    The tremendous need for bone tissue in numerous clinical situations and the limited availability of suitable bone grafts are driving the development of tissue engineering approaches to bone repair. In order to engineer viable bone grafts, one needs to understand the mechanisms of native bone development and fracture healing, as these processes should ideally guide the selection of optimal conditions for tissue culture and implantation. Engineered bone grafts have been shown to have capacity for osteogenesis, osteoconduction, osteoinduction and osteointegration - functional connection between the host bone and the graft. Cells from various anatomical sources in conjunction with scaffolds and osteogenic factors have been shown to form bone tissue in vitro. The use of bioreactor systems to culture cells on scaffolds before implantation further improved the quality of the resulting bone grafts. Animal studies confirmed the capability of engineered grafts to form bone and integrate with the host tissues. However, the vascularization of bone remains one of the hurdles that need to be overcome if clinically sized, fully viable bone grafts are to be engineered and implanted. We discuss here the biological guidelines for tissue engineering of bone, the bioreactor cultivation of human mesenchymal stem cells on three-dimensional scaffolds, and the need for vascularization and functional integration of bone grafts following implantation. PMID:19075755

  15. Tissue Factor Activity in Dialysis Access Grafts.

    PubMed

    Hasenstab, David; Kirkman, Thomas R; Clowes, Alexander W; Kohler, Ted R

    2016-02-01

    Intimal hyperplasia at the venous anastomosis of dialysis grafts causes early failure. We developed a sheep model of arteriovenous prosthetic grafts that fail rapidly due to intimal hyperplasia with histologic features nearly identical to human access grafts. A prominent feature of lesion development in this model is formation of luminal thrombus that becomes organized into stenosing lesions by macrophage and myofibroblast infiltration. To better understand this process, we examined the presence and activity of tissue factor (TF) in this system. This protein is the physiological initiator of coagulation in vivo and is known to contribute to development of intimal hyperplasia after vascular injury. Expanded polytetrafluorethylene (ePTFE) grafts were placed between the carotid artery and external jugular vein in sheep. Grafts were examined for luminal TF activity using a novel ex vivo assay. In a separate series of grafts, immunohistochemistry was used to localize smooth muscle cells, monocytes, and TF protein. At 2 days, luminal TF activity already was higher in the venous and arterial end of the graft than in the adventitia. This high level of activity persisted at 8 weeks. TF activity was higher in the venous end of the grafts than in the arterial end at 2 and 8 weeks (40% and 47% increase, n = 5, n = 3, respectively, P < 0.05). Immunohistochemistry revealed TF protein localized in regions with or adjacent to fibrin accumulation and often in regions close to the lumen. This study further examines the development of intimal hyperplasia in ePTFE dialysis access grafts. In this model, TF levels on the luminal surface were increased throughout the arteriovenous grafts and the adjacent vessels as early as 2 days after engraftment and for as long as 8 weeks thereafter. The highest levels of activity were found in the venous end of the graft, where hyperplasia is most robust. Increased activity of TF is associated with luminal thrombus, which provides a

  16. [Chondral and osteochondral fractures].

    PubMed

    Kayaoğlu, E Esin; Binnet, Mehmet S

    2007-01-01

    The incidence of traumatic chondral and osteochondral fractures and their role in the development of joint degeneration are not fully elucidated. While assessing traumatic knee injuries, one important criterion for the diagnosis of chondral fractures is to remember the possibility of a chondral or osteochondral fracture. Symptoms in osteochondral fractures are more obvious and cause severe pain and difficulty in movement of knee with hemarthrosis. The presence of hemarthrosis facilitates the diagnosis of an osteochondral fracture. Chondral and osteochondral fractures may be associated with other intra-articular pathologies. There are two main mechanisms of these fractures, including a direct effect causing avulsion or impaction and, a more common mechanism, flexion-rotation force to the knee, which is also the mechanism for an acute patellar dislocation. It is known that arthroscopic treatment is the best method for the diagnosis and treatment of chondral and osteochondral fractures. In osteochondral lesions, the aim of treatment is to restore the congruity of articular surfaces. In agreement with literature data, our clinical experience favors internal fixation as the most effective method for the treatment of osteochondral fractures.

  17. Optical methods for diagnostic of cell-tissue grafts

    NASA Astrophysics Data System (ADS)

    Timchenko, P. E.; Timchenko, E. V.; Volova, L. T.; Boltovskaya, V. V.; Zherdeva, L. A.; Belousov, N. V.; Pershutkina, S. V.

    2015-08-01

    In this work the results of cell-tissue grafts research with a complex of optical methods - confocal fluorescent microscopy and Raman spectroscopy are presented. It was established that coefficient M scatter is related to irregularity of demineralization process. It was microscopically shown that the quantity of integrated cells into these types of transplants amounts to 20% of its surface.

  18. Additive Manufacturing of Vascular Grafts and Vascularized Tissue Constructs.

    PubMed

    Elomaa, Laura; Yang, Yunzhi Peter

    2017-01-10

    There is a great need for engineered vascular grafts among patients with cardiovascular diseases who are in need of bypass therapy and lack autologous healthy blood vessels. In addition, because of the severe worldwide shortage of organ donors, there is an increasing need for engineered vascularized tissue constructs as an alternative to organ transplants. Additive manufacturing (AM) offers great advantages and flexibility of fabrication of cell-laden, multimaterial, and anatomically shaped vascular grafts and vascularized tissue constructs. Various inkjet-, extrusion-, and photocrosslinking-based AM techniques have been applied to the fabrication of both self-standing vascular grafts and porous, vascularized tissue constructs. This review discusses the state-of-the-art research on the use of AM for vascular applications and the key criteria for biomaterials in the AM of both acellular and cellular constructs. We envision that new smart printing materials that can adapt to their environment and encourage rapid endothelialization and remodeling will be the key factor in the future for the successful AM of personalized and dynamic vascular tissue applications.

  19. Tissue-Engineered Autologous Grafts for Facial Bone Reconstruction

    PubMed Central

    Bhumiratana, Sarindr; Bernhard, Jonathan C.; Alfi, David M.; Yeager, Keith; Eton, Ryan E.; Bova, Jonathan; Shah, Forum; Gimble, Jeffrey M.; Lopez, Mandi J.; Eisig, Sidney B.; Vunjak-Novakovic, Gordana

    2016-01-01

    Facial deformities require precise reconstruction of the appearance and function of the original tissue. The current standard of care—the use of bone harvested from another region in the body—has major limitations, including pain and comorbidities associated with surgery. We have engineered one of the most geometrically complex facial bones by using autologous stromal/stem cells, without bone morphogenic proteins, using native bovine bone matrix and a perfusion bioreactor for the growth and transport of living grafts. The ramus-condyle unit (RCU), the most eminent load-bearing bone in the skull, was reconstructed using an image-guided personalized approach in skeletally mature Yucatan minipigs (human-scale preclinical model). We used clinically approved decellularized bovine trabecular bone as a scaffolding material, and crafted it into an anatomically correct shape using image-guided micromilling, to fit the defect. Autologous adipose-derived stromal/stem cells were seeded into the scaffold and cultured in perfusion for 3 weeks in a specialized bioreactor to form immature bone tissue. Six months after implantation, the engineered grafts maintained their anatomical structure, integrated with native tissues, and generated greater volume of new bone and greater vascular infiltration than either non-seeded anatomical scaffolds or untreated defects. This translational study demonstrates feasibility of facial bone reconstruction using autologous, anatomically shaped, living grafts formed in vitro, and presents a platform for personalized bone tissue engineering. PMID:27306665

  20. Aspects of Magnetic Resonance in the surgical treatment of osteochondral lesions of the knee.

    PubMed

    Macarini, Luca; Murrone, Mario; Marini, Stefania; Moretti, Biagio; Patella, Vittorio

    2003-01-01

    To assess the magnetic resonance (MR) appearance of knee cartilage chondroplasty procedures and their evolution in order to evaluate the usefulness of the method in monitoring postoperative rehabilitation. Sixty-two patients treated with knee chondroplasty for high-grade cartilage injuries (Noyes' stages II and III) were examined with MR. Forty patients were treated with abrasion chondroplasty, fifteen with osteochondral graft in the injury site and seven with the matrix-induced autologous chondrocyte transplant technique. All patients were operated on by the same orthopaedic team and examined with the same MR protocol. The MR follow-up was performed six months and one year after surgery in the patients treated with abrasion chondroplasty and osteochondral graft, and one week, three months and one year after surgery in the patients treated with cartilage transplant. In the patients treated with abrasion chondroplasty we assessed the fibrocartilage repair and the subchondral bone features, in the patients treated with osteochondral graft we examined the cartilage, the subchondral bone and the graft borders, while in the patients treated with cartilage transplant we evaluated the features and the evolution of the transplant and the subchondral bone. Arthrosynovitis was assessed in all patients. In seven patients a cartilage repair biopsy was performed in arthroscopy. In all the patients MR imaging proved useful in monitoring the chondroplasty. In the patients treated with abrasion chondroplasty the cartilage repair appeared as a hypointense non-homogeneous irregular strip of tissue that replaced the articular surface. The subchondral bone was sclerotic with some geodes. In the later examination the repair was unchanged. In the patients treated with osteochondral graft the articular cartilage was similar to the adjacent hyaline cartilage, although more non-homogeneous. The subchondral bone was sclerotic and in three cases oedematous. In four cases the graft extended

  1. Cartilage repair with autogenic perichondrium cell and polylactic acid grafts.

    PubMed

    Dounchis, J S; Bae, W C; Chen, A C; Sah, R L; Coutts, R D; Amiel, D

    2000-08-01

    The repair of articular cartilage injuries remains a challenge, with many of the current therapeutic strategies based on the grafting or recruitment of chondrogenic tissues or cells. This 1-year study compared the repair of a 3.7-mm diameter by 3-mm deep osteochondral defect in the medial femoral condyle of 24 New Zealand White rabbits; the defect was obtained using an autogenic perichondrium cell polylactic acid composite graft with a contralateral control in which the osteochondral defect remained empty. To elucidate the effect of host immune responses on the repair process after perichondrium cell transplantation, the results of the autogenic perichondrium cell polylactic acid graft group were compared with those obtained in the authors' previous 1-year study of allogenic perichondrium cell polylactic acid composite grafts implanted in a similar model. One year after surgery, the repair site underwent gross inspection and histologic, histomorphometric, biochemical, and biomechanical analyses. The autogenic perichondrium cell polylactic acid graft group (92%) and the control group in which the osteochondral defect remained empty (88%) resulted in a high percentage of grossly acceptable repairs. The autogenic grafts appeared to augment the intrinsic healing capacity of the animals (as compared with the animals in the No Implant Group). The autogenic perichondrium cell polylactic and grafts improved the histologic appearance and percentage of Type II collagen of the cartilaginous repair tissue. Compared with allogenic grafts, the autogenic grafts had better reconstitution of the subchondral bone. However, the results of this experimental model suggest a suboptimal concentration of glycosaminoglycans in the neocartilage matrix, a depressed surface of the repair tissue, a histologic appearance that was not equivalent to that of normal articular cartilage, and reduced biomechanical properties for the repair tissue. The future application of growth factors to this

  2. Patient outcomes following subepithelial connective tissue graft and free gingival graft procedures.

    PubMed

    Wessel, Jeffrey R; Tatakis, Dimitris N

    2008-03-01

    Subepithelial connective tissue grafts (CTGs) and free gingival grafts (FGGs) are common periodontal procedures with similar indications; however, they may differ regarding patient outcomes. Reports on postoperative periodontal patient outcomes are limited. The aim of this observational trial was to compare patient-based outcomes for CTGs and FGGs. Patients who received CTG or FGG completed postoperative questionnaires at 3 days and 3 weeks to assess pain, number of analgesic pills taken, and number of days pills were taken. Postoperative pain was assessed using a visual analog scale (VAS). Twenty-three subjects (12 CTGs and 11 FGGs) completed the study. Differences between CTG and FGG groups in VAS pain scores at 3 days did not reach statistical significance. The proportion of subjects reporting pain in the palate at 3 days was significantly greater for FGG (P <0.05). There were no significant intergroup differences at 3 weeks. For the FGG group, 3-week VAS pain scores were less than the 3-day ones (P <0.01). For the entire study population, the number of days analgesic pills were taken, total number of analgesic pills taken, and number of pills taken from day 3 to the end of the study correlated with the 3-week pain scores. FGG is associated with a greater incidence of donor site pain compared to CTG at the early postoperative period. Longer-term pain after soft tissue grafting is associated with greater analgesic usage. There is an opportunity to improve the postoperative protocols of soft tissue grafting, particularly for FGG.

  3. A preliminary study of osteochondral regeneration using a scaffold-free three-dimensional construct of porcine adipose tissue-derived mesenchymal stem cells.

    PubMed

    Murata, Daiki; Tokunaga, Satoshi; Tamura, Tadashi; Kawaguchi, Hiroaki; Miyoshi, Noriaki; Fujiki, Makoto; Nakayama, Koichi; Misumi, Kazuhiro

    2015-03-18

    Osteoarthritis (OA) is a major joint disease in humans and many other animals. Consequently, medical countermeasures for OA have been investigated diligently. This study was designed to examine the regeneration of articular cartilage and subchondral bone using three-dimensional (3D) constructs of adipose tissue-derived mesenchymal stem cells (AT-MSCs). AT-MSCs were isolated and expanded until required for genetical and immunological analysis and construct creation. A construct consisting of about 760 spheroids that each contained 5.0 × 10(4) autologous AT-MSCs was implanted into an osteochondral defect (diameter: 4 mm; depth: 6 mm) created in the femoral trochlear groove of two adult microminipigs. After implantation, the defects were monitored by computed tomography every month for 6 months in animal no. 1 and 12 months in animal no. 2. AT-MSCs were confirmed to express the premature genes and to be positive for CD90 and CD105 and negative for CD34 and CD45. Under specific nutrient conditions, the AT-MSCs differentiated into osteogenic, chondrogenic, and adipogenic lineages, as evidenced by the expressions of related marker genes and the production of appropriate matrix molecules. A radiopaque area emerged from the boundary between the bone and the implant and increased more steadily upward and inward for the implants in both animal no. 1 and animal no. 2. The histopathology of the implants after 6 months revealed active endochondral ossification underneath the plump fibrocartilage in animal no. 1. The histopathology after 12 months in animal no. 2 showed not only that the diminishing fibrocartilage was as thick as the surrounding normal cartilage but also that massive subchondral bone was present. The present results suggest that implantation of a scaffold-free 3D construct of AT-MSCs into an osteochondral defect may induce regeneration of the original structure of the cartilage and subchondral bone over the course of 1 year, although more

  4. Bone cysts after osteochondral allograft repair of cartilage defects in goats suggest abnormal interaction between subchondral bone and overlying synovial joint tissues.

    PubMed

    Pallante-Kichura, Andrea L; Cory, Esther; Bugbee, William D; Sah, Robert L

    2013-11-01

    The efficacy of osteochondral allografts (OCAs) may be affected by osseous support of the articular cartilage, and thus affected by bone healing and remodeling in the OCA and surrounding host. Bone cysts, and their communication pathways, may be present in various locations after OCA insertion and reflect distinct pathogenic mechanisms. Previously, we analyzed the effect of OCA storage (FRESH, 4°C/14d, 4°C/28d, FROZEN) on cartilage quality in fifteen adult goats after 12months in vivo. The objectives of this study were to further analyze OCAs and contralateral non-operated (Non-Op) CONTROLS from the medial femoral condyle to (1) determine the effect of OCA storage on local subchondral bone (ScB) and trabecular bone (TB) structure, (2) characterize the location and structure of bone cysts and channels, and (3) assess the relationship between cartilage and bone properties. (1) Overall bone structure after OCAs was altered compared to Non-Op, with OCA samples displaying bone cysts, ScB channels, and ScB roughening. ScB BV/TV in FROZEN OCAs was lower than Non-Op and other OCAs. TB BV/TV in FRESH, 4°C/14d, and 4°C/28d OCAs did not vary compared to Non-Op, but BS/TV was lower. (2) OCAs contained "basal" cysts, localized to deeper regions, some "subchondral" cysts, localized near the bone-cartilage interface, and some ScB channels. TB surrounding basal cysts exhibited higher BV/TV than Non-Op. (3) Basal cysts occurred (a) in isolation, (b) with subchondral cysts and ScB channels, (c) with ScB channels, or (d) with subchondral cysts, ScB channels, and ScB erosion. Deterioration of cartilage gross morphology was strongly associated with abnormal μCT bone structure. Evidence of cartilage-bone communication following OCA repair may favor fluid intrusion as a mechanism for subchondral cyst formation, while bone resorption at the graft-host interface without affecting overall bone and cartilage structure may favor bony contusion mechanism for basal cyst formation. These

  5. Engineered Composite Tissue as a Bioartificial Limb Graft

    PubMed Central

    Jank, Bernhard J.; Xiong, Linjie; Moser, Philipp T.; Guyette, Jacques P.; Ren, Xi; Leonard, David A.; Fernandez, Leopoldo; Ott, Harald C.

    2015-01-01

    The loss of an extremity is a disastrous injury with tremendous impact on a patient’s life. Current mechanical prostheses are technically highly sophisticated, but only partially replace physiologic function and aesthetic appearance. As a biologic alternative, approximately 70 patients have undergone allogeneic hand transplantation to date worldwide. While outcomes are favorable, risks and side effects of transplantation and long-term immunosuppression pose a significant ethical dilemma. An autologous, bio-artificial graft based on native extracellular matrix and patient derived cells could be produced on demand and would not require immunosuppression after transplantation. To create such a graft, we decellularized rat and primate forearms by detergent perfusion and yielded acellular scaffolds with preserved composite architecture. We then repopulated muscle and vasculature with cells of appropriate phenotypes, and matured the composite tissue in a perfusion bioreactor under electrical stimulation in vitro. After confirmation of composite tissue formation, we transplanted the resulting bio-composite grafts to confirm perfusion in vivo. PMID:26004237

  6. Qualitative assessment of connective tissue graft with epithelial component. A microsurgical periodontal plastic surgical technique for soft tissue esthetics.

    PubMed

    Rossi, Roberto; Pilloni, Andrea; Morales, Regina Santos

    2009-01-01

    Connective tissue grafts have been used successfully in the treatment of gingival recession. In the mid 80s and late 90s, the periodontal literature presented various techniques such as free gingival grafts, pedicle flaps, subepithelial connective tissue grafts, acellular dermal matrix grafts, and guided tissue regeneration to cover denuded root surfaces. Currently, connective tissue grafting is a reliable treatment for esthetic root coverage. This paper presents a qualitative assessment of a surgical technique that uses a connective tissue graft, including a portion of epithelium in the shape of the defect. This procedure enhances the healing of the covered root surface, increases the thickness of the soft tissue and improves esthetics. The criteria used for evaluation were: color, volume, texture, and blending. This evaluation demonstrated encouraging results from an esthetic viewpoint.

  7. Electrospun scaffolds for tissue engineering of vascular grafts.

    PubMed

    Hasan, Anwarul; Memic, Adnan; Annabi, Nasim; Hossain, Monowar; Paul, Arghya; Dokmeci, Mehmet R; Dehghani, Fariba; Khademhosseini, Ali

    2014-01-01

    There is a growing demand for off-the-shelf tissue engineered vascular grafts (TEVGs) for the replacement or bypass of damaged arteries in various cardiovascular diseases. Scaffolds from the decellularized tissue skeletons to biopolymers and biodegradable synthetic polymers have been used for fabricating TEVGs. However, several issues have not yet been resolved, which include the inability to mimic the mechanical properties of native tissues, and the ability for long-term patency and growth required for in vivo function. Electrospinning is a popular technique for the production of scaffolds that has the potential to address these issues. However, its application to human TEVGs has not yet been achieved. This review provides an overview of tubular scaffolds that have been prepared by electrospinning with potential for TEVG applications. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  8. Electrospun Scaffolds for Tissue Engineering of Vascular Grafts

    PubMed Central

    Hasan, Anwarul; Memic, Adnan; Annabi, Nasim; Hossain, Monowar; Paul, Arghya; Dokmeci, Mehmet R.; Dehghani, Fariba; Khademhosseini, Ali

    2013-01-01

    There is a growing demand for off-the-shelf tissue engineered vascular grafts (TEVGs) for replacement or bypass of damaged arteries in various cardiovascular diseases. Scaffolds from the decellularized tissue skeletons to biopolymers and biodegradable synthetic polymers have been used for fabricating TEVGs. However, several issues have not yet been resolved, which include the inability to mimic the mechanical properties of native tissues, and the ability for long term patency and growth required for in vivo function. Electrospinning is a popular technique for the production of scaffolds that has the potential to address these issues. However, its application to human TEVGs has not yet been achieved. This review provides an overview of tubular scaffolds that have been prepared by electrospinning with potential for TEVG applications. PMID:23973391

  9. Donor-site giant cell reaction following backfill with synthetic bone material during osteochondral plug transfer.

    PubMed

    Fowler, Donald E; Hart, Joseph M; Hart, Jennifer A; Miller, Mark D

    2009-10-01

    Osteochondral defects are common in younger, active patients. Multiple strategies have been used to treat these lesions, including microfracture and osteochondral plug transfer. We describe a patient experiencing chronic knee pain and a full-thickness cartilage defect on the lateral femoral condyle. After failing conservative management and microfracture surgery, the patient underwent osteochondral autograft plug transfer, with backfilling of the donor sites using synthetic bone graft substitute. Initial recovery was uncomplicated until the patient experienced pain following a twist of the knee. Magnetic resonance imaging for the subsequent knee injury revealed poor healing at the donor sites. The donor sites were debrided, and specimens revealed a foreign body giant cell reaction. Donor-site morbidity is of primary concern during osteochondral plug transfer; however, insufficient data exist to support the use of synthetic bone graft material. Our results indicate that off-label use of synthetic bone graft substitute during a primary procedure requires further investigation.

  10. Cylindrical Costal Osteochondral Autograft for Reconstruction of Large Defects of the Capitellum Due to Osteochondritis Dissecans

    PubMed Central

    Shimada, Kozo; Tanaka, Hiroyuki; Matsumoto, Taiichi; Miyake, Junichi; Higuchi, Haruhisa; Gamo, Kazushige; Fuji, Takeshi

    2012-01-01

    Background: There is a need to clarify the usefulness of and problems associated with cylindrical costal osteochondral autograft for reconstruction of large defects of the capitellum due to osteochondritis dissecans. Methods: Twenty-six patients with advanced osteochondritis dissecans of the humeral capitellum were treated with use of cylindrical costal osteochondral autograft. All were males with elbow pain and full-thickness articular cartilage lesions of ≥15 mm in diameter. Clinical, radiographic, and magnetic resonance imaging outcomes were evaluated at a mean follow-up of thirty-six months (range, twenty-four to fifty-one months). Results: All patients had rapid functional improvement after treatment with costal osteochondral autograft and returned to their former activities, including sports. Five patients needed additional minor surgical procedures, including screw removal, loose body removal, and shaving of protruded articular cartilage. Mean elbow function, assessed with use of the clinical rating system of Timmerman and Andrews, was 111 points preoperatively and improved to 180 points at the time of follow-up and to 190 points after the five patients underwent the additional operations. Mean elbow motion was 126° of flexion with 16° of extension loss preoperatively and improved to 133° of flexion with 3° of extension loss at the time of follow-up. Osseous union of the graft on radiographs was obtained within three months in all patients. Revascularization of the graft depicted on T1-weighted magnetic resonance imaging and congruity of the reconstructed articular surface depicted on T2-weighted or short tau inversion recovery imaging were assessed at twelve and twenty-four months postoperatively. Functional recovery was good, and all patients were satisfied with the final outcomes. Conclusions: Cylindrical costal osteochondral autograft was useful for the treatment of advanced osteochondritis dissecans of the humeral capitellum. Functional recovery

  11. Evaluation and analysis of graft hypertrophy by means of arthroscopy, biochemical MRI and osteochondral biopsies in a patient following autologous chondrocyte implantation for treatment of a full-thickness-cartilage defect of the knee.

    PubMed

    Niemeyer, Philipp; Uhl, Markus; Salzmann, Gian M; Morscheid, Yannik P; Südkamp, Norbert P; Madry, Henning

    2015-06-01

    Graft hypertrophy represents a characteristic complication following autologous chondrocyte implantation (ACI) for treatment of cartilage defects. Although some epidemiological data suggest that incidence is associated with first-generation ACI using autologous chondrocyte implantation, it has also been reported in other technical modifications of ACI using different biomaterials. Nevertheless, it has not been described in autologous, non-periosteum, implant-free associated ACI. In addition, little is known about histological and T2-relaxation appearance of graft hypertrophy. The present case report provides a rare case of extensive graft hypertrophy following ACI using an autologous spheres technique with clinical progression over time. Detailed clinical, MR tomographic and histological evaluation has been performed, which demonstrates a high quality of repair tissue within the hypertrophic as well as non-hypertrophic transplanted areas of the repair tissue. No expression of collagen type X (a sign of chondrocyte hypertrophy), only slight changes of the subchondral bone and a nearly normal cell-matrix ratio suggest that tissue within the hypertrophic area does not significantly differ from intact and high-quality repair tissue and therefore seems not to cause graft hypertrophy. This is in contrast to the assumption that histological hypertrophy might cause or contribute to an overwhelming growth of the repair tissue within the transplantation site. Data presented in this manuscript might contribute to further explain the etiology of graft hypertrophy following ACI.

  12. Orthopaedic Interface Tissue Engineering for the Biological Fixation of Soft Tissue Grafts

    PubMed Central

    Moffat, Kristen L .; Wang, I-Ning Elaine; Rodeo, Scott A.; Lu, Helen H.

    2012-01-01

    Interface tissue engineering is a promising new strategy aimed at the regeneration of tissue interfaces and ultimately enabling the biological fixation of soft tissue grafts utilized in orthopaedic repair and sports medicine. Many ligaments and tendons with direct insertions into subchondral bone exhibit a complex enthesis consisting of several distinct yet continuous regions of soft tissue, noncalcified fibrocartilage, calcified fibrocartilage and bone. Regeneration of this multi-tissue interface will be critical for functional graft integration and improving long term clinical outcome. This review will highlight current knowledge of the structure-function relationship at the interface, the mechanism of interface regeneration, and the strategic biomimicry implemented in stratified scaffold design for interface tissue engineering and multi-tissue regeneration. Potential challenges and future directions in this emerging field will also be discussed. It is anticipated that interface tissue engineering will lead to the design of a new generation of integrative fixation devices for soft tissue repair, and it will be instrumental for the development of integrated musculoskeletal tissue systems with biomimetic complexity and functionality. PMID:19064172

  13. Establishing proof of concept: Platelet-rich plasma and bone marrow aspirate concentrate may improve cartilage repair following surgical treatment for osteochondral lesions of the talus.

    PubMed

    Smyth, Niall A; Murawski, Christopher D; Haleem, Amgad M; Hannon, Charles P; Savage-Elliott, Ian; Kennedy, John G

    2012-07-18

    Osteochondral lesions of the talus are common injuries in the athletic patient. They present a challenging clinical problem as cartilage has a poor potential for healing. Current surgical treatments consist of reparative (microfracture) or replacement (autologous osteochondral graft) strategies and demonstrate good clinical outcomes at the short and medium term follow-up. Radiological findings and second-look arthroscopy however, indicate possible poor cartilage repair with evidence of fibrous infill and fissuring of the regenerative tissue following microfracture. Longer-term follow-up echoes these findings as it demonstrates a decline in clinical outcome. The nature of the cartilage repair that occurs for an osteochondral graft to become integrated with the native surround tissue is also of concern. Studies have shown evidence of poor cartilage integration, with chondrocyte death at the periphery of the graft, possibly causing cyst formation due to synovial fluid ingress. Biological adjuncts, in the form of platelet-rich plasma (PRP) and bone marrow aspirate concentrate (BMAC), have been investigated with regard to their potential in improving cartilage repair in both in vitro and in vitro settings. The in vitro literature indicates that these biological adjuncts may increase chondrocyte proliferation as well as synthetic capability, while limiting the catabolic effects of an inflammatory joint environment. These findings have been extrapolated to in vitro animal models, with results showing that both PRP and BMAC improve cartilage repair. The basic science literature therefore establishes the proof of concept that biological adjuncts may improve cartilage repair when used in conjunction with reparative and replacement treatment strategies for osteochondral lesions of the talus.

  14. The effects of low-dose radiotherapy on fresh osteochondral allografts: An experimental study in rabbits.

    PubMed

    Gönç, Uğur; Çetinkaya, Mehmet; Atabek, Mesut

    2016-10-01

    The aim of this study was to investigate the effects of low-dose fractionated radiotherapy on cartilage degeneration after distal femoral fresh massive osteochondral allograft transplantation. Twenty-four New Zealand White rabbits were divided into three groups of 8 rabbits each. All rabbits underwent distal femoral medial condyle fresh massive osteochondral allograft transplantation from California rabbits. The group 1 underwent transplantation without any preliminary process. The group 2 underwent fractionated local radiotherapy of 100 cGy for five days starting on the transplantation day. The group 3 included the rabbits to which the grafts transplanted after radiating in vitro by a single dose radiation of 1500 cGy. The hosts were sacrificed twelve weeks later. Anteroposterior and lateral radiographs were taken. Synovial tissue, cartilaginous tissue, and subchondral bone were assessed histopathologically. Nonunion was present in three cases of group 2 and one of group 3 in which cartilage degeneration was more severe. Synovial hypertrophy and pannus formation were more obvious in non-radiated rabbits. Hypocellularity and necrosis of the subchondral bone were rare in group 2. More cartilage tissue impairment was present in group 3 compared to group 1. In osteochondral massive allograft transplantations, the immune reaction of the host could be precluded with radiotherapy, and the side-effects can be prevented by low-dose fractionated regimen. The total dose of fractionated radiotherapy for an immune suppression should be adjusted not to damage the cartilage tissue, but to avoid articular degeneration in the long term. Copyright © 2016 Turkish Association of Orthopaedics and Traumatology. Production and hosting by Elsevier B.V. All rights reserved.

  15. Matrix generation within a macroporous non-degradable implant for osteochondral defects is not enhanced with partial enzymatic digestion of the surrounding tissue: evaluation in an in vivo rabbit model.

    PubMed

    Krych, Aaron J; Wanivenhaus, Florian; Ng, Kenneth W; Doty, Stephen; Warren, Russell F; Maher, Suzanne A

    2013-10-01

    Articular cartilage defects are a significant source of pain, have limited ability to heal, and can lead to the development of osteoarthritis. However, a surgical solution is not available. To tackle this clinical problem, non-degradable implants capable of carrying mechanical load immediately after implantation and for the duration of implantation, while integrating with the host tissue, may be viable option. But integration between articular cartilage and non-degradable implants is not well studied. Our objective was to assess the in vivo performance of a novel macroporous, nondegradable, polyvinyl alcohol construct. We hypothesized that matrix generation within the implant would be enhanced with partial digestion of the edges of articular cartilage. Our hypothesis was tested by randomizing an osteochondral defect created in the trochlea of 14 New Zealand white rabbits to treatment with: (i) collagenase or (ii) saline, prior to insertion of the implant. At 1 and 3-month post-operatively, the gross morphology and histologic appearance of the implants and the surrounding tissue were assessed. At 3 months, the mechanical properties of the implant were also quantified. Overall, the hydrogel implants performed favorably; at all time-points and in all groups the implants remained well fixed, did not cause inflammation or synovitis, and did not cause extensive damage to the opposing articular cartilage. Regardless of treatment with saline or collagenase, at 1 month post-operatively implants from both groups had a contiguous interface with adjacent cartilage and were populated with chondrocyte-like cells. At 3 months fibrous encapsulation of all implants was evident, there was no difference between area of aggrecan staining in the collagenase versus saline groups, and implant modulus was similar in both groups; leading us to reject our hypothesis. In summary, a porous PVA osteochondral implant remained well fixed in a short term in vivo osteochondral defect model

  16. Development of tissue-engineered self-expandable aortic stent grafts (Bio stent grafts) using in-body tissue architecture technology in beagles.

    PubMed

    Kawajiri, Hidetake; Mizuno, Takeshi; Moriwaki, Takeshi; Ishibashi-Ueda, Hatsue; Yamanami, Masashi; Kanda, Keiichi; Yaku, Hitoshi; Nakayama, Yasuhide

    2015-02-01

    In this study, we aimed to describe the development of tissue-engineered self-expandable aortic stent grafts (Bio stent graft) using in-body tissue architecture technology in beagles and to determine its mechanical and histological properties. The preparation mold was assembled by insertion of an acryl rod (outer diameter, 8.6 mm; length, 40 mm) into a self-expanding nitinol stent (internal diameter, 9.0 mm; length, 35 mm). The molds (n = 6) were embedded into the subcutaneous pouches of three beagles for 4 weeks. After harvesting and removing each rod, the excessive fragile tissue connected around the molds was trimmed, and thus tubular autologous connective tissues with the stent were obtained for use as Bio stent grafts (outer diameter, approximately 9.3 mm in all molds). The stent strut was completely surrounded by the dense collagenous membrane (thickness, ∼150 µm). The Bio stent graft luminal surface was extremely flat and smooth. The graft wall of the Bio stent graft possessed an elastic modulus that was almost two times higher than that of the native beagle abdominal aorta. This Bio stent graft is expected to exhibit excellent biocompatibility after being implanted in the aorta, which may reduce the risk of type 1 endoleaks or migration.

  17. Refixation of osteochondral fractures by ultrasound-activated, resorbable pins

    PubMed Central

    Neumann, H.; Schulz, A. P.; Gille, J.; Klinger, M.; Jürgens, C.; Reimers, N.; Kienast, B.

    2013-01-01

    Objectives Osteochondral injuries, if not treated adequately, often lead to severe osteoarthritis. Possible treatment options include refixation of the fragment or replacement therapies such as Pridie drilling, microfracture or osteochondral grafts, all of which have certain disadvantages. Only refixation of the fragment can produce a smooth and resilient joint surface. The aim of this study was the evaluation of an ultrasound-activated bioresorbable pin for the refixation of osteochondral fragments under physiological conditions. Methods In 16 Merino sheep, specific osteochondral fragments of the medial femoral condyle were produced and refixed with one of conventional bioresorbable pins, titanium screws or ultrasound-activated pins. Macro- and microscopic scoring was undertaken after three months. Results The healing ratio with ultrasound-activated pins was higher than with conventional pins. No negative heat effect on cartilage has been shown. Conclusion As the material is bioresorbable, no further surgery is required to remove the implant. MRI imaging is not compromised, as it is with implanted screws. The use of bioresorbable pins using ultrasound is a promising technology for the refixation of osteochondral fractures. PMID:23610699

  18. [Long-term follow-up of osteochondritis dissecans].

    PubMed

    Gudas, Rimtautas; Kunigiskis, Giedrius; Kalesinskas, Romas Jonas

    2002-01-01

    Fifty-two patients with osteochondritis dissecans lesions were evaluated after 7-25 years after excision of a partially detached (grade III) fragment or loose (grade IV) fragment from the medial femoral condyles. Average follow-up time was 17.2 (range 7-25 years). Two homogenic groups based on special inclusion criteria were formed; 31 patient was in the first and 21--in the second group. The only difference between the groups was the age; the age average in the first group was--25.6 years (range 15-35 years), and -45.2 years (range 35-55 years) in the second group. Patients were evaluated through ICRS (International Cartilage Repair Society), modified HSS and KOOS (Knee injury and osteoarthritis Outcome score) scales, and with X-rays. Evaluation with the ICRS, modified HSS and KOOS rating scales for osteochondritis dissecans revealed in 9 cases (17%) good results, 32 cases (62%)--fair, and 11 cases (21%)--failure results. Final ICRS and modified HSS evaluation showed statistically significantly better results in the younger patient group at the 21 years (p < 0.04). At an average 17.2 year follow-up X-rays and KOOS evaluation form showed initial and second-degree (according to Ahlbäck) osteoarthritis signs in the knees. The long-term results of the natural history of osteochondritis dissecans are extremely poor. Consequently, we recommend autologous osteochondral grafting for the replacement of the osteochondritis dissecans defects in the knee joint.

  19. In Vitro and In Vivo Studies of Alar-Nasal Cartilage Using Autologous Micro-Grafts: The Use of the Rigenera® Protocol in the Treatment of an Osteochondral Lesion of the Nose

    PubMed Central

    Ceccarelli, Gabriele; Gentile, Pietro; Marcarelli, Marco; Balli, Martina; Ronzoni, Flavio Lorenzo; Benedetti, Laura; Cusella De Angelis, Maria Gabriella

    2017-01-01

    Cartilage defects represent a serious problem due to the poor regenerative properties of this tissue. Regarding the nose, nasal valve collapse is associated with nasal blockage and persistent airway obstruction associated with a significant drop in the quality of life for patients. In addition to surgical techniques, several cell-based tissue-engineering strategies are studied to improve cartilage support in the nasal wall, that is, to ameliorate wall insufficiency. Nevertheless, there are no congruent data available on the benefit for patients during the follow-up time. In this manuscript, we propose an innovative approach in the treatment of cartilage defects in the nose (nasal valve collapse) based on autologous micro-grafts obtained by mechanical disaggregation of a small portion of cartilage tissue (Rigenera® protocol). In particular, we first analyzed in vitro murine and human cartilage micro-grafts; secondly, we analyzed the clinical results of a patient with pinched nose deformity treated with autologous micro-grafts of chondrocytes obtained by Rigenera® protocol. The use of autologous micro-graft produced promising results in surgery treatment of cartilage injuries and could be safely and easily administrated to patients with cartilage tissue defects. PMID:28608799

  20. Mechanical and morphological evaluation of osteochondral implants in dogs.

    PubMed

    Bavaresco, Vanessa P; Garrido, Luiz; Batista, Nilza A; Malmonge, Sônia M; Belangero, William D

    2008-04-01

    The mechanical behavior of osteochondral defects was evaluated in this study with the intention of developing alternative procedures. Cylindrical pins (5.00 mm in diameter and in height) made of pHEMA hydrogel covered ultra-high molecular weight polyethylene (UHMWPE) or beta-tricalcium phosphate (beta-TCP) matrix were used. Ostoechondral defects were caused in the knees of adult dogs and the evaluation was carried out after a 9-month follow-up period. The mechanical behavior of the implants was evaluated by means of an indentation creep test that showed that the UHMWPE matrix maintained its viscoelastic behavior even after follow-up time, while the beta-TCP matrix osteochondral implants presented significant alterations. It is believed that the beta-TCP osteochondral implants were unable to withstand the load applied, causing an increase of complacency when compared to the UHMWPE osteochondral implants. Based on micro and macroscopic analysis, no significant wear was observed in either of the osteochondral implants when compared to the controls. However, morphological alterations, with fragmentation indices in the patella, were observed either due to friction with the hydrogel in the first postoperative months or due to forming of a dense conjunctive tissue. This wear mechanism caused on the counterface of the implant (patella) was observed, notwithstanding the osteochondral implant studied.

  1. A new surgical technique to facilitate osteochondral autograft transfer in osteochondral defects of the capitellum: a case report.

    PubMed

    Bilsel, Kerem; Demirhan, Mehmet; Atalar, Ata Can; Akkaya, Semih

    2010-01-01

    A 17-year-old boy who was engaged in amateur weightlifting and body building presented with complaints of right elbow pain and limitation in elbow range of motion. Plain x-rays and magnetic resonance imaging showed an osteochondral defect in the medial third of the capitellum. At surgery, as a new technique, the lateral collateral ligament was detached from the humeral attachment to provide access to the capitellum with a clear and perpendicular exposure. Following removal of loose fragments within the joint, an osteochondral graft harvested from the lateral femoral condyle was implanted to the defect area of the capitellum. Postoperative radiologic controls showed that the defect was entirely filled by the graft with appropriate graft height. On follow-up examination at 12 months, the patient did not have any complaint about his elbow, and had no limitation of movement compared to the left elbow. Magnetic resonance imaging showed that the graft was successfully adapted to the recipient site without any sign of loosening. At final follow-up 40 months after surgery, the surface of the articular cartilage appeared normal. The range of elbow motion was preserved and the patient had no restriction in daily and sports activities. Considering technical difficulties posed by the narrow and complex structure of the elbow joint, this new technique involving detachment of the lateral collateral ligament facilitates perpendicular implantation of the graft. In our opinion, utilization of this new technique will improve functional and radiological results of osteochondral autograft transfer.

  2. Reconstruction of the proximal humerus by combined use of extracorporeally-irradiated osteochondral graft and free vascularized fibula following resection of Ewing sarcoma.

    PubMed

    Muramatsu, Keiichi; Fukano, Reiji; Ihara, Koichiro; Iwanaga, Ryuta; Taguchi, Toshihiko

    2010-12-01

    Reconstruction of the proximal humerus following limb-saving resection of malignant bone tumor is extremely challenging. We describe here a novel anatomical reconstruction technique in a young patient. A 6-year-old girl with Ewing sarcoma of the proximal humerus was treated by wide excision of the tumor followed by reconstruction with extracorporeally-irradiated osteoarticular autograft combined with an intramedullary inserted free vascularized fibula graft. Proper alignment of the shoulder joint was maintained with no osteoarthritic changes after 16 months. The resulting limb function was satisfactory. This biological reconstruction method was safe and without serious complication. It is indicated for the reconstruction of non-weight-bearing joints and is ideal for the proximal humerus.

  3. [Soft tissues volumes changing in malar and cheek area after fat grafting].

    PubMed

    Nadtochiy, A G; Grischenko, S V; Malitskaya, O A

    2016-01-01

    To improve the predictability of facial soft tissues fat grafting results tissue thickness dynamics before and 1 year postoperatively was assessed by means of ultrasonic method in 58 patients under standardized position of the ultrasonic transducer, physical and technical scanning conditions. The study revealed direct correlation of soft tissues thickness increase after fat grafting with the initial thickness of recipient area tissues. One year after fat grafting 60-65% of additional thickness remained in the lower regions of malar-cheek area (with the greatest soft tissues thickness), and only 25-27% preserved in the upper regions with the minimal initial thickness of soft tissues. I.e. to achieve necessary correction volume in a zone with small initial soft tissues thickness it is necessary to increase the amount of fat grafting stages. As the rates of soft tissues thickness in correction area change during 3-4 months after fat grafting remaining stable after this period it is expedient to assess postoperative results and to carry out repeated fat grafting not earlier than 4 months after operation.

  4. Validation of tissue quality parameters for donor corneas, designated for emergency cases: corneal graft survival.

    PubMed

    Rijneveld, Wilhelmina J; Wolff, Rachel; Völker-Dieben, Hennie J M; Pels, Elisabeth

    2011-12-01

    To validate tissue quality parameters for donor corneas designated for emergency grafting for corneal graft survival. In a longitudinal cohort follow-up study, 131 emergency penetrating grafts were studied. Grafts were performed with a pool of organ-cultured donor corneas designated for emergency grafting and prepared for immediate use with all safety tests performed. Assignation criteria were: corneas with a small superficial stromal opacity but meeting all selection criteria for PKP tissue and corneas without stromal opacity, but an endothelial cell density from 1800 to 2300 cells/mm(2) or mild polymegathism or pleomorphism. Cox multivariate regression analysis, Kaplan-Meier survival and log rank test were applied. Of the 131 keratoplasties, 115 could be followed. One eye was lost during surgery because of an expulsive bleeding. In 15 cases, a conjunctival transplantation finished off the penetrating graft. Corneal graft survival was not significantly related to the presence of PKP quality of the donor endothelium, neither with a cloudy graft nor with endothelial decompensation as the cause of failure. Main risk factors for a failed graft were vascularization of the host cornea (p=0.0001), the presence of a systemic auto immune disease in the recipient (p=0.003) and the disease leading to the (imminent) perforation and emergency graft (p=0.021). A selected pool of donor corneas designated for emergency grafting that does not interfere with the scheduled procedures allows more efficient and safe use of donor tissue in case of a(n) (imminent) perforation. Corneal graft survival rates justify the criteria for selection. © 2009 The Authors. Journal compilation © 2009 Acta Ophthalmol.

  5. Osteochondral Allograft of the Talus

    PubMed Central

    Bisicchia, Salvatore; Rosso, Federica; Amendola, Annunziato

    2014-01-01

    Osteochondral lesions of the talus are being recognized as an increasingly common injury. They are most commonly located postero-medially or antero-laterally, while centrally located lesions are uncommon. Large osteochondral lesions have significant biomechanical consequences and often require resurfacing with osteochondral autograft transfer, mosaicplasty, autologous chondrocyte implantation (or similar methods) or osteochondral allograft transplantation. Allograft procedures have become popular due to inherent advantages over other resurfacing techniques. Cartilage viability is one of the most important factors for successful clinical outcomes after transplantation of osteochondral allografts and is related to storage length and intra-operative factors. While there is abundant literature about osteochondral allograft transplantation in the knee, there are few papers about this procedure in the talus. Failure of non-operative management, initial debridement, curettage or microfractures are an indication for resurfacing. Patients should have a functional ankle motion, closed growth plates, absence of cartilage lesions on the tibial side. This paper reviews the published literature about osteochondral allograft transplantation of the talus focusing on indications, pre-operative planning, surgical approaches, postoperative management, results and complications of this procedure. PMID:25328456

  6. Polyethylene glycol-grafted bovine pericardium: a novel hybrid tissue resistant to calcification.

    PubMed

    Vasudev, S C; Chandy, T

    1999-02-01

    Calcification is a frequent cause of the clinical failure of bioprosthetic heart valves fabricated from glutaraldehyde pretreated bovine pericardium (GATBP). An investigation was made of the grafting of different molecular weight polyethylene glycol (PEG 600, 1500, 4000 and 6000) via glutaraldehyde (GA) linkages to bovine pericardium (BP) and of their stability and calcification. The process of the calcification profile was studied by in vitro experiments via incubating pericardial samples in a metastable solution of calcium phosphate. Calcification of bovine pericardium grafted with PEG 6000 was significantly decreased compared to low molecular weight PEG grafts or Sodium dodecyl sulphate- (SDS) and GA-treated tissues. The mechanical properties of these modified tissues after enzyme (Trypsin) digestion and calcification were investigated. The biocompatibility aspects of grafted tissues were also established by monitoring the platelet adhesion, octane contact angle and water of hydration. PEG 6000-grafted tissues retained the maximum strength in trypsin buffer and calcium phosphate solutions. Scanning electron micrographs revealed that the PEG-grafted bovine pericardium had substantially inhibited the platelet-surface attachment and their spreading. It is conceivable that high molecular weight polyethylene glycol-grafted pericardium (a hybrid tissue) may be a suitable calcium-resistant material for developing prosthetic valves due to their stability and biocompatibility.

  7. Engineering the mechanical and biological properties of nanofibrous vascular grafts for in situ vascular tissue engineering.

    PubMed

    Henry, Jeffrey J D; Yu, Jian; Wang, Aijun; Lee, Randall; Fang, Jun; Li, Song

    2017-08-17

    Synthetic small diameter vascular grafts have a high failure rate, and endothelialization is critical for preventing thrombosis and graft occlusion. A promising approach is in situ tissue engineering, whereby an acellular scaffold is implanted and provides stimulatory cues to guide the in situ remodeling into a functional blood vessel. An ideal scaffold should have sufficient binding sites for biomolecule immobilization and a mechanical property similar to native tissue. Here we developed a novel method to blend low molecular weight (LMW) elastic polymer during electrospinning process to increase conjugation sites and to improve the mechanical property of vascular grafts. LMW elastic polymer improved the elasticity of the scaffolds, and significantly increased the amount of heparin conjugated to the micro/nanofibrous scaffolds, which in turn increased the loading capacity of vascular endothelial growth factor (VEGF) and prolonged the release of VEGF. Vascular grafts were implanted into the carotid artery of rats to evaluate the in vivo performance. VEGF treatment significantly enhanced endothelium formation and the overall patency of vascular grafts. Heparin coating also increased cell infiltration into the electrospun grafts, thus increasing the production of collagen and elastin within the graft wall. This work demonstrates that LMW elastic polymer blending is an approach to engineer the mechanical and biological property of micro/nanofibrous vascular grafts for in situ vascular tissue engineering.

  8. Phase II Clinical Trial of Intraoral Grafting of Human Tissue-Engineered Oral Mucosa

    DTIC Science & Technology

    2016-10-01

    graft contracture and Wound Healing Index; and ancillary outcome measures of tissue perfusion measured graft color and laser Doppler flowmetry, and...infection, fluid loss, and foreign material contamination and relapse secondary to wound contracture. Oral mucosa is in limited supply for use in...reconstructive procedures in the oral cavity. This is especially prevalent after large avulsed soft tissue wounds involving the mouth and lips seen in

  9. Removal of an amalgam tattoo using a subepithelial connective tissue graft and laser deepithelialization.

    PubMed

    Campbell, Casey M; Deas, David E

    2009-05-01

    A 56-year-old female presented for periodontal treatment with a large amalgam tattoo located in alveolar mucosa on the facial aspect of her maxillary central incisors. The lesion had been present for 42 years since having endodontic surgery at teeth #8 and #9 after a traumatic childhood incident. A two-stage surgical approach was used to eliminate the lesion, beginning with a subepithelial connective tissue graft to increase tissue thickness subjacent to the amalgam tattoo. After 6 weeks of healing, the overlying pigmented tissue was removed using laser surgery to expose the underlying grafted connective tissue. After 2 months of healing following laser surgery, the amalgam pigmentation was completely removed, with good color match and an increased width of keratinized tissue at the surgical site. A relatively large amalgam tattoo in the esthetic zone can be adequately removed by a two-stage procedure using grafted palatal connective tissue and laser deepithelialization.

  10. Arthroscopically assisted autologous osteochondral transplantation for osteochondral lesions of the talar dome: an MRI and clinical follow-up study.

    PubMed

    Assenmacher, J A; Kelikian, A S; Gottlob, C; Kodros, S

    2001-07-01

    Osteochondral Lesions of the Talar Dome (OLT) are common problems encountered in orthopedics. Although the etiology remains uncertain, a myriad of treatment options exists. The authors describe arthroscopically assisted autologous osteochondral graft (OCG) transplantation procedures in the treatment of unstable OLTs in nine patients. The patients underwent standard preoperative MRI examination to assess fragment stability (using De Smet criteria for stability). Intraoperative arthroscopy was used to correlate the preoperative MRI assessment (using Cheng/Ferkel grading). After transplantation procedures, MRI (using De Smet criteria for stability) assessed graft incorporation for stability at an average of 9.3 months after the procedure. Preoperative MRI correlated highly with arthroscopic findings of OLT instability (sensitivity = 1.0). This has been demonstrated in the current orthopedic literature. The post transplantation MRI demonstrated stable graft osteointegration by De Smet criteria in all patients. Postoperative visual analogue pain scales showed significant improvement from preoperative assessment. Postoperative AOFAS Ankle-Hindfoot scores averaged 80.2 (S.D. +/- 18.9). Our favorable early results and those of other authors using similar techniques may validate OCG transplantation as a viable alternative for treating unstable osteochondral defects in the talus that are refractive to more commonly used surgical techniques.

  11. How I Manage Osteochondritis Dissecans.

    ERIC Educational Resources Information Center

    DiStefano, Vincent J.

    1986-01-01

    Osteochondritis dissecans, a lesion found most often on the femur at the knee joint, occurs most frequently in active adolescents. This article describes treatment for preadolescents, adolescents, and adults. Osteochondritus dissecans of the patella is also presented. (MT)

  12. How I Manage Osteochondritis Dissecans.

    ERIC Educational Resources Information Center

    DiStefano, Vincent J.

    1986-01-01

    Osteochondritis dissecans, a lesion found most often on the femur at the knee joint, occurs most frequently in active adolescents. This article describes treatment for preadolescents, adolescents, and adults. Osteochondritus dissecans of the patella is also presented. (MT)

  13. Implantation study of a tissue-engineered self-expanding aortic stent graft (bio stent graft) in a beagle model.

    PubMed

    Kawajiri, Hidetake; Mizuno, Takeshi; Moriwaki, Takeshi; Iwai, Ryosuke; Ishibashi-Ueda, Hatsue; Yamanami, Masashi; Kanda, Keiichi; Yaku, Hitoshi; Nakayama, Yasuhide

    2015-03-01

    The use of stent grafts for endovascular aortic repair has become an important treatment option for aortic aneurysms requiring surgery. This treatment has achieved excellent outcomes; however, problems like type 1 endoleaks and stent graft migration remain. Bio stent grafts (BSGs), which are self-expanding stents covered with connective tissue, were previously developed using "in-body tissue architecture" technology. We assessed their early adaptation to the aorta after transcatheter implantation in a beagle model. BSGs were prepared by subcutaneous embedding of acryl rods mounted with self-expanding nitinol stents in three beagles for 4 weeks (n = 3/dog). The BSGs were implanted as allografts into infrarenal abdominal aortas via the femoral artery of three other beagles. After 1 month of implantation, aortography revealed no stenosis or aneurysmal changes. The luminal surface of the BSGs was completely covered with neointimal tissue, including endothelialization, without any thrombus formation. The cover tissue could fuse the luminal surface of the native aorta with tight conjunctions even at both ends of the stents, resulting in complete impregnation of the strut into the reconstructed vascular wall, which is expected to prevent endoleaks and migration in clinical applications.

  14. Cadmium Telluride Quantum Dots as a Fluorescence Marker for Adipose Tissue Grafts.

    PubMed

    Deglmann, Claus J; Błażków-Schmalzbauer, Katarzyna; Moorkamp, Sarah; Susha, Andrei S; Herrler, Tanja; Giunta, Riccardo E; Wagner, Ernst; Rogach, Andrey L; Baumeister, Ruediger G; Ogris, Manfred

    2017-02-01

    Plastic and reconstructive surgeons increasingly apply adipose tissue grafting in a clinical setting, although the anticipation of graft survival is insecure. There are only few tools for tracking transplanted fat grafts in vivo.Murine adipose tissue clusters were incubated with negatively charged, mercaptoproprionic acid-coated cadmium telluride quantum dots (QDs) emitting in the dark red or near infrared. The intracellular localization of QDs was studied by confocal laser scanning microscopy.As a result, the adipose tissue clusters showed a proportional increase in fluorescence with increasing concentrations (1, 10, 16, 30, 50 nM) of cadmium telluride QDs. Laser scanning microscopy demonstrated a membrane bound localization of QDs. Vacuoles and cell nuclei of adipocytes were spared by QDs. We conclude that QDs were for the first time proven intracellular in adult adipocytes and demonstrate a strong fluorescence signal. Therefore, they may play an essential role for in vivo tracking of fat grafts.

  15. Augmentation of the rat jaw with autogeneic cortico-cancellous bone grafts and guided tissue regeneration.

    PubMed

    Donos, Nikolaos; Kostopoulos, Lambros; Karring, Thorkild

    2002-04-01

    The aim of the present study was to evaluate the effect of augmenting the maxillary alveolar ridge and the lateral aspect of the mandible with onlay autogeneic cortico-cancellous bone grafts that were covered with e-PTFE membranes. The experiment was carried out in 51 rats. In 15 rats, the edentulous maxillary jaw between the incisor and the first molar was augmented by means of an autogeneic ischiac bone graft that was fixed with a gold-coated microimplant. In one side, the graft was covered with an e-PTFE membrane, while the other side, which served as control, was treated without a membrane. In the other 36 rats, the lateral aspect of the mandible was augmented in both sides by means of an autogeneic ischiac bone graft that was fixed with a gold-coated or a titanium microimplant. In one side, the augmented area was covered with an e-PTFE membrane, while the contralateral side was treated without a membrane. Histological analysis at 60, 120 and 180 days after augmentation of the maxilla showed that, in the case of the test sites (where most of the membranes were either exposed or lost), the bone grafts presented extensive resorption and there was a lack of bone continuity between the graft and the recipient site. Similar findings were made at the non-membrane-treated control sides. In the case of augmentation of the mandible with membranes, the bone grafts were not resorbed, but were integrated into newly formed bone at the recipient site. In the control sides, the grafts presented varying degrees of resorption and integration into the recipient bone. It is concluded that, in comparison to bone grafting alone, onlay ischiac bone grafting combined with guided tissue regeneration eliminates the risk of bone graft resorption and ensures integration of the graft into newly formed bone at the recipient site, provided that closure of the operated area can be maintained during healing.

  16. Treatment of osteochondral injuries. Genetic engineering.

    PubMed

    Martinek, V; Fu, F H; Lee, C W; Huard, J

    2001-04-01

    Articular cartilage injuries are commonly encountered problems in sports medicine and orthopaedics. The treatment of chondral and osteochondral lesions, which possess only a very limited potential for healing, still represents a great challenge to clinicians and to scientists. Experimental investigations reported over the last 20 years have shown that a variety of methods, including implantation of periosteum, perichondrium, artificial matrices, growth factors, and transplanted cells, can stimulate formation of new cartilage. Genetic engineering--a combination of gene transfer techniques and tissue engineering--will facilitate new approaches to the treatment of articular cartilage injuries.

  17. Osteochondral Autograft from the Ipsilateral Femoral Head by Surgical Dislocation for Treatment of Femoral Head Fracture Dislocation: A Case Report.

    PubMed

    Won, Yougun; Lee, Gi Soo; Kim, Sang Bum; Kim, Sun Joong; Yang, Kyu Hyun

    2016-11-01

    As anatomical reduction of the articular surface of femoral head fractures and restoration of damaged cartilage are essential for good long-term results, many treatment options have been suggested, including fixation of the fracture using various surgical exposures and implants, as well as arthroscopic irrigation and debridement, bone marrow stimulating techniques, osteochondral allograft, autograft, and autogenous chondrocyte implantation. We report a case of osteochondral autograft harvested from its own femoral articular surface through surgical hip dislocation. The osteochondral graft was harvested from the inferior non-weight-bearing articular surface and grafted to the osteochondral defect. One year later, the clinical and radiological results were good, without the collapse of the femoral head or arthritic change. This procedure introduced in our case is considered convenient and able to lessen surgical time without morbidity of the donor site associated with the harvest.

  18. Fabrication of engineered heart tissue grafts from alginate/collagen barium composite microbeads.

    PubMed

    Bai, X P; Zheng, H X; Fang, R; Wang, T R; Hou, X L; Li, Y; Chen, X B; Tian, W M

    2011-08-01

    Cardiac tissue engineering holds great promise for the treatment of myocardial infarction. However, insufficient cell migration into the scaffolds used and inflammatory reactions due to scaffold biodegradation remain as issues to be addressed. Engineered heart tissue (EHT) grafts fabricated by means of a cell encapsulation technique provide cells with a tissue-like environment, thereby potentially enhancing cellular processes such as migration, proliferation, and differentiation, and tissue regeneration. This paper presents a study on the fabrication and characterization of EHT grafts from novel alginate/collagen composite microbeads by means of cell encapsulation. Specifically, the microbeads were fabricated from alginate and collagen by barium ion cross-linking, with neonatal rat cardiomyocytes encapsulated in the composite microbeads during the fabrication of the EHT grafts. To evaluate the suitablity of these EHT grafts for heart muscle repair, the growth of cardiac cells in the microbeads was examined by means of confocal microscopy and staining with DAPI and F-actin. The EHT grafts were analyzed by scanning electron microscopy and transmission electron microscopy, and the contractile function of the EHT grafts monitored using a digital video camera at different time points. The results show the proliferation of cardiac cells in the microbeads and formation of interconnected multilayer heart-like tissues, the presence of well-organized and dense cell structures, the presence of intercalated discs and spaced Z lines, and the spontaneous synchronized contractility of EHT grafts (at a rate of 20-30 beats min(-1) after two weeks in culture). Taken together, these observations demonstrate that the novel alginate/collagen composite microbeads can provide a tissue-like microenvironment for cardiomyocytes that is suitable for fabricating native heart-like tissues.

  19. Plasma-modified and polyethylene glycol-grafted polymers for potential tissue engineering applications.

    PubMed

    Svorcík, V; Makajová, Z; Kasálková-Slepicková, N; Kolská, Z; Bacáková, L

    2012-08-01

    Modified and grafted polymers may serve as building blocks for creating artificial bioinspired nanostructured surfaces for tissue engineering. Polyethylene (PE) and polystyrene (PS) were modified by Ar plasma and the surface of the plasma activated polymers was grafted with polyethylene glycol (PEG). The changes in the surface wettability (contact angle) of the modified polymers were examined by goniometry. Atomic Force Microscopy (AFM) was used to determine the surface roughness and morphology and electrokinetical analysis (Zeta potential) characterized surface chemistry of the modified polymers. Plasma treatment and subsequent PEG grafting lead to dramatic changes in the polymer surface morphology, roughness and wettability. The plasma treated and PEG grafted polymers were seeded with rat vascular smooth muscle cells (VSMCs) and their adhesion and proliferation were studied. Biological tests, performed in vitro, show increased adhesion and proliferation of cells on modified polymers. Grafting with PEG increases cell proliferation, especially on PS. The cell proliferation was shown to be an increasing function of PEG molecular weight.

  20. Repair of porcine articular cartilage defect with a biphasic osteochondral composite.

    PubMed

    Jiang, Ching-Chuan; Chiang, Hongsen; Liao, Chun-Jen; Lin, Yu-Ju; Kuo, Tzong-Fu; Shieh, Chang-Shun; Huang, Yi-You; Tuan, Rocky S

    2007-10-01

    Autologous chondrocyte implantation (ACI) has been recently used to treat cartilage defects. Partly because of the success of mosaicplasty, a procedure that involves the implantation of native osteochondral plugs, it is of potential significance to consider the application of ACI in the form of biphasic osteochondral composites. To test the clinical applicability of such composite construct, we repaired osteochondral defect with ACI at low cell-seeding density on a biphasic scaffold, and combined graft harvest and implantation in a single surgery. We fabricated a biphasic cylindrical porous plug of DL-poly-lactide-co-glycolide, with its lower body impregnated with beta-tricalcium phosphate as the osseous phase. Osteochondral defects were surgically created at the weight-bearing surface of femoral condyles of Lee-Sung mini-pigs. Autologous chondrocytes isolated from the cartilage were seeded into the upper, chondral phase of the plug, which was inserted by press-fitting to fill the defect. Defects treated with cell-free plugs served as control. Outcome of repair was examined 6 months after surgery. In the osseous phase, the biomaterial retained in the center and cancellous bone formed in the periphery, integrating well with native subchondral bone with extensive remodeling, as depicted on X-ray roentgenography by higher radiolucency. In the chondral phase, collagen type II immunohistochemistry and Safranin O histological staining showed hyaline cartilage regeneration in the experimental group, whereas only fibrous tissue formed in the control group. On the International Cartilage Repair Society Scale, the experimental group had higher mean scores in surface, matrix, cell distribution, and cell viability than control, but was comparable with the control group in subchondral bone and mineralization. Tensile stress-relaxation behavior determined by uni-axial indentation test revealed similar creep property between the surface of the experimental specimen and native

  1. Alveolar ridge augmentation by connective tissue grafting using a pouch method and modified connective tissue technique: A prospective study

    PubMed Central

    Agarwal, Ashish; Gupta, Narinder Dev

    2015-01-01

    Background: Localized alveolar ridge defect may create physiological and pathological problems. Developments in surgical techniques have made it simpler to change the configuration of a ridge to create a more aesthetic and more easily cleansable shape. The purpose of this study was to compare the efficacy of alveolar ridge augmentation using a subepithelial connective tissue graft in pouch and modified connective tissue graft technique. Materials and Methods: In this randomized, double blind, parallel and prospective study, 40 non-smoker individuals with 40 class III alveolar ridge defects in maxillary anterior were randomly divided in two groups. Group I received modified connective tissue graft, while group II were treated with subepithelial connective tissue graft in pouch technique. The defect size was measured in its horizontal and vertical dimension by utilizing a periodontal probe in a stone cast at base line, after 3 months, and 6 months post surgically. Analysis of variance and Bonferroni post-hoc test were used for statistical analysis. A two-tailed P < 0.05 was considered to be statistically significant. Results: Mean values in horizontal width after 6 months were 4.70 ± 0.87 mm, and 4.05 ± 0.89 mm for group I and II, respectively. Regarding vertical heights, obtained mean values were 4.75 ± 0.97 mm and 3.70 ± 0.92 mm for group I and group II, respectively. Conclusion: Within the limitations of this study, connective tissue graft proposed significantly more improvement as compare to connective tissue graft in pouch. PMID:26759591

  2. Tissue Responses to Endovascular Stent Grafts for Saccular Abdominal Aortic Aneurysms in a Canine Model

    PubMed Central

    Kim, Hyun Beom; Choi, Young Ho; So, Young Ho; Min, Seung-Kee; Kim, Hyo-Cheol; Kim, Young Il; Park, Jae Hyung

    2012-01-01

    We investigated tissue responses to endoskeleton stent grafts for saccular abdominal aortic aneurysms (AAAs) in canines. Saccular AAAs were made with Dacron patch in 8 dogs, and were excluded by endoskeleton stent grafts composed of nitinol stent and expanded polytetrafluoroethylene graft. Animals were sacrificed at 2 months (Group 1; n = 3) or 6 months (Group 2; n = 5) after the placement, respectively. The aortas embedding stent grafts were excised en bloc for gross inspection and sliced at 5 to 8 mm intervals for histopathologic evaluation. Stent grafts were patent in all except a dog showing a thrombotic occlusion in Group 2. In the 7 dogs with patent lumen, the graft overhanging the saccular aneurysm was covered by thick or thin thrombi with no endothelial layer, and the graft over the aortic wall was completely covered by neointima with an endothelial layer. Transgraft cell migration was less active at an aneurysm than at adjacent normal aorta. In conclusion, endoskeleton stent grafts over saccular aneurysms show no endothelial coverage and poor transgraft cell migration in a canine model. PMID:23091313

  3. Tissue-engineered bone grafts for osteoplasty in patients with cleft alveolus.

    PubMed

    Pradel, Winnie; Lauer, Günter

    2012-11-01

    Alveolar bone grafting is an integral part of the treatment concept in cleft palate patients. As an alternative to autogenous bone, tissue-engineered grafts have found some clinical application. The aim of the present study has been to compare ossification in the cleft area using tissue-engineered grafts in a case series of patients with ossification after transplantation of autogenous spongious bone as the gold standard in alveoloplasty. Eight children with complete cleft lips and cleft palates were included in the study. In four children (group A), the cleft defect was filled with tissue-engineered bone (autogenous osteoblasts cultured on demineralized bone matrix Osteovit(®)); as control in another 4 children (group B), the alveoloplasty was performed using spongious iliac bone. Preoperative and 6 months postoperative cone-beam computed tomography was performed, and volumes of the remaining cleft defects were calculated using 3D navigation software. Wound healing was uneventful in both groups. Six months postoperatively the mean volume of the cleft was 0.55±0.24cm(3) after grafting of tissue-engineered bone (group A) and 0.59±0.23cm(3) after transplantation of autogenous spongiosa. In group A, 40.9% of the cleft defect was ossified; in the control group (group B), 36.6%. Tissue-engineered bone is a promising alternative in alveolar bone grafting and no disadvantages were observed in comparison to the gold standard. Copyright © 2012 Elsevier GmbH. All rights reserved.

  4. Infiltrating cells from host brain restore the microglial population in grafted cortical tissue

    PubMed Central

    Wang, Cong; Tao, Sijue; Fang, Yukun; Guo, Jing; Zhu, Lirui; Zhang, Shengxiang

    2016-01-01

    Transplantation of embryonic cortical tissue is considered as a promising therapy for brain injury. Grafted neurons can reestablish neuronal network and improve cortical function of the host brain. Microglia is a key player in regulating neuronal survival and plasticity, but its activation and dynamics in grafted cortical tissue remain unknown. Using two-photon intravital imaging and parabiotic model, here we investigated the proliferation and source of microglia in the donor region by transplanting embryonic cortical tissue into adult cortex. Live imaging showed that the endogenous microglia of the grafted tissue were rapidly lost after transplantation. Instead, host-derived microglia infiltrated and colonized the graft. Parabiotic model suggested that the main source of infiltrating cells is the parenchyma of the host brain. Colonized microglia proliferated and experienced an extensive morphological transition and eventually differentiated into resting ramified morphology. Collectively, these results demonstrated that donor tissue has little contribution to the activated microglia and host brain controls the microglial population in the graft. PMID:27615195

  5. Osseous differentiation of human fat tissue grafts: From tissue engineering to tissue differentiation

    PubMed Central

    Bondarava, Maryna; Cattaneo, Chiara; Ren, Bin; Thasler, Wolfgang E.; Jansson, Volkmar; Müller, Peter E.; Betz, Oliver B.

    2017-01-01

    Conventional bone tissue engineering approaches require isolation and in vitro propagation of autologous cells, followed by seeding on a variety of scaffolds. Those protracted procedures impede the clinical applications. Here we report the transdifferentiation of human fat tissue fragments retrieved from subcutaneous fat into tissue with bone characteristics in vitro without prior cell isolation and propagation. 3D collagen-I cultures of human fat tissue were cultivated either in growth medium or in osteogenic medium (OM) with or without addition of Bone Morphogenetic Proteins (BMPs) BMP-2, BMP-7 or BMP-9. Ca2+ depositions were observed after two weeks of osteogenic induction which visibly increased when either type of BMP was added. mRNA levels of alkaline phosphatase (ALP) and osteocalcin (OCN) increased when cultured in OM alone but addition of BMP-2, BMP-7 or BMP-9 caused significantly higher expression levels of ALP and OCN. Immunofluorescent staining for OCN, osteopontin and sclerostin supported the observed real-time-PCR data. BMP-9 was the most effective osteogenic inducer in this system. Our findings reveal that tissue regeneration can be remarkably simplified by omitting prior cell isolation and propagation, therefore removing significant obstacles on the way to clinical applications of much needed regeneration treatments. PMID:28054585

  6. Effect of connective tissue grafting on peri-implant tissue in single immediate implant sites: a RCT.

    PubMed

    Zuiderveld, Elise G; Meijer, Henny J A; den Hartog, Laurens; Vissink, Arjan; Raghoebar, Gerry M

    2017-09-23

    To assess the effect of connective tissue grafting on the mid-buccal mucosal level (MBML) of immediately placed and provisionalized single implants in the maxillofacial esthetic zone. Sixty patients with a failing tooth were provided with an immediately placed and provisionalized implant. During implant placement, patients randomly received either a connective tissue graft from the maxillary tuberosity (n=30, test group) or no graft (n=30, control group). Follow-up visits were at one (T1 ) and twelve months (T12 ) after final crown placement. The primary outcome measure was any change in MBML compared to the pre-operative situation. In addition, gingival biotype, esthetics (using the Pink Esthetic Score-White Esthetic Score), marginal bone level, soft tissue peri-implant parameters and patient satisfaction were assessed. The mean MBML change at T12 was -0.5±1.1mm in the control group and 0.1±0.8mm in the test group (p=0.03). No significant differences regarding other outcome variables were observed, neither was gingival biotype associated with a gain or loss in MBML. This one-year study shows that connective tissue grafting in single, immediately placed and provisionalized implants leads to less recession of the peri-implant soft tissue at the mid-buccal aspect, irrespective of the gingival biotype (www.trialregister.nl: TC3815). This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  7. Grinding and polishing instead of sectioning for the tissue samples with a graft: Implications for light and electron microscopy.

    PubMed

    Mukhamadiyarov, Rinat A; Sevostyanova, Victoria V; Shishkova, Daria K; Nokhrin, Andrey V; Sidorova, Olga D; Kutikhin, Anton G

    2016-06-01

    A broad use of the graft replacement requires a detailed investigation of the host-graft interaction, including both histological examination and electron microscopy. A high quality sectioning of the host tissue with a graft seems to be complicated; in addition, it is difficult to examine the same tissue area by both of the mentioned microscopy techniques. To solve these problems, we developed a new technique of epoxy resin embedding with the further grinding, polishing, and staining. Graft-containing tissues prepared by grinding and polishing preserved their structure; however, sectioning frequently required the explantation of the graft and led to tissue disintegration. Moreover, stained samples prepared by grinding and polishing may then be assessed by both light microscopy and backscattered scanning electron microscopy. Therefore, grinding and polishing outperform sectioning when applied to the tissues with a graft.

  8. The use of embryonic cells in the treatment of osteochondral defects of the knee: an ovine in vivo study

    PubMed Central

    MANUNTA, ANDREA FABIO; ZEDDE, PIETRO; PILICCHI, SUSANNA; ROCCA, STEFANO; POOL, ROY R.; DATTENA, MARIA; MASALA, GEROLAMO; MARA, LAURA; CASU, SARA; SANNA, DANIELA; MANUNTA, MARIA LUCIA; PASSINO, ERALDO SANNA

    2016-01-01

    Purpose the aim of this study was to determine whether local delivery of embryonic stem-like (ESL) cells into osteochondral defects in the femoral condyles of sheep would enhance regeneration of hyaline articular cartilage. Methods male ESL cells embedded in fibrin glue were engrafted into osteochondral defects in the medial condyles (ESL-M) of the left femur in 22 ewes. An identical defect was created in the medial condyle of the contralateral stifle joint and left untreated as a control (empty defect, ED). The ewes were divided into 5 groups. Four sheep each were euthanized at 1, 2, 6, and 12 months from surgery, and 6 ewes were euthanized 24 months post-implantation. To study the effect of varying loads on the long-term regeneration process, an identical defect was also created and ESL cell engraftment performed in the lateral condyle (ESL-L) of the left stifle joint of the animals in the 12- and 24-month groups. The evaluation of regenerated tissue was performed by biomechanical, macroscopic, histological, immunohistochemical (collagen type II) and fluorescent in situ hybridization (FISH) assays. Results no significant differences were found between treated and control sites in the biomechanical assays at any time point. ESL cell grafts showed significantly greater macroscopic evidence of regeneration as compared to controls at 24 months after surgery; significantly better histological evidence of repair in ESL-M samples versus controls was found throughout the considered period. At 24 months from surgery there was significantly improved integration of graft edges with the host tissue in the ESL-M as compared to the ESL-L samples, demonstrating that load bearing positively affects the long-term regeneration process. Conclusions ESL cells enhanced the regeneration of hyaline cartilage. FISH confirmed that the regenerative tissue originated from ESL cells. Clinical Relevance ESL cells are able to self-renew for prolonged periods without differentiation and, most

  9. In vivo applications of electrospun tissue-engineered vascular grafts: a review.

    PubMed

    Rocco, Kevin A; Maxfield, Mark W; Best, Cameron A; Dean, Ethan W; Breuer, Christopher K

    2014-12-01

    There is great clinical demand for synthetic vascular grafts with improved long-term efficacy. The ideal vascular conduit is easily implanted, nonthrombogenic, biocompatible, resists aneurysmal dilatation, and ultimately degrades or is assimilated as the patient remodels the graft into tissue resembling native vessel. The field of vascular tissue engineering offers an opportunity to design the ideal synthetic graft, and researchers have evaluated a variety of methods and materials for use in graft construction. Electrospinning is one method that has received considerable attention within tissue engineering for constructing so-called tissue scaffolds. Tissue scaffolds are temporary, porous structures which are commonly composed of bioresorbable polymers that promote native tissue ingrowth and have degradation kinetics compatible with a patient's rate of extracellular matrix production in order to successfully transit from synthetic conduits into neovessels. In this review, we summarize the history of tissue-engineered vascular grafts (TEVG), focusing on scaffolds generated by the electrospinning process, and discuss in vivo applications. We review the materials commonly employed in this approach and the preliminary results after implantation in animal models in order to gauge clinical viability of the electrospinning process for TEVG construction. Scientists have studied electrospinning technology for decades, but only recently has it been orthotopically evaluated in animal models such as TEVG. Advantages of electrospun TEVG include ease of construction, favorable cellular interactions, control of scaffold features such as fiber diameter and pore size, and the ability to choose from a variety of polymers possessing a range of mechanical and chemical properties and degradation kinetics. Given its advantages, electrospinning technology merits investigation for use in TEVG, but an emphasis on long-term in vivo evaluation is required before its role in clinical vascular

  10. Arthroscopic Curettage and Bone Grafting of Bone Cysts of the Talar Body.

    PubMed

    Lui, Tun Hing

    2017-02-01

    Talar bone cysts can develop as a result of osteochondral lesions of the talus. This can be a source of deep ankle pain. Open debridement and bone grafting of the bone cysts requires extensive soft tissue dissection and malleolar osteotomy. Removal of normal cartilage of the talus is frequently required to approach the bone cysts. Alternatively, the cysts can be grafted arthroscopically with minimal disruption of the normal cartilage surface. The key to success is careful preoperative planning with a computed tomogram of the ankle. Bone cyst of the posterior half of the talar body can be grafted via posterior ankle endoscopy. Bone cyst of the anterior half of the talar body can be debrided and grafted via anterior talar osseous portals. The purpose of this technical note is to describe a minimally invasive approach of curettage and bone grafting of the talar bone cysts with preservation of the articular surfaces.

  11. Multilayer scaffolds in orthopaedic tissue engineering.

    PubMed

    Atesok, Kivanc; Doral, M Nedim; Karlsson, Jon; Egol, Kenneth A; Jazrawi, Laith M; Coelho, Paulo G; Martinez, Amaury; Matsumoto, Tomoyuki; Owens, Brett D; Ochi, Mitsuo; Hurwitz, Shepard R; Atala, Anthony; Fu, Freddie H; Lu, Helen H; Rodeo, Scott A

    2016-07-01

    The purpose of this study was to summarize the recent developments in the field of tissue engineering as they relate to multilayer scaffold designs in musculoskeletal regeneration. Clinical and basic research studies that highlight the current knowledge and potential future applications of the multilayer scaffolds in orthopaedic tissue engineering were evaluated and the best evidence collected. Studies were divided into three main categories based on tissue types and interfaces for which multilayer scaffolds were used to regenerate: bone, osteochondral junction and tendon-to-bone interfaces. In vitro and in vivo studies indicate that the use of stratified scaffolds composed of multiple layers with distinct compositions for regeneration of distinct tissue types within the same scaffold and anatomic location is feasible. This emerging tissue engineering approach has potential applications in regeneration of bone defects, osteochondral lesions and tendon-to-bone interfaces with successful basic research findings that encourage clinical applications. Present data supporting the advantages of the use of multilayer scaffolds as an emerging strategy in musculoskeletal tissue engineering are promising, however, still limited. Positive impacts of the use of next generation scaffolds in orthopaedic tissue engineering can be expected in terms of decreasing the invasiveness of current grafting techniques used for reconstruction of bone and osteochondral defects, and tendon-to-bone interfaces in near future.

  12. The role of tissue adaptation and graft size in immune tolerance.

    PubMed

    Hauben, Ehud; Roncarolo, Maria Grazia; Draghici, Elena; Nevo, Uri

    2007-11-01

    Understanding how immune tolerance is induced and maintained is critical for our approach to immune-related diseases. Ecoimmunity is a new theory that views the immune system-tissue interaction as a co-adapting predator-prey system. Ecoimmunity suggests that tissues adapt to the selective immune pressure during ontogeny and throughout life. Therefore, immune tolerance towards 'self' represents a symmetric balance between the propensity of the immune system to prey on 'self' cells, and the tissue's specific capacity to undergo phenotypic adaptations in order to avoid destructive immune interaction. According to this theory, we hypothesized that tissues of adult immune-deficient mice, which are not exposed to selective immune pressure, will not withstand immune activity and will therefore display higher susceptibility to graft rejection. To test this prediction, C57Bl/6 wild type female mice were rendered diabetic by streptozotocin and transplanted with syngeneic pancreatic islets isolated from either immune-deficient C57Bl/6 SCID or wild type females. Remarkably, recipients of islet grafts from immune-deficient syngeneic donors displayed significantly impaired glucose homeostasis compared to mice transplanted with islets of wild type donors (p<0.001, two way repeated measures ANOVA). The severity of this impairment was correlated with islet graft size, suggesting a capacity of transplanted islets to gradually acquire a tolerogenic phenotype. These findings support the view of graft survival that is based on 'natural selection' of tissue cells. In addition, we describe a new experimental system for molecular characterization of self-tolerance.

  13. Tissue specificity in rat peripheral nerve regeneration through combined skeletal muscle and vein conduit grafts.

    PubMed

    Tos, P; Battiston, B; Geuna, S; Giacobini-Robecchi, M G; Hill, M A; Lanzetta, M; Owen, E R

    2000-01-01

    Diffusible factors from the distal stumps of transected peripheral nerves exert a neurotropic effect on regenerating nerves in vivo (specificity). This morphological study was designed to investigate the existence of tissue specificity in peripheral nerve fiber regeneration through a graft of vein filled with fresh skeletal muscle. This tubulization technique demonstrated experimental and clinical results similar to those obtained with traditional autologous nerve grafts. Specifically, we used Y-shaped grafts to assess the orientation pattern of regenerating axons in the distal stump tissue. Animal models were divided into four experimental groups. The proximal part of the Y-shaped conduit was sutured to a severed tibial nerve in all experiments. The two distal stumps were sutured to different targets: group A to two intact nerves (tibial and peroneal), group B to an intact nerve and an unvascularized tendon, group C to an intact nerve and a vascularized tendon, and group D to a nerve graft and an unvascularized tendon. Morphological evaluation by light and electron microscopy was conducted in the distal forks of the Y-shaped tube. Data showed that almost all regenerating nerve fibers spontaneously oriented towards the nerve tissue (attached or not to the peripheral innervation field), showing a good morphological pattern of regeneration in both the early and late phases of regeneration. When the distal choice was represented by a tendon (vascularized or not), very few nerve fibers were detected in the corresponding distal fork of the Y-shaped graft. These results show that, using the muscle-vein-combined grafting technique, regenerating axons are able to correctly grow and orientate within the basement membranes of the graft guided by the neurotropic lure of the distal nerve stump. Copyright 2000 Wiley-Liss, Inc.

  14. Diagnosing, planning and evaluating osteochondral ankle defects with imaging modalities

    PubMed Central

    van Bergen, Christiaan JA; Gerards, Rogier M; Opdam, Kim TM; Terra, Maaike P; Kerkhoffs, Gino MMJ

    2015-01-01

    This current concepts review outlines the role of different imaging modalities in the diagnosis, preoperative planning, and follow-up of osteochondral ankle defects. An osteochondral ankle defect involves the articular cartilage and subchondral bone (usually of the talus) and is mostly caused by an ankle supination trauma. Conventional radiographs are useful as an initial imaging tool in the diagnostic process, but have only moderate sensitivity for the detection of osteochondral defects. Computed tomography (CT) and magnetic resonance imaging (MRI) are more accurate imaging modalities. Recently, ultrasonography and single photon emission CT have been described for the evaluation of osteochondral talar defects. CT is the most valuable modality for assessing the exact location and size of bony lesions. Cartilage and subchondral bone damage can be visualized using MRI, but the defect size tends to be overestimated due to bone edema. CT with the ankle in full plantar flexion has been shown a reliable tool for preoperative planning of the surgical approach. Postoperative imaging is useful for objective assessment of repair tissue or degenerative changes of the ankle joint. Plain radiography, CT and MRI have been used in outcome studies, and different scoring systems are available. PMID:26716090

  15. Reconstruction of interdental papilla using autogenous bone and connective tissue grafts

    PubMed Central

    Muthukumar, Santhanakrishnan; Ajit, Pooja; Sundararajan, Shiyamali; Rao, Suresh Ranga

    2016-01-01

    Previous studies have reported the management of Class I and II papillary defects, but knowledge on Class III defects, estimated to have a poor periodontal prognosis, remains minimal. In this case report, a Class III papillary defect reconstruction was attempted mainly since the patient reported with difficulty in phonetics. In Stage I, autogenous bone graft from the maxillary tuberosity and subepithelial connective tissue graft was augmented to decrease the distance between the interdental bone crest and contact point, simultaneously achieving a switch in the periodontal biotype. In Stage II, subepithelial connective tissue graft was augmented to achieve papillary fill. To avoid manual errors associated with quantifying the posttreatment outcomes, image data processing ImageJ software was used to assess the length, perimeter, and surface area of papillary loss using the preoperative images.

  16. Tissue-engineered skin preserving the potential of epithelial cells to differentiate into hair after grafting.

    PubMed

    Larouche, Danielle; Cuffley, Kristine; Paquet, Claudie; Germain, Lucie

    2011-03-01

    The aim of this study was to evaluate whether tissue-engineered skin produced in vitro was able to sustain growth of hair follicles in vitro and after grafting. Different tissues were designed. Dissociated newborn mouse keratinocytes or newborn mouse hair buds (HBs) were added onto dermal constructs consisting of a tissue-engineered cell-derived matrix elaborated from either newborn mouse or adult human fibroblasts cultured with ascorbic acid. After 7-21 days of maturation at the air-liquid interface, no hair was noticed in vitro. Epidermal differentiation was observed in all tissue-engineered skin. However, human fibroblast-derived tissue-engineered dermis (hD) promoted a thicker epidermis than mouse fibroblast-derived tissue-engineered dermis (mD). In association with mD, HBs developed epithelial cyst-like inclusions presenting outer root sheath-like attributes. In contrast, epidermoid cyst-like inclusions lined by a stratified squamous epithelium were present in tissues composed of HBs and hD. After grafting, pilo-sebaceous units formed and hair grew in skin elaborated from HBs cultured 10-26 days submerged in culture medium in association with mD. However, the number of normal hair follicles decreased with longer culture time. This hair-forming capacity after grafting was not observed in tissues composed of hD overlaid with HBs. These results demonstrate that epithelial stem cells can be kept in vitro in a permissive tissue-engineered dermal environment without losing their potential to induce hair growth after grafting.

  17. Bioactive glass and connective tissue graft used to treat intrabony periodontal defects.

    PubMed

    Deliberador, Tatiana Miranda; Trotta, Daniel Rizzo; Klug, Luis Gustavo; Zielak, Joao Cesar; Giovanini, Allan Fernando

    2013-07-01

    Different techniques and materials can be used to treat intrabony periodontal defects caused by periodontal diseases. This case report presents an intrabony periodontal defect with bioactive glass and connective tissue graft used as a barrier. Probing depth and clinical attachment gain were reduced at 6 and 12 months post-treatment.

  18. Tissue-engineered dermo-epidermal skin grafts prevascularized with adipose-derived cells.

    PubMed

    Klar, Agnieszka S; Güven, Sinan; Biedermann, Thomas; Luginbühl, Joachim; Böttcher-Haberzeth, Sophie; Meuli-Simmen, Claudia; Meuli, Martin; Martin, Ivan; Scherberich, Arnaud; Reichmann, Ernst

    2014-06-01

    The major problem in skin grafting is that tissue-engineered skin grafts after their transplantation are initially entirely dependent on diffusion. Since this process is slow and inefficient, nutrients, growth factors, and oxygen will insufficiently be supplied and the regenerating graft will undergo a physiological crisis, resulting in scar-like dermal structures and shrinkage. The tissue-engineering of a vascular network in human dermo-epidermal skin substitutes (DESS) is a promising approach to overcome this limitation. Here we report, for the first time, on the use of the adipose stromal vascular fraction (SVF)-derived endothelial cell population to tissue-engineer DESS containing a highly efficient capillary plexus. To develop vascular networks in vitro, we employed optimized 3D fibrin or collagen type I hydrogel systems. Upon transplantation onto immune-deficient rats, these pre-formed vascular networks anastomosed to the recipient's vasculature within only four days. As a consequence, the neo-epidermis efficiently established tissue homeostasis, the dermis underwent almost no contraction, and showed sustained epidermal coverage in vivo. Overall, the here described rapid and efficient perfusion of SVF-based skin grafts opens new perspectives for the treatment of hitherto unmet clinical needs in burn/plastic surgery and dermatology. Copyright © 2014 Elsevier Ltd. All rights reserved.

  19. The effect of supracrestal tricalcium phosphate ceramic-microfibrillar collagen grafting on postsurgical soft tissue levels.

    PubMed

    Blumenthal, N M

    1988-01-01

    In this clinical study, a combined tricalcium phosphate ceramic (TCP) microfibrillar collagen supracrestal graft was used in conjunction with pocket elimination surgery as a soft tissue space filler to maintain marginal and interproximal tissue heights. Fifteen volunteer adult patients (8 male, 7 female) aged 37 to 48 years were selected. Standardized clinical probing measurements were used to evaluate soft tissue recession, clinical attachment, and probing pocket depths. Intraoral photographs and radiographs supplemented documentation. Twenty-five paired test and control sites were evaluated. Following flap surgery and minor osteoplasty, test defects received supracrestal grafts of TCP microfibrillar collagen prior to flap closure. Intrabony defects were excluded. Treatment modalities were compared by analyzing changes in clinical probing measurements over four months using the students t test for paired samples at the 0.01 level of significance. Supracrestal grafting resulted in 16.74% less soft tissue recession than the control sites. In addition, there was a significant gain in clinical attachment levels compared with the controls. All implanted sites demonstrated an advantage in the use of supracrestal TCP-collagen grafts compared with the controls. This treatment modality may have value in reducing esthetic deformities, food impaction areas, and root exposure that often occur following periodontal surgery.

  20. Evaluation of a hybrid scaffold/cell construct in repair of high-load-bearing osteochondral defects in rabbits.

    PubMed

    Shao, Xin Xin; Hutmacher, Dietmar W; Ho, Saey Tuan; Goh, James C H; Lee, Eng Hin

    2006-03-01

    The objective of this study was to evaluate the feasibility and potential of a hybrid scaffold system in large- and high-load-bearing osteochondral defects repair. The implants were made of medical-grade PCL (mPCL) for the bone compartment whereas fibrin glue was used for the cartilage part. Both matrices were seeded with allogenic bone marrow-derived mesenchymal cells (BMSC) and implanted in the defect (4 mm diameter x 5 mm depth) on medial femoral condyle of adult New Zealand White rabbits. Empty scaffolds were used at the control side. Cell survival was tracked via fluorescent labeling. The regeneration process was evaluated by several techniques at 3 and 6 months post-implantation. Mature trabecular bone regularly formed in the mPCL scaffold at both 3 and 6 months post-operation. Micro-Computed Tomography showed progression of mineralization from the host-tissue interface towards the inner region of the grafts. At 3 months time point, the specimens showed good cartilage repair. In contrast, the majority of 6 months specimens revealed poor remodeling and fissured integration with host cartilage while other samples could maintain good cartilage appearance. In vivo viability of the transplanted cells was demonstrated for the duration of 5 weeks. The results demonstrated that mPCL scaffold is a potential matrix for osteochondral bone regeneration and that fibrin glue does not inherit the physical properties to allow for cartilage regeneration in a large and high-load-bearing defect site.

  1. Effect of different cryoprotectant agents on spermatogenesis efficiency in cryopreserved and grafted neonatal mouse testicular tissue.

    PubMed

    Yildiz, Cengiz; Mullen, Brendan; Jarvi, Keith; McKerlie, Colin; Lo, Kirk C

    2013-08-01

    Restoration of male fertility associated with use of the cryopreserved testicular tissue would be a significant advance in human and animal assisted reproductive technology. The purpose of this study was to test the effects of four different cryoprotectant agents (CPA) on spermatogenesis and steroidogenesis in cryopreserved and allotransplanted neonatal mouse testicular tissue. Hank's balanced salt solution (HBSS) with 5% fetal bovine serum including either 0.7 M dimethyl sulfoxide (DMSO), 0.7 M propylene glycol (PrOH), 0.7 M ethylene glycol (EG), or glycerol was used as the cryoprotectant solution. Donor testes were collected and dissected from neonatal pups of CD-1 mice (one day old). Freezing and seeding of the testicular whole tissues was performed using an automated controlled-rate freezer. Four fresh (non-frozen) or frozen-thawed pieces of testes were subcutaneously grafted onto the hind flank of each castrated male NCr nude recipient mouse and harvested after 3 months. Fresh neonatal testes grafts recovered from transplant sites had the most advanced rate of spermatogenesis with elongated spermatid and spermatozoa in 46.6% of seminiferous tubules and had higher levels of serum testosterone compared to all other frozen-thawed-graft groups (p<0.05). Fresh grafts and frozen-thawed grafts in the DMSO group had the highest rate of tissue survival compared to PrOH, EG, and glycerol after harvesting (p>0.05). The most effective CPA for the freezing and thawing of neonatal mouse testes was DMSO in comparison with EG (p<0.05) in both pre-grafted and post-grafted tissues based on histopathological evaluation. Likewise, the highest level of serum testosterone was obtained from the DMSO CPA group compared to all other cryoprotectants evaluated (p<0.05). The typical damage observed in the frozen-thawed grafts included disruption of the interstitial stroma, intercellular connection ruptures, and detachment of spermatogonia from the basement membrane. These findings

  2. Investigation of a hybrid method of soft tissue graft fixation for anterior cruciate ligament reconstruction.

    PubMed

    Au, Anthony G; Otto, David D; Raso, V James; Amirfazli, Alidad

    2005-04-01

    To increase knee stability following anterior cruciate ligament (ACL) reconstruction, development of increasingly stronger and stiffer fixation is required. This study assessed the initial pullout force, stiffness of fixation, and failure modes for a novel hybrid fixation method combining periosteal and direct fixation using porcine femoral bone. A soft tissue graft was secured by combining both an interference screw and an EndoButton (Smith and Nephew Endoscopy, Andover, MA). The results were compared with the traditional direct fixation method using a titanium interference screw. Twenty porcine hindlimbs were divided into two groups. Specimens were loaded in line with the bone tunnel on a materials testing machine. Maximum pullout force of the hybrid fixation (588+/-37 N) was significantly greater than with an interference screw alone (516+/-37 N). The stiffness of the hybrid fixation (52.1+/-12.8 N/mm) was similar to that of screw fixation (56.5+/-10.2 N/mm). Graft pullout was predominant for screw fixation, whereas a combination of graft pullout and graft failure was seen for hybrid fixation. These results indicate that initial pullout force of soft tissue grafts can be increased by using the suggested novel hybrid fixation method.

  3. Autogenous onlay grafting for enhancement of extracortical tissue formation over porous-coated segmental replacement prostheses.

    PubMed

    Virolainen, P; Inoue, N; Nagao, M; Ohnishi, I; Frassica, F; Chao, E Y

    1999-04-01

    Prosthetic reconstruction with extracortical bone-bridging is an effective method of limb salvage after resection of a malignant or locally invasive benign bone tumor. Use of cancellous bone graft alone is less effective in achieving extracortical bone-bridging. The present study was performed to investigate the effects of a corticocancellous onlay graft on bone and soft-tissue formation over a porous-coated replacement prosthesis in the mid-diaphyseal region of canine femora. Bilateral resection of a six-centimeter segment of the femoral diaphysis and reconstruction with a porous-coated segmental prosthesis was performed in six mongrel dogs. In one limb (the experimental side), eight strips of corticocancellous bone were evenly placed around the junctions between the femur and the prosthetic surface. Cancellous bone was placed under and between the strips of cortical bone. No graft was used in the other limb (the control side). The animals were followed for twelve weeks, with sequential assessments of load-bearing and radiographic evaluation. Biomechanical, histological, and microradiographic analyses of the specimens were performed after death. On the control side, load-bearing at four weeks postoperatively was significantly decreased compared with the preoperative value (p<0.05); no difference in these values could be detected on the experimental side. Both the area of the callus and the contact area between the bone and the prosthetic shoulder were greater on the experimental side (p<0.05). The mechanical stiffness and the maximum torque at failure of the extracortical bridging tissue across the junction between the bone and the prosthetic shoulder were eighteen (p<0.007) and five times greater (p<0.05), respectively, on the experimental side. Extracortical bone-bridging was accomplished with corticocancellous onlay bone-grafting. Without bone-grafting, bone formed only occasionally. Bone-grafting also enhanced the formation of a soft-tissue capsule around the

  4. Design and optimization of a tissue-engineered bone graft substitute

    NASA Astrophysics Data System (ADS)

    Shimko, Daniel Andrew

    2004-12-01

    In 2000, 3.1 million surgical procedures on the musculoskeletal system were reported in the United States. For many of these cases, bone grafting was essential for successful fracture stabilization. Current techniques use intact bone obtained either from the patient (autograft) or a cadaver (allograft) to repair large defects, however, neither source is optimal. Allografts suffer integration problems, and for autografts, the tissue supply is limited. Because of these shortcomings, and the high demand for graft tissues, alternatives are being explored. To successfully engineer a bone graft replacement, one must employ a three pronged research approach, addressing (1) the cells that will inhabit the new tissue, (2) the culture environment that these cells will be exposed to, and (3) the scaffold in which these cells will reside. The work herein examines each of these three aspects in great detail. Both adult and embryonic stem cells (ESCs) were considered for the tissue-engineered bone graft. Both exhibited desirable qualities, however, neither were optimal in all categories examined. In the end, the possibility of teratoma formation and ethical issues surrounding ESCs, made the use of adult marrow-derived stem cells in the remaining experiments obligatory. In subsequent experiments, the adult stem cells' ability to form bone was optimized. Basic fibroblast growth factor, fetal bovine serum, and extracellular calcium supplementation studies were all performed. Ultimately, adult stem cells cultured in alpha-MEM supplemented with 10% fetal bovine serum, 10mM beta-glycerophosphate, 10nM dexamethasone, 50mug/ml ascorbic acid, 1%(v/v) antibiotic/antimycotic, and 10.4mM CaCl2 performed the best, producing nearly four times more mineral than any other medium formulation. Several scaffolds were then investigated including those fabricated from poly(alpha-hydroxy esters), tantalum, and poly-methylmethacrylate. In the final study, the most appealing cell type, medium

  5. Meiosis in autologous ectopic transplants of immature testicular tissue grafted to Callithrix jacchus.

    PubMed

    Wistuba, Joachim; Luetjens, C Marc; Wesselmann, Ramona; Nieschlag, Eberhard; Simoni, Manuela; Schlatt, Stefan

    2006-04-01

    Grafting of immature testicular tissue provides a tool to examine testicular development and may offer a perspective for preservation of fertility in prepubertal patients. Successful xenografting in mice, resulting in mature spermatids, has been performed in several species but has failed with testicular tissues from the common marmoset, Callithrix jacchus. Previous data indicate that the hormonal milieu provided by the mouse host might cause this failure. We conducted autologous ectopic transplantation of testicular fragments under the back skin in newborn marmoset monkeys. Seventeen months after transplantation, we found viable transplants in 2 out of the 4 grafted animals. In the transplants, tubules developed up to a state intermediate between the pregraft situation and adult controls. Dividing spermatogonia and primary spermatocytes were present. Boule-like positivity and CDC25A negativity indicated that spermatogenesis was arrested at early meiosis. Immunohistochemistry revealed normal maturation of Sertoli cells, Leydig cells, and peritubular cells. Serum testosterone values were not restored to the normal range and bioactive chorionic gonadotropin levels increased to castrate levels. Meiotic arrest could have occurred in the grafts because of lack of sufficient testosterone or because of hyperthermia caused by the ectopic position of the grafts. We conclude that autologous transplants of immature testicular tissues in the marmoset can mature up to meiosis but that normal serum testosterone levels are not restored. Further studies have to be performed to overcome the meiotic arrest to explore the model further and to develop therapeutic options.

  6. Nanoassemblies of Tissue-Reactive, Polyoxazoline Graft-Copolymers Restore the Lubrication Properties of Degraded Cartilage.

    PubMed

    Morgese, Giulia; Cavalli, Emma; Müller, Mischa; Zenobi-Wong, Marcy; Benetti, Edmondo M

    2017-03-13

    Osteoarthritis leads to an alteration in the composition of the synovial fluid, which is associated with an increase in friction and the progressive and irreversible destruction of the articular cartilage. In order to tackle this degenerative disease, there has been a growing interest in the medical field to establish effective, long-term treatments to restore cartilage lubrication after damage. Here we develop a series of graft-copolymers capable of assembling selectively on the degraded cartilage, resurfacing it, and restoring the lubricating properties of the native tissue. These comprise a polyglutamic acid backbone (PGA) coupled to brush-forming, poly-2-methyl-2-oxazoline (PMOXA) side chains, which provide biopassivity and lubricity to the surface, and to aldehyde-bearing tissue-reactive groups, for the anchoring on the degenerated cartilage via Schiff bases. Optimization of the graft-copolymer architecture (i.e., density and length of side chains and amount of tissue-reactive functions) allowed a uniform passivation of the degraded cartilage surface. Graft-copolymer-treated cartilage showed very low coefficients of friction within synovial fluid, reestablishing and in some cases improving the lubricating properties of the natural cartilage. Due to these distinctive properties and their high biocompatibility and stability under physiological conditions, cartilage-reactive graft-copolymers emerge as promising injectable formulations to slow down the progression of cartilage degradation, which characterizes the early stages of osteoarthritis.

  7. Integrating biologically inspired nanomaterials and table-top stereolithography for 3D printed biomimetic osteochondral scaffolds.

    PubMed

    Castro, Nathan J; O'Brien, Joseph; Zhang, Lijie Grace

    2015-09-07

    The osteochondral interface of an arthritic joint is notoriously difficult to regenerate due to its extremely poor regenerative capacity and complex stratified architecture. Native osteochondral tissue extracellular matrix is composed of numerous nanoscale organic and inorganic constituents. Although various tissue engineering strategies exist in addressing osteochondral defects, limitations persist with regards to tissue scaffolding which exhibit biomimetic cues at the nano to micro scale. In an effort to address this, the current work focused on 3D printing biomimetic nanocomposite scaffolds for improved osteochondral tissue regeneration. For this purpose, two biologically-inspired nanomaterials have been synthesized consisting of (1) osteoconductive nanocrystalline hydroxyapatite (nHA) (primary inorganic component of bone) and (2) core-shell poly(lactic-co-glycolic) acid (PLGA) nanospheres encapsulated with chondrogenic transforming growth-factor β1 (TGF-β1) for sustained delivery. Then, a novel table-top stereolithography 3D printer and the nano-ink (i.e., nHA + nanosphere + hydrogel) were employed to fabricate a porous and highly interconnected osteochondral scaffold with hierarchical nano-to-micro structure and spatiotemporal bioactive factor gradients. Our results showed that human bone marrow-derived mesenchymal stem cell adhesion, proliferation, and osteochondral differentiation were greatly improved in the biomimetic graded 3D printed osteochondral construct in vitro. The current work served to illustrate the efficacy of the nano-ink and current 3D printing technology for efficient fabrication of a novel nanocomposite hydrogel scaffold. In addition, tissue-specific growth factors illustrated a synergistic effect leading to increased cell adhesion and directed stem cell differentiation.

  8. Integrating biologically inspired nanomaterials and table-top stereolithography for 3D printed biomimetic osteochondral scaffolds

    NASA Astrophysics Data System (ADS)

    Castro, Nathan J.; O'Brien, Joseph; Zhang, Lijie Grace

    2015-08-01

    The osteochondral interface of an arthritic joint is notoriously difficult to regenerate due to its extremely poor regenerative capacity and complex stratified architecture. Native osteochondral tissue extracellular matrix is composed of numerous nanoscale organic and inorganic constituents. Although various tissue engineering strategies exist in addressing osteochondral defects, limitations persist with regards to tissue scaffolding which exhibit biomimetic cues at the nano to micro scale. In an effort to address this, the current work focused on 3D printing biomimetic nanocomposite scaffolds for improved osteochondral tissue regeneration. For this purpose, two biologically-inspired nanomaterials have been synthesized consisting of (1) osteoconductive nanocrystalline hydroxyapatite (nHA) (primary inorganic component of bone) and (2) core-shell poly(lactic-co-glycolic) acid (PLGA) nanospheres encapsulated with chondrogenic transforming growth-factor β1 (TGF-β1) for sustained delivery. Then, a novel table-top stereolithography 3D printer and the nano-ink (i.e., nHA + nanosphere + hydrogel) were employed to fabricate a porous and highly interconnected osteochondral scaffold with hierarchical nano-to-micro structure and spatiotemporal bioactive factor gradients. Our results showed that human bone marrow-derived mesenchymal stem cell adhesion, proliferation, and osteochondral differentiation were greatly improved in the biomimetic graded 3D printed osteochondral construct in vitro. The current work served to illustrate the efficacy of the nano-ink and current 3D printing technology for efficient fabrication of a novel nanocomposite hydrogel scaffold. In addition, tissue-specific growth factors illustrated a synergistic effect leading to increased cell adhesion and directed stem cell differentiation.

  9. In vitro skin models and tissue engineering protocols for skin graft applications.

    PubMed

    Naves, Lucas B; Dhand, Chetna; Almeida, Luis; Rajamani, Lakshminarayanan; Ramakrishna, Seeram

    2016-11-30

    In this review, we present a brief introduction of the skin structure, a concise compilation of skin-related disorders, and a thorough discussion of different in vitro skin models, artificial skin substitutes, skin grafts, and dermal tissue engineering protocols. The advantages of the development of in vitro skin disorder models, such as UV radiation and the prototype model, melanoma model, wound healing model, psoriasis model, and full-thickness model are also discussed. Different types of skin grafts including allografts, autografts, allogeneic, and xenogeneic are described in detail with their associated applications. We also discuss different tissue engineering protocols for the design of various types of skin substitutes and their commercial outcomes. Brief highlights are given of the new generation three-dimensional printed scaffolds for tissue regeneration applications. © 2016 The Author(s). published by Portland Press Limited on behalf of the Biochemical Society.

  10. Magnetic resonance imaging of hyaline cartilage regeneration in neocartilage graft implantation.

    PubMed

    Tan, C F; Ng, K K; Ng, S H; Cheung, Y C

    2003-12-01

    The purpose of this study was to investigate the regenerative potential of hyaline cartilage in a neocartilage graft implant with the aid of MR cartilage imaging using a rabbit model. Surgical osteochondral defects were created in the femoral condyles of 30 mature New Zealand rabbits. The findings of neocartilage in autologous cartilage grafts packed into osteochondral defects were compared with control group of no implant to the osteochondral defect. The outcome of the implantations was correlated with histologic and MR cartilage imaging findings over a 3-month interval. Neocartilage grafts packed into osteochondral defects showed regeneration of hyaline cartilage at the outer layer of the implant using MR cartilage imaging. Fibrosis of fibrocartilage developed at the outer layer of the autologous cartilage graft together with an inflammatory reaction within the osteochondral defect. This animal study provides evidence of the regenerative ability of hyaline cartilage in neocartilage transplants to repair articular cartilage.

  11. A novel approach to regenerating periodontal tissue by grafting autologous cultured periosteum.

    PubMed

    Mizuno, Hirokazu; Hata, Ken-Ichiro; Kojima, Koji; Bonassar, Lawrence J; Vacanti, Charles A; Ueda, Minoru

    2006-05-01

    In the field of oral and maxillofacial surgery, tissue-engineering techniques have been found useful in regenerating lost tissues. Periodontal disease causes severe destruction of periodontal tissue, including the alveolar bone. In this study we attempted to regenerate canine periodontal tissue defects by grafting autologous cultured membrane derived from the periosteum. Under appropriate culture conditions, periosteal cells produce enough extracellular matrix to form sheets. Periosteum specimens were peeled from the mandibular body of adult hybrid dogs and were cultured until cells formed membrane. ALP activity was measured to determine an optimal time for grafting. The cultured periosteum (CP) was grafted and sutured on a mechanically made Class III furcation defect in the 4th mandibular premolars. After 3 months, the samples were harvested and observed radiologically and histologically. In cases of CP, the bone defects were regenerated and filled with newly formed hard tissue, whereas in the controls the defects remained. These results show that our novel treatment is effective in regenerating alveolar bone for the treatment of periodontal disease.

  12. Mechanical integrity of subchondral bone in osteochondral autografts and allografts

    PubMed Central

    Wohl, Greg; Goplen, Gordon; Ford, Jason; Novak, Kelli; Hurtig, Mark; McPherson, Roger; McGann, Locksley; Schachar, Norman; Zernicke, Ronald F.

    1998-01-01

    Objective To assess the influence of osteochondral graft preservation techniques on post-transplant biomechanics of graft and host subchondral bone in the knee joint. Design An experimental animal model (sheep), specifically the weight-bearing articular surface of the medial femoral condyle of the knee joints. Intervention Each sheep received, in the ipsilateral knee, an allograft that was (a) frozen without dimethyl sulfoxide (DMSO), (b) snap-frozen in liquid nitrogen or (c) frozen with DMSO. The contralateral knee received an autograft that was (a) snap-frozen, (b) treated with DMSO or (c) left untreated (fresh). Main outcome measures Mechanical and material properties of bone, including maximal compression stress, modulus of elasticity and bone mineral ash content of subchondral bone cores (from the graft centre and surrounding host bone). Results No significant differences were found in the mechanical properties of the subchondral bone under the graft, but there were significant changes in surrounding bone. Bone surrounding the grafts that were snap-frozen or frozen without DMSO was significantly stronger than the normal control bone. However, bone surrounding fresh autografts and cryoprotected allografts was not significantly different from normal control bone. Conclusions The changes in the mechanical behaviour of the host bone may be associated with graft cell viability. The greater stiffness of the subchondral host bone may have consequences for long-term graft integrity and for the development of degenerative osteoarthritis. PMID:9627549

  13. Recent Advances in Egypt for Treatment of Talar Osteochondral Lesions.

    PubMed

    Haleem, Amgad M; AbouSayed, Mostafa M; Gomaa, Mohammed

    2016-06-01

    Treatment of osteochondral defects (OCLs) of the talus is a challenging orthopedic surgery. Treatment of talar OCLs has evolved through the 3 "R" paradigm: reconstruction, repair, and replacement. This article highlights current state-of-the-art techniques and reviews recent advances in the literature about articular cartilage repair using various novel tissue engineering approaches, including various scaffolds, growth factors, and cell niches; which include chondrocytes and culture-expanded bone marrow-derived mesenchymal stem cells.

  14. Bipolar fresh osteochondral allograft of the ankle.

    PubMed

    Giannini, Sandro; Buda, Roberto; Grigolo, Brunella; Bevoni, Roberto; Di Caprio, Francesco; Ruffilli, Alberto; Cavallo, Marco; Desando, Giovanna; Vannini, Francesca

    2010-01-01

    Severe post-traumatic ankle arthritis poses a reconstructive challenge in the young and active patient. Bipolar fresh osteochondral allograft (BFOA) may represent an intriguing alternative to arthrodesis and prosthetic replacement. The aim of this study was to describe a lateral trans-malleolar technique for BFOA, and to evaluate the results in a case series. From 2004 to 2006, 32 patients, mean age of 36.8 +/- 8.4 years, affected by ankle arthritis underwent BFOA with a mean followup of 31.2 months. The graft was prepared by specifically designed jigs, including the talus and the tibia with the medial malleolus. The host surfaces were prepared by the same jigs through a lateral approach. The graft was placed and fixed with twist-off screws. Patients were evaluated clinically and radiographically at 2, 4, and 6 month after operation, and at a minimum 24 months followup. A biopsy of the grafted areas was obtained from 7 patients at 1-year followup for histological and immunohistochemical examination. Preoperative AOFAS score was 33.1 +/- 10.9 and postoperatively 69.5 +/- 19.4 (p < 0.0005). Six failures occurred. Cartilage harvests showed hyaline-like histology with a normal collagen component but low proteoglycan presence and a disorganized structure. Samples were positive for MMP-1, MMP-13 and Capsase-3. The use of BFOA represents an intriguing alternative to arthrodesis or arthroplasty. We believe precise allograft sizing, stable fitting and fixation and delayed weightbearing were key factors for a successful outcome. Further research regarding the immunological behavior of transplanted cartilage is needed.

  15. Tissue-engineered oral mucosa grafts for intraoral lining reconstruction of the maxilla and mandible with a fibula flap.

    PubMed

    Sieira Gil, Ramón; Pagés, Carles Martí; Díez, Eloy García; Llames, Sara; Fuertes, Ada Ferrer; Vilagran, Jesús Lopez

    2015-01-01

    Many types of soft tissue grafts have been used for grafting or prelaminating bone flaps for intraoral lining reconstruction. The best results are achieved when prelaminating free flaps with mucosal grafts. We suggest a new approach to obtain keratinized mucosa over a fibula flap using full-thickness, engineered, autologous oral mucosa. We report on a pilot study for grafting fibula flaps for mandibular and maxilla reconstruction with full-thickness tissue-engineered autologous oral mucosa. We describe 2 different techniques: prelaminating the fibula flap and second-stage grafting of the fibula after mandibular reconstruction. Preparation of the full-thickness tissue-engineered oral mucosa is also described. The clinical outcome of the tissue-engineered intraoral lining reconstruction and response after implant placement are reported. A peri-implant granulation tissue response was not observed when prelaminating the fibula, and little response was observed when intraoral grafting was performed. Tissue engineering represents an alternative method by which to obtain sufficient autologous tissue for reconstructing mucosal oral defects. The full-thickness engineered autologous oral mucosa offers definite advantages in terms of reconstruction planning, donor site morbidity, and quality of the intraoral soft tissue reconstruction, thereby restoring native tissue and avoiding peri-implant tissue complications. Copyright © 2015 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.

  16. Design of a Novel 3D Printed Bioactive Nanocomposite Scaffold for Improved Osteochondral Regeneration.

    PubMed

    Castro, Nathan J; Patel, Romil; Zhang, Lijie Grace

    2015-09-01

    Chronic and acute osteochondral defects as a result of osteoarthritis and trauma present a common and serious clinical problem due to the tissue's inherent complexity and poor regenerative capacity. In addition, cells within the osteochondral tissue are in intimate contact with a 3D nanostructured extracellular matrix composed of numerous bioactive organic and inorganic components. As an emerging manufacturing technique, 3D printing offers great precision and control over the microarchitecture, shape and composition of tissue scaffolds. Therefore, the objective of this study is to develop a biomimetic 3D printed nanocomposite scaffold with integrated differentiation cues for improved osteochondral tissue regeneration. Through the combination of novel nano-inks composed of organic and inorganic bioactive factors and advanced 3D printing, we have successfully fabricated a series of novel constructs which closely mimic the native 3D extracellular environment with hierarchical nanoroughness, microstructure and spatiotemporal bioactive cues. Our results illustrate several key characteristics of the 3D printed nanocomposite scaffold to include improved mechanical properties as well as excellent cytocompatibility for enhanced human bone marrow-derived mesenchymal stem cell adhesion, proliferation, and osteochondral differentiation in vitro. The present work further illustrates the effectiveness of the scaffolds developed here as a promising and highly tunable platform for osteochondral tissue regeneration.

  17. Polymers, scaffolds and bioactive molecules with therapeutic properties in osteochondral pathologies: what's new?

    PubMed

    López-Ruiz, Elena; Jiménez, Gema; García, María Ángel; Antich, Cristina; Boulaiz, Houria; Marchal, Juan Antonio; Perán, Macarena

    2016-08-01

    Despite clinical efforts, treatments to heal osteochondral lesions remain inefficient and frequently result, long-term, in joint arthroplasty. The complex structure of cartilage tissue, composed of a highly hydrated extracellular matrix (ECM), an avascular nature, and slow cellular turnover, hamper tissue regeneration after trauma or disease. Tissue engineering provides new promising alternatives to current treatments designed to regenerate osteochondral defects. This review describes current and recent strategies of enhancing osteochondral repair through the use of cells, scaffolds, and bioactive molecules. Here, we review the latest (2011-2015) innovative patents in osteochondral regeneration, emphasizing novel strategies for articular cartilage repair. Finally, we present a summary of ongoing clinical trials that are testing innovative engineered products. Promising tissue engineering based procedures have emerged as a therapeutic option for the treatment of osteochondral lesions. The development of multilayer scaffolds and the controlled release of bioactive molecules to promote in situ regeneration of both cartilage and bone are some of the latest technologies that intended to improve on the available traditional treatments. To confirm the potential of these novel approaches, long-term evaluation is necessary with special focus on studying the biological and mechanical proprieties of the synthesized tissues.

  18. Design of a Novel 3D Printed Bioactive Nanocomposite Scaffold for Improved Osteochondral Regeneration

    PubMed Central

    Castro, Nathan J.; Patel, Romil; Zhang, Lijie Grace

    2015-01-01

    Chronic and acute osteochondral defects as a result of osteoarthritis and trauma present a common and serious clinical problem due to the tissue's inherent complexity and poor regenerative capacity. In addition, cells within the osteochondral tissue are in intimate contact with a 3D nanostructured extracellular matrix composed of numerous bioactive organic and inorganic components. As an emerging manufacturing technique, 3D printing offers great precision and control over the microarchitecture, shape and composition of tissue scaffolds. Therefore, the objective of this study is to develop a biomimetic 3D printed nanocomposite scaffold with integrated differentiation cues for improved osteochondral tissue regeneration. Through the combination of novel nano-inks composed of organic and inorganic bioactive factors and advanced 3D printing, we have successfully fabricated a series of novel constructs which closely mimic the native 3D extracellular environment with hierarchical nanoroughness, microstructure and spatiotemporal bioactive cues. Our results illustrate several key characteristics of the 3D printed nanocomposite scaffold to include improved mechanical properties as well as excellent cytocompatibility for enhanced human bone marrow-derived mesenchymal stem cell adhesion, proliferation, and osteochondral differentiation in vitro. The present work further illustrates the effectiveness of the scaffolds developed here as a promising and highly tunable platform for osteochondral tissue regeneration. PMID:26366231

  19. Osteochondral autologous transplantation for the treatment of full-thickness articular cartilage defects of the shoulder.

    PubMed

    Scheibel, M; Bartl, C; Magosch, P; Lichtenberg, S; Habermeyer, P

    2004-09-01

    We performed eight osteochondral autologous transplantations from the knee joint to the shoulder. All patients (six men, two women; mean age 43.1 years) were documented prospectively. In each patient the stage of the osteochondral lesion was Outerbridge grade IV with a mean size of the affected area of 150 mm2. All patients were assessed by using the Constant score for the shoulder and the Lysholm score for the knee. Standard radiographs, magnetic resonance imaging and second-look arthroscopy were used to assess the presence of glenohumeral osteoarthritis and the integrity of the grafts. After a mean of 32.6 months (8 to 47), the mean Constant score increased significantly. Magnetic resonance imaging revealed good osseointegration of the osteochondral plugs and congruent articular cartilage at the transplantation site in all but one patient. Second-look arthroscopy performed in two cases revealed a macroscopically good integration of the autograft with an intact articular surface. Osteochondral autologous transplantation in the shoulder appears to offer good clinical results for treating full-thickness osteochondral lesions of the glenohumeral joint. However, our study suggests that the development of osteoarthritis and the progression of pre-existing osteoarthritic changes cannot be altered by this technique.

  20. Osteochondral diseases and fibrodysplasia ossificans progressiva.

    PubMed

    Morales-Piga, Antonio; Kaplan, Frederick S

    2010-01-01

    Osteochondrodysplasias like thanatophoric dysplasia, osteogenesis imperfecta, achondroplasia, and other genetic skeletal disorders like fibrodysplasia ossificans progressiva are infrequently seen in clinical practice. In cases of sporadic achondroplasia as well as in fibrodysplasia ossificans progressiva, there is a strong association with paternal age, a relationship that is less evident in other genetic osteochondral diseases. No other constitutional or environmental factor has proven to be associated with these disorders. The use of prenatal ultrasonography as a routine component of prenatal care is crucial in the early suspicion of osteochondrodysplasias whereas definitive diagnosis is usually obtained by pre-natal molecular analysis. In the case of fibrodysplasia ossificans progressiva, recognition of congenital great toe malformations associated with rapidly-appearing soft tissue swelling is sufficient to make the proper clinical diagnosis, which can be confirmed by genetic testing. Large regional centres will improve diagnosis performance, provide accurate genetic counselling, and ensure an integral assistance for these often severe and incapacitating conditions.

  1. Osteochondral Diseases and Fibrodysplasia Ossificans Progressiva

    PubMed Central

    Kaplan, Frederick S.

    2016-01-01

    Osteochondrodysplasias like thanatophoric dysplasia, osteogenesis imperfecta, achondroplasia, and other genetic skeletal disorders like fibrodysplasia ossificans progressiva are infrequently seen in clinical practice. In cases of sporadic achondroplasia as well as in fibrodysplasia ossificans progressiva, there is a strong association with paternal age, a relationship that is less evident in other genetic osteochondral diseases. No other constitutional or environmental factor has proven to be associated with these disorders. The use of prenatal ultrasonography as a routine component of prenatal care is crucial in the early suspicion of osteochondrodysplasias whereas definitive diagnosis is usually obtained by pre-natal molecular analysis. In the case of fibrodysplasia ossificans progressiva, recognition of congenital great toe malformations associated with rapidly–appearing soft tissue swelling is sufficient to make the proper clinical diagnosis, which can be confirmed by genetic testing. Large regional centres will improve diagnosis performance, provide accurate genetic counselling, and ensure an integral assistance for these often severe and incapacitating conditions. PMID:20824454

  2. The effects of prolonged deep freezing on the biomechanical properties of osteochondral allografts.

    PubMed

    Rozen, Benjamin; Brosh, Tamar; Salai, Moshe; Herman, Amir; Dudkiewicz, Israel

    2009-02-01

    Musculo-skeletal allografts sterilized and deep frozen are among the most common human tissue to be preserved and utilized in modern medicine. The effects of a long deep freezing period on cortical bone has already been evaluated and found to be insignificant. However, there are no reports about the influences of a protracted deep freezing period on osteochondral allografts. One hundred osteochondral cylinders were taken from a fresh specimen and humeral heads of 1 year, 2 years, 3 years and 4 year old bones. Twenty chips from each period, with a minimum of 3 chips per humeral head. Each was mechanically tested by 3 point compression. The fresh osteochondral allografts were significantly mechanically better than the deep frozen osteochondral allografts. There was no statistical significant time dependent difference between the deep frozen groups in relation to the freezing period. Therefore, we conclude that, from the mechanical point of view deep freezing of osteochondral allografts over a period of 4 years, is safe without further deterioration of the biomechanical properties of the osteochondral allografts.

  3. Tissue-engineered tracheal reconstruction using three-dimensionally printed artificial tracheal graft: preliminary report.

    PubMed

    Chang, Jae Won; Park, Su A; Park, Ju-Kyeong; Choi, Jae Won; Kim, Yoo-Suk; Shin, Yoo Seob; Kim, Chul-Ho

    2014-06-01

    Three-dimensional printing has come into the spotlight in the realm of tissue engineering. We intended to evaluate the plausibility of 3D-printed (3DP) scaffold coated with mesenchymal stem cells (MSCs) seeded in fibrin for the repair of partial tracheal defects. MSCs from rabbit bone marrow were expanded and cultured. A half-pipe-shaped 3DP polycaprolactone scaffold was coated with the MSCs seeded in fibrin. The half-pipe tracheal graft was implanted on a 10 × 10-mm artificial tracheal defect in four rabbits. Four and eight weeks after the operation, the reconstructed sites were evaluated bronchoscopically, radiologically, histologically, and functionally. None of the four rabbits showed any sign of respiratory distress. Endoscopic examination and computed tomography showed successful reconstruction of trachea without any collapse or blockage. The replaced tracheas were completely covered with regenerated respiratory mucosa. Histologic analysis showed that the implanted 3DP tracheal grafts were successfully integrated with the adjacent trachea without disruption or granulation tissue formation. Neo-cartilage formation inside the implanted graft was sufficient to maintain the patency of the reconstructed trachea. Scanning electron microscope examination confirmed the regeneration of the cilia, and beating frequency of regenerated cilia was not different from those of the normal adjacent mucosa. The shape and function of reconstructed trachea using 3DP scaffold coated with MSCs seeded in fibrin were restored successfully without any graft rejection. Copyright © 2014 International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

  4. Combination of negative pressure wound therapy with open bone grafting for bone and soft tissue defects.

    PubMed

    Deng, Kai; Yu, Ai-Xi; Xia, Cheng-Yan; Li, Zong-Huan; Wang, Wei-Yang

    2013-08-01

    The aim of this study was to investigate the efficiency of negative pressure wound therapy (NPWT) combined with open bone graft (OBG; NPWT-OBG) for the treatment of bone and soft tissue defects with polluted wounds in an animal model. All rabbits with bone and soft tissue defects and polluted wounds were randomly divided into two groups, the experimental group (NPWT with bone graft) and the control group (OBG). The efficacy of the treatment was assessed by the wound conditions and healing time. Bacterial bioburdens and bony calluses were evaluated by bacteria counting and X-rays, respectively. Furthermore, granulation tissue samples from the wounds on days 0, 3, 7 and 14 of healing were evaluated for blood vessels and vascular endothelial growth factor (VEGF) levels. Wounds in the experimental group tended to have a shorter healing time, healthier wound conditions, lower bacterial bioburden, improvement of the bony calluses and an increased blood supply compared with those in the control group. With NPWT, wound infection was effectively controlled. For wounds with osseous and soft tissue defects, NPWT combined with bone grafting was demonstrated to be more effective than an OBG.

  5. Follicle development in cryopreserved bitch ovarian tissue grafted to immunodeficient mouse.

    PubMed

    Commin, L; Buff, S; Rosset, E; Galet, C; Allard, A; Bruyere, P; Joly, T; Guérin, P; Neto, V

    2012-01-01

    The present study evaluated: (1) in vivo follicular development in canine ovarian tissue after slow freezing and xenotransplantation; and (2) the use of erythropoietin (EPO) as an angiogenic factor to optimise the transplantation procedure. Frozen-thawed ovarian tissue from five bitches was grafted into severe combined immunodeficient (SCID) mice (n=47) treated with or without EPO (500 IU kg(-1), once daily for 3 days) (Groups A and B, respectively) and analysed after 0, 1, 8 or 16 weeks. Follicle grade, follicle density, follicle morphology and stromal cells density were assessed by histological analysis, whereas vascularisation of the graft was quantified by immunohistochemistry with anti-α-smooth muscle actin antibody. Despite a massive loss of follicles after grafting, secondary follicle density was higher at 8 and 16 weeks than at 1 week regardless of EPO treatment. EPO significantly improved early follicle morphology and stromal cell density after 8 weeks and blood vessel density at 16 weeks after transplantation (P<0.05). Intact secondary follicles with more than three granulosa cells layers were observed 16 weeks after transplantation. The results suggest that canine ovarian tissue can be successfully preserved by our slow-freezing protocol because the tissue showed follicular growth after xenotransplantation. EPO treatment did not lessen the massive loss of follicles after transplantation.

  6. Fragmented Adipose Tissue Graft for Bone Healing: Histological and Histometric Study in Rabbits’ Calvaria

    PubMed Central

    Oliveira, Lidiane C.; Giovanini, Allan F.; Abuabara, Allan; Klug, Luiz G.; Gonzaga, Carla C.; Zielak, João C.; Urban, Cícero A.

    2013-01-01

    Objective The adipose tissue represents an important reservoir of stem cells. There are few studies in the literature with which to histologically evaluate whether or not the adipose tissue graft is really a safe option to achieve bone repair. This study histologically analyzed the effect of fragmented autogenous adipose tissue grafts on bone healing in surgically created, critical-size defects (CSD) in a rabbit’s calvaria. Study design Forty-two New Zealand rabbits were used in this study. CSD that were 15 mm in diameter were created in the calvarium of each animal. The defects were randomly divided into two groups: in Group C (control), the defect was filled only by a blood clot and, in Group FAT (i.e., fragmented adipose tissue), the defect was filled with fragmented autogenous adipose tissue grafts. The groups were divided into subgroups (n = 7) for euthanasia at 7, 15, and 40 days after the procedure had been conducted. Histologic and histometric analyses were performed. Data were statistically analysed with ANOVA and Tukey’s tests (p < 0.05). Results The amount of bone formation did not show statistically significant differences seven days after the operation, which indicates that the groups had similar amounts of mineral deposition in the earlier period of the repair. Conversely, a significant of amount of bone matrix deposition was identified in the FAT group at 15 and 40 days following the operation, both on the border and in the body of the defect. Such an outcome was not found in the control group. Conclusion In this study, an autologous adipose tissue graft may be considered as likely biomaterial for bone regeneration, since it positively affected the amount of bone formation in surgically created CSD in the rabbits’ calvaria 40 days after the procedure had been performed. Further investigations with a longer time evaluation are warranted to determine the effectiveness of autologous adipose tissue graft in the bone healing. Key words

  7. Technology insight: the evolution of tissue-engineered vascular grafts--from research to clinical practice.

    PubMed

    L'Heureux, Nicolas; Dusserre, Nathalie; Marini, Alicia; Garrido, Sergio; de la Fuente, Luis; McAllister, Todd

    2007-07-01

    There is a considerable clinical need for alternatives to the autologous vein and artery tissues used for vascular reconstructive surgeries such as CABG, lower limb bypass, arteriovenous shunts and repair of congenital defects to the pulmonary outflow tract. So far, synthetic materials have not matched the efficacy of native tissues, particularly in small diameter applications. The development of cardiovascular tissue engineering introduced the possibility of a living, biological graft that might mimic the functional properties of native vessels. While academic research in the field of tissue engineering in general has been active, as yet there has been no clear example of clinical and commercial success. The recent transition of cell-based therapies from experimental to clinical use has, however, reinvigorated the field of cardiovascular tissue engineering. Here, we discuss the most promising approaches specific to tissue-engineered blood vessels and briefly introduce our recent clinical results. The unique regulatory, reimbursement and production challenges facing personalized medicine are also discussed.

  8. Osteochondritis dissecans of the talus

    PubMed Central

    ZANON, GIACOMO; DI VICO, GIOVANNI; MARULLO, MATTEO

    2014-01-01

    Osteochondritis dissecans (OCD) is an acquired idiopathic lesion of subchondral bone that can produce delamination and sequestration with or without articular cartilage involvement and instability. The cause of OCD is still debated: the most recognized etiology is the occurrence of repetitive micro-traumas associated with vascular impairment, causing progressive ankle pain and dysfunction in skeletally immature and young adult patients. Ankle OCD is classically located in the medial part of the talus, while lateral and posterior involvement is less frequent. Diagnosis of OCD, based on MRI findings, is quite straightforward; MRI examination can also be very useful for dating the defect and obtaining information about the associated bone bruise. Osteochondritis dissecans, if not recognized and treated appropriately, may lead to secondary osteoarthritis with pain and functional limitation. Surgical treatment is mandatory especially in young patients with unstable cartilage fragments. There are various surgical options: fixation, microfracture, or substitution using autologous chondrocyte implantation techniques. PMID:25606554

  9. Current trends in safety assurance for tissue grafts used in burn treatment.

    PubMed

    Mericka, P

    2006-01-01

    The author presents a summary of current safety standards for allogeneic and xenogeneic biological skin grafts. The fundamental document relevant to allogeneic transplants, establishing the minimal level of safety guaranteed in European Union states, is the European Parliament and Council Directive (2004/23/EC) from March 31st 2004. This Directive determines that grafts will be prepared by a licensed or accredited tissue bank, and that this arrangement must be put in place by the member states within 2 years. In the Czech Republic licensing of tissue banks took place immediately after issuance of the Directive. Licensing was also a condition for product reimbursement by insurance companies. To gain a licence, tissue banks had to fulfil many safety criteria associated with screening of living or deceased donors for health suitability, providing traceability of the donor-recipient route, prevention of secondary and cross-contamination during processing and storage of the harvested tissues, proof of product microbiology check up, and cold chain control. The Tissue Bank of the Faculty Hospital in Hradec Králové is one of the two tissue banks that gained the broader type of 'multifunctional' licence and was granted registration number MTB 006. Obtaining the licence was facilitated by completion of a new workplace project conceived as a combination of cryogenic and clean-room technology. Currently, this tissue bank prepares cryopreserved dermoepidermal and dermal grafts as well as amnion and chorioamnion grafts. All tissue banks will have to renew their licences again according to the conditions established by a new law about human tissues and cells which is currently in preparation. Neither the Directive of the European Parliament nor the Transplantation Law of the Czech Republic regulates the issue of xenografts. Since availability of allogeneic biological covers is limited, it is significant that the WHO perspective on the use of xenogeneic biological covers, as

  10. Changes of the peri-implant soft tissue thickness after grafting with a collagen matrix

    PubMed Central

    Zafiropoulos, Gregory-George; Deli, Giorgio; Hoffmann, Oliver; John, Gordon

    2016-01-01

    Background: The aim of this study was to determine the treatment outcome of the use of a porcine monolayer collagen matrix (mCM) to increase soft-tissue volume as a part of implant site development. Materials and Methods: Implants were placed in single sites in 27 patients. In the test group, mCM was used for soft-tissue augmentation. No graft was placed in the control group. Soft-tissue thickness (STTh) was measured at the time of surgery (T0) and 6 months postoperatively (T1) at two sites (STTh 1, 1 mm below the gingival margin; STTh 2, 3 mm below the mucogingival margin). Results: Significant increases (P < 0.001) in STTh (STTh 1 = 1.06 mm, 117%; STTh 2 = 0.89 mm, 81%) were observed in the test group. Biopsy results showed angiogenesis and mature connective tissue covered by keratinized epithelium. Conclusions: Within the limitations of this study, it could be concluded that mCM leads to a significant increase of peri-implant soft-tissue thickness, with good histological integration and replacement by soft tissue and may serve as an alternative to connective tissue grafting. PMID:28298828

  11. Adipose tissue-derived stem cell-seeded small intestinal submucosa for tunica albuginea grafting and reconstruction.

    PubMed

    Ma, Limin; Yang, Yijun; Sikka, Suresh C; Kadowitz, Philip J; Ignarro, Louis J; Abdel-Mageed, Asim B; Hellstrom, Wayne J G

    2012-02-07

    Porcine small intestinal submucosa (SIS) has been widely used in tunica albuginea (TA) reconstructive surgery. Adipose tissue-derived stem cells (ADSCs) can repair damaged tissue, augment cellular differentiation, and stimulate release of multiple growth factors. The aim of this rat study was to assess the feasibility of seeding ADSCs onto SIS grafts for TA reconstruction. Here, we demonstrate that seeding syngeneic ADSCs onto SIS grafts (SIS-ADSC) resulted in significant cavernosal tissue preservation and maintained erectile responses, similar to controls, in a rat model of bilateral incision of TA, compared with sham-operated animals and rats grafted with SIS graft (SIS) alone. In addition to increased TGF-β1 and FGF-2 expression levels, cross-sectional studies of the rat penis with SIS and SIS-ADSC revealed mild to moderate fibrosis and an increase of 30% and 40% in mean diameter in flaccid and erectile states, respectively. SIS grafting induced transcriptional up-regulation of iNOS and down-regulation of endothelial NOS, neuronal NOS, and VEGF, an effect that was restored by seeding ADCSs on the SIS graft. Taken together, these data show that rats undergoing TA incision with autologous SIS-ADSC grafts maintained better erectile function compared with animals grafted with SIS alone. This study suggests that SIS-ADSC grafting can be successfully used for TA reconstruction procedures and can restore erectile function.

  12. Adipose tissue-derived stem cell-seeded small intestinal submucosa for tunica albuginea grafting and reconstruction

    PubMed Central

    Ma, Limin; Yang, Yijun; Sikka, Suresh C.; Kadowitz, Philip J.; Ignarro, Louis J.; Abdel-Mageed, Asim B.; Hellstrom, Wayne J. G.

    2012-01-01

    Porcine small intestinal submucosa (SIS) has been widely used in tunica albuginea (TA) reconstructive surgery. Adipose tissue-derived stem cells (ADSCs) can repair damaged tissue, augment cellular differentiation, and stimulate release of multiple growth factors. The aim of this rat study was to assess the feasibility of seeding ADSCs onto SIS grafts for TA reconstruction. Here, we demonstrate that seeding syngeneic ADSCs onto SIS grafts (SIS-ADSC) resulted in significant cavernosal tissue preservation and maintained erectile responses, similar to controls, in a rat model of bilateral incision of TA, compared with sham-operated animals and rats grafted with SIS graft (SIS) alone. In addition to increased TGF-β1 and FGF-2 expression levels, cross-sectional studies of the rat penis with SIS and SIS-ADSC revealed mild to moderate fibrosis and an increase of 30% and 40% in mean diameter in flaccid and erectile states, respectively. SIS grafting induced transcriptional up-regulation of iNOS and down-regulation of endothelial NOS, neuronal NOS, and VEGF, an effect that was restored by seeding ADCSs on the SIS graft. Taken together, these data show that rats undergoing TA incision with autologous SIS-ADSC grafts maintained better erectile function compared with animals grafted with SIS alone. This study suggests that SIS-ADSC grafting can be successfully used for TA reconstruction procedures and can restore erectile function. PMID:22308363

  13. Intrathecal grafting of unencapsulated adrenal medullary tissue can bring CD4 T lymphocytes into CSF: a potentially deleterious event for the graft.

    PubMed

    Tkaczuk, J; Bes, J C; Duplan, H; Sallerin, B; Tafani, M; Charlet, J P; Abbal, M; Lazorthes, Y; Ohayon, E

    2000-01-01

    Adrenal medullary tissue including chromaffin cells was grafted intrathecally in cancer patients to relieve intractable pain. The central nervous system (CNS) is considered an immune privileged site. Therefore, non-HLA-matched and unencapsulated tissue was grafted in 15 patients and 1 sham control in a series of at least 20 grafts. We observed an increase in CSF lymphocyte counts in 15/20 allografts (75%). In contrast to peripheral blood, CD4 T cells predominated in the CSF, but failed to exhibit an activated phenotype (CD25+ CD45RO+ HLA-DR+). The positive effect of graft on pain, the high met-enkephalin levels, the absence of any increase in CSF cytokine levels particularly for IFN-gamma or IL-2 (but not IL-10 and IL-6), indirectly indicated that the graft was tolerated despite the presence of CSF lymphocytes. The single treatment failure and three of four cases of partial efficacy occurred in grafts where CSF lymphocytes were present. Moreover, when assayed (n = 7), the CD4+ CSF lymphocytes still retained the capacity to exhibit ex vivo a normal or enhanced frequency of T CD4 cells producing IFN-gamma and IL-2. Taken together, our observations indicate that impairment of the local immunosuppressive balance can lead to activation of those CSF CD4 T cells and drive a rejection process. This study suggests further work on the purification and/or the immunoisolation of tissues grafted in the CNS will be necessary, particularly when the possibility of long-term and repeated grafting is considered.

  14. Skeletal Muscle Regeneration on Protein-Grafted and Microchannel-Patterned Scaffold for Hypopharyngeal Tissue Engineering

    PubMed Central

    Shen, Zhisen; Guo, Shanshan; Ye, Dong; Chen, Jingjing; Kang, Cheng; Qiu, Shejie; Lu, Dakai; Li, Qun; Xu, Kunjie; Lv, Jingjing

    2013-01-01

    In the field of tissue engineering, polymeric materials with high biocompatibility like polylactic acid and polyglycolic acid have been widely used for fabricating living constructs. For hypopharynx tissue engineering, skeletal muscle is one important functional part of the whole organ, which assembles the unidirectionally aligned myotubes. In this study, a polyurethane (PU) scaffold with microchannel patterns was used to provide aligning guidance for the seeded human myoblasts. Due to the low hydrophilicity of PU, the scaffold was grafted with silk fibroin (PU-SF) or gelatin (PU-Gel) to improve its cell adhesion properties. Scaffolds were observed to degrade slowly over time, and their mechanical properties and hydrophilicities were improved through the surface grafting. Also, the myoblasts seeded on PU-SF had the higher proliferative rate and better differentiation compared with those on the control or PU-Gel. Our results demonstrate that polyurethane scaffolds seeded with myoblasts hold promise to guide hypopharynx muscle regeneration. PMID:24175281

  15. Use of Sertoli cell transplants to provide local immunoprotection for tissue grafts.

    PubMed

    Halberstadt, Craig; Emerich, Dwaine F; Gores, Paul

    2004-06-01

    The recent success of allogeneic islet transplantation for the treatment of type I diabetes has renewed interest in cell therapy for diseases of secretory cell dysfunction. Unfortunately, widespread clinical use of cell transplantation is limited by tissue availability and the need for long-term immunosuppresion. Testicular Sertoli cells can confer local immunoprotection for co-transplanted cells and may provide a means of overcoming the obstacles associated with cell transplantation. Sertoli cell grafts protect islets in animal models of diabetes and can be transplanted into the brain to enhance regeneration and promote the survival of co-grafted tissues. This review describes the role that Sertoli cells normally play in testicular immunology, details the preclinical data using transplanted Sertoli cells in models of diabetes and Parkinson's disease and discusses some of the possible mechanisms involved in this phenomena, as well as the future of this technology.

  16. Skeletal muscle regeneration on protein-grafted and microchannel-patterned scaffold for hypopharyngeal tissue engineering.

    PubMed

    Shen, Zhisen; Guo, Shanshan; Ye, Dong; Chen, Jingjing; Kang, Cheng; Qiu, Shejie; Lu, Dakai; Li, Qun; Xu, Kunjie; Lv, Jingjing; Zhu, Yabin

    2013-01-01

    In the field of tissue engineering, polymeric materials with high biocompatibility like polylactic acid and polyglycolic acid have been widely used for fabricating living constructs. For hypopharynx tissue engineering, skeletal muscle is one important functional part of the whole organ, which assembles the unidirectionally aligned myotubes. In this study, a polyurethane (PU) scaffold with microchannel patterns was used to provide aligning guidance for the seeded human myoblasts. Due to the low hydrophilicity of PU, the scaffold was grafted with silk fibroin (PU-SF) or gelatin (PU-Gel) to improve its cell adhesion properties. Scaffolds were observed to degrade slowly over time, and their mechanical properties and hydrophilicities were improved through the surface grafting. Also, the myoblasts seeded on PU-SF had the higher proliferative rate and better differentiation compared with those on the control or PU-Gel. Our results demonstrate that polyurethane scaffolds seeded with myoblasts hold promise to guide hypopharynx muscle regeneration.

  17. Bilateral vestibuloplasty utilizing palatal soft tissue grafts in an HIV-positive patient.

    PubMed

    Kolhatkar, Shilpa; Mason, Suzanne; Winkler, James R; Bhola, Monish

    2009-07-01

    The dental treatment of HIV-positive individuals has undergone a change from the management of HIV-associated oral lesions to routine comprehensive dental care. To the authors' knowledge, this is the first report in which palatal soft tissue grafts were used for vestibuloplasty in an HIV-positive patient with a shallow mandibular vestibule. No adverse sequelae were seen during follow-up.

  18. Preliminary study of coconut water for graft tissues preservation in transplantation.

    PubMed

    César, Jorge Miguel Schettino; Petroianu, Andy; Vasconcelos, Leonardo de Souza; Cardoso, Valbert Nascimento; Mota, Luciene das Graças; Barbosa, Alfredo José Afonso; Soares, Cristina Duarte Vianna; de Oliveira, Amanda Lima

    2015-01-01

    to verify the effectiveness of coconut water in preserving tissues for transplant. Fifty male Wistar rats were randomly distributed in five groups, according to the following preservation solutions for tissue grafts: Group 1: Lactated Ringer; Group 2: Belzer solution; Group 3: mature coconut water; Group 4: green coconut water; Group 5: modified coconut water. In Group 5, the green coconut water has been modified like the Belzer solution. From each animal we harvested the spleen, ovaries and skin of the back segment. These tissues were preserved for six hours in one of the solutions. Then, the grafts were reimplanted. The recovery of the function of the implanted tissues was assessed 90 days after surgery, by splenic scintigraphy and blood exam. The implanted tissues were collected for histopathological examination. The serum levels did not differ among groups, except for the animals in Group 5, which showed higher levels of IgG than Group 1, and differences in relation to FSH between groups 1 and 2 (p <0.001), 4 and 2 (p = 0.03) and 5 and 2 (p = 0.01). The splenic scintigraphy was not different between groups. The ovarian tissue was better preserved in mature coconut water (p <0.007). the coconut water-based solutions preserves spleen, ovary, and rat skin for six hours, maintaining their normal function.

  19. Connective tissue graft vs. emdogain: A new approach to compare the outcomes

    PubMed Central

    Sayar, Ferena; Akhundi, Nasrin; Gholami, Sanaz

    2013-01-01

    Background: The aim of this clinical trial study was to clinically evaluate the use of enamel matrix protein derivative combined with the coronally positioned flap to treat gingival recession compared to the subepithelial connective tissue graft by a new method to obtain denuded root surface area. Materials and Methods: Thirteen patients, each with two or more similar bilateral Miller class I or II gingival recession (40 recessions) were randomly assigned to the test (enamel matrix protein derivative + coronally positioned flap) or control group (subepithelial connective tissue graft). Recession depth, width, probing depth, keratinized gingival, and plaque index were recorded at baseline and at one, three, and six months after treatment. A stent was used to measure the denuded root surface area at each examination session. Results were analyzed using Kolmogorov-Smirnov, Wilcoxon, Friedman, paired-sample t test. Results: The average percentages of root coverage for control and test groups were 63.3% and 55%, respectively. Both groups showed significant keratinized gingival increase (P < 0.05). Recession depth decreased significantly in both groups. Root surface area was improved significantly from baseline with no significant difference between the two study groups (P > 0.05). The results of Friedman test were significant for clinical indices (P < 0.05), except for probing depth in control group (P = 0.166). Conclusion: Enamel matrix protein derivative showed the same results as subepithelial connective tissue graft with relatively easy procedure to perform and low patient morbidity. PMID:23878562

  20. 3D printing of novel osteochondral scaffolds with graded microstructure.

    PubMed

    Nowicki, Margaret A; Castro, Nathan J; Plesniak, Michael W; Zhang, Lijie Grace

    2016-10-14

    Osteochondral tissue has a complex graded structure where biological, physiological, and mechanical properties vary significantly over the full thickness spanning from the subchondral bone region beneath the joint surface to the hyaline cartilage region at the joint surface. This presents a significant challenge for tissue-engineered structures addressing osteochondral defects. Fused deposition modeling (FDM) 3D bioprinters present a unique solution to this problem. The objective of this study is to use FDM-based 3D bioprinting and nanocrystalline hydroxyapatite for improved bone marrow human mesenchymal stem cell (hMSC) adhesion, growth, and osteochondral differentiation. FDM printing parameters can be tuned through computer aided design and computer numerical control software to manipulate scaffold geometries in ways that are beneficial to mechanical performance without hindering cellular behavior. Additionally, the ability to fine-tune 3D printed scaffolds increases further through our investment casting procedure which facilitates the inclusion of nanoparticles with biochemical factors to further elicit desired hMSC differentiation. For this study, FDM was used to print investment-casting molds innovatively designed with varied pore distribution over the full thickness of the scaffold. The mechanical and biological impacts of the varied pore distributions were compared and evaluated to determine the benefits of this physical manipulation. The results indicate that both mechanical properties and cell performance improve in the graded pore structures when compared to homogeneously distributed porous and non-porous structures. Differentiation results indicated successful osteogenic and chondrogenic manipulation in engineered scaffolds.

  1. 3D printing of novel osteochondral scaffolds with graded microstructure

    NASA Astrophysics Data System (ADS)

    Nowicki, Margaret A.; Castro, Nathan J.; Plesniak, Michael W.; Zhang, Lijie Grace

    2016-10-01

    Osteochondral tissue has a complex graded structure where biological, physiological, and mechanical properties vary significantly over the full thickness spanning from the subchondral bone region beneath the joint surface to the hyaline cartilage region at the joint surface. This presents a significant challenge for tissue-engineered structures addressing osteochondral defects. Fused deposition modeling (FDM) 3D bioprinters present a unique solution to this problem. The objective of this study is to use FDM-based 3D bioprinting and nanocrystalline hydroxyapatite for improved bone marrow human mesenchymal stem cell (hMSC) adhesion, growth, and osteochondral differentiation. FDM printing parameters can be tuned through computer aided design and computer numerical control software to manipulate scaffold geometries in ways that are beneficial to mechanical performance without hindering cellular behavior. Additionally, the ability to fine-tune 3D printed scaffolds increases further through our investment casting procedure which facilitates the inclusion of nanoparticles with biochemical factors to further elicit desired hMSC differentiation. For this study, FDM was used to print investment-casting molds innovatively designed with varied pore distribution over the full thickness of the scaffold. The mechanical and biological impacts of the varied pore distributions were compared and evaluated to determine the benefits of this physical manipulation. The results indicate that both mechanical properties and cell performance improve in the graded pore structures when compared to homogeneously distributed porous and non-porous structures. Differentiation results indicated successful osteogenic and chondrogenic manipulation in engineered scaffolds.

  2. Evaluation of small intestine grafts decellularization methods for corneal tissue engineering.

    PubMed

    Oliveira, Ana Celeste; Garzón, Ingrid; Ionescu, Ana Maria; Carriel, Victor; Cardona, Juan de la Cruz; González-Andrades, Miguel; Pérez, María del Mar; Alaminos, Miguel; Campos, Antonio

    2013-01-01

    Advances in the development of cornea substitutes by tissue engineering techniques have focused on the use of decellularized tissue scaffolds. In this work, we evaluated different chemical and physical decellularization methods on small intestine tissues to determine the most appropriate decellularization protocols for corneal applications. Our results revealed that the most efficient decellularization agents were the SDS and triton X-100 detergents, which were able to efficiently remove most cell nuclei and residual DNA. Histological and histochemical analyses revealed that collagen fibers were preserved upon decellularization with triton X-100, NaCl and sonication, whereas reticular fibers were properly preserved by decellularization with UV exposure. Extracellular matrix glycoproteins were preserved after decellularization with SDS, triton X-100 and sonication, whereas proteoglycans were not affected by any of the decellularization protocols. Tissue transparency was significantly higher than control non-decellularized tissues for all protocols, although the best light transmittance results were found in tissues decellularized with SDS and triton X-100. In conclusion, our results suggest that decellularized intestinal grafts could be used as biological scaffolds for cornea tissue engineering. Decellularization with triton X-100 was able to efficiently remove all cells from the tissues while preserving tissue structure and most fibrillar and non-fibrillar extracellular matrix components, suggesting that this specific decellularization agent could be safely used for efficient decellularization of SI tissues for cornea TE applications.

  3. Novel β-TCP/PVA bilayered hydrogels with considerable physical and bio-functional properties for osteochondral repair.

    PubMed

    Yao, Hang; Kang, Junpei; Li, Weichang; Liu, Jian; Xie, Renjian; Wang, Yingjun; Liu, Sa; Wang, Dong An; Ren, Li

    2017-08-09

    Cartilage repairing graft had been widely studied and the osteochondral replacement hydrogels were proved to be an excellent method in research and clinical. However, it was difficult to solve three main issues in osteochondral replacement preparation at one time: surface lubrication, overall mechanical support and good simulations of cell regeneration. A novel integrated bilayered hydrogel osteochondral replacement was constructed by blending polyvinyl alcohol (PVA) and β-tricalcium phosphate (β-TCP) in this study. Separated Nano-ball milling with ultrasound dispersion prepared β-TCP showed suitable properties of tiny particle size, high purity and ideal distribution, which made the novel integrated hydrogel acquired improved mechanical properties, cartilage-like lubrication effect and high biocompatibility including cytocompatibility and osteogenesis. The reinforcement of β-TCP and integrated molding technology made the hydrogel with excellent component compatibility and with good bonding exist between the two layers, which promoted the compression modulus and tensile modulus for up to 3 times strengthen by the mechanical test. The surface lubrication properties of the novel osteochondral hydrogel were similar to the natural cartilage by friction coefficient characterization. The two layer of novel integrated graft provided a considerable bio-function by co-cultured with chondrocytes and synovium mesenchymal stem cells: chondrocytes promoted adherence achieved by upper density layer and better osteogenesis performed by porous down layer. The design of bilayered β-TCP/PVA osteochondral hydrogel would be hopeful in articular cartilage repair. . © 2017 IOP Publishing Ltd.

  4. Porous tantalum biocomposites for osteochondral defect repair

    PubMed Central

    Mrosek, E. H.; Chung, H-W.; Fitzsimmons, J. S.; Reinholz, G. G.; Schagemann, J. C.

    2016-01-01

    Objectives We sought to determine if a durable bilayer implant composed of trabecular metal with autologous periosteum on top would be suitable to reconstitute large osteochondral defects. This design would allow for secure implant fixation, subsequent integration and remodeling. Materials and Methods Adult sheep were randomly assigned to one of three groups (n = 8/group): 1. trabecular metal/periosteal graft (TMPG), 2. trabecular metal (TM), 3. empty defect (ED). Cartilage and bone healing were assessed macroscopically, biochemically (type II collagen, sulfated glycosaminoglycan (sGAG) and double-stranded DNA (dsDNA) content) and histologically. Results At 16 weeks post-operatively, histological scores amongst treatment groups were not statistically different (TMPG: overall 12.7, cartilage 8.6, bone 4.1; TM: overall 14.2, cartilage 9.5, bone 4.9; ED: overall 13.6, cartilage 9.1, bone 4.5). Metal scaffolds were incorporated into the surrounding bone, both in TM and TMPG. The sGAG yield was lower in the neo-cartilage regions compared with the articular cartilage (AC) controls (TMPG 20.8/AC 39.5, TM 25.6/AC 33.3, ED 32.2/AC 40.2 µg sGAG/1 mg respectively), with statistical significance being achieved for the TMPG group (p < 0.05). Hypercellularity of the neo-cartilage was found in TM and ED, as the dsDNA content was significantly higher (p < 0.05) compared with contralateral AC controls (TM 126.7/AC 71.1, ED 99.3/AC 62.8 ng dsDNA/1 mg). The highest type II collagen content was found in neo-cartilage after TM compared with TMPG and ED (TM 60%/TMPG 40%/ED 39%). Inter-treatment differences were not significant. Conclusions TM is a highly suitable material for the reconstitution of osseous defects. TM enables excellent bony ingrowth and fast integration. However, combined with autologous periosteum, such a biocomposite failed to promote satisfactory neo-cartilage formation. Cite this article: E. H. Mrosek, H-W. Chung, J. S. Fitzsimmons, S. W. O’Driscoll, G. G

  5. From natural bone grafts to tissue engineering therapeutics: Brainstorming on pharmaceutical formulative requirements and challenges.

    PubMed

    Baroli, Biancamaria

    2009-04-01

    Tissue engineering is an emerging multidisciplinary field of investigation focused on the regeneration of diseased or injured tissues through the delivery of appropriate molecular and mechanical signals. Therefore, bone tissue engineering covers all the attempts to reestablish a normal physiology or to speed up healing of bone in all musculoskeletal disorders and injuries that are lashing modern societies. This article attempts to give a pharmaceutical perspective on the production of engineered man-made bone grafts that are described as implantable tissue engineering therapeutics, and to highlight the importance of understanding bone composition and structure, as well as osteogenesis and bone healing processes, to improve the design and development of such implants. In addition, special emphasis is given to pharmaceutical aspects that are frequently minimized, but that, instead, may be useful for formulation developments and in vitro/in vivo correlations.

  6. Construction of tissue-engineered small-diameter vascular grafts in fibrin scaffolds in 30 days.

    PubMed

    Gui, Liqiong; Boyle, Michael J; Kamin, Yishai M; Huang, Angela H; Starcher, Barry C; Miller, Cheryl A; Vishnevetsky, Michael J; Niklason, Laura E

    2014-05-01

    Tissue-engineered small-diameter vascular grafts have been developed as a promising alternative to native veins or arteries for replacement therapy. However, there is still a crucial need to improve the current approaches to render the tissue-engineered blood vessels more favorable for clinical applications. A completely biological blood vessel (3-mm inner diameter) was constructed by culturing a 50:50 mixture of bovine smooth muscle cells (SMCs) with neonatal human dermal fibroblasts in fibrin gels. After 30 days of culture under pulsatile stretching, the engineered blood vessels demonstrated an average burst pressure of 913.3±150.1 mmHg (n=6), a suture retention (53.3±15.4 g) that is suitable for implantation, and a compliance (3.1%±2.5% per 100 mmHg) that is comparable to native vessels. These engineered grafts contained circumferentially aligned collagen fibers, microfibrils and elastic fibers, and differentiated SMCs, mimicking a native artery. These promising mechanical and biochemical properties were achieved in a very short culture time of 30 days, suggesting the potential of co-culturing SMCs with fibroblasts in fibrin gels to generate functional small-diameter vascular grafts for vascular reconstruction surgery.

  7. Construction of Tissue-Engineered Small-Diameter Vascular Grafts in Fibrin Scaffolds in 30 Days

    PubMed Central

    Gui, Liqiong; Boyle, Michael J.; Kamin, Yishai M.; Huang, Angela H.; Starcher, Barry C.; Miller, Cheryl A.; Vishnevetsky, Michael J.

    2014-01-01

    Tissue-engineered small-diameter vascular grafts have been developed as a promising alternative to native veins or arteries for replacement therapy. However, there is still a crucial need to improve the current approaches to render the tissue-engineered blood vessels more favorable for clinical applications. A completely biological blood vessel (3-mm inner diameter) was constructed by culturing a 50:50 mixture of bovine smooth muscle cells (SMCs) with neonatal human dermal fibroblasts in fibrin gels. After 30 days of culture under pulsatile stretching, the engineered blood vessels demonstrated an average burst pressure of 913.3±150.1 mmHg (n=6), a suture retention (53.3±15.4 g) that is suitable for implantation, and a compliance (3.1%±2.5% per 100 mmHg) that is comparable to native vessels. These engineered grafts contained circumferentially aligned collagen fibers, microfibrils and elastic fibers, and differentiated SMCs, mimicking a native artery. These promising mechanical and biochemical properties were achieved in a very short culture time of 30 days, suggesting the potential of co-culturing SMCs with fibroblasts in fibrin gels to generate functional small-diameter vascular grafts for vascular reconstruction surgery. PMID:24320793

  8. Characterization of evolving biomechanical properties of tissue engineered vascular grafts in the arterial circulation.

    PubMed

    Udelsman, Brooks V; Khosravi, Ramak; Miller, Kristin S; Dean, Ethan W; Bersi, Matthew R; Rocco, Kevin; Yi, Tai; Humphrey, Jay D; Breuer, Christopher K

    2014-06-27

    We used a murine model to assess the evolving biomechanical properties of tissue engineered vascular grafts (TEVGs) implanted in the arterial circulation. The initial polymeric tubular scaffold was fabricated from poly(lactic acid)(PLA) and coated with a 50:50 copolymer of poly(caprolactone) and poly(lactic acid)(P[PC/LA]). Following seeding with syngeneic bone marrow derived mononuclear cells, TEVGs (n=50) were implanted as aortic interposition grafts in wild-type mice and monitored serially using ultrasound. A custom biaxial mechanical testing device was used to quantify the in vitro circumferential and axial mechanical properties of grafts explanted at 3 or 7 months. At both times, TEVGs were much stiffer than native tissue in both directions. Repeated mechanical testing of some TEVGs treated with elastase or collagenase suggested that elastin did not contribute significantly to the overall stiffness whereas collagen did contribute. Traditional histology and immunostaining revealed smooth muscle cell layers, significant collagen deposition, and increasing elastin production in addition to considerable scaffold at both 3 and 7 months, which likely dominated the high stiffness seen in mechanical testing. These results suggest that PLA has inadequate in vivo degradation, which impairs cell-mediated development of vascular neotissue having properties closer to native arteries. Assessing contributions of individual components, such as elastin and collagen, to the developing neovessel is needed to guide computational modeling that may help to optimize the design of the TEVG. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Optimized preservation of extracellular matrix in cardiac tissues: implications for long-term graft durability.

    PubMed

    Schenke-Layland, Katja; Xie, Jiansong; Heydarkhan-Hagvall, Sepideh; Hamm-Alvarez, Sarah F; Stock, Ulrich A; Brockbank, Kelvin G M; MacLellan, W Robb

    2007-05-01

    Cryopreservation of human tissues, particularly heart valves, is widespread in clinical practice although the effects of this process on underlying tissue structures and its potential impact on valve durability have been poorly studied. Multiphoton imaging and second-harmonic generation (SHG) microscopy permit high-resolution, noninvasive analysis of living tissues at a subcellular level. In the present study we used these novel imaging modalities to compare the effects of vitreous and frozen cryopreservation on the extracellular matrix (ECM) of cardiac tissues. Conventional histology, electron microscopy, and multiphoton imaging to obtain autofluorescence and SHG images were performed on cardiac tissues to characterize the ECM in fresh, vitrified, and frozen cryopreserved tissues. Autofluorescence and particularly SHG images revealed that conventional frozen cryopreservation of cardiac valves, when compared with fresh or vitrified tissues, leads to the loss of normal ECM structures in valve leaflets. Similar results were found in all other cardiac tissues suggesting that structural deterioration of the ECM is a common consequence of frozen cryopreservation. Our results demonstrate that conventional cryopreservation, when compared with fresh or vitrified tissues, causes more destruction of normal ECM structure, which might contribute to eventual graft dysfunction. Whether vitrification preservation will translate into greater durability or less valve failure will need to be determined.

  10. Intravascular Ultrasound Characterization of a Tissue-Engineered Vascular Graft in an Ovine Model.

    PubMed

    Pepper, Victoria K; Clark, Elizabeth S; Best, Cameron A; Onwuka, Ekene A; Sugiura, Tadahisa; Heuer, Eric D; Moko, Lilamarie E; Miyamoto, Shinka; Miyachi, Hideki; Berman, Darren P; Cheatham, Sharon L; Chisolm, Joanne L; Shinoka, Toshiharu; Breuer, Christopher K; Cheatham, John P

    2017-04-01

    Patients who undergo implantation of a tissue-engineered vascular graft (TEVG) for congenital cardiac anomalies are monitored with echocardiography, followed by magnetic resonance imaging or angiography when indicated. While these methods provide data regarding the lumen, minimal information regarding neotissue formation is obtained. Intravascular ultrasound (IVUS) has previously been used in a variety of conditions to evaluate the vessel wall. The purpose of this study was to evaluate the utility of IVUS for evaluation of TEVGs in our ovine model. Eight sheep underwent implantation of TEVGs either unseeded or seeded with bone marrow-derived mononuclear cells. Angiography, IVUS, and histology were directly compared. Endothelium, tunica media, and graft were identifiable on IVUS and histology at multiple time points. There was strong agreement between IVUS and angiography for evaluation of luminal diameter. IVUS offers a valuable tool to evaluate the changes within TEVGs, and clinical translation of this application is warranted.

  11. Perivenous application of cyanoacrylate tissue sealants reduces intimal and medial thickening of the vein graft and inflammatory responses in a rabbit model of carotid artery bypass grafting.

    PubMed

    Dai, Longsheng; Gao, Mingxin; Gu, Chengxiong; Zhang, Fan; Yu, Yang

    2016-02-01

    Effective therapies to prevent vein graft failure after coronary artery bypass grafting (CABG) are still lacking. α-Cyanoacrylate (α-CA, 99% n-octyl-α-cyanoacrylate + n-butyl-α-cyanoacrylate) has been increasingly used as a tissue sealant for wound closure because of its bacteriostatic, biodegradable and haemostatic properties. As a strong tissue adhesive, α-CA might prevent an arterial circulation-induced mechanical stretch on vein graft to attenuate intimal hyperplasia. Here, we investigated the effects of perivenous application of α-CA on the vein graft in a rabbit model of carotid artery bypass grafting. Healthy New Zealand white rabbits were randomized into no graft, graft or graft + α-CA group (n = 10 per group). Rabbit carotid artery was bypassed with the jugular vein. α-CA sealants were sprayed on the entire jugular graft including both anastomotic sites after completion of anastomoses. Blood flow parameters and histological characteristics of the vein grafts including vessel wall thickness, number of medial elastic lamina and proliferation index were evaluated 4 weeks after the surgery. The mRNA or protein levels of proinflammatory factors, chemokine (C-C motif) ligand-2 (CCL-2) and tumour necrosis factor-α (TNF-α) were measured 4 weeks after the operation by quantitative reverse transcription polymerase chain reaction or enzyme-linked immunosorbent assay. Compared with the untreated vein grafts at Week 4 after the operation, the α-CA spray significantly improved graft flow (39.4 ± 1.5 vs 27.8 ± 2.9 ml/min, P < 0.01), attenuated intimal and medial thickening (116.3 ± 1.0 vs 159.7 ± 0.9 μm, P < 0.01), reduced anti-proliferating cell nuclear antigen proliferation index of the vein grafts (15.0 ± 0.4 vs 23.6 ± 0.4%, P < 0.01), decreased the mRNA levels of plasminogen activator inhibitor-1 and CCL-2, and reduced the serum levels of TNF-α (92.9 ± 1.7 vs 102.7 ± 1.8 pg/ml, P < 0.01). Perivenous application of α-CA sealants exerts short

  12. Drilling techniques for osteochondritis dissecans.

    PubMed

    Heyworth, Benton E; Edmonds, Eric W; Murnaghan, M Lucas; Kocher, Mininder S

    2014-04-01

    Although the advanced stages of osteochondritis dissecans remain challenging to treat, most early-stage lesions in skeletally immature patients, if managed appropriately, can be stimulated to heal. For stable lesions that do not demonstrate adequate healing with nonoperative measures, such as activity modification, weight-bearing protection, or bracing, drilling of the subchondral bone has emerged as the gold standard of management. Several techniques of drilling exist, including transarticular drilling, retroarticular drilling, and notch drilling. Although each technique has been shown to be effective in small retrospective studies, higher-powered prospective comparative studies are needed to better elucidate their relative advantages and disadvantages.

  13. Stem cell attraction via SDF-1α expressing fat tissue grafts.

    PubMed

    Zwingenberger, Stefan; Yao, Zhenyu; Jacobi, Angela; Vater, Corina; Valladares, Roberto D; Li, Chenguang; Nich, Christophe; Rao, Allison J; Christman, Jane E; Antonios, Joseph K; Gibon, Emmanuel; Schambach, Axel; Mätzig, Tobias; Günther, Klaus-Peter; Goodman, Stuart B; Stiehler, Maik

    2013-07-01

    Mesenchymal stromal cell (MSCs) are key cellular components for site-specific tissue regeneration. The chemokine stromal derived factor 1 alpha (SDF-1α) is known to attract stem cells via the C-X-C chemokine receptor-4 (CXCR4) receptor. The aim of the study was to develop a model for stem cell attraction using SDF-1α overexpressing fat tissue grafts. Murine MSCs were lentiviral transduced to express the genes for enhanced green fluorescent protein, firefly luciferace, and human CXCR4 (hCXCR4). Murine fat tissue was adenoviral transduced to express SDF-1α and red fluorescent protein transgenes. MSCs were cultured on transwells with SDF-1α containing supernatants from transduced fat tissue. The numbers of migrated MSCs in four groups (with hCXCR4 positive (+) or hCXCR4 negative (-) MSCs with or without SDF-1α containing supernatant) were investigated. After 36 h of culture, 9025 ± 925 cells migrated through the membrane of the transwells in group 1 (CXCR4+/SDF-1α+), 4817 ± 940 cells in group 2 (CXCR4-/SDF-1α+), 2050 ± 766 cells in group 3 (CXCR4+/SDF-1α-), and 2108 ± 426 cells in group 4 (CXCR4-/SDF-1α-). Both, the presence of SDF-1α and the expression of hCXCR4 significantly increased the migration rates (p < 0.0001). MSCs overexpressing the CXCR4 receptor by lentiviral transduction are highly attracted by medium from SDF-1α expressing fat tissue in vitro. Thus, SDF-1α activated tissue grafts may be a strategy to enhance site-specific musculoskeletal tissue regeneration. Copyright © 2012 Wiley Periodicals, Inc.

  14. Stem cell attraction via SDF-1α expressing fat tissue grafts

    PubMed Central

    Zwingenberger, Stefan; Yao, Zhenyu; Jacobi, Angela; Vater, Corina; Valladares, Roberto D.; Li, Chenguang; Nich, Christophe; Rao, Allison J.; Christman, Jane E.; Antonios, Joseph K.; Gibon, Emmanuel; Schambach, Axel; Mätzig, Tobias; Günther, Klaus-Peter; Goodman, Stuart B.; Stiehler, Maik

    2014-01-01

    Mesenchymal stromal cell (MSCs) are key cellular components for site-specific tissue regeneration. The chemokine stromal derived factor 1 alpha (SDF-1α) is known to attract stem cells via the C-X-C chemokine receptor-4 (CXCR4) receptor. The aim of the study was to develop a model for stem cell attraction using SDF-1α overexpressing fat tissue grafts. Murine MSCs were lentiviral transduced to express the genes for enhanced green fluorescent protein, firefly luciferace, and human CXCR4 (hCXCR4). Murine fat tissue was adenoviral transduced to express SDF-1α and red fluorescent protein transgenes. MSCs were cultured on transwells with SDF-1α containing supernatants from transduced fat tissue. The numbers of migrated MSCs in four groups (with hCXCR4 positive (+) or hCXCR4 negative (−) MSCs with or without SDF-1α containing supernatant) were investigated. After 36 h of culture, 9025 ± 925 cells migrated through the membrane of the transwells in group 1 (CXCR4+/SDF-1α+), 4817 ± 940 cells in group 2 (CXCR4-/SDF-1α+), 2050 ± 766 cells in group 3 (CXCR4+/SDF-1α−), and 2108 ± 426 cells in group 4 (CXCR4-/SDF-1α-). Both, the presence of SDF-1α and the expression of hCXCR4 significantly increased the migration rates (p < 0.0001). MSCs overexpressing the CXCR4 receptor by lentiviral transduction are highly attracted by medium from SDF-1α expressing fat tissue in vitro. Thus, SDF-1α activated tissue grafts may be a strategy to enhance site-specific musculoskeletal tissue regeneration. PMID:23281045

  15. [Tissue-engineered graft constructed by bone marrow mononuclear cells and heterogeneous acellularized tissue matrix: an animal experiment].

    PubMed

    Huang, Hui-min; Ma, Liang-long; Ren, Hong; Wu, Shao-feng; Jiang, Zu-ming

    2007-12-25

    To construct a tissue-engineered graft by using bone marrow cells as seeding cells and heterogeneous acellularized matrix as scaffold. Mononuclear cells were isolated from the bone marrow from piglets and cultured in different mediums including either vessel endothelial growth factor (VEGF) or platelet derived growth factor BB (PDGF-BB) to observe the differentiation of cells. Immunoassay was used to detect the expression of specific markers of endothelial cells or specific markers of smooth muscle cells. Adult dogs were killed and their thoracic or abdominal aortas were taken out and processed by multi-step decellularizing technique to remove the original cells while the elastic and collagen fibers were preserved. The undifferentiated bone marrow mononuclear cells were seeded onto the acellularized matrix and incubated in vitro. The cell-seeded grafts were then transplanted to the bone marrow donating piglets to substitute part of the native pulmonary artery. Three weeks later right ventriculography was performed. 100 days later the piglets were sacrificed. The transplanted vessels and the nearby tissues of native pulmonary vessels were excised for inspection. The mononuclear cells cultured in the medium including VEGF showed the morphological features of endothelial cells and were positive of the specific markers of endothelial cells: platelet-endothelial cell adhesion molecule-l, vascular endothelial growth factor receptor Flk-1, VE-cadherin, and plasma factor VIII. The cells cultured in the medium including PDGF-BB showed morphological feature of smooth muscle cells and were positive of he specific marks of smooth muscle cells: alpha-SMA and calponin. One hundred days after transplantation, the inner surfaces of the grafts were smooth without thrombosis, calcification, and aneurysm. The maximal load was 2.76 N and the maximal elongation was 20.31 mm. Under the microscopy a great number of growing endothelial cells and smooth muscle cells could be seen and

  16. Osteochondral tissue formation through adipose-derived stromal cell differentiation on biomimetic polycaprolactone nanofibrous scaffolds with graded insulin and Beta-glycerophosphate concentrations.

    PubMed

    Erisken, Cevat; Kalyon, Dilhan M; Wang, Hongjun; Ornek-Ballanco, Ceren; Xu, Jiahua

    2011-05-01

    The ability to fabricate tissue engineering scaffolds containing systematic gradients in the distributions of stimulators provides additional means for the mimicking of the important gradients observed in native tissues. Here the concentration distributions of two bioactive agents were varied concomitantly for the first time (one increasing, whereas the other decreasing monotonically) in between the two sides of a nanofibrous scaffold. This was achieved via the application of a new processing method, that is, the twin-screw extrusion and electrospinning method, to generate gradients of insulin, a stimulator of chondrogenic differentiation, and β-glycerophosphate (β-GP), for mineralization. The graded poly(ɛ-caprolactone) mesh was seeded with human adipose-derived stromal cells and cultured over 8 weeks. The resulting tissue constructs were analyzed for and revealed indications of selective differentiation of human adipose-derived stromal cells toward chondrogenic lineage and mineralization as functions of position as a result of the corresponding concentrations of insulin and β-GP. Chondrogenic differentiation of the stem cells increased at insulin-rich locations and mineralization increased at β-GP-rich locations.

  17. Clinical application of subdermal areolar tissue and superficial fascia graft as a new material for coverage of small exposure of bone cortex or orthopaedic fixation device.

    PubMed

    Gondo, Masahide; Matsumura, Hajime; Watanabe, Katsueki

    2017-08-01

    The presence of cortical bone, tendon that has been exposed by defects, may result in infection or osteomyelitis. In such cases, perifascial areolar tissue grafting (PATG) may be performed as a minimally invasive surgical procedure. However, perifascial areolar tissue (PAT) is located deep in the subcutaneous layer. It was considered that grafting of the superficial vascular network might enable less invasive surgery. This study reports use of subdermal areolar tissue (SAT) and superficial fascia (SF) to close avascular areas. This study treated eight areas of exposed bone, tendon, or orthopaedic fixation device in seven patients treated in the department between 2010-2013. The patients included five men and two women aged 15-80 years. Subdermal areolar tissue grafting (SATG) was performed on four areas, and superficial fascia grafting (SFG) on the remaining four areas. In all cases, split thickness skin graft (STSG) was used to cover the grafted tissue in a single procedure. The tissue grafted successfully in seven areas, and primary engraftment of the skin grafts was also achieved in three areas treated with SATG and one area treated with SFG. Additional skin grafting was performed to achieve closure in one site treated with SATG and two sites treated with SFG. The tissue graft became necrotic in one site treated with SFG. There were no problems at any donor sites and no graft site infections or other complications. SATG and SFG achieved good clinical results for the closure of exposed avascular tissue or artifacts.

  18. Fluorescence characteristics of human Barrett tissue specimens grafted on chick chorioallantoic membrane.

    PubMed

    Holz, Jasmin A; Boerwinkel, David F; Meijer, Sybren L; Visser, Mike; van Leeuwen, Ton G; Bergman, Jacques J G H M; Aalders, Maurice C G

    2016-01-01

    To improve (pre)malignant lesion identification in Barrett's esophagus (BE), recent research focuses on new developments in fluorescence imaging and spectroscopy to enhance tissue contrast. Our aim was to validate the chorioallantoic membrane (CAM) model as a preclinical tool to study the fluorescence characteristics such as autofluorescence and exogenously induced fluorescence of human Barrett's tissue. Therefore, esophageal biopsy specimens from Barrett's patients were freshly grafted onto the CAM of fertilized hen's eggs to simulate the in vivo situation. The BE biopsy specimens stayed between 1 and 9 days on the CAM to study the persistence of vitality. Fluorescence spectroscopy was performed using six excitation wavelengths (369, 395, 400, 405, 410, 416 nm). Obtained autofluorescence spectra were compared with in vivo spectra of an earlier study. Exogenous administration of 5-aminolevulinic-acid to the biopsy specimens was followed by fluorescence spectroscopy at several time points. Afterwards, the biopsy specimens were harvested and histologically evaluated. In total, 128 biopsy specimens obtained from 34 patients were grafted on the CAM. Biopsy specimens which stayed on average 1.7 days on the CAM were still vital. Autofluorescence spectra of the specimens correlated well with in vivo spectra. Administered 5-aminolevulinic-acid to the biopsy specimens showed conversion into protoporphyrin-IX. In conclusion, we showed that grafting freshly collected human BE biopsy specimens on the CAM is feasible. Our results suggest that the CAM model might be used to study the fluorescence behavior of human tissue specimens. Therefore, the CAM model might be a preclinical research tool for new photosensitizers.

  19. Dimensional changes during early healing after a subepithelial connective tissue graft procedure.

    PubMed

    Rotenberg, Shaun A; Tatakis, Dimitris N

    2014-07-01

    The subepithelial connective tissue graft (CTG) is a popular means to treat gingival recession and augment keratinized tissue. Studies exist that examine long-term outcomes of this procedure; however, changes in tissue dimensions during early healing (0 to 21 days postoperatively) are unknown. The aim of this study is to examine bucco-lingual tissue dimension (gingival tissue thickness [GT]) changes during early CTG healing using a non-invasive technique. Thirteen patients who had treatment planned for CTG on a single tooth were recruited for the study. Using a customized acrylic stent, GT was measured preoperatively, at surgery completion, and at 3, 7, 14, and 21 days postoperatively. CTG was performed using an envelope technique. GT changes were analyzed by repeated-measures analysis of variance. All CTG procedures were considered successful with no postoperative complications. GT increased 1.5 mm immediately after surgery (baseline) compared to the preoperative measurement. GT increased on average 96%, 47%, and 2% compared to baseline at days 3, 7, and 14, respectively. Day 3 and day 7 measurements were significantly different from baseline (P <0.001). At day 21, GT decreased 15% compared to baseline, with an average increase of 1.29 mm from preoperative measurements. The early postoperative healing of CTGs used for root coverage exhibits a significant but transient increase in bucco-lingual tissue dimension. The observed increase in bucco-lingual tissue dimension subsides by the end of the second postoperative week.

  20. Posterosuperior osteochondritis of the calcaneus.

    PubMed

    Cugat, Ramón; Cuscó, Xavier; García, Montserrat; Samitier, Gonzalo; Seijas, Roberto

    2007-09-01

    Osteochondritis of the posterosuperior area of the talocalcaneal surface is a relatively uncommon injury, and only 1 case has been described in the literature. We present a 37-year-old man who complained of pain in the tarsal canal area during walking and when standing up. The magnetic resonance imaging study showed an osteochondral signal in the posterosuperior medial area of the calcaneus on the talocalcaneal surface. The persistence of pain and lack of improvement with conservative treatment made arthroscopic debridement of the injury necessary. The arthroscopic procedure was performed through 2 medial portals, made under fluoroscopy, marked with needles, and dissected with mosquito clamps, and the affected surface could be fully visualized, showing a chondral lesion. Debridement of the osteonecrotic area was performed, and the Steadman technique was used on the injured bone surface. The patient was pain-free, and limited activity (i.e., standing up and walking without symptoms) was allowed. After 24 months, the patient remains asymptomatic with weight-bearing working activities and when standing. Arthroscopic curettage and scission of the injury have been shown to yield good or excellent outcomes in 75% to 80% of patients with regard to the talar surface.

  1. EHTIC study: Evaluation of a new hemostatic agent based on tissue factor in skin grafting procedures.

    PubMed

    Rojas, Santiago; Pérez Del Caz, M Dolores; Esteban Vico, Juan Ramón; Villaverde, Eloísa; Llinas, Abel; Martínez, José Ramón; Brage, Carlos; Valero, Javier; González Rodríguez, Alba; Garcia Barreiro, Juan; López-Suso, Eugenia; Fernandez-Cañamaque, Jose Luis; López, Ramón; Murat, Jesus

    2017-06-01

    Excessive bleeding is a major concern in scar debridement and grafting procedures. TT-173 is a new topical hemostatic agent based on recombinant human tissue factor that has shown promising results in patients who underwent tooth extraction. EHTIC study sought to evaluate the efficacy and safety of TT-173 to reduce the bleeding in donor sites of skin grafting procedures. EHTIC study was a phase II, randomized, parallel, double blind, placebo controlled trial. Patients received TT-173 (n=38) or placebo (n=33) sprayed over donor site after graft harvest. Time to hemostasis and incidence of adverse events were recorded. Systemic absorption of the product and its immunogenicity were also measured during the follow up of the subjects. Treatment with TT-173 significantly reduced the bleeding time from 7 to 3min (Log-Rank p<0.0001). Moreover, bleeding stopped within the 10min of evaluation period in all the patients that received TT-173. In contrast, 24.24% of patients from placebo group required additional measures to arrest hemorrhage (Fisher p=0.0013). Product related adverse events, systemic absorption into blood stream, interferences with the healing of the donor site or immunogenic reaction against TT-173 were not observed. The new hemostatic agent TT-173 has proven efficacious and safe to reduce the bleeding from donor site. This study paves the way for further investigation of the product as topical hemostatic treatment in plastic surgery and other surgical indications. Copyright © 2017 Elsevier Ltd and ISBI. All rights reserved.

  2. Alveolar Ridge Contouring with Free Connective Tissue Graft at Implant Placement: A 5-Year Consecutive Clinical Study.

    PubMed

    Hanser, Thomas; Khoury, Fouad

    2016-01-01

    This study evaluated volume stability after alveolar ridge contouring with free connective tissue grafts at implant placement in single-tooth gaps. A total of 52 single-tooth gaps with labial volume deficiencies in the maxilla (incisors, canines, and premolars) were consecutively treated with implants and concomitant free palatal connective tissue grafts in 46 patients between 2006 and 2009. Implants had to be covered with at least 2 mm peri-implant local bone after insertion. At implant placement, a free connective tissue graft from the palate was fixed inside a labial split-thickness flap to form an existing concave buccal alveolar ridge contour due to tissue volume deficiency into a convex shape. Standardized volumetric measurements of the labial alveolar contour using a template were evaluated before connective tissue grafting and at 2 weeks, 1 year, and 5 years after implantprosthetic incorporation. Tissue volume had increased significantly (P < .05) in all six reference points representing the outer alveolar soft tissue contour of the implant before connective tissue grafting to baseline (2 weeks after implant-prosthetic incorporation). Statistically, 50% of the reference points (P > .05) kept their volume from baseline to 1 year after prosthetic incorporation and from baseline to 5 years after prosthetic incorporation, respectively, whereas reference points located within the area of the implant sulcus showed a significant (P < .05) decrease in volume. Clinically, 5 years after prosthetic incorporation the originally concave buccal alveolar contour was still convex in all implants, leading to a continuous favorable anatomical shape and improved esthetic situation. Intraoral radiographs confirmed osseointegration and stable peri-implant parameters with a survival rate of 100% after a follow-up of approximately 5 years. Implant placement with concomitant free connective tissue grafting appears to be an appropriate long-term means to contour preexisting buccal

  3. Computationally Optimizing the Compliance of a Biopolymer Based Tissue Engineered Vascular Graft

    PubMed Central

    Harrison, Scott; Tamimi, Ehab; Uhlorn, Josh; Leach, Tim; Vande Geest, Jonathan P.

    2016-01-01

    Coronary heart disease is a leading cause of death among Americans for which coronary artery bypass graft (CABG) surgery is a standard surgical treatment. The success of CABG surgery is impaired by a compliance mismatch between vascular grafts and native vessels. Tissue engineered vascular grafts (TEVGs) have the potential to be compliance matched and thereby reduce the risk of graft failure. Glutaraldehyde (GLUT) vapor-crosslinked gelatin/fibrinogen constructs were fabricated and mechanically tested in a previous study by our research group at 2, 8, and 24 hrs of GLUT vapor exposure. The current study details a computational method that was developed to predict the material properties of our constructs for crosslinking times between 2 and 24 hrs by interpolating the 2, 8, and 24 hrs crosslinking time data. matlab and abaqus were used to determine the optimal combination of fabrication parameters to produce a compliance matched construct. The validity of the method was tested by creating a 16-hr crosslinked construct of 130 μm thickness and comparing its compliance to that predicted by the optimization algorithm. The predicted compliance of the 16-hr construct was 0.00059 mm Hg−1 while the experimentally determined compliance was 0.00065 mm Hg−1, a relative difference of 9.2%. Prior data in our laboratory has shown the compliance of the left anterior descending porcine coronary (LADC) artery to be 0.00071 ± 0.0003 mm Hg−1. Our optimization algorithm predicts that a 258-μm-thick construct that is GLUT vapor crosslinked for 8.1 hrs would match LADC compliance. This result is consistent with our previous work demonstrating that an 8-hr GLUT vapor crosslinked construct produces a compliance that is not significantly different from a porcine coronary LADC. PMID:26593773

  4. Computationally Optimizing the Compliance of a Biopolymer Based Tissue Engineered Vascular Graft.

    PubMed

    Harrison, Scott; Tamimi, Ehab; Uhlorn, Josh; Leach, Tim; Vande Geest, Jonathan P

    2016-01-01

    Coronary heart disease is a leading cause of death among Americans for which coronary artery bypass graft (CABG) surgery is a standard surgical treatment. The success of CABG surgery is impaired by a compliance mismatch between vascular grafts and native vessels. Tissue engineered vascular grafts (TEVGs) have the potential to be compliance matched and thereby reduce the risk of graft failure. Glutaraldehyde (GLUT) vapor-crosslinked gelatin/fibrinogen constructs were fabricated and mechanically tested in a previous study by our research group at 2, 8, and 24 hrs of GLUT vapor exposure. The current study details a computational method that was developed to predict the material properties of our constructs for crosslinking times between 2 and 24 hrs by interpolating the 2, 8, and 24 hrs crosslinking time data. matlab and abaqus were used to determine the optimal combination of fabrication parameters to produce a compliance matched construct. The validity of the method was tested by creating a 16-hr crosslinked construct of 130 μm thickness and comparing its compliance to that predicted by the optimization algorithm. The predicted compliance of the 16-hr construct was 0.00059 mm Hg-1 while the experimentally determined compliance was 0.00065 mm Hg-1, a relative difference of 9.2%. Prior data in our laboratory has shown the compliance of the left anterior descending porcine coronary (LADC) artery to be 0.00071 ± 0.0003 mm Hg-1. Our optimization algorithm predicts that a 258-μm-thick construct that is GLUT vapor crosslinked for 8.1 hrs would match LADC compliance. This result is consistent with our previous work demonstrating that an 8-hr GLUT vapor crosslinked construct produces a compliance that is not significantly different from a porcine coronary LADC.

  5. The role of myeloid cell-derived PDGF-B in neotissue formation in a tissue-engineered vascular graft.

    PubMed

    Onwuka, Ekene; Best, Cameron; Sawyer, Andrew; Yi, Tai; Heuer, Eric; Sams, Malik; Wiet, Matthew; Zheng, Hong; Kyriakides, Themis; Breuer, Christopher

    2017-04-01

    Inflammatory myeloid lineage cells mediate neotissue formation in tissue-engineered vascular grafts, but the molecular mechanism is not completely understood. We examined the role of vasculogenic PDGF-B in tissue-engineered vascular graft neotissue development. Myeloid cell-specific PDGF-B knockout mice (PDGF-KO) were generated using bone marrow transplantation, and scaffolds were implanted as inferior vena cava interposition grafts in either PDGF-KO or wild-type mice. After 2 weeks, grafts from PDGF-KO mice had more remaining scaffold polymer and less intimal neotissue development. Increased macrophage apoptosis, decreased smooth muscle cell proliferation and decreased collagen content was also observed. Myeloid cell-derived PDGF contributes to vascular neotissue formation by regulating macrophage apoptosis, smooth muscle cell proliferation and extracellular matrix deposition.

  6. Acellular dermal allograft for vestibuloplasty--an alternative to autogenous soft tissue grafts in preprosthetic surgical procedures: a clinical report.

    PubMed

    Bhola, Monish; Newell, Donald H; Hancock, Everett B

    2003-06-01

    Various vestibular extension procedures have been described in the literature over the past 6 decades, including the use of free gingival grafts. An acellular dermal allograft has been used as a substitute for autogenous soft tissue grafts in root coverage procedures. This clinical report describes the use of such an allograft as a substitute for palatal donor tissue in the vestibular extension of an edentulous maxillary arch with multiple frenum attachments before fabrication of a complete denture. In this patient, healing was uneventful, and 6-month clinical observations demonstrated an apical positioning of the mucogingival junction with an increase in vestibular depth, and the absence of multiple frenae. The acellular dermal allograft appears to be a useful substitute for autogenous palatal grafts in preprosthetic surgery. This approach has many advantages over the free gingival graft, including no donor site morbidity, unlimited availability, and better color match.

  7. Arthroscopic Particulated Juvenile Cartilage Allograft Transplantation for the Treatment of Osteochondral Lesions of the Talus

    PubMed Central

    Adams, Samuel B.; Demetracopoulos, Constantine A.; Parekh, Selene G.; Easley, Mark E.; Robbins, Justin

    2014-01-01

    Several options exist for the treatment of osteochondral lesions of the talus. Particulated juvenile cartilage allograft transplantation (PJCAT) has become a viable treatment modality for osteochondral lesions of the talus that are not amenable to microfracture or for which microfracture has failed. Arthroscopic placement of this type of graft obviates the need for osteotomy or plafondplasty and does not prevent additional procedures from being performed through an anterior approach. Special instrumentation and setup are not required to perform this procedure. Our arthroscopic technique for placement of particulated juvenile cartilage into osteochondral lesions of the talus is described. Case series and outcomes after arthroscopic ankle PJCAT are currently not reported within the literature; however, it is believed that the outcomes are at least similar to those of open ankle PJCAT. PMID:25264516

  8. The Use of Autologous Fat Grafting for Treatment of Scar Tissue and Scar-Related Conditions: A Systematic Review.

    PubMed

    Negenborn, Vera L; Groen, Jan-Willem; Smit, Jan Maerten; Niessen, Frank B; Mullender, Margriet G

    2016-01-01

    Scar tissue can cause cosmetic impairments, functional limitations, pain, and itch. It may also cause emotional, social, and behavioral problems, especially when it is located in exposed areas. To date, no gold standard exists for the treatment of scar tissue. Autologous fat grafting has been introduced as a promising treatment option for scar tissue-related symptoms. However, the scientific evidence for its effectiveness remains unclear. This systematic review aims to evaluate the available evidence regarding the effectiveness of autologous fat grafting for the treatment of scar tissue and scar-related conditions. A systematic literature review was performed using MEDLINE, Cochrane Library, EMBASE, and Web of Science. No language restrictions were imposed. Twenty-six clinical articles were included, reporting on 905 patients in total. Meta-analysis was not performed because of the heterogeneous methodology demonstrated among the articles. Main outcome measures were scar appearance and skin characteristics, restoration of volume and/or (three-dimensional) contour, itch, and pain. All publications report a beneficial effect of autologous fat grafting on scar tissue. There is statistical significant improvement of the scar appearance, skin characteristics, and pain. Itch and restoration of volume and three-dimensional contour also improved. Autologous fat grafting is used to improve a variety of symptoms related to scar tissue. This systematic review suggests that autologous fat grafting provides beneficial effects with limited side effects. However, the level of evidence and methodological quality are quite low. Future randomized controlled trials with a methodologically strong design are necessary to confirm the effects of autologous fat grafting on scar tissue and scar-related conditions.

  9. Nanopatterning of collagen scaffolds improve the mechanical properties of tissue engineered vascular grafts.

    PubMed

    Zorlutuna, P; Elsheikh, A; Hasirci, V

    2009-04-13

    Tissue engineered constructs with cells growing in an organized manner have been shown to have improved mechanical properties. This can be especially important when constructing tissues that need to perform under load, such as cardiac and vascular tissue. Enhancement of mechanical properties of tissue engineered vascular grafts via orientation of smooth muscle cells by the help of topographical cues have not been reported yet. In the present study, collagen scaffolds with 650, 500, and 332.5 nm wide nanochannels and ridges were designed and seeded with smooth muscle cells isolated from the human saphenous vein. Cell alignment on the construct was shown by SEM and fluorescence microscopy. The ultimate tensile strength (UTS) and Young's modulus of the scaffolds were determined after 45 and 75 days. Alamar Blue assay was used to determine the number of viable cells on surfaces with different dimensioned patterns. Presence of nanopatterns increased the UTS from 0.55 +/- 0.11 to as much as 1.63 +/- 0.46 MPa, a value within the range of natural arteries and veins. Similarly, Young's modulus values were found to be around 4 MPa, again in the range of natural vessels. The study thus showed that nanopatterns as small as 332.5 nm could align the smooth muscle cells and that alignment significantly improved mechanical properties, indicating that nanopatterned collagen scaffolds have the potential for use in the tissue engineering of small diameter blood vessels.

  10. Biphasic nanofibrous constructs with seeded cell layers for osteochondral repair.

    PubMed

    Jin, Guang-Zhen; Kim, Jung-Ju; Park, Jeong-Hui; Seo, Seog-Jin; Kim, Joong-Hyun; Lee, Eun-Jung; Kim, Hae-Won

    2014-11-01

    Biphasic scaffolds have gained increasing attention for the regeneration of osteochondral interfacial tissue because they are expected to effectively define the interfacial structure of tissue that comprises stratified cartilage with a degree of calcification. Here, we propose a biphasic nanofiber construct made of poly(lactide-co-caprolactone) (PLCL) and its mineralized form (mPLCL) populated with cells. Primary rat articular chondrocytes (ACs) and bone marrow-derived mesenchymal stem cells (MSCs) were cultured on the layers of bare PLCL and mPLCL nanofibers, respectively, for 7 days, and the biphasic cell-nanofiber construct was investigated at 4 weeks after implantation into nude mice. Before implantation, the ACs and MSCs grown on each layer of PLCL and mPLCL nanofibers exhibited phenotypes typical of chondrocytes and osteoblasts, respectively, under proper culture conditions, as analyzed by electron microscopy, histological staining, cell growth kinetics, and real-time polymerase chain reaction. The biphasic constructs also showed the development of a possible formation of cartilage and bone tissue in vivo. Results demonstrated that the cell-laden biphasic nanofiber constructs may be useful for the repair of osteochondral interfacial tissue structure.

  11. In vivo tissue-engineered small-caliber arterial graft prosthesis consisting of autologous tissue (biotube).

    PubMed

    Nakayama, Yasuhide; Ishibashi-Ueda, Hatsue; Takamizawa, Keiichi

    2004-01-01

    In this study, vascular-like tubular tissues called biotubes, consisting of autologous tissues, were prepared using in vivo tissue engineering. Their mechanical properties were evaluated for application as a small-caliber artificial vascular prosthesis. The biotubes were prepared by embedding six kinds of polymeric rods [poly(ethylene) (PE), poly(fluoroacetate) (PFA), poly(methyl methacrylate) (PMMA), segmented poly(urethane) (PU), poly(vinyl chloride) (PVC), and silicone (Si)] as a mold in six subcutaneous pouches in the dorsal skin of New Zealand White rabbits. For rods apart from PFA, biotubes were constructed after 1 month of implantation by encapsulation around the polymeric implants. The wall thickness of the biotubes ranged from about 50 to 200 microm depending on the implant material and were in the order PFA < PVC < PMMA < PU < PE. As for PE, PMMA, and PVC, the thickness increased after 3 months of implantation and ranged from 1.5-to 2-fold. None of the biotubes were ruptured when a hydrostatic pressure was gradually applied to their lumen up to 200 mmHg. The relationship between the intraluminal pressure and the external diameter, which was highly reproducible, showed a "J"-shaped curve similar to the native artery. The tissue mostly consisted of collagen-rich extracellular matrices and fibroblasts. Generally, the tissue was relatively firm and inelastic for Si and soft for PMMA. For PMMA, PE, and PVC the stiffness parameter (beta value; one of the indexes for compliance) of the biotubes obtained was similar to those of the human coronary, femoral, and carotid arteries, respectively. Biotubes, which possess the ability for wide adjustments in their matrices, mechanics, shape, and luminal surface design, can be applied for use as small-caliber blood vessels and are an ideal implant because they avoid immunological rejection.

  12. Long-term storage and preservation of tissue engineered articular cartilage.

    PubMed

    Nover, Adam B; Stefani, Robert M; Lee, Stephanie L; Ateshian, Gerard A; Stoker, Aaron M; Cook, James L; Hung, Clark T

    2016-01-01

    With limited availability of osteochondral allografts, tissue engineered cartilage grafts may provide an alternative treatment for large cartilage defects. An effective storage protocol will be critical for translating this technology to clinical use. The purpose of this study was to evaluate the efficacy of the Missouri Osteochondral Allograft Preservation System (MOPS) for room temperature storage of mature tissue engineered grafts, focusing on tissue property maintenance during the current allograft storage window (28 days). Additional research compares MOPS to continued culture, investigates temperature influence, and examines longer-term storage. Articular cartilage constructs were cultured to maturity using adult canine chondrocytes, then preserved with MOPS at room temperature, in refrigeration, or kept in culture for an additional 56 days. MOPS storage maintained desired chondrocyte viability for 28 days of room temperature storage, retaining 75% of the maturity point Young's modulus without significant decline in biochemical content. Properties dropped past this time point. Refrigeration maintained properties similar to room temperature at 28 days, but proved better at 56 days. For engineered grafts, MOPS maintained the majority of tissue properties for the 28-day window without clearly extending that period as it had for native grafts. These results are the first evaluating engineered cartilage storage.

  13. [The future of treatment for chondral and osteochondral lesions].

    PubMed

    Cirpar, Meriç; Korkusuz, Feza

    2007-01-01

    The population of patients with symptomatic focal or generalized cartilage lesions is growing due to prolongation of life expectancy and to increasing frequency of sports injuries. Cartilage tissue lesions which were defined as untreatable in the past have now become treatable thanks to advances in basic scientific research. With the development of technologies regarding biomaterial, cell and local regulators, and with the introduction of new surgical techniques, it is estimated that, in the near future, clinical applications of cartilage tissue engineering will also receive particular attention in our country. Currently, all alternatives used in the treatment of cartilage lesions have merits and demerits, including arthroscopic debridement and lavage, mesenchymal stem cell stimulation, osteochondral replacement techniques, and autologous chondrocyte transplantation. Preliminary results of experimental cartilage tissue engineering are encouraging for the replacement of disrupted tissue with that having mechanical properties of hyaline cartilage. Clinical applications of cartilage tissue engineering include bioabsorbable scaffolds as extracellular collagen, hyaluronic acid matrices, and genetically engineered bioactive materials.

  14. Endoscopic management of tissue-engineered tracheal graft stenosis in an ovine model.

    PubMed

    Pepper, Victoria K; Onwuka, Ekene A; Best, Cameron A; King, Nakesha; Heuer, Eric; Johnson, Jed; Breuer, Christopher K; Grischkan, Jonathan M; Chiang, Tendy

    2017-10-01

    To evaluate the safety and efficacy of bronchoscopic interventions in the management of tissue-engineered tracheal graft (TETG) stenosis. Animal research study. TETGs were constructed with seeded autologous bone marrow-derived mononuclear cells on a bioartificial graft. Eight sheep underwent tracheal resection and orthotopic implantation of this construct. Animals were monitored by bronchoscopy and fluoroscopy at 3 weeks, 6 weeks, 3 months, and 4 months. Bronchoscopic interventions, including dilation and stenting, were performed to manage graft stenosis. Postdilation measurements were obtained endoscopically and fluoroscopically. Seven dilations were performed in six animals. At the point of maximal stenosis, the lumen measured 44.6 ± 8.4 mm(2) predilation and 50.7 ± 14.1 postdilation by bronchoscopy (P = 0.3517). By fluoroscopic imaging, the airway was 55.9 ± 12.9 mm(2) predilation and 65.9 ± 22.4 mm(2) postdilation (P = 0.1303). Stents were placed 17 times in six animals. Pre- and poststenting lumen sizes were 62.8 ± 38.8 mm(2) and 80.1 ± 54.5 mm(2) by bronchoscopy (P = 0.6169) and 77.1 ± 38.9 mm(2) and 104 ± 60.7 mm(2) by fluoroscopy (P = 0.0825). Mortality after intervention was 67% with dilation and 0% with stenting (P = 0.0004). The average days between bronchoscopy were 8 ± 2 for the dilation group and 26 ± 17 in the stenting group (P = 0.05). One hundred percent of dilations and 29% of stent placements required urgent follow-up bronchoscopy (P = 0.05). Dilation has limited efficacy for managing TETG stenosis, whereas stenting has a more lasting clinical effect. NA. Laryngoscope, 127:2219-2224, 2017. © 2017 The American Laryngological, Rhinological and Otological Society, Inc.

  15. The innate immune system contributes to tissue-engineered vascular graft performance.

    PubMed

    Hibino, Narutoshi; Mejias, Dane; Pietris, Nicholas; Dean, Ethan; Yi, Tai; Best, Cameron; Shinoka, Toshiharu; Breuer, Christopher

    2015-06-01

    The first clinical trial of tissue-engineered vascular grafts (TEVGs) identified stenosis as the primary cause of graft failure. In this study, we aimed to elucidate the role of the host immune response in the development of stenosis using a murine model of TEVG implantation. We found that the C.B-17 wild-type (WT) mouse (control) undergoes a dramatic stenotic response, which is nearly completely abolished in the immunodeficient SCID/beige (bg) variant. SCID mice, which lack an adaptive immune system due to the absence of T and B lymphocytes, experienced rates of stenosis comparable to WT controls (average luminal diameter, WT: 0.071 ± 0.035 mm, SCID: 0.137 ± 0.032 mm, SCID/bg: 0.804 ± 0.039 mm; P < 0.001). The bg mutation is characterized by NK cell and platelet dysfunction, and systemic treatment of WT mice with either NK cell-neutralizing (anti-NK 1.1 antibody) or antiplatelet (aspirin/Plavix [clopidogrel bisulfate]; Asp/Pla) therapy achieved nearly half the patency observed in the SCID/bg mouse (NK Ab: 0.356 ± 0.151 mm, Asp/Pla: 0.452 ± 0.130 mm). Scaffold implantation elicited a blunted immune response in SCID/bg mice, as demonstrated by macrophage number and mRNA expression of proinflammatory cytokines in TEVG explants. Implicating the initial innate immune response as a critical factor in graft stenosis may provide a strategy for prognosis and therapy of second-generation TEVGs.

  16. Nasal spreader grafts: a comparison of medpor to autologous tissue reconstruction.

    PubMed

    Reiffel, Alyssa J; Cross, Kevin J; Spinelli, Henry M

    2011-01-01

    Reconstruction of the damaged nasal vault is challenging. Limited available autologous tissue has lead surgeons to pursue alloplastic alternatives. A retrospective review comparing 18 patients who underwent secondary rhinoplasty with internal nasal valve reconstruction with spreader graft (SG) implants using either autologous tissue or high-density porous polyethylene (Medpor) was performed. All underwent bilateral SG reconstruction of the internal nasal valve with Medpor (10 cases) or autologous cartilage (8 cases). Mean follow-up was 26 months for the autologous group and 29 months for the Medpor group. Functional performance and aesthetic results were identical. Complications were few: 1 case of unilateral infection in the Medpor group treated with partial excision, and 1 case of erythema at the auricular donor site for the autologous tissue group. For patients who have exhausted autologous tissue options or are unwilling to tolerate potential donor-site morbidity, the Medpor SG is an appropriate option that allows for excellent aesthetic and functional results that remains stable over time.

  17. Midterm results of osteochondral allograft transplantation to the humeral head.

    PubMed

    Riff, Andrew J; Yanke, Adam B; Shin, Jason J; Romeo, Anthony A; Cole, Brian J

    2017-07-01

    This study evaluated clinical outcomes of osteochondral allograft (OCA) transplantation for humeral head osteochondral defects. We hypothesized that patients with isolated humeral head disease would achieve favorable results and that patients with bipolar disease would experience inferior outcomes. We identified patients who underwent humeral head OCA transplantation. Subjective questionnaire data were obtained preoperatively and at a minimum of 2 years postoperatively. Radiographs were evaluated for graft incorporation. Failure was defined by conversion to shoulder arthroplasty, American Shoulder and Elbow Surgeons score <50, or dissatisfaction with the surgical result. Twenty patients (65% male) met inclusion criteria. Patients were an average age of 24.8 ± 8.1 years. Eleven patients underwent concomitant glenoid surgery (microfracture or meniscal allograft resurfacing). Follow-up was available for 18 patients (90%) at mean of 67 months. All grafts incorporated except 2. Four patients underwent shoulder arthroplasty at mean of 25 months postoperatively (all after pain pump chondrolysis). Eleven of the 20 patients were satisfied (all dissatisfied patients underwent glenoid surgery). Significant improvements (P < .001) were seen for the visual analog scale (from 6.1 to 1.5), Simple Shoulder Test (from 32 to 73), American Shoulder and Elbow Surgeons score (from 39 to 76), and the physical component of the 12-Item Short Form Survey (from 38 to 48). Pain pump patients who did not progress to arthroplasty experienced inferior satisfaction (40% vs. 87.5%, P = .04) and a trend toward inferior outcomes compared with the rest of the cohort. OCA transplantation is a viable option for young patients with isolated humeral chondral injury. Patients with bipolar disease or a history of intra-articular pain pump have increased failure and decreased subjective outcomes. Copyright © 2016 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier

  18. Graft-induced behavioral recovery from subcallosal septohippocampal damage in rats depends on maturity stage of donor tissue.

    PubMed

    Cassel, J C; Kelche, C; Peterson, G M; Ballough, G P; Goepp, I; Will, B

    1991-01-01

    Long-Evans female rats sustained electrolytic lesions of the fimbria and the dorsal fornix and, 10-14 days later, received intrahippocampal suspension grafts of septal-diagonal band tissue from either 14-day-old (Group S14, n = 8) or 16-day-old fetuses (Group S16, n = 10), or of parietal cortex from 16-day-old fetuses (Group Cx, n = 10). Sham-operated (Group S, n = 10) and lesion-only (Group Fifo, n = 21) rats served as non-grafted controls. Spontaneous alternation was assessed in a T-maze at three weeks and two months post-grafting. Home cage and open field activity as well as radial maze learning were assessed from two months post-grafting onwards. Fimbria-fornix lesions induced lasting hyperactivity in both the open field and the home cage, impaired radial maze learning and transiently reduced spontaneous alternation rates. Neither type of graft significantly affected home cage activity. Septal-diagonal band grafts improved open field habituation (within trial decline of ambulatory activity) and radial maze learning; the former was observed only in S16 rats, whereas the latter was observed only in S14 rats. Acetylcholinesterase histochemistry revealed an initial lesion-induced depletion of hippocampal acetylcholinesterase (eight days post-surgery) which was no longer observed at the end of the experiment. Acetylcholinesterase positivity was similar in S14 and S16 grafts, which also contained many choline acetyltransferase-positive neurons. Cortical grafts were found to be almost devoid of acetylcholinesterase positivity and no well-stained choline acetyltransferase-positive neurons could be identified. Septal-diagonal band grafts from 14-day-old fetuses and cortical grafts contained more parvalbumin-positive neurons than septal-diagonal band grafts provided by 16-day-old fetuses. These results suggest that grafts rich in cholinergic neurons may promote behavioral recovery from fimbria-fornix lesion-induced deficits. However, such a recovery may concern different

  19. Chondrocyte viability in fresh and frozen large human osteochondral allografts: effect of cryoprotective agents.

    PubMed

    Judas, F; Rosa, S; Teixeira, L; Lopes, C; Ferreira Mendes, A

    2007-10-01

    Chondrocyte survival is a major goal for the effective storage and clinical performance of human osteochondral allografts. The majority of animal and human cryopreservation studies conducted so far have been performed in small osteochondral cylinders. Using human tibial plateaus as a model for large osteochondral pieces, this work sought to evaluate the cryoprotective efficiency of glycerol and dimethylsulfoxide (DMSO), and to identify cryopreservation conditions suitable for use in tissue banks. Human tibial plateaus harvested from 7 cadaveric tissue donors were incubated in the presence or absence of cryoprotective agents (CPA): 10% or 15% glycerol and 10% DMSO in a Ham F-12 nutrient mixture. Chondrocyte viability was assessed immediately after thawing, using the MTT reduction assay and a fluorescence microscopic method. The tibial plateaus frozen in the absence of CPA showed a significant decrease in chondrocyte viability. The use of CPA significantly increased chondrocyte viability compared with cartilage frozen without CPA (nearly 50% versus 80% living chondrocytes with 10% glycerol versus 10% DMSO, respectively) relative to that in fresh cartilage. In this regard, 10% DMSO was slightly more effective than either 10% or 15% glycerol, eliciting the recovery of approximately 15% relative to the living chondrocyte content in fresh cartilage. In all conditions, fluorescence microscopic studies showed that surviving chondrocytes were restricted to the superficial cartilage layer. Human tibial plateaus seemed to be a good experimental model to establish cryopreservation methods applicable to large human osteochondral pieces in tissue banks.

  20. Open mosaicplasty in osteochondral lesions of the talus: a prospective study.

    PubMed

    Emre, Tuluhan Yunus; Ege, Tolga; Cift, Hakan Turan; Demircioğlu, Demet Tekdöş; Seyhan, Bahadir; Uzun, Macit

    2012-01-01

    Osteochondral lesions of the talus present with symptoms of pain and painful motion, affecting the quality of the patient's daily life. We evaluated the 2-year short-term outcomes of patients whose large osteochondral lesions of the talus were treated with medial malleolar osteotomy and a mosaic graft harvested from the knee on the same side. A total of 32 patients who had cartilage lesions due to osteochondritis dissecans in the medial aspect of the talus underwent mosaicplasty after medial malleolar osteotomy. The patients were followed up for a mean period of 16.8 (range 12 to 24) months. The staging and treatment plan of the osteochondral lesions of the talus were made according to the Bristol classification. The follow-up protocol for the patients included direct radiography and magnetic resonance imaging. The American Orthopaedic Foot and Ankle Society scoring system was used to assess the patients during the pre- and postoperative periods. Of the 32 patients, 3 (9.4%) were female and 29 (90.6%) male, with a mean age of 27.5 (range 20 to 47) years. The mean preoperative American Orthopaedic Foot and Ankle Society score was 59.12 ± 7.72 but had increased to 87.94 ± 3.55 during the postoperative 2 years. The increase in American Orthopaedic Foot and Ankle Society score was statistically significant (p < .05). We have concluded that open mosaicplasty is a reliable and effective method for the treatment of osteochondral lesions with subchondral cyst formation in the talus, exceeding 1.5 cm in diameter.

  1. Hybrid Matrix Grafts to Favor Tissue Regeneration in Rabbit Femur Bone Lesions

    PubMed Central

    Goy, Dante Pascual; Gorosito, Emmanuel; Costa, Hermes S; Mortarino, Pablo; Pedemonte, Noelia Acosta; Toledo, Javier; Mansur, Herman S; Pereira, Marivalda M; Battaglino, Ricardo; Feldman, Sara

    2012-01-01

    At present, typical approaches employed to repair fractures and other bone lesions tend to use matrix grafts to promote tissue regeneration. These grafts act as templates, which promote cellular adhesion, growth and proliferation, osteoconduction, and even osteoinduction, which commonly results in de novo osteogenesis. The present work aimed to study the bone-repairing ability of hybrid matrixes (HM) prepared with polyvinyl alcohol (PVA) and bioactive glass in an experimental rabbit model. The HM were prepared by combining 30% bioactive glass (nominal composition of 58% SiO2 -33 % CaO - 9% P2O5) and 70% PVA. New Zealand rabbits were randomly divided into the control group (C group) and two groups with bone lesions, in which one received a matrix implant HM (Implant group), while the other did not (no Implant group). Clinical monitoring showed no altered parameters from either the Implant or the no Implant groups as compared to the control group, for the variables of diet grades, day and night temperatures and hemograms. In the Implant group, radiologic and tomographic studies showed implanted areas with clean edges in femoral non-articular direction, and radio-dense images that suggest incipient integration. Minimum signs of phlogosis could be observed, whereas no signs of rejection at this imaging level could be identified. Histological analysis showed evidence of osteo-integration, with the formation of a trabecular bone within the implant. Together, these results show that implants of hybrid matrixes of bioactive glass are capable of promoting bone regeneration. PMID:22848334

  2. Synthesis of biodegradable and electroactive tetraaniline grafted poly(ester amide) copolymers for bone tissue engineering.

    PubMed

    Cui, Haitao; Liu, Yadong; Deng, Mingxiao; Pang, Xuan; Zhang, Peibiao; Wang, Xianhong; Chen, Xuesi; Wei, Yen

    2012-09-10

    Biodegradable poly(ester amide)s have recently been used as biomaterials due to their desirable chemical and biological characteristics as well as their mechanical properties, which are amendable for material processing. In this study, electroactive tetraaniline (TA) grafted poly(ester amide)s were successfully synthesized and characterized. The poly(ester amide)s-graft-tetraaniline copolymers (PEA-g-TA) exhibited good electroactivity, mechanical properties, and biodegradability. The biocompatibility of the PEA-g-TA copolymers in vitro was systematically studied, which demonstrated that they were nontoxic and led to favorable adhesion and proliferation of mouse preosteoblastic MC3T3-E1 cells. Moreover, the PEA-g-TA copolymers stimulated by pulsed electrical signal could serve to promote the differentiation of MC3T3-E1 cells compared with TCPs. Hence, the biodegradable and electroactive PEA-g-TA copolymers possessed the properties in favor of the long-time potential application in vivo (electrical stimulation directly to the desired area) as bone repair scaffold materials in tissue engineering.

  3. Clinical Comparison of Full and Partial Double Pedicle Flaps with Connective Tissue Grafts for Treatment of Gingival Recession

    PubMed Central

    Ranjbari, Ardeshir; Gholami, Gholam Ali; Amid, Reza; Kadkhodazadeh, Mahdi; Youssefi, Navid; Mehdizadeh, Amir Reza; Aghaloo, Maryam

    2016-01-01

    Statement of the Problem Gingival recession has been considered as the most challenging issue in the field of periodontal plastic surgery. Purpose The purpose of this study was to evaluate the clinical efficacy of root coverage procedures by using partial thickness double pedicle graft and compare it with full thickness double pedicle graft. Materials and Method Eight patients, aged 15 to 58 years including 6 females and 2 males with 20 paired (mirror image) defects with class I and II gingival recession were randomly assigned into two groups. Clinical parameters such as recession depth, recession width, clinical attachment level, probing depth, and width of keratinized tissue were measured at the baseline and 6 months post-surgery. A mucosal double papillary flap was elevated and the respective root was thoroughly planed. The connective tissue graft was harvested from the palate, and then adapted over the root. The pedicle flap was secured over the connective tissue graft and sutured. The surgical technique was similar in the control group except for the prepared double pedicle graft which was full thickness. Results The mean root coverage was 88.14% (2.83 mm) in the test group and 85.7% (2.75 mm) in the control group. No statistical differences were found in the mean reduction of vertical recession, width of recession, or probing depth between the test and control groups. In both procedures, the width of keratinized tissue increased after three months and the difference between the two groups was not statistically significant in this respect. Conclusion Connective tissue with partial and full thickness double pedicle grafts can be successfully used for treatment of marginal gingival recession. PMID:27602394

  4. Reverse lateral supramalleolar adipofascial flap and skin grafting for one-stage soft tissue reconstruction of foot and ankle joint.

    PubMed

    Lee, Jae-Hoon; Chung, Duke-Whan

    2010-09-01

    The aim of this report is to present the clinical result and efficacy of reverse lateral supramalleolar adipofascial flap and skin grafting for one stage soft tissue reconstruction of the foot and ankle joints. Reconstruction using a reverse lateral supramalleolar adipofascial flap and skin grafting was performed in eight cases between January 2005 and March 2009. All the subjects were male with a mean age of 53 years. The mean follow-up period was 20 months. The reasons for soft tissue defects were diabetic foot, infected bursitis, open injuries of the foot, and chronic osteomyelitis. The mean size of the flaps was 3.5 (3-4) × 4.5 (4-6) cm. The flaps were elevated in the form of an adipofascial flap and split-thickness skin grafting was performed over the flaps and adjoining raw areas. Flaps survived in all cases. The implantation of the split-thickness skin graft over the flap was also successful in all cases. Neither partial necrosis in the adipofascial flap nor venous congestion was observed. At the last follow-up, there were no limited motions in the ankle and the toe. No cases complained of inconveniences in ambulation or had difficulties in selecting footwear. In cases that require a flap for the exposed bone or tendon of the foot with a small-sized defect, reverse lateral supramalleolar adipofascial flap and skin grafting is considered a useful method as it lowers the morbidity rate of the donor site and reconstructs soft tissues.

  5. Sporicidal efficacy of genipin: a potential theoretical alternative for biomaterial and tissue graft sterilization.

    PubMed

    Reich, Michael S; Akkus, Ozan

    2013-09-01

    Terminal sterilization of musculoskeletal allografts by gamma radiation minimizes the risk of disease transmission but impairs allograft mechanical properties. Commonly employed crosslinking agents can sterilize tissues without affecting mechanical properties adversely; however, these agents are toxic. Genipin is reported to be a benign crosslinking agent that strengthens mechanical properties of tissues; however, the antimicrobial capacity of genipin is largely unknown. The present study's aims were: (1) to assess the sporicidal potential of genipin, (2) to improve antimicrobial capacity by changing chemical and physical treatment conditions. To establish genipin's sterilization potential Bacillus subtilis var. niger spore strips were treated with 0-10% genipin in PBS or in 1:1 DMSO:PBS up to 72 h at room temperature (RT). Sterilizing doses and concentrations of genipin were used to treat B. pumilus and Geobacillus stearothermophilus spores to assess broader spectrum sporicidal activity of genipin. Scanning electron microscopy (SEM) was performed to evaluate gross morphological changes after genipin treatment. Optimal sterilization conditions were determined by evaluating the effects of temperature (RT-50 °C), DMSO:PBS ratio (0:100-100:0), and treatment duration (24-72 h) on B. subtilis. Genipin penetration of full thickness bovine patellar tendon and cortical bone specimens was observed to assess the feasibility of the agent for treating grafts. Initial studies showed that after 72 h of treatment at RT with 0.63-10% genipin/DMSO:PBS B. subtilis spore strips were sterilized; 0.63% genipin/PBS did not sterilize spore strips at 72 h at RT. Genipin doses and concentrations that sterilized B. subtilis spore strips sterilized B. pumilus and G. stearothermophilus spore strips. SEM revealed no gross morphological differences between untreated and treated spores. Treatment optimization resulted in sterilization within 24 h with 100% PBS, and DMSO facilitated sporicidal

  6. Nonsuture closure of arterial defect by vein graft using isobutyl-2-cyanoacrylate as a tissue adhesive.

    PubMed

    Celik, H; Caner, H; Tahta, K; Ozcan, O E; Erbengi, A; Onol, B

    1991-01-01

    Nonsuture repair of intracranial blood vessel is still a challenging issue in neurosurgery. In this experimental study isobutyl-2-cyanoacrylate was used for nonsuture repairing of carotid artery to observe its histotoxicity. Isobutyl-2-cyanoacrylate was dropped over the edge of defect on carotid artery of rat under the surgical microscope and jugular vein was placed to cover the defect. Rats were divided in 6 group according to postoperative survival period (2, 5, 7, 14, 21, 28 days respectively). The carotid arteries were patent of all rats according to angiography and autopsy specimen. Histopathological results showed that: the changes are mainly inflammatory respons +, there was no necrotic tissue at any time the healing progress and no significant differences in the vein graft. Therefore isobutyl-2-cyanoacrylate is considered helpful and safe adhesive material for the nonsuture repair of blood vessel.

  7. Electrospinning of aniline pentamer-graft-gelatin/PLLA nanofibers for bone tissue engineering.

    PubMed

    Liu, Yadong; Cui, Haitao; Zhuang, Xiuli; Wei, Yen; Chen, Xuesi

    2014-12-01

    Blends of aniline pentamer-graft-gelatin (AP-g-GA) and poly(l-lactide) (PLLA) were electrospun to prepare uniform nanofibers as biomimetic scaffolds. The nanofibers exhibited good electroactivity, thermal stability and biodegradability. The biocompatibility of the nanofibers in vitro was evaluated by the adhesion and proliferation of mouse preosteoblastic MC3T3-E1 cells. The cellular elongation was significantly greater on electroactive AP-g-GA/PLLA nanofibers than on PLLA nanofibers. Moreover, the AP-g-GA/PLLA nanofibers stimulated by an electrical pulsed signal could promote the differentiation of MC3T3-E1 cells compared with pure PLLA nanofibers. Our results demonstrated that the biodegradable and electroactive AP-g-GA/PLLA nanofibers had potential application in vivo as bone repair scaffold materials in tissue engineering.

  8. Haemostatic effects of laser tissue solder as a reinforcement to anastomoses with PTFE grafts

    NASA Astrophysics Data System (ADS)

    Birch, Jeremy F.; Seifalian, Alexander M.; Bell, Peter R.

    2003-06-01

    Laser activated tissue solders have been used for sutureless anastomosis in various contexts. Solders were initially developed in response to the finding that the use of lasers alone caused vessel damage resulting in aneurysm formation and medical damage. Many reports exist of the use of lasers to perform micro-anastomoses, but little has been reported on the use of laser tissue solder in the formation of medium sized vessel anastomoses or in vivo. This group has recently developed a methylene blue based albumin solder for use in vascular anastomoses. The early work concentrated on a rabbit carotid end-to-end model. More recently this has progressed into its application in medium sized vessels. The use of PTFE is common in clinical practice particularly relating to peripheral vascular reconstruction or vascular access surgery. In these instances conventional surgical techniques applied to PTFE will result in excessive bleeding at the site of the anastomosis. Suture materials commonly used such as polypropylene or polyamide leave holes in such prostheses. To compound the problem patients are often anticoagulated or suffer impaired platelet function, improving the chances of graft survival, but increasing bleeding time, the time required to achieve haemostasis and also the post operative complications related to bleeding such as haematoma formation. It was therefore intended to apply the techniques of soldered vascular anastomoses to such a scenario, by reinforcing the anastomotic suture line of grafts placed in an animal model, with MB based solder. The bleeding times, overall operating times and postoperative complications were then analyzed and compared to sutured controls.

  9. Achilles tendon repair with acellular tissue graft augmentation in neglected ruptures.

    PubMed

    Lee, Daniel K

    2007-01-01

    Neglected Achilles tendon rupture injuries present surgical challenges because of the quality and quantity of tendon tissue during repair combined with the magnitude of mechanical forces placed on this tendon. The purpose of this study was to evaluate the effects of an acellular human dermal tissue matrix, GRAFTJACKET, as an augmentation material in neglected Achilles tendon repair. Nine patients with neglected Achilles tendon ruptures were evaluated and followed up for a minimum of 20 months. Primary repair was followed by augmentation with the graft and suturing circumferentially around the tendon. Patients were placed in an early, functional rehabilitation program with postoperative evaluation at 3, 6, and 12 months. Outcome scores were calculated based on the American Orthopaedic Foot and Ankle Society ankle-hindfoot scoring system. At 20 to 30 months postoperative follow-up range, there has been no incidence of re-rupture or recurrent pain. The average return-to-activity time was 15.2 +/- 1.7 weeks. The results from this retrospective clinical series suggest that using an acellular human dermal tissue matrix to augment neglected Achilles tendon rupture primary repair offers desirable return-to-activity time points and viable surgical alternative over previously reported surgical options.

  10. Tissue engineering of acellular vascular grafts capable of somatic growth in young lambs

    PubMed Central

    Syedain, Zeeshan; Reimer, Jay; Lahti, Matthew; Berry, James; Johnson, Sandra; Bianco, Richard; Tranquillo, Robert T.

    2016-01-01

    Treatment of congenital heart defects in children requiring right ventricular outflow tract reconstruction typically involves multiple open-heart surgeries because all existing graft materials have no growth potential. Here we present an ‘off-the-shelf' vascular graft grown from donor fibroblasts in a fibrin gel to address this critical unmet need. In a proof-of-concept study, the decellularized grafts are implanted as a pulmonary artery replacement in three young lambs and evaluated to adulthood. Longitudinal ultrasounds document dimensional growth of the grafts. The lambs show normal growth, increasing body weight by 366% and graft diameter and volume by 56% and 216%, respectively. Explanted grafts display physiological strength and stiffness, complete lumen endothelialization and extensive population by mature smooth muscle cells. The grafts also show substantial elastin deposition and a 465% increase in collagen content, without signs of calcification, aneurysm or stenosis. Collectively, our data support somatic growth of this completely biological graft. PMID:27676438

  11. [Use of vacuum sealing drainage and mesh grafting in treating defects of skin and soft tissue in foot].

    PubMed

    Ye, Zhou; Zhan, Bei-Lei; Zhan, Yun-Zhong

    2010-03-01

    To explore the therapeutic effectiveness of vacuum sealing drainage (VSD) technique and mesh grafting in treating defects of skin and soft tissues in foot. A retrospective analysis was done on 17 cases (11 male and 6 female) suffering from defects of skin and soft tissues in foot, which were treated by vacuum sealing drainage and mesh grafting. The age of patients was from 18 to 67 years with an average of 43 years. The wound surface was filled with polyvinyl alcohol gelatin sponge after debridement and continuous negative pressure drainage was taken for 24 h. After 7 days, granulation tissue growing mesh grafting was performed and to observe the skin colour, survival rate and feet function. All the infection of wounds was controlled with VSD for 1 to 3 times. Skin survival rate of 14 cases more than 98%, 2 cases more than 95%; skin edge of 1 case had little necrosis, but foot function obtained rehabilitation after dress-changing. Vacuum sealing drainage (VSD) technique and mesh grafting is effective methods for the treatment of defects of skin and soft tissues in foot and is worthy generalization and application.

  12. The tent pole splint: a bone-supported stereolithographic surgical splint for the soft tissue matrix expansion graft procedure.

    PubMed

    Cillo, Joseph E; Theodotou, Nicholas; Samuels, Marc; Krajekian, Joseph

    2010-06-01

    This report details the use of computer-aided planning and intraoperative stereolithographic direct-bone-contact surgical splints for the accurate extraoral placement of dental implants in the soft tissue matrix expansion (tent pole) graft of the severely resorbed mandible.

  13. Induction of tolerance to parental parathyroid grafts using allogeneic thymus tissue in patients with DiGeorge anomaly.

    PubMed

    Chinn, Ivan K; Markert, M Louise

    2011-06-01

    DiGeorge anomaly can affect both thymic and parathyroid function. Although athymia is corrected by allogeneic thymus transplantation, treatment options for hypoparathyroidism have been unsatisfactory. Parathyroid transplantation offers the potential for definitive cure but remains challenging because of graft rejection. Some allogeneic parathyroid grafts have functioned in adult recipients in the context of immunosuppression for renal transplantation. Other efforts have attempted to reduce the allogenicity of the parathyroid grafts through manipulation of the parathyroid tissues before transplantation (by using encapsulation or special culture techniques). Recently, we demonstrated the efficacy of parental parathyroid transplantation when combined with allogeneic thymus transplantation in an infant with complete DiGeorge anomaly. The recipient developed tolerance toward the parathyroid donor. The parathyroid graft has functioned for 5 years after transplantation without the need for continued immunosuppression or calcium supplementation. We observed that matching of the allogeneic thymus graft to the parathyroid donor HLA class II alleles that are unshared with the recipient appears to be associated with the induction of tolerance toward the parathyroid graft. Further work is needed to determine the optimal means for using combined allogeneic thymus and parental parathyroid transplantation to correct hypoparathyroidism in patients with both complete and partial DiGeorge anomaly.

  14. Dimensional changes in soft tissues around dental implants following free gingival grafting: an experimental study in dogs.

    PubMed

    Bengazi, Franco; Lang, Niklaus P; Caroprese, Marino; Urbizo Velez, Joaquin; Favero, Vittorio; Botticelli, Daniele

    2015-02-01

    To study the buccal dimensional tissue changes at oral implants following free gingival grafting, with or without including the keratin layer, performed at the time of implant installation into alveolar mucosa. The mandibular premolars and first molars were extracted bilaterally in six Beagle dogs. In the right side of the mandible (Test), flaps were first elevated, and the buccal as well as part of the lingual masticatory mucosa was removed. An incision of the periosteum at the buccal aspect was performed to allow the flap to be coronally repositioned. Primary wound closure was obtained. In the left side, the masticatory (keratinized) mucosa was left in situ, and no sutures were applied (Control). After 3 months of healing, absence of keratinized mucosa was confirmed at the test sites. Two recipient sites were prepared at each side of the mandible in the region of the third and fourth premolars. All implants were installed with the shoulder placed flush with the buccal alveolar bony crest, and abutments were connected to allow a non-submerged healing. Two free gingival mucosal grafts were harvested from the buccal region of the maxillary canines. One graft was left intact (gingival mucosal graft), while for the second, the epithelial layer was removed (gingival connective tissue graft). Subsequently, the grafts were fixed around the test implants in position of the third and fourth premolars, respectively. After 3 months, the animals were euthanized and ground sections obtained. Similar bony crest resorption and coronal extension of osseointegration were found at test and control sites. Moreover, similar dimensions of the peri-implant soft tissues were obtained at test and control sites. The increase in the alveolar mucosal thickness by means of a gingival graft affected the peri-implant marginal bone resorption and soft tissue recession around implants. This resulted in outcomes that were similar to those at implants surrounded by masticatory mucosa, indicating

  15. Reconstruction of the interdental papilla with an underlying subepithelial connective tissue graft: technical considerations and case reports.

    PubMed

    Carranza, Nelson; Zogbi, Carim

    2011-01-01

    This article introduces a surgical technique developed to achieve soft tissue augmentation of the interproximal space. The technique was designed to minimize surgical trauma and blockage of blood supply to the existing papilla by accessing the papillary area through vertical incisions and by elevating a single full-thickness flap without disrupting the papillary bridge. A free connective tissue graft was placed beneath the undermined papilla and secured with sutures. Advantages and variations of the technique are discussed.

  16. The rationale for soft-tissue grafting and vestibuloplasty in association with endosseous implants: a literature review.

    PubMed

    Hoelscher, D C; Simons, A M

    1994-01-01

    Significant soft-tissue complications have been reported around endosseous implant permucosal abutments. Peri-implantitis with associated bone loss can have a negative effect on the long-term prognosis of the implant reconstruction. The rationale for soft-tissue grafting and vestibuloplasty techniques is presented in the form of a literature review. The implant sulcular epithelium, permucosal seal, and the peri-implant connective tissues are discussed. The etiology of soft-tissue complications as well as the significance of attached gingiva surrounding implant abutments are presented along with techniques for surgical intervention.

  17. The rationale for soft-tissue grafting and vestibuloplasty in association with endosseous implants: a literature review.

    PubMed

    Hoelscher, D C; Simons, A M

    1996-03-01

    Significant soft-tissue complications have been reported around endosseous implant permucosal abutments. Peri-implants with associated bone loss can have a negative effect on the long-term prognosis of the implant reconstruction. The rationale for soft-tissue grafting and vestibuloplasty techniques is presented in the form of a literature review. The implant sulcular epithelium, permucosal seal, and the peri-implant connective tissues are discussed. The etiology of soft-tissue complications as well as the significance of attached gingiva surrounding implant abutments are presented along with techniques for surgical intervention.

  18. Osteochondral Allografts in the Ankle Joint

    PubMed Central

    Vannini, Francesca; Buda, Roberto; Ruffilli, Alberto; Cavallo, Marco; Giannini, Sandro

    2013-01-01

    Purpose: The aim of this systematic review is to report about the clinical use of partial and total fresh osteochondral allograft in the ankle joint. The state of the art of allografts with regard to basic science, procurement and storage methods, immunogenicity, generally accepted indications and contraindications, and the rationale of the allografting procedure have been described. Methods: All studies published in PubMed from 2000 to January 2012 addressing fresh osteochondral allograft procedures in the ankle joint were identified, including those that fulfilled the following criteria: (a) level I-IV evidence addressing the areas of interest outlined above; (b) measures of functional, clinical, or imaging outcome; and (c) outcome related to ankle cartilage lesions or ankle arthritis treated by allografts. Results: The analysis showed a progressively increasing number of articles from 2000. The number of selected articles was 14; 9 of those focused on limited dimension allografts (plugs, partial) and 5 on bipolar fresh osteochondral allografts. The evaluation of evidence level showed 14 case series and no randomized studies. Conclusions: Fresh osteochondral allografts are now a versatile and suitable option for the treatment of different degrees of osteochondral disease in the ankle joint and may even be used as total joint replacement. Fresh osteochondral allografts used for total joint replacement are still experimental and might be considered as a salvage procedure in otherwise unsolvable situations. A proper selection of the patients is therefore a key point. Moreover, the patients should be adequately informed about the possible risks, benefits, and alternatives to the allograft procedure. PMID:26069666

  19. Bone marrow aspiration concentrate and platelet rich plasma for osteochondral repair in a porcine osteochondral defect model.

    PubMed

    Betsch, Marcel; Schneppendahl, Johannes; Thuns, Simon; Herten, Monika; Sager, Martin; Jungbluth, Pascal; Hakimi, Mohssen; Wild, Michael

    2013-01-01

    Bone marrow aspiration concentrate (BMAC) may possess a high potency for cartilage and osseous defect healing because it contains stem cells and multiple growth factors. Alternatively, platelet rich plasma (PRP), which contains a cocktail of multiple growth factors released from enriched activated thrombocytes may potentially stimulate the mesenchymal stem cells (MSCs) in bone marrow to proliferate and differentiate. A critical size osteochondral defect (10×6 mm) in both medial femoral condyles was created in 14 Goettinger mini-pigs. All animals were randomized into the following four groups: biphasic scaffold alone (TRUFIT BGS, Smith & Nephew, USA), scaffold with PRP, scaffold with BMAC and scaffold in combination with BMAC and PRP. After 26 weeks all animals were euthanized and histological slides were cut, stained and evaluated using a histological score and immunohistochemistry. The thrombocyte number was significantly increased (p = 0.049) in PRP compared to whole blood. In addition the concentration of the measured growth factors in PRP such as BMP-2, BMP-7, VEGF, TGF-β1 and PDGF were significantly increased when compared to whole blood (p<0.05). In the defects of the therapy groups areas of chondrogenic tissue were present, which stained blue with toluidine blue and positively for collagen type II. Adding BMAC or PRP in a biphasic scaffold led to a significant improvement of the histological score compared to the control group, but the combination of BMAC and PRP did not further enhance the histological score. The clinical application of BMAC or PRP in osteochondral defect healing is attractive because of their autologous origin and cost-effectiveness. Adding either PRP or BMAC to a biphasic scaffold led to a significantly better healing of osteochondral defects compared with the control group. However, the combination of both therapies did not further enhance healing.

  20. Bone Marrow Aspiration Concentrate and Platelet Rich Plasma for Osteochondral Repair in a Porcine Osteochondral Defect Model

    PubMed Central

    Betsch, Marcel; Schneppendahl, Johannes; Thuns, Simon; Herten, Monika; Sager, Martin; Jungbluth, Pascal; Hakimi, Mohssen; Wild, Michael

    2013-01-01

    Background Bone marrow aspiration concentrate (BMAC) may possess a high potency for cartilage and osseous defect healing because it contains stem cells and multiple growth factors. Alternatively, platelet rich plasma (PRP), which contains a cocktail of multiple growth factors released from enriched activated thrombocytes may potentially stimulate the mesenchymal stem cells (MSCs) in bone marrow to proliferate and differentiate. Methods A critical size osteochondral defect (10×6 mm) in both medial femoral condyles was created in 14 Goettinger mini-pigs. All animals were randomized into the following four groups: biphasic scaffold alone (TRUFIT BGS, Smith & Nephew, USA), scaffold with PRP, scaffold with BMAC and scaffold in combination with BMAC and PRP. After 26 weeks all animals were euthanized and histological slides were cut, stained and evaluated using a histological score and immunohistochemistry. Results The thrombocyte number was significantly increased (p = 0.049) in PRP compared to whole blood. In addition the concentration of the measured growth factors in PRP such as BMP-2, BMP-7, VEGF, TGF-β1 and PDGF were significantly increased when compared to whole blood (p<0.05). In the defects of the therapy groups areas of chondrogenic tissue were present, which stained blue with toluidine blue and positively for collagen type II. Adding BMAC or PRP in a biphasic scaffold led to a significant improvement of the histological score compared to the control group, but the combination of BMAC and PRP did not further enhance the histological score. Conclusions The clinical application of BMAC or PRP in osteochondral defect healing is attractive because of their autologous origin and cost-effectiveness. Adding either PRP or BMAC to a biphasic scaffold led to a significantly better healing of osteochondral defects compared with the control group. However, the combination of both therapies did not further enhance healing. PMID:23951201

  1. ABO antigen expression in graft tissue: is titration against donor erythrocytes relevant?

    PubMed

    Rydberg, Lennart; Skogsberg, Ulrika; Mölne, Johan

    2007-12-27

    ABO-incompatible living donor renal transplantation has become an accepted treatment for end-stage renal disease. Two main factors appear to be important when crossing the ABO barrier, the donor organ A/B antigen expression and the amount of recipient anti-A/B antibody. Antigen expression depends on the ABO blood group and subgroup and may vary in different tissues and cells. The amount of recipient anti-A/B antibody, determined by titration, is very variable. One major drawback with titration is the lack of conformity between different laboratories, making comparisons difficult. For clinical use, the anti-A/B antibody titration technique has to be simple, rapid, and cheap, in addition to being accurate. Although there is a need for more standardized procedures for determination of ABO antibodies, existing techniques are sufficient in the clinical care of patients. To illustrate the variation in susceptibility of different graft tissues to ABO antibodies, in this paper we describe a case of an ABO-incompatible combined liver and kidney transplantation.

  2. In vitro and in vivo bioluminescent imaging of hypoxia in tissue-engineered grafts.

    PubMed

    Liu, Jun; Barradas, Ana; Fernandes, Hugo; Janssen, Frank; Papenburg, Bernke; Stamatialis, Dimitrios; Martens, Anton; van Blitterswijk, Clemens; de Boer, Jan

    2010-06-01

    Survival and growth of cellular grafts in tissue engineering (TE) are limited by the rate of oxygen (O(2)) and nutrient diffusion. As such, monitoring the levels of nutrients and O(2) available to the cells is essential to assess the physiology of the cells and to evaluate strategies aiming at improving nutrient availability. In this article, a reporter system containing the luciferase gene driven by a hypoxia responsive promoter was used to monitor cellular hypoxia in a TE context. We report that luciferase activity correlates with the O(2) tension in the cell culture medium. When transgenic cells were seeded onto scaffolds and implanted in immune-deficient mice subcutaneously, luciferase activity was detected. To validate the response to O(2) levels of this reporter system, we cultured transgenic cells on biomaterials in a flow perfusion bioreactor and observed that cells in the bioreactor displayed a drastically lower luciferase activity than conventional static culture, and that higher luciferase activity is observed in the interior of a tissue-engineered construct, illustrating the uneven O(2) distribution in three-dimensional constructs under conventional static culture. We conclude that this reporter system is a versatile tool to investigate cellular O(2) availability in TE both in vitro and in vivo.

  3. A collagen/smooth muscle cell-incorporated elastic scaffold for tissue-engineered vascular grafts.

    PubMed

    Park, In Su; Kim, Sang-Heon; Kim, Young Ha; Kim, Ik Hwan; Kim, Soo Hyun

    2009-01-01

    Biodegradable tubular scaffolds have been developed for vascular graft application. This study was focused to improve the adhesion and proliferation of vascular smooth muscle cells (SMCs) in a tubular scaffold. Tubular scaffolds (ID 4 mm, OD 6 mm) were fabricated from a biodegradable elastic polymer, poly(L-lactide-co-epsilon-caprolactone) (PLCL) (50:50, M(n) 1.58 x 10(5)), by an extrusion/particulate leaching method. SMCs suspended in a collagen solution were infiltrated in tubular PLCL scaffolds under vacuum and incubated for 1 h at 37 degrees C to form a collagenous gel. Results from SEM image analysis showed that collagen was infiltrated into the inside of the scaffolds. Cell adhesion and proliferation rate increased in collagen/SMC-incorporated tubular PLCL scaffolds as compared with the scaffolds in which only SMCs were seeded. From SEM image and histological analysis, we further found that SMCs grew on the inside as well as on the surface of collagen/SMCs-incorporated scaffolds and the cells continued to grow as a monolayer on collagen fibers. In particular, cell proliferation and elastin contents were the highest in a PLCL scaffold with 50-100 microm pore size than any other scaffolds used in this experiment. A collagen/SMC-incorporated PLCL scaffold may support SMC growth and functions and can be used as a scaffold for tissue engineering to facilitate small-diameter vascular-tissue formation.

  4. Polymer brush: a promising grafting approach to scaffolds for tissue engineering

    PubMed Central

    Kim, Woonjung; Jung, Jongjin

    2016-01-01

    Polymer brush is a soft material unit tethered covalently on the surface of scaffolds. It can induce functional and structural modification of a substrate’s properties. Such surface coating approach has attracted special attentions in the fields of stem cell biology, tissue engineering, and regenerative medicine due to facile fabrication, usability of various polymers, extracellular matrix (ECM)-like structural features, and in vivo stability. Here, we summarized polymer brush-based grafting approaches comparing self-assembled monolayer (SAM)-based coating method, in addition to physico-chemical characterization techniques for surfaces such as wettability, stiffness/elasticity, roughness, and chemical composition that can affect cell adhesion, differentiation, and proliferation. We also reviewed recent advancements in cell biological applications of polymer brushes by focusing on stem cell differentiation and 3D supports/implants for tissue formation. Understanding cell behaviors on polymer brushes in the scale of nanometer length can contribute to systematic understandings of cellular responses at the interface of polymers and scaffolds and their simultaneous effects on cell behaviors for promising platform designs. PMID:27697112

  5. Pre-implanted Sensory Nerve Could Enhance the Neurotization in Tissue-Engineered Bone Graft.

    PubMed

    Wu, Yan; Jing, Da; Ouyang, Hongwei; Li, Liang; Zhai, Mingming; Li, Yan; Bi, Long; Guoxian, Pei

    2015-08-01

    In our previous study, it was found that implanting the sensory nerve tract into the tissue-engineered bone to repair large bone defects can significantly result in better osteogenesis effect than tissue-engineered bone graft (TEBG) alone. To study the behavior of the preimplanted sensory nerve in the TEBG, the TEBG was constructed by seeding bone mesenchymal stem cells into β-tricalcium phosphate scaffold with (treatment group) or without (blank group) implantation of the sensory nerve. The expression of calcitonin gene-related peptide (CGRP), which helps in the healing of bone defect in the treatment group was significantly higher than the blank group at 4, 8, and 12 weeks. The expression of growth-associated protein 43 (GAP43), which might be expressed during nerve healing in the treatment group, was significantly higher than the blank group at 4 and 8 weeks. The nerve tracts of the preimplanted sensory nerve were found in the scaffold by the nerve tracing technique. The implanted sensory nerve tracts grew into the pores of scaffolds much earlier than the vascular. The implanted sensory nerve tracts traced by Dil could be observed at 4 weeks, but at the same time, no vascular was observed. In conclusion, the TEBG could be benefited from the preimplanted sensory nerve through the healing behavior of the sensory nerve. The sensory nerve fibers could grow into the pores of the TEBG rapidly, and increase the expression of CGRP, which is helpful in regulating the bone formation and the blood flow.

  6. Repair of osteochondral defects with in vitro engineered cartilage based on autologous bone marrow stromal cells in a swine model

    PubMed Central

    He, Aijuan; Liu, Lina; Luo, Xusong; Liu, Yu; Liu, Yi; Liu, Fangjun; Wang, Xiaoyun; Zhang, Zhiyong; Zhang, Wenjie; Liu, Wei; Cao, Yilin; Zhou, Guangdong

    2017-01-01

    Functional reconstruction of large osteochondral defects is always a major challenge in articular surgery. Some studies have reported the feasibility of repairing articular osteochondral defects using bone marrow stromal cells (BMSCs) and biodegradable scaffolds. However, no significant breakthroughs have been achieved in clinical translation due to the instability of in vivo cartilage regeneration based on direct cell-scaffold construct implantation. To overcome the disadvantages of direct cell-scaffold construct implantation, the current study proposed an in vitro cartilage regeneration strategy, providing relatively mature cartilage-like tissue with superior mechanical properties. Our strategy involved in vitro cartilage engineering, repair of osteochondral defects, and evaluation of in vivo repair efficacy. The results demonstrated that BMSC engineered cartilage in vitro (BEC-vitro) presented a time-depended maturation process. The implantation of BEC-vitro alone could successfully realize tissue-specific repair of osteochondral defects with both cartilage and subchondral bone. Furthermore, the maturity level of BEC-vitro had significant influence on the repaired results. These results indicated that in vitro cartilage regeneration using BMSCs is a promising strategy for functional reconstruction of osteochondral defect, thus promoting the clinical translation of cartilage regeneration techniques incorporating BMSCs. PMID:28084417

  7. Efficacy of tissue engineered bone grafts containing mesenchymal stromal cells for cleft alveolar osteoplasty in a rat model.

    PubMed

    Korn, P; Schulz, M C; Range, U; Lauer, G; Pradel, W

    2014-10-01

    The development of sufficient tissue engineered bone grafts for alveolar cleft osteoplasty could reduce the necessity of autogenous bone grafts and its donor site morbidity. The aim of the study was to evaluate tissue engineered bone grafts in an artificially created bone defect. Bone grafts were created in vitro colonizing a synthetic hydroxyapatite-tricalciumphosphate scaffold (BONITmatrix(®)) with either undifferentiated mesenchymal stromal cells (group 1) or osteogenic differentiated mesenchymal stromal cells (group 2). Cells were multiplied from bone marrow of donor rats. Unmodified scaffolds (group 3) and the tissue engineered bone grafts were inserted into artificial maxillary defects of 54 Lewis rats. In 18 animals the defects remained unfilled (control). After one, three and six weeks the rats were sacrificed. The defect was evaluated radiologically and histologically with regard to the remaining defect volume and diameter. Statistical analysis followed. The bone grafts led to a specific bone formation at the defect margin. No complete reunion of any defect was observed within the healing time. After six weeks, the remaining defect volume was 6.86 ± 3.21 mm(3) (control), 4.08 ± 1.36 mm(3) (group 1), 5.00 ± 0.84 mm(3) (group 2) 5.50 ± 1.05 mm(3) (group 3). The remaining defect diameter measured 2.63 ± 0.52 mm (control), 2.39 ± 0.23 mm (group 1), 2.53 ± 0.22 mm (group 2) and 2.70 ± 0.66 mm (group 3). In all experimental groups the defect volume and diameter decreased over time, which was significant for group 1 (p = 0.014), group 2 (p = 0.025) and group 3 (p = 0.048). The defect volume and width was significantly reduced for bone grafts containing undifferentiated cells compared to control (p = 0.035) or scaffolds only (p = 0.05). Tissue engineered bone grafts induce a pronounced bone formation in artificial bone defects compared to unfilled controls or scaffolds only. Copyright © 2014 European Association for Cranio-Maxillo-Facial Surgery

  8. Effects of sterilization and storage on the properties of ALP-grafted biomaterials for prosthetic and bone tissue engineering applications.

    PubMed

    Ferraris, S; Pan, G; Cassinelli, C; Mazzucco, L; Vernè, E; Spriano, S

    2012-10-01

    Grafting of the biomaterial surfaces with biomolecules is nowadays a challenging research field for prosthetic and bone tissue engineering applications. On the other hand, very few research works investigate the effect of the sterilization processes on the properties of functionalized biomaterials. In this study, the effects of different sterilization techniques (e.g. gamma and electron beam irradiation, ethylene oxide) on the enzymatic activity of bioactive glasses and Ti6Al4V grafted with alkaline phosphatase (ALP) have been analyzed. Sterility maintenance and in vitro bioactivity of the sterilized surfaces have also been investigated. Finally the effect of packaging and storage conditions has been considered.

  9. Surgical treatment of an osteochondral lesion associated with stress fracture of the tarsal navicular: a case report.

    PubMed

    Kanazawa, Kazuki; Yoshimura, Ichiro; Shiokawa, Teruaki; Hagio, Tomonobu; Naito, Masatoshi

    2013-01-01

    We surgically treated an osteochondral lesion associated with a stress fracture of the tarsal navicular. The surgical procedure involved the confirmation and complete resection of the lesion under direct vision, followed by the transplantation of block-shaped iliac bone grafts. The postoperative computed tomography scan showed that the lesions had disappeared, the grafted bone had fused, and the stress fracture had healed. However, the tarsal navicular joint surface was slightly irregular. The patient was able to resume her sports activities 15 weeks after surgery. We have described a novel method to reconstruct the tarsal navicular after osteochondral lesion resection. Copyright © 2013 American College of Foot and Ankle Surgeons. Published by Elsevier Inc. All rights reserved.

  10. Fresh-Stored Osteochondral Allograft for Treatment of Osteochondritis Dissecans the Femoral Head

    DTIC Science & Technology

    2010-01-01

    longevity and sur- vivorship [7, 30, 31]. A rotational osteotomy is another viable option because it is a joint-preserving procedure and delays the need...Osteochondral defects of the femoral head are exceedingly rare, with limited treatment options. Restora- tion procedures for similar defects involving the...develop secondary to trauma and underwent subsequent treatment using a fresh-stored osteochondral allograft via a trochanteric osteotomy . At the 1-year

  11. The Augmentation of a Collagen/Glycosaminoglycan Biphasic Osteochondral Scaffold with Platelet-Rich Plasma and Concentrated Bone Marrow Aspirate for Osteochondral Defect Repair in Sheep

    PubMed Central

    Henson, Frances; Skelton, Carrie; Herrera, Emilio; Brooks, Roger; Fortier, Lisa A.; Rushton, Neil

    2012-01-01

    Objective: This study investigates the combination of platelet-rich plasma (PRP) or concentrated bone marrow aspirate (CBMA) with a biphasic collagen/glycosaminoglycan (GAG) osteochondral scaffold for the treatment of osteochondral defects in sheep. Design: Acute osteochondral defects were created in the medial femoral condyle (MFC) and the lateral trochlea sulcus (LTS) of 24 sheep (n = 6). Defects were left empty or filled with a 6 × 6-mm scaffold, either on its own or in combination with PRP or CBMA. Outcome measures at 6 months included mechanical testing, International Cartilage Repair Society (ICRS) repair score, modified O’Driscoll histology score, qualitative histology, and immunohistochemistry for type I, II, and VI collagen. Results: No differences in mechanical properties, ICRS repair score, or modified O’Driscoll score were detected between the 4 groups. However, qualitative assessments of the histological architecture, Safranin O content, and collagen immunohistochemistry indicated that in the PRP/scaffold groups, there was a more hyaline cartilage–like tissue repair. In addition, the addition of CBMA and PRP to the scaffold reduced cyst formation in the subchondral bone of healed lesions. Conclusion: There was more hyaline cartilage–like tissue formed in the PRP/scaffold group and less subchondral cystic lesion formation in the CBMA and PRP/scaffold groups, although there were no quantitative differences in the repair tissue formed. PMID:26069645

  12. Design of a multiphase osteochondral scaffold. I. Control of chemical composition.

    PubMed

    Lynn, Andrew K; Best, Serena M; Cameron, Ruth E; Harley, Brendan A; Yannas, Ioannis V; Gibson, Lorna J; Bonfield, William

    2010-03-01

    This is the first in a series of articles that describe the design and development of a family of osteochondral scaffolds based on collagen-glycosaminoglycan (collagen-GAG) and calcium phosphate technologies, engineered for the regenerative repair of defects in articular cartilage. The osteochondral scaffolds consist of two layers: a mineralized type I collagen-GAG scaffold designed to regenerate the underlying subchondral bone and a nonmineralized type II collagen-GAG scaffold designed to regenerate cartilage. The subsequent articles in this series describe the fabrication and properties of a mineralized scaffold as well as a two-layer (one mineralized, the other not) osteochondral scaffold for regeneration of the underlying bone and cartilage, respectively. This article describes a technology through which the chemical composition-particularly the calcium phosphate mass fraction-of triple coprecipitated nanocomposites of collagen, glycosaminoglycan, and calcium phosphate can be accurately and reproducibly varied without the need for titrants or other additives. Here, we describe how the mineral:organic ratio can be altered over a range that includes that for articular cartilage (0 wt % mineral) and for bone (75 wt % mineral). This technology achieves the objective of mimicking the composition of two main tissue types found in articular joints, with particular emphasis on the osseous compartment of an osteochondral scaffold. Exclusion of titrants avoids the formation of potentially harmful contaminant phases during freeze-drying steps crucial for scaffold fabrication, ensuring that the potential for binding growth factors and drugs is maintained.

  13. Rational design of an improved tissue-engineered vascular graft: determining the optimal cell dose and incubation time.

    PubMed

    Lee, Yong-Ung; Mahler, Nathan; Best, Cameron A; Tara, Shuhei; Sugiura, Tadahisa; Lee, Avione Y; Yi, Tai; Hibino, Narutoshi; Shinoka, Toshiharu; Breuer, Christopher

    2016-03-01

    We investigated the effect of cell seeding dose and incubation time on tissue-engineered vascular graft (TEVG) patency. Various doses of bone marrow-derived mononuclear cells (BM-MNCs) were seeded onto TEVGs, incubated for 0 or 12 h, and implanted in C57BL/6 mice. Different doses of human BM-MNCs were seeded onto TEVGs and measured for cell attachment. The incubation time showed no significant effect on TEVG patency. However, TEVG patency was significantly increased in a dose-dependent manner. In the human graft, more bone marrow used for seeding resulted in increased cell attachment in a dose-dependent manner. Increasing the BM-MNC dose and reducing incubation time is a viable strategy for improving the performance and utility of the graft.

  14. Repair Mechanism of Osteochondral Defect Promoted by Bioengineered Chondrocyte Sheet

    PubMed Central

    Kamei, Naosuke; Adachi, Nobuo; Hamanishi, Michio; Kamei, Goki; Mahmoud, Elhussein Elbadry; Nakano, Tomohiro; Iwata, Takanori; Yamato, Masayuki; Okano, Teruo; Ochi, Mitsuo

    2015-01-01

    Cell sheet engineering has developed as a remarkable method for cell transplantation. In the field of cartilage regeneration, several studies previously reported that cartilage defects could be regenerated by transplantation of a chondrocyte sheet using cell sheet engineering. However, it remains unclear how such a thin cell sheet could repair a deep cartilage defect. We, therefore, focused on the mechanism of cartilage repair using cell sheet engineering in this study. Chondrocyte sheets and synovial cell sheets were fabricated using cell sheet engineering, and these allogenic cell sheets were transplanted to cover an osteochondral defect in a rat model. Macroscopic and histological evaluation was performed at 4 and 12 weeks after transplantation. Analysis of the gene expression of each cell sheet and of the regenerated tissue at 1 week after transplantation was performed. In addition, green fluorescent protein (GFP) transgenic rats were used as donors (transplanted chondrocyte sheets) or recipients (osteochondral defect models) to identify the cell origin of regenerated cartilage. Cartilage repair was significantly better in the group implanted with a chondrocyte sheet than in that with a synovial cell sheet. The results of gene expression analysis suggest that the possible factor contributing to cartilage repair might be TGFβ1. Cell tracking experiments using GFP transgenic rats showed that the regenerated cartilage was largely composed of cells derived from the transplanted chondrocyte sheets. PMID:25396711

  15. Composite graft including bone tissue: a case report of successful reattachment of multiple fingertip oblique amputation.

    PubMed

    Lee, Kyung Suk; Lim, Yun sub; Choi, Jaehoon; Kim, Nam Gyun; Kim, Jun Sik

    2013-02-01

    A composite graft for reattachment of an amputated fingertip is a very controversial and challenging procedure. An osteocutaneous composite graft is rarely conducted and has a low success rate following fingertip amputation. A 21-year-old male patient was referred to our emergency clinic with dorsal oblique amputation of the middle, ring and small fingers of the left hand through the distal interphalangeal joint and middle phalanx. The amputated parts of the middle and ring fingers were reattached with osteocutaneous composite grafts. The amputated part of the small finger was revascularised to the ulnar palmar digital artery of the small finger. The composite grafts of the middle and ring fingers and the revascularised small finger survived completely. We suggest that careful patient selection will allow an osteocutaneous composite graft to become an acceptable method for the treatment of fingertip amputation. A large-scale study of osteocutaneous graft of amputated fingertips is required to improve the survival rate.

  16. Clinical evaluation of expanded mesh connective tissue graft in the treatment for multiple adjacent gingival recessions in the esthetic zone

    PubMed Central

    Shanmugam, M.; Shivakumar, B.; Meenapriya, B.; Anitha, V.; Ashwath, B.

    2015-01-01

    Background: Multiple approaches have been used to replace lost, damaged or diseased gingival tissues. The connective tissue graft (CTG) procedure is the golden standard method for root coverage. Although multiple sites often need grafting, the palatal mucosa supplies only a limited area of grafting material. To overcome this limitation, expanded mesh graft provides a method whereby a graft can be stretched to cover a large area. The aim of this study was to evaluate the effectiveness and the predictability of expanded mesh CTG (e-MCTG) in the treatment of adjacent multiple gingival recessions. Materials and Methods: Sixteen patients aged 20–50 years contributed to 55 sites, each site falling into at least three adjacent Miller's Class 1 or Class 2 gingival recession. The CTG obtained from the palatal mucosa was expanded to cover the recipient bed, which was 1.5 times larger than the graft. Clinical measurements were recorded at baseline and 3 months, 12 months postoperatively. Results: A mean coverage of 1.96 mm ± 0.66 mm and 2.22 mm ± 0.68 mm was obtained at the end of 3rd and 12th month, respectively. Twelve months after surgery a statistically significant increase in CAL (2.2 mm ± 0.68 mm, P < 0.001) and increasing WKT (1.75 ± 0.78, P < 0.001) were obtained. In 80% of the treated sites, 100% root coverage was achieved (mean 93.5%). Conclusions: The results of this study demonstrated that multiple adjacent recessions were treated by using e-MCTG technique can be applied and highly predictable root coverage can be achieved. PMID:26321829

  17. Molecular, cellular and pharmaceutical aspects of autologous grafts for peri-implant hard and soft tissue defects.

    PubMed

    Lu, Jiayu; Hao, Yongming; Zhao, Wei; Lv, Chengqi; Zou, Derong

    2016-12-01

    The lack of supporting hard and soft tissues always prevents the rehabilitation with dental implants. Among various hard and soft tissue augmentation procedures, autologous grafts have been considered to be the gold standard. Autologous mesenchymal stem cells (MSCs) from bone marrow, dental tissue and adipose tissue have been described as promising alternatives for bone regeneration in the field of dental implantation. Mucosal cells, gingival fibroblasts and dental progenitor cells (DPS) can enhance peri-implant soft tissue augmentation and regenerate periodontal tissues around dental implants. Obtained from patients, platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) are enriched in autologous platelets, which contain a great deal of growth factors and cytokines that are conducive to the regeneration of both hand and soft tissues around dental implants. Pharmaceutical treatments for osteoporosis and diabetes should be locally applied with implant procedures to restrict the resorption of autologous bone grafts and reduction of bone volume. Although autografts hold great potentials for dental implants, new approaches should also be explored with minimally invasion donor sites methods such as tissue engineering combined with autologous three factors and bio-3D printing involving self-assembling cell aggregates.

  18. Clinical Trial of Intraoral Grafting of Human Tissue-Engineered Oral Mucosa, Phase 2

    DTIC Science & Technology

    2014-10-01

    study will determine differences in the primary efficacy measure of increased keratinized mucosa; secondary measures of graft contracture and Wound...palatal oral mucosa (POM) graft (Group 2). The study population will include non-smoking adults (ages 18 and older) in need of additional keratinized ...POM), Keratinized mucosa, graft contracture 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF PAGES 19a. NAME OF RESPONSIBLE

  19. Comparison of glenohumeral contact pressures and contact areas after glenoid reconstruction with latarjet or distal tibial osteochondral allografts.

    PubMed

    Bhatia, Sanjeev; Van Thiel, Geoffrey S; Gupta, Deepti; Ghodadra, Neil; Cole, Brian J; Bach, Bernard R; Shewman, Elizabeth; Wang, Vincent M; Romeo, Anthony A; Verma, Nikhil N; Provencher, Matthew T

    2013-08-01

    Glenoid reconstruction with distal tibial allografts offers the theoretical advantage over Latarjet reconstruction of improved joint congruity and a cartilaginous articulation for the humeral head. Hypothesis/ To investigate changes in the magnitude and location of glenohumeral contact areas, contact pressures, and peak forces after (1) the creation of a 30% anterior glenoid defect and subsequent glenoid bone augmentation with (2) a flush Latarjet coracoid graft or (3) a distal tibial osteochondral allograft. It was hypothesized that the distal tibial bone graft would best normalize glenohumeral contact areas, contact pressures, and peak forces. Controlled laboratory study. Eight cadaveric shoulder specimens were dissected free of all soft tissues and randomly tested in 3 static positions of humeral abduction with a 440-N compressive load: 30°, 60°, and 60° of abduction with 90° of external rotation (ABER). Glenohumeral contact area, contact pressure, and peak force were determined sequentially using a digital pressure mapping system for (1) the intact glenoid, (2) the glenoid with a 30% anterior bone defect, and (3) the glenoid after reconstruction with a distal tibial allograft or a Latarjet bone block. Glenoid reconstruction with distal tibial allografts resulted in significantly higher glenohumeral contact areas than reconstruction with Latarjet bone blocks in 60° of abduction (4.87 vs. 3.93 cm2, respectively; P < .05) and the ABER position (3.98 vs. 2.81 cm2, respectively; P < .05). Distal tibial allograft reconstruction also demonstrated significantly lower peak forces than Latarjet reconstruction in the ABER position (2.39 vs. 2.61 N, respectively; P < .05). Regarding the bone loss model, distal tibial allograft reconstruction exhibited significantly higher contact areas and significantly lower contact pressures and peak forces than the 30% defect model at all 3 abduction positions. Latarjet reconstruction also followed this same pattern, but

  20. Arthroscopically Assisted Treatment of Navicular Osteochondral Defect Using Flowable Collagen, Iliac Crest Bone Marrow Aspirate and Fibrin Glue: A Case Report.

    PubMed

    Keller, Thomas C; Dempsey, Ian J; Park, Joseph S

    2015-10-01

    A 32-year-old male recreational athlete presented with activity-related chronic dorsal midfoot pain. Conservative treatment, including a prolonged period of immobilization, physical therapy, nonsteroidal anti-inflammatory drugs, and use of a bone stimulator, failed to resolve his symptoms. Computed tomography and magnetic resonance imaging demonstrated a cystic appearing focus within the navicular in conjunction with a osteochondral lesion within the proximal articular surface of the navicular. This case report presents an arthroscopically assisted treatment of a navicular osteochondral lesion using curettage and backfilling with fibrin glue, flowable collagen, and autogenous bone grafting. Therapeutic, Level IV. © 2014 The Author(s).

  1. Genetics Home Reference: familial osteochondritis dissecans

    MedlinePlus

    ... Familial osteochondritis dissecans Seattle Children's TeensHealth from Nemours: Knee Injuries University of Connecticut Health Center Patient Support and Advocacy Resources (1 link) American College of Rheumatology: Osteoarthritis ClinicalTrials.gov (1 link) ClinicalTrials.gov Scientific Articles ...

  2. Targeted SPECT/CT Imaging of Matrix Metalloproteinase Activity in the Evaluation of Remodeling Tissue-Engineered Vascular Grafts Implanted in a Growing Lamb Model

    PubMed Central

    Stacy, Mitchel R.; Naito, Yuji; Maxfield, Mark W.; Kurobe, Hirotsugu; Tara, Shuhei; Chan, Chung; Rocco, Kevin A.; Shinoka, Toshiharu; Sinusas, Albert J.; Breuer, Christopher K.

    2014-01-01

    Objective(s) The clinical translation of tissue-engineered vascular grafts has been demonstrated in children. The remodeling of biodegradable, cell-seeded scaffolds to functional neovessels is partially attributed to matrix metalloproteinases. Noninvasive assessment of matrix metalloproteinase activity may indicate graft remodeling and elucidate the progression of neovessel formation. Therefore, matrix metalloproteinase activity was evaluated in grafts implanted in lambs using in vivo and ex vivo hybrid imaging. Graft growth and remodeling was quantified using in vivo X-ray computed tomography angiography. Methods Cell-seeded and unseeded scaffolds were implanted in lambs (n=5) as inferior vena cava interposition grafts. At 2 and 6 months post-implantation, in vivo angiography assessed graft morphology. In vivo and ex vivo single photon emission tomography/X-ray computed tomography imaging was performed with a radiolabeled compound targeting matrix metalloproteinase activity at 6 months. Neotissue was examined at 6 months using qualitative histologic and immunohistochemical staining and quantitative biochemical analysis. Results Seeded grafts demonstrated significant luminal and longitudinal growth from 2 to 6 months. In vivo imaging revealed subjectively higher matrix metalloproteinase activity in grafts vs. native tissue. Ex vivo imaging confirmed a quantitative increase in matrix metalloproteinase activity and demonstrated higher activity in unseeded vs. seeded grafts. Glycosaminoglycan content was increased in seeded grafts vs. unseeded grafts, without significant differences in collagen content. Conclusions Matrix metalloproteinase activity remains elevated in tissue-engineered grafts 6 months post-implantation and may indicate remodeling. Optimization of in vivo imaging to noninvasively evaluate matrix metalloproteinase activity may assist in serial assessment of vascular graft remodeling. PMID:24952823

  3. Chitosan-Based Bilayer Hydroxyapatite Nanorod Composite Scaffolds for Osteochondral Regeneration

    NASA Astrophysics Data System (ADS)

    Swanson, Shawn

    Osteochondral defects involve injury to bone and cartilage. As articular cartilage is worn down, bone in the joint begins to rub together, causing bone spurs. This is known as osteoarthritis, and is a common issue among the aging population. This problem presents an interesting opportunity for tissue engineering. Tissue engineering is an approach to treatment of tissue defects where synthetic, three dimensional (3-D) scaffolds are implanted in a defect to facilitate healing. The osteochondral scaffold consists of two regions in the form of a bilayer scaffold- one to mimic bone with osteoconductive properties, and one to mimic cartilage with biomimetic properties. One approach to improving the osteoconductivity of tissue engineering scaffolds is the addition of hydroxyapatite (HAp), the main mineral phase in bone. HAp with nanorod morphology is desirable because it is biomimetic for the calcium phosphate found in bone. Incorporating HAp nanorods in bone tissue engineering scaffolds to form a composite material may increase scaffold osteoconductivity. The cartilage scaffold is fabricated from chitosan and hyaluronic acid (HA). HA is a known component of cartilage and thus is biomimetic. The bilayer scaffolds were seeded with osteoblast-like MG-63 cells to investigate cell migration and were evaluated with Alamar Blue proliferation assay. The cells successfully migrated to the bone region of the scaffold, indicating that the bilayer scaffold provides a promising osteochondral scaffold.

  4. Hemi-Descemet membrane endothelial keratoplasty transplantation: a potential method for increasing the pool of endothelial graft tissue.

    PubMed

    Lam, Fook Chang; Baydoun, Lamis; Dirisamer, Martin; Lie, Jessica; Dapena, Isabel; Melles, Gerrit R J

    2014-12-01

    This study evaluates the technical feasibility and clinical outcomes of a Descemet membrane endothelial keratoplasty (DMEK) technique that could increase the availability of donor tissue for DMEK. To evaluate the clinical outcome of using a semicircular, large-diameter Descemet membrane graft in DMEK (hemi-DMEK), potentially allowing the use of a single donor cornea for 2 DMEK procedures. A prospective, interventional case series was conducted at a tertiary referral center. Three eyes of 3 patients with Fuchs endothelial dystrophy received a hemi-DMEK. Transplantation of a semicircular, large-diameter hemi-DMEK graft. Best-corrected visual acuity, endothelial cell density, pachymetry, and intraoperative and postoperative complications. The patients' best-corrected visual acuity at 6 months was 0.70 (Snellen equivalent, 20/29), 0.50 (20/40 [amblyopic eye]), and 1.20 (20/17). At 1 month, endothelial cell density decreased by 49%, 31%, and 39%, respectively, and endothelial cell migration appeared to continue for up to 6 months. Central corneal thicknesses decreased from 682, 707, and 681 μm before surgery to 523, 534, and 489 μm, respectively, at 6 months. No intraoperative or postoperative complications were seen. Hemi-DMEK (using half-moon-shaped grafts) is technically feasible and may provide visual outcomes similar to those obtained with routine DMEK (full-moon-shaped graft). If so, this technique may have the potential to double the availability of donor endothelial tissue for DMEK surgery.

  5. Osteointegration of soft tissue grafts within the bone tunnels in anterior cruciate ligament reconstruction can be enhanced.

    PubMed

    Kuang, Guan-Ming; Yau, W P; Lu, William W; Chiu, K Y

    2010-08-01

    Anterior cruciate ligament reconstruction with a soft tissue autograft (hamstring autograft) has grown in popularity in the last 10 years. However, the issues of a relatively long healing time and an inferior histological healing result in terms of Sharpey-like fibers connection in soft tissue grafts are still unsolved. To obtain a promising outcome in the long run, prompt osteointegration of the tendon graft within the bone tunnel is essential. In recent decades, numerous methods have been reported to enhance osteointegration of soft tissue graft in the bone tunnel. In this article, we review the current literature in this research area, mainly focusing on strategies applied to the local bone tunnel environment. Biological strategies such as stem cell and gene transfer technology, as well as the local application of specific growth factors have been reported to yield exciting results. The use of biological bone substitute and physical stimulation also obtained promising results. Artificially engineered tissue has promise as a solution to the problem of donor site morbidity. Despite these encouraging results, the current available evidence is still experimental. Further clinical studies in terms of randomized control trial in the future should be conducted to extrapolate these basic science study findings into clinical practice.

  6. A histologic evaluation of various stages of palatal healing following subepithelial connective tissue grafting procedures: a comparison of eight cases.

    PubMed

    Soileau, Kristi M; Brannon, Robert B

    2006-07-01

    It is often necessary to procure tissue from the same area of the palate in cases where connective autogenous grafting procedures are warranted due to limitations caused by anatomical features. The purpose of this study was to determine whether the length of time between a first procurement and a second would have any bearing on the quality of tissue available for recipient sites. Eight patients requiring more than one grafting procedure underwent surgery at baseline and again at various intervals ranging from 6 weeks to 11 months. Specimens were taken from the palate and evaluated microscopically, and photographs were taken for purposes of visual comparison. The 6.9- to 7.7-week specimens exhibited complete reepithelialization. The lamina propriae were composed of a cellular proliferation of fibroblasts with loosely arranged collagen deposition and an occasional thin vascular channel. However, remodeling of the wound appeared complete in the specimens removed at the 9-week interval and beyond. The lamina propria was, in general, composed of thick, dense, interlacing bundles of collagen. Small-caliber blood vessels were interspersed throughout the fibrous element. Reharvesting of tissue performed earlier than at 9 weeks may result in poorer autogenous graft quality due to indications that remodeling of the connective tissue is still progressing and not as mature as specimens noted at weeks 9 to 47.

  7. Graft of autologous fibroblasts in gingival tissue in vivo after culture in vitro. Preliminary study on rats.

    PubMed

    Simain-Sato, F; Lahmouzi, J; Heinen, E; Defresne, M P; De Pauw-Gillet, M C; Grisar, T; Legros, J J; Legrand, R

    1999-08-01

    Several grafting techniques and guided tissue regeneration techniques (GTR) have been well-developed in periodontal surgery. However, these techniques could induce pain and side effects, such as a gingival recession during the healing period following the therapy. The graft of a small autologous connective tissue, using non-invasive surgical techniques could yield several benefits for the patients. Our preliminary study explores the feasibility of collecting healthy gingival tissues, culturing them in vitro to amplify rat gingival fibroblasts (RGF) and inoculating the obtained cells into autologous rat gingival tissues in vivo. Gingival tissues samples were cultured as explants as described by Freshney et al. and Adolphe. Confluent cells surrounding explants were detached after 7 d of culture from Petri dishes using 0.05% trypsin and designated "first transferred cells" (T1). At the third passage (T3), cells cultured as monolayer were either examined under microscopy--phase contrast, scanning, or transmission electron--or numerated after trypan blue exclusion test. Autologous RGF labelled with fluorochrome were inoculated at the vestibular and palatine site of gingival tissue close to the superior incisors. In this preliminary study, 12 Wistar rats were used; for each, 2 biopsies were dissected and fixed for phase contrast or fluorescence microscopy. On d 1, 3 and 7 after injection in rat gingival tissues, fluorochrome-labelled cells could be detected in all these.

  8. Graft-versus-host disease reduces regulatory T-cell migration into the tumour tissue

    PubMed Central

    Dürr, Christoph; Follo, Marie; Idzko, Marco; Reichardt, Wilfried; Zeiser, Robert

    2012-01-01

    The therapeutic principle of allogeneic haematopoietic cell transplantation (allo-HCT) is based on an active donor immune system that eliminates host-derived tumour cells. We hypothesized that in addition to the alloantigen-driven anti-tumour response, disruption of the immunological microenvironment within the tumour is responsible for its elimination after allo-HCT. We observed that induction of graft-versus-host disease (GvHD) significantly reduced the abundance of luc+ FoxP3+ regulatory T (Treg) cells in the tumour tissue, which is indicative of impaired or over-ridden tumour recruitment signals towards Treg cells. Analysis of the intestines and liver revealed chemokines and purine nucleotides as candidates for attracting Treg to these sites of inflammation. Despite its expression on tissue-residing Treg cells, the chemokine receptor CCR3 was not critical for Treg-cell function following allo-HCT. Extracellular ATP can attract immune cells via P2Y2. P2Y2 was found to be expressed on Treg cells, and we found a partial reduction of GvHD prevention when P2Y2−/− rather than P2Y2+/+ Treg cells were given. Exogenous local inflammation reduced Treg-cell accumulation in the tumour, suggesting a potential clinical approach to prevent Treg-cell-mediated tumour escape. In conclusion, we demonstrate that GvHD-related inflammation reduced Treg-cell numbers at the tumour sites, which may in turn help to explain the observation that patients with GvHD have a lower risk of tumour relapse. PMID:22681312

  9. Blocking of tumor necrosis factor activity promotes natural repair of osteochondral defects in rabbit knee

    PubMed Central

    2009-01-01

    Background and purpose Osteochondral defects have a limited capacity for repair. We therefore investigated the effects of tumor necrosis factor (TNF) signal blockade by etanercept (human recombinant soluble TNF receptor) on the repair of osteochondral defects in rabbit knees. Material and methods Osteochondral defects (5 mm in diameter) were created in the femoral patellar groove in rabbits. Soon after the procedure, a first subcutaneous injection of etanercept was performed. This single injection or, alternatively, 4 injections in total (twice a week for 2 weeks) were given. Each of these 2 groups was divided further into 3 subgroups: a low-dose group (0.05 μg/kg), an intermediate-dose group (0.4 μ g/kg), and a high-dose group (1.6 μ g /kg) with 19 rabbits in each. As a control, 19 rabbits were injected with water alone. The rabbits in each subgroup were killed 4 weeks (6 rabbits), 8 weeks (6 rabbits), or 24 weeks (7 rabbits) after surgery and repair was assessed histologically. Results Histological examination revealed that the natural process of repair of the osteochondral defects was promoted by 4 subcutaneous injections of intermediate-dose etanercept and by 1 or 4 injections of high-dose etanercept at the various time points examined postoperatively (4, 8, and 24 weeks). Western blot showed that rabbit TNFα had a high affinity for etanercept. Interpretation Blocking of TNF by etanercept enabled repair of osteochondral defects in rabbit knee. Anti-TNF therapy could be a strategy for the use of tissue engineering for bone and cartilage repair. PMID:19916697

  10. A multilayer biomaterial for osteochondral regeneration shows superiority vs microfractures for the treatment of osteochondral lesions in a multicentre randomized trial at 2 years.

    PubMed

    Kon, Elizaveta; Filardo, Giuseppe; Brittberg, Mats; Busacca, Maurizio; Condello, Vincenzo; Engebretsen, Lars; Marlovits, Stefan; Niemeyer, Philipp; Platzer, Patrik; Posthumus, Michael; Verdonk, Peter; Verdonk, Renè; Victor, Jan; van der Merwe, Willem; Widuchowski, Wojciech; Zorzi, Claudio; Marcacci, Maurilio

    2017-09-14

    The increasing awareness on the role of subchondral bone in the etiopathology of articular surface lesions led to the development of osteochondral scaffolds. While safety and promising results have been suggested, there are no trials proving the real potential of the osteochondral regenerative approach. Aim was to assess the benefit provided by a nanostructured collagen-hydroxyapatite (coll-HA) multilayer scaffold for the treatment of chondral and osteochondral knee lesions. In this multicentre randomized controlled clinical trial, 100 patients affected by symptomatic chondral and osteochondral lesions were treated and evaluated for up to 2 years (51 study group and 49 control group). A biomimetic coll-HA scaffold was studied, and bone marrow stimulation (BMS) was used as reference intervention. Primary efficacy measurement was IKDC subjective score at 2 years. Secondary efficacy measurements were: KOOS, IKDC Knee Examination Form, Tegner and VAS Pain scores evaluated at 6, 12 and 24 months. Tissue regeneration was evaluated with MRI MOCART scoring system at 6, 12 and 24 months. An external independent agency was involved to ensure data correctness and objectiveness. A statistically significant improvement of all clinical scores was obtained from basal evaluation to 2-year follow-up in both groups, although no overall statistically significant differences were detected between the two treatments. Conversely, the subgroup of patients affected by deep osteochondral lesions (i.e. Outerbridge grade IV and OCD) showed a statistically significant better IKDC subjective outcome (+12.4 points, p = 0.036) in the coll-HA group. Statistically significant better results were also found for another challenging group: sport active patients (+16.0, p = 0.027). Severe adverse events related to treatment were documented only in three patients in the coll-HA group and in one in the BMS group. The MOCART score showed no statistical difference between the two groups. This

  11. Tectal Tissue Grafted to the Midbrain of Newborn Rats: Effect of Donor Age on the Survival, Growth and Connectivity of Transplants

    PubMed Central

    Majda, Bernadette T.; Harvey, Alan R.

    1989-01-01

    Tectal tissue from fetal (E15, E18, E20) and newborn (P0) rats was transplanted to the midbrain of newborn rats. Graft survival and size decreased with increasing donor age. Host retinal input was found in E15, E18 and E20 grafts; the specific pattern of retinal innervation was similar for all fetal donor ages. PMID:2485121

  12. Surgical treatment of localized gingival recessions using coronally advanced flaps with or without subepithelial connective tissue graft

    PubMed Central

    Bellver-Fernández, Ricardo; Martínez-Rodriguez, Ana-María; Gioia-Palavecino, Claudio; Caffesse, Raul-Guillermo

    2016-01-01

    Background A coronally advanced flap with subepithelial connective tissue graft is the gold standard surgical treatment of gingival recessions, since it offers a higher probability of achieving complete root coverage compared with other techniques. However, optimum short- and middle-term clinical results have also been obtained with coronally advanced flaps alone. The aim of the present study was to evaluate the results obtained by the surgical treatment of localized gingival recessions using coronally advanced flaps with or without subepithelial connective tissue graft. Material and Methods The reduction of recession height was assessed, together with the gain in gingival attachment apical to the recession, and total reduction of recession, in a comparative study of two techniques. Twenty-two gingival recessions were operated upon: 13 in the control group (coronally advanced flap) and 9 in the test group (coronally advanced flap associated to subepithelial connective tissue graft). Results After 18 months, the mean reduction of recession height was 2.2 ± 0.8 mm in the control group and 2.3 ± 0.7 mm in the test group, with a mean gain in gingival attachment of 1.3 ± 0.9 mm and 2.3 ± 1.3 mm, respectively. In percentage terms, the mean reduction of recession height was 84.6 ± 19.6% in the control group and 81.7 ± 17.8% in the test group, with a mean gain in gingival attachment of 20.5 ± 37.4% and 184.4 ± 135.5%, respectively. Conclusions Significant reduction of gingival recession was achieved with both techniques, though the mean gain in gingival attachment (in mm and as a %) was greater in test group. Key words:Gingival recession, coronally advanced flap, subepthelial connective tissue graft. PMID:26595836

  13. Histological advantages of the tumor graft: a murine model involving transplantation of human pancreatic cancer tissue fragments.

    PubMed

    Akashi, Yoshimasa; Oda, Tatsuya; Ohara, Yusuke; Miyamoto, Ryoichi; Hashimoto, Shinji; Enomoto, Tsuyoshi; Yamada, Keiichi; Kobayashi, Akihiko; Fukunaga, Kiyoshi; Ohkochi, Nobuhiro

    2013-11-01

    Experimental data based on cell line-derived xenograft models (cell xenograft) seldom reproduce the clinical situation, and therefore we demonstrated here the superiority of a murine model involving transplantation of human pancreatic cancer tissue fragments (tumor graft), focusing on the histological features and drug delivery characteristics. Tumor pieces from 10 pancreatic cancer patients were transplanted into SCID (severe combined immunodeficient) mice. Histological characteristics of tumor grafts, including morphology, desmoplastic reaction, and vascularization, were compared with those of cell xenografts. Drug delivery was evaluated by quantifying the concentrations of injected drug, and the results were compared with its histological features. Eight of the 10 transplanted tumors successfully engrafted. Histological comparisons between tumor grafts and cell xenografts revealed the following: the amount of stroma was more (22.9% ± 11.8% vs 10.8% ± 5.4%; P < 0.05), vessel-cancer cell distance was longer (35.3 ± 39.0 vs 3.9 ± 3.1 μm; P < 0.001), and microvessel density was lower (6.8 ± 1.9 vs 10.8 ± 2.1 vessels/0.4 mm(2); P < 0.05) in tumor grafts. Drug concentrations in tumor grafts were lower than those in cell xenografts (3.3 ± 1.2 vs 6.0±0.2 μg/mL; P = 0.003), and the differences were correlated with the histological differences. Pancreatic tumor grafts better reproduce the histological nature of clinical cancer and thus provide a more realistic model that is applicable for pharmacokinetic studies.

  14. Tissue engineering of autologous cartilage grafts in three-dimensional in vitro macroaggregate culture system.

    PubMed

    Naumann, Andreas; Dennis, James E; Aigner, Joachim; Coticchia, James; Arnold, James; Berghaus, Alexander; Kastenbauer, Ernst R; Caplan, Arnold I

    2004-01-01

    and auricular macroaggregates were negative for anti-elastin antibody (interterritorially). The measurement of total GAG content demonstrated higher GAG content for reformed nasoseptal cartilage compared with elastic auricular cartilage. However, the total GAG content of engineered macroaggregates was lower than that of native cartilage. In spite of the mechanical stability of the auricular macroaggregates, there was no equilibrium of indentation. The histomorphological and immunohistochemical results demonstrate successful cartilage engineering without the use of biomaterials, and identify characteristics unique to hyaline as well as elastic cartilage. The GAG content of engineered cartilage was lower than in native cartilage and the biomechanical properties were not determinable by indentation assay. This study illustrates a novel in vitro macroaggregate culture system as a promising technique for tissue engineering of cartilage grafts. Further long-term in vitro and in vivo studies must be done before this method can be applied to reconstructive surgery of the nose or auricle.

  15. Effects of expanded human adipose tissue-derived mesenchymal stem cells on the viability of cryopreserved fat grafts in the nude mouse.

    PubMed

    Ko, Myung-Soon; Jung, Ji-Youl; Shin, Il-Seob; Choi, Eun-Wha; Kim, Jae-Hoon; Kang, Sung Keun; Ra, Jeong Chan

    2011-03-14

    Adipose-derived mesenchymal stem cells (AdMSCs) augment the ability to contribute to microvascular remodeling in vivo and to modulate vascular stability in fresh fat grafts. Although cryopreserved adipose tissue is frequently used for soft tissue augmentation, the viability of the fat graft is poor. The effects of culture-expanded human adipose tissue-derived mesenchymal stem cells (hAdMSCs) on the survival and quality of the cryopreserved fat graft were determined. hAdMSCs from the same donor were mixed with fat tissues cryopreserved at -70 °C for 8 weeks and injected subcutaneously into 6-week-old BALB/c-nu nude mice. Graft volume and weight were measured, and histology was evaluated 4 and 15 weeks post-transplantation. The hAdMSC-treated group showed significantly enhanced graft volume and weight. The histological evaluation demonstrated significantly better fat cell integrity compared with the vehicle-treated control 4 weeks post-transplantation. No significant difference in graft weight, volume, or histological parameters was found among the groups 15 weeks post-transplantation. The hAdMSCs enhanced the survival and quality of transplanted cryopreserved fat tissues. Cultured and expanded hAdMSCs have reconstructive capacity in cryopreserved fat grafting by increasing the number of stem cells.

  16. Effects of Expanded Human Adipose Tissue-Derived Mesenchymal Stem Cells on the Viability of Cryopreserved Fat Grafts in the Nude Mouse

    PubMed Central

    Ko, Myung-Soon; Jung, Ji-Youl; Shin, Il-Seob; Choi, Eun-Wha; Kim, Jae-Hoon; Kang, Sung Keun; Ra, Jeong Chan

    2011-01-01

    Adipose-derived mesenchymal stem cells (AdMSCs) augment the ability to contribute to microvascular remodeling in vivo and to modulate vascular stability in fresh fat grafts. Although cryopreserved adipose tissue is frequently used for soft tissue augmentation, the viability of the fat graft is poor. The effects of culture-expanded human adipose tissue-derived mesenchymal stem cells (hAdMSCs) on the survival and quality of the cryopreserved fat graft were determined. hAdMSCs from the same donor were mixed with fat tissues cryopreserved at -70°C for 8 weeks and injected subcutaneously into 6-week-old BALB/c-nu nude mice. Graft volume and weight were measured, and histology was evaluated 4 and 15 weeks post-transplantation. The hAdMSC-treated group showed significantly enhanced graft volume and weight. The histological evaluation demonstrated significantly better fat cell integrity compared with the vehicle-treated control 4 weeks post-transplantation. No significant difference in graft weight, volume, or histological parameters was found among the groups 15 weeks post-transplantation. The hAdMSCs enhanced the survival and quality of transplanted cryopreserved fat tissues. Cultured and expanded hAdMSCs have reconstructive capacity in cryopreserved fat grafting by increasing the number of stem cells. PMID:21448310

  17. Tissue-engineered rhesus monkey nerve grafts for the repair of long ulnar nerve defects: similar outcomes to autologous nerve grafts

    PubMed Central

    Jiang, Chang-qing; Hu, Jun; Xiang, Jian-ping; Zhu, Jia-kai; Liu, Xiao-lin; Luo, Peng

    2016-01-01

    Acellular nerve allografts can help preserve normal nerve structure and extracellular matrix composition. These allografts have low immunogenicity and are more readily available than autologous nerves for the repair of long-segment peripheral nerve defects. In this study, we repaired a 40-mm ulnar nerve defect in rhesus monkeys with tissue-engineered peripheral nerve, and compared the outcome with that of autograft. The graft was prepared using a chemical extract from adult rhesus monkeys and seeded with allogeneic Schwann cells. Pathomorphology, electromyogram and immunohistochemistry findings revealed the absence of palmar erosion or ulcers, and that the morphology and elasticity of the hypothenar eminence were normal 5 months postoperatively. There were no significant differences in the mean peak compound muscle action potential, the mean nerve conduction velocity, or the number of neurofilaments between the experimental and control groups. However, outcome was significantly better in the experimental group than in the blank group. These findings suggest that chemically extracted allogeneic nerve seeded with autologous Schwann cells can repair 40-mm ulnar nerve defects in the rhesus monkey. The outcomes are similar to those obtained with autologous nerve graft. PMID:28123431

  18. Fabrication of tissue-engineered vascular grafts with stem cells and stem cell-derived vascular cells

    PubMed Central

    Wang, Lunchang; Hu, Jiang; Sorek, Claire E; Chen, Eugene Y; Ma, Peter X; Yang, Bo

    2016-01-01

    Introduction Cardiovascular disease is the leading cause of mortality worldwide. Current surgical treatments for cardiovascular disease include vascular bypass grafting and replacement with autologous blood vessels or synthetic vascular grafts. However, there is a call for better alternative biological grafts. Areas covered Tissue-engineered vascular grafts (TEVGs) are promising novel alternatives to replace diseased vessels. However, obtaining enough functional and clinically usable vascular cells for fabrication of TEVGs remains a major challenge. New findings in adult stem cells and recent advances in pluripotent stem cells have opened a new avenue for stem cell-based vascular engineering. In this review, recent advances on stem cell sourcing for TEVGs including the use of adult stem cells and pluripotent stem cells, and advantages, disadvantages, and possible future implementations of different types of stem cells will be discussed. In addition, current strategies used during the fabrication of TEVGs will be highlighted. Expert opinion The application of patient-specific TEVGs constructed with vascular cells derived from immune-compatible stem cells possesses huge clinical potential. Advances in lineage-specific differentiation approaches and innovative vascular engineering strategies will promote the vascular regeneration field from bench to bedside. PMID:26560995

  19. A Bi-Layered Elastomeric Scaffold for Tissue Engineering of Small-Diameter Vascular Grafts

    PubMed Central

    Soletti, Lorenzo; Hong, Yi; Guan, Jianjun; Stankus, John J.; El-Kurdi, Mohammed S.; Wagner, William R.; Vorp, David A.

    2011-01-01

    A major barrier in the development of a clinically-useful small-diameter tissue engineered vascular graft (TEVG) is the scaffold component. Scaffold requirements include matching the mechanical and structural properties with those of native vessels and optimizing the microenvironment to foster cell integration, adhesion, and growth. We have developed a small-diameter, bi-layered, biodegradable, elastomeric scaffold based on a synthetic, biodegradable elastomer. The scaffold incorporates a highly porous inner layer, allowing cell integration and growth, and an external, fibrous reinforcing layer deposited by electrospinning. Scaffold morphology and mechanical properties were assessed, quantified, and compared to those of native vessels. Scaffolds were then seeded with adult stem cells via a rotational vacuum seeding device to obtain a TEVG, cultured in dynamic conditions for 7 days, and evaluated for cellularity. The scaffold showed a firm integration of the two polymeric layers with no delaminations. Mechanical properties were physiologically-consistent showing anisotropy, elastic modulus (1.4±0.4 MPa), and ultimate tensile stress (8.3±1.7 MPa) comparable with native vessels. Compliance and suture retention force were 4.6±0.5×10−4 mmHg−1 and 3.4±0.3 N, respectively. Seeding resulted in a rapid, uniform, bulk integration of cells, with a seeding efficiency of 92±1%. The scaffolds maintained a high level of cellular density throughout dynamic culture. This approach, combining artery-like mechanical properties and a rapid and efficient cellularization, might contribute to the future clinical translation of TEVGs. PMID:19540370

  20. Biomechanical Comparison of an Intramedullary and Extramedullary Free-Tissue Graft Reconstruction of the Acromioclavicular Joint Complex

    PubMed Central

    Garg, Rishi; Javidan, Pooya; Lee, Thay Q.

    2013-01-01

    Background Several different surgical techniques have been described to address the coracoclavicular (CC) ligaments in acromioclavicular (AC) joint injuries. However, very few techniques focus on reconstructing the AC ligaments, despite its importance in providing stability. The purpose of our study was to compare the biomechanical properties of two free-tissue graft techniques that reconstruct both the AC and CC ligaments in cadaveric shoulders, one with an extramedullary AC reconstruction and the other with an intramedullary AC reconstruction. We hypothesized intramedullary AC reconstruction will provide greater anteroposterior translational stability and improved load to failure characteristics than an extramedullary technique. Methods Six matched cadaveric shoulders underwent translational testing at 10 N and 15 N in the anteroposterior and superoinferior directions, under AC joint compression loads of 10 N, 20 N, and 30 N. After the AC and CC ligaments were transected, one of the specimens was randomly assigned the intramedullary free-tissue graft reconstruction while its matched pair received the extramedullary graft reconstruction. Both reconstructed specimens then underwent repeat translational testing, followed by load to failure testing, via superior clavicle distraction, at a rate of 50 mm/min. Results Intramedullary reconstruction provided significantly greater translational stability in the anteroposterior direction than the extramedullary technique for four of six loading conditions (p < 0.05). There were no significant differences in translational stability in the superoinferior direction for any loading condition. The intramedullary reconstructed specimens demonstrated improved load to failure characteristics with the intramedullary reconstruction having a lower deformation at yield and a higher ultimate load than the extramedullary reconstruction (p < 0.05). Conclusions Intramedullary reconstruction of the AC joint provides greater stability in the

  1. In vitro characterization of design and compressive properties of 3D-biofabricated/decellularized hybrid grafts for tracheal tissue engineering.

    PubMed

    Johnson, Christopher; Sheshadri, Priyanka; Ketchum, Jessica M; Narayanan, Lokesh K; Weinberger, Paul M; Shirwaiker, Rohan A

    2016-06-01

    Infection or damage to the trachea, a thin walled and cartilage reinforced conduit that connects the pharynx and larynx to the lungs, leads to serious respiratory medical conditions which can often prove fatal. Current clinical strategies for complex tracheal reconstruction are of limited availability and efficacy, but tissue engineering and regenerative medicine approaches may provide viable alternatives. In this study, we have developed a new "hybrid graft" approach that utilizes decellularized tracheal tissue along with a resorbable polymer scaffold, and holds promise for potential clinical applications. First, we evaluated the effect of our decellularization process on the compression properties of porcine tracheal segments, and noted approximately 63% decrease in resistance to compression following decellularization. Next we developed four C-shape scaffold designs by varying the base geometry and thickness, and fabricated polycaprolactone scaffolds using a combination of 3D-Bioplotting and thermally-assisted forming. All scaffolds designs were evaluated in vitro under three different environmental testing conditions to determine the design that offered the best resistance to compression. These were further studied to determine the effect of gamma radiation sterilization and cyclic compression loading. Finally, hybrid grafts were developed by securing these optimal design scaffolds to decellularized tracheal segments and evaluated in vitro under physiological testing conditions. Results show that the resistance to compression offered by the hybrid grafts created using gamma radiation sterilized scaffolds was comparable to that of fresh tracheal segments. Given that current clinical attempts at tracheal transplantation using decellularized tissue have been fraught with luminal collapse and complications, our data support the possibility that future embodiments using a hybrid graft approach may reduce the need for intraluminal stenting in tracheal transplant

  2. Changes in the osteochondral unit during osteoarthritis: structure, function and cartilage-bone crosstalk.

    PubMed

    Goldring, Steven R; Goldring, Mary B

    2016-11-01

    In diarthrodial joints, the articular cartilage, calcified cartilage, and subchondral cortical and trabecular bone form a biocomposite - referred to as the osteochondral unit - that is uniquely adapted to the transfer of load. During the evolution of the osteoarthritic process the compositions, functional properties, and structures of these tissues undergo marked alterations. Although pathological processes might selectively target a single joint tissue, ultimately all of the components of the osteochondral unit will be affected because of their intimate association, and thus the biological and physical crosstalk among them is of great importance. The development of targeted therapies against the osteoarthritic processes in cartilage or bone will, therefore, require an understanding of the state of these joint tissues at the time of the intervention. Importantly, these interventions will not be successful unless they are applied at the early stages of disease before considerable structural and functional alterations occur in the osteochondral unit. This Review describes the changes that occur in bone and cartilage during the osteoarthritic process, and highlights strategies for how this knowledge could be applied to develop new therapeutic interventions for osteoarthritis.

  3. Phase 2 Clinical Trial of Intraoral Grafting of Human Tissue-Engineered Oral Mucosa

    DTIC Science & Technology

    2013-10-01

    differences in the primary efficacy measure of increased keratinized mucosa; secondary measures of graft contracture and Wound Healing Index; and...per treatment group, will be randomized to receive either the experimental treatment , EVPOME (Group 1), or standard of care, the palatal oral mucosa...Keratinized mucosa, graft contracture 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF PAGES 19a. NAME OF RESPONSIBLE

  4. [Marketing role of corneal graft tissue donation to an eye bank and donors' socioeconomic profile].

    PubMed

    Farias, Roberta Jansen de Mello; Sousa, Luciene Barbosa de

    2008-01-01

    Penetrating keratoplasty has been the leading and the most successful type of transplant in the world, however corneal deficiency is a commom problem usually presented to corneal surgeons. Impact evaluation of the number of corneal graft donations to the Sorocaba Eye Bank after the implementation of a corneal graft procurement system; to draw the socioeconomic profile of corneal graft donors of the Sorocaba Eye Bank (SEB). Retrospective study on donations to SEB from its creation and after the development of media marketing. Prospective analysis of the socioeconomic profile of corneal graft donors by a questionnaire sent as letters to the families of the donors in a certain month. SEB began its work in 1971 by spreading need of organ donation through lectures in churches, shopping malls, community meetings, radio programs, television programs, etc. In the 70s, the number of retrieved corneal grafts was 1 or 2/month. Between 1984 - 1989 a procurement coordination team was trained to act in mortuaries and by 2000 they also began to work in public hospitals. In 1984 only 260 corneal grafts were retrieved. This number has been increasing to 2,778 corneal graft donations in 2004. The questionnaire was answered by 76 of the 93 donor families, with a response rate of 81.7%. Donor age had a mean of 65.1 +/- 14.7 y/o, forty-two (55.3%) were men. Educational level of the donor families was an important factor for organ donation, once 36.8% had concluded high school and 34.2% completed university. The great majority, sixty-three (82.9%) of the corneal grafts were donated through the efforts of the procurement coordination team. The role of the media and institutional credibility are mandatory for public commitment to organ donation. The proficiency of the procurement coordination team requires intensive training, as the results show that 82.9% donations were made thanks to their efforts.

  5. A gingival cul-de-sac following a root coverage procedure with a subepithelial connective tissue submerged graft.

    PubMed

    Wei, Pein-Chi; Geivelis, Milton

    2003-09-01

    The subepithelial connective tissue graft (SCTG) used as a submerged graft in combination with a partial thickness advanced flap or rotated flap is a predictable technique for achieving coverage of the denuded root surface and/or for increasing the width of attached gingiva in Miller's class I and II marginal tissue recessions. However, even with a successful result, complications may occasionally occur. A 4-mm marginal tissue recession with an insufficient zone of attached gingiva on the facial aspect of a mandibular left lateral incisor (#23) was covered with a submerged SCTG and an envelope partial thickness flap. The mucogingival defect was successfully corrected. Nevertheless, the facial gingiva, specifically between teeth #22 (mandibular left canine) and #23, remained bulky and was reshaped 6 months postoperatively. A 5-mm long gingival cul-de-sac with an intermittent thick white discharge was detected 3 months following the gingivoplasty procedure. With a periodontal probe kept in the tract, the lining of the cul-de-sac was exposed to the oral cavity by making an incision on the facial gingiva, along the axis of the probe, through to the lumen. The gingival tissue was then removed with a rotary bur until only a thin layer of periosteum remained. The mucosal defect was subsequently repaired by grafting with a non-submerged SCTG. The gingival contour was significantly improved and no sign of recurrence was noted up to 4 years later. The existence of a cyst cannot be verified without a histological evaluation. However, the development of a gingival cyst should be suspected with persistent tissue bulkiness and/or emergence of a thick white discharge from a site where a submerged SCTG procedure was performed. In addition, the ill-circumscribed border around the lesion makes complete elimination of the pathosis relatively hard to achieve by a "superficial" gingivoplasty procedure.

  6. Free bone graft reconstruction of irradiated facial tissue: Experimental effects of basic fibroblast growth factor stimulation

    SciTech Connect

    Eppley, B.L.; Connolly, D.T.; Winkelmann, T.; Sadove, A.M.; Heuvelman, D.; Feder, J. )

    1991-07-01

    A study was undertaken to evaluate the potential utility of basic fibroblast growth factor in the induction of angiogenesis and osseous healing in bone previously exposed to high doses of irradiation. Thirty New Zealand rabbits were evaluated by introducing basic fibroblast growth factor into irradiated mandibular resection sites either prior to or simultaneous with reconstruction by corticocancellous autografts harvested from the ilium. The fate of the free bone grafts was then evaluated at 90 days postoperatively by microangiographic, histologic, and fluorochrome bone-labeling techniques. Sequestration, necrosis, and failure to heal to recipient osseous margins was observed both clinically and histologically in all nontreated irradiated graft sites as well as those receiving simultaneous angiogenic stimulation at the time of graft placement. No fluorescent activity was seen in these graft groups. In the recipient sites pretreated with basic fibroblast growth factor prior to placement of the graft, healing and reestablishment of mandibular contour occurred in nearly 50 percent of the animals. Active bone formation was evident at cortical margins adjacent to the recipient sites but was absent in the more central cancellous regions of the grafts.

  7. Osteochondral defects in the ankle: why painful?

    PubMed Central

    Reilingh, Mikel L.; Zengerink, Maartje; van Bergen, Christiaan J. A.

    2010-01-01

    Osteochondral defects of the ankle can either heal and remain asymptomatic or progress to deep ankle pain on weight bearing and formation of subchondral bone cysts. The development of a symptomatic OD depends on various factors, including the damage and insufficient repair of the subchondral bone plate. The ankle joint has a high congruency. During loading, compressed cartilage forces its water into the microfractured subchondral bone, leading to a localized high increased flow and pressure of fluid in the subchondral bone. This will result in local osteolysis and can explain the slow development of a subchondral cyst. The pain does not arise from the cartilage lesion, but is most probably caused by repetitive high fluid pressure during walking, which results in stimulation of the highly innervated subchondral bone underneath the cartilage defect. Understanding the natural history of osteochondral defects could lead to the development of strategies for preventing progressive joint damage. PMID:20151110

  8. Osteochondritis of the Distal Tibial Epiphysis

    PubMed Central

    EL Hajj, Firass; Sebaaly, Amer; Kharrat, Khalil; Ghanem, Ismat

    2012-01-01

    Osteochondritis of the distal tibial epiphysis is a very rare entity. 9 cases have been described in 7 articles and 8 other cases have been mentioned in textbooks. This paper describes the 10th case of osteochondritis of the distal tibial epiphysis and summarizes the clinical and radiological presentations of the 9 other cases. The etiology of this entity is well debated in the literature. We believe that it results from a vascular abnormality in the distal tibial epiphysis associated with a mechanical stress (trauma, excessive overload, etc.). Since it is a self-limited disease, the prognosis is good and the younger the patient is the better the prognosis will be. In general, this entity responds well to conservative treatment. PMID:23193412

  9. Osteochondral defects in the ankle: why painful?

    PubMed

    van Dijk, C Niek; Reilingh, Mikel L; Zengerink, Maartje; van Bergen, Christiaan J A

    2010-05-01

    Osteochondral defects of the ankle can either heal and remain asymptomatic or progress to deep ankle pain on weight bearing and formation of subchondral bone cysts. The development of a symptomatic OD depends on various factors, including the damage and insufficient repair of the subchondral bone plate. The ankle joint has a high congruency. During loading, compressed cartilage forces its water into the microfractured subchondral bone, leading to a localized high increased flow and pressure of fluid in the subchondral bone. This will result in local osteolysis and can explain the slow development of a subchondral cyst. The pain does not arise from the cartilage lesion, but is most probably caused by repetitive high fluid pressure during walking, which results in stimulation of the highly innervated subchondral bone underneath the cartilage defect. Understanding the natural history of osteochondral defects could lead to the development of strategies for preventing progressive joint damage.

  10. Effect of repeated subcutaneous injections of carbon dioxide (CO2) on inflammation linked to hypoxia in adipose tissue graft.

    PubMed

    Nisi, G; Barberi, L; Ceccaccio, L; Cuomo, R; Sisti, A; Castagna, A; Zerini, I; Brandi, C; Grimaldi, L; D'Aniello, C

    2015-12-01

    The purposes of this study was to assess the effect of repeated subcutaneous injections of CO2 on adipose tissue graft survival in immunosuppressed female nude mice. The authors designed an experimental study using volume measures, histopathological analysis and nuclear magnetic resonance of fat graft. The effect of repeated subcutaneous injection of CO2 is not yet investigated Approximately 0.5 ml of human fat were transplanted in a group of female nude mice. The mice were treated with 3 injections of 80 µl each carbon dioxide (total 240 µl) for 7 weeks. Initially, in vivo measurements were conducted and subsequently a comprehensive histopathological analysis was performed. The presence of inflammation was graded absent to minimal in animals treated with CO2 while a minimal to moderate grade was assigned to the control group. CO2 injection enhances the inflammatory response of the implanted tissue and reduces the reabsorption rate. The treatment may improve the graft survival in a more prolonged time-frame.

  11. Comparison of a Closed System to a Standard Open Technique for Preparing Tissue-Engineered Vascular Grafts

    PubMed Central

    Kurobe, Hirotsugu; Maxfield, Mark W.; Naito, Yuji; Cleary, Muriel; Stacy, Mitchel R.; Solomon, Daniel; Rocco, Kevin A.; Tara, Shuhei; Lee, Avione Y.; Sinusas, Albert J.; Snyder, Edward L.; Shinoka, Toshiharu

    2015-01-01

    We developed a prototype for a closed apparatus for assembling tissue-engineered vascular grafts (TEVGs) with the goal of creating a simple operator-independent method for making TEVGs to optimize safety and enable widespread application of this technology. The TEVG is made by seeding autologous bone marrow-derived mononuclear cells onto a biodegradable tubular scaffold and is the first man-made vascular graft to be successfully used in humans. A critical barrier, which has prevented the widespread clinical adoption of the TEVG, is that cell isolation, scaffold seeding, and incubation are performed using an open method. To reduce the risk of contamination, the TEVG is assembled in a clean room. Clean rooms are expensive to build, complex to operate, and are not available in most hospitals. In this investigation, we used an ovine model to compare the safety and efficacy of TEVGs created using either a standard density centrifugation-based open method or the new filter-based closed system. We demonstrated no graft-related complications and maintenance of growth capacity in TEVGs created using the closed apparatus. In addition, the use of the closed system reduced the amount of time needed to assemble the TEVG by ∼50%. Adaptation of similar methodologies may facilitate the safe translation and the widespread use of other tissue engineering technologies. PMID:24866863

  12. Cell Magnetic Targeting System for Repair of Severe Chronic Osteochondral Defect in a Rabbit Model.

    PubMed

    Mahmoud, Elhussein Elbadry; Kamei, Goki; Harada, Yohei; Shimizu, Ryo; Kamei, Naosuke; Adachi, Nobuo; Misk, Nabil Ahmed; Ochi, Mitsuo

    The aim of this study was to investigate a cell delivery system for repair of severe chronic osteochondral defects using magnetically labeled mesenchymal stem cells (m-MSCs), with the aid of an external magnetic device, through the accumulation of a small number of m-MSCs into a desired area and to detect the suitable number of autologous m-MSCs needed for repair of the defect. Twenty-six male Japanese white rabbits aged 6 months were used. An osteochondral defect was created bilaterally at the weight-bearing surface of the medial femoral condyle of the rabbits' knees (3 mm diameter; 4 mm depth). At 4 weeks after creation of the defect, autogenic transplantation of the m-MSCs into the defect area was performed, followed by 10-min exposure to an external magnetic device, where animals were divided into four groups: high (1 × 10(6) m-MSCs), medium (2 × 10(5) m-MSCs), low (4 × 10(4) m-MSCs), and control (PBS injection). At 4 and 12 weeks posttransplantation of m-MSCs, repaired tissue was assessed histologically using the Fortier score with toluidine blue staining. Transplantation of a low number of m-MSCs was not enough to improve osteogenesis and chondrogenesis, but the medium and high groups improved repair of the chronic defect with chondrogenic tissues and showed histologically significantly better results than the control and low groups. The use of a magnetic targeting system for delivering m-MSCs has the potential to overcome the clinical hurdles for repair of the severe chronic osteochondral defect. Furthermore, this system is predicted to produce good clinical outcomes for humans, not only to repair osteochondral defects but also to repair a variety of damaged tissues.

  13. Bioactive and metal uptake studies of carboxymethyl chitosan-graft-D-glucuronic acid membranes for tissue engineering and environmental applications.

    PubMed

    Jayakumar, R; Rajkumar, M; Freitas, H; Sudheesh Kumar, P T; Nair, S V; Furuike, T; Tamura, H

    2009-08-01

    Carboxymethyl chitosan-graft-D-glucuronic acid (CMCS-g-D-GA) was prepared by grafting D-GA onto CMCS in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and then the membranes were made from it. In this work, the bioactivity studies of CMCS-g-D-GA membranes were carried out and then characterized by SEM, CLSM, XRD and FT-IR. The CMCS-g-D-GA membranes were found to be bioactive. The adsorption of Ni2+, Zn2+ and Cu2+ ions onto CMCS-g-D-GA membranes has also been investigated. The maximum adsorption capacity of CMCS-g-D-GA for Ni2+, Zn2+ and Cu2+ was found to be 57, 56.4 and 70.2 mg/g, respectively. Hence, these membranes were useful for tissue engineering, environmental and water purification applications.

  14. The Long-Term Clinical Outcomes Following Autogenous Bone Grafting for Large-Volume Defects of the Knee

    PubMed Central

    Delano, Mark; Spector, Myron; Pittsley, Andrew; Gottschalk, Alexander

    2014-01-01

    Objective: We report the long-term clinical outcomes of patients who underwent autogenous bone grafting of large-volume osteochondral defects of the knee due to osteochondritis dessicans (OCD) and osteonecrosis (ON). This is the companion report to one previous published on the biological response. We hypothesized that these grafts would integrate with host bone and the articular surface would form fibrocartilage providing an enduring clinical benefit. Design: Three groups (patients/knees) were studied: OCD without a fragment (n = 12/13), OCD with a partial fragment (n = 14/16), and ON (n = 25/26). Twenty-five of 52 patients were available for clinical follow-up between 12 and 21 years. Electronic medical records provided comparison clinical information. In addition, there were plain film radiographs, MRIs, plus repeat arthroscopy and biopsy on 14 patients. Results: Autogenous bone grafts integrated with the host bone. MRI showed soft tissue covering all the grafts at long-term follow-up. Biopsy showed initial surface fibrocartilage that subsequently converted to fibrocartilage and hyaline cartilage at 20 years. OCD patients had better clinical outcomes than ON patients. No OCD patients were asymptomatic at anytime following surgery. Half of the ON patients came to total knee replacement within 10 years. Conclusions: Autogenous bone grafting provides an alternative biological matrix to fill large-volume defects in the knee as a singular solution integrating with host bone and providing an enduring articular cartilage surface. The procedure is best suited for those with OCD. The treatment for large-volume articular defects by this method remains salvage in nature and palliative in outcome. PMID:26069688

  15. Microsphere-based gradient implants for osteochondral regeneration: a long-term study in sheep

    PubMed Central

    Mohan, Neethu; Gupta, Vineet; Sridharan, Banu Priya; Mellott, Adam J; Easley, Jeremiah T; Palmer, Ross H; Galbraith, Richard A; Key, Vincent H; Berkland, Cory J; Detamore, Michael S

    2015-01-01

    Background: The microfracture technique for cartilage repair has limited ability to regenerate hyaline cartilage. Aim: The current study made a direct comparison between microfracture and an osteochondral approach with microsphere-based gradient plugs. Materials & methods: The PLGA-based scaffolds had opposing gradients of chondroitin sulfate and β-tricalcium phosphate. A 1-year repair study in sheep was conducted. Results: The repair tissues in the microfracture were mostly fibrous and had scattered fissures with degenerative changes. Cartilage regenerated with the gradient plugs had equal or superior mechanical properties; had lacunated cells and stable matrix as in hyaline cartilage. Conclusion: This first report of gradient scaffolds in a long-term, large animal, osteochondral defect demonstrated potential for equal or better cartilage repair than microfracture. PMID:26418471

  16. Evaluation of oriented electrospun fibers for periosteal flap regeneration in biomimetic triphasic osteochondral implant.

    PubMed

    Liu, Xudong; Liu, Shen; Liu, Shenghe; Cui, Wenguo

    2014-10-01

    Osteochondral defects represent a serious clinical problem. Although the cell-scaffold complexes have been reported to be effective for repairing osteochondral defects, a periosteal flap is frequently needed to arrest leakage of the implanted cells into the defect and to contribute to the secretion of cytokines to stimulate cartilage repair. The electrospun mesh mimicking the function of the flap assists tissue regeneration by preventing cell leakage and merits favorable outcomes in the cartilaginous region. In this study, an oriented poly(ε-caprolactone) (PCL) fibrous membrane (OEM) was fabricated by electrospinning as a periosteal scaffold and then freeze-dried with a collagen type I and hyaluronic acid cartilage scaffold (CH) and finally, freeze-dried with a tricalcium phosphate (TCP) bone substratum. Scanning electron microscopic images show obvious microstructure formation of the trilayered scaffolds, and electrospun fibrous membranes have an oriented fibrous network structure for the periosteal phase. Also shown are opened and interconnected pores with well designed three-dimensional structure, able to be bound in the CH (chondral phase) and TCP (osseous phase) scaffolds. In vitro results showed that the OEM can promote the orientation of bone marrow mesenchymal stem cell (BMSCs) and BMSCs can penetrate into the CH and TCP. After successfully combining the BMSCs, the tissue-engineered cartilage which contained the OEM and TCP complex was successfully used to regenerate the osteochondral defects in the rabbit model with greatly improved repair effects.

  17. Kinetic study of the replacement of porcine small intestinal submucosa grafts and the regeneration of meniscal-like tissue in large avascular meniscal defects in dogs.

    PubMed

    Cook, J L; Tomlinson, J L; Arnoczky, S P; Fox, D B; Reeves Cook, C; Kreeger, J M

    2001-06-01

    Porcine small intestinal submucosa (SIS) was used to replace large, avascular defects in the medial menisci of dogs. Twelve dogs received SIS grafts and 3 dogs were left untreated as controls. Dogs were evaluated at 4, 8, and 12 weeks by means of lameness scoring and ultrasonography. Dogs were sacrificed at 1, 6, or 12 weeks after implantation, and the tissue at the site of meniscal resection was evaluated for gross and histologic appearance, cross-sectional and surface area, and collagen types I and II. The femoral and tibial condyles were assessed for articular cartilage damage. Control dogs were significantly more lame than grafted dogs 8 and 12 weeks after instrumentation. Grafted dogs' replacement tissue appeared meniscal-like when evaluated grossly and ultrasonographically 12 weeks after instrumentation. The amount of replacement tissue was significantly greater in both cross-sectional and surface area for grafted dogs than for controls at all time points. Histologically, the SIS biomaterial could be identified in all grafted dogs at 1 week post-implantation, but in none at 6 weeks post-implantation. Subjectively, grafted dogs' replacement tissue was histologically superior to that of controls with respect to tissue type, organization, and architecture. Collagen types I and II immunoreactivity in grafted menisci were similar to that of normal menisci. Control dogs had significantly more articular cartilage damage than grafted dogs. SIS appears to induce regeneration of meniscal-like tissue in large, avascular meniscal defects in dogs, resulting in superior clinical function and articular cartilage protection compared to ungrafted controls.

  18. Dorsoproximal proximal phalanx osteochondral fragmentation in 117 Warmblood horses.

    PubMed

    Declercq, J; Martens, A; Maes, D; Boussauw, B; Forsyth, R; Boening, K J

    2009-01-01

    The objective of the present study was to determine clinical and arthroscopic characteristics associated with dorsoproximal proximal phalanx (P1) fragments in Warmblood horses, as well as to examine their histopathological appearance. One hundred sixty-eight fragments were removed from 150 fetlocks of 117 Warmblood horses. Details of signalment and results of clinical examination were collected prior to surgery. After arthroscopic fragment removal and joint evaluation for synovial and/or cartilage abnormalities, the fragments were measured and evaluated histopathologically. The vast majority of the fragments (95.2%) were found medially, without predilection for front or hind limbs. In 10% of the joints, more than one fragment was present. The mean size of the fragments was 6.8 +/- 2.6 mm. Only eight horses presented fetlock-related lameness. Horses of seven years of age and older (OR = 13.32; p = 0.033) and the presence of more than one fragment (OR = 11.12; p = 0.016) were significantly associated with lameness. Arthroscopic evaluation revealed one or more abnormalities in 50.7% of the joints. On histopathology, osteochondral fragments presented as a bony center covered with smooth hyaline cartilage on one side and some fibrous tissue on the other side. No clear histopathological signs were indicating precisely their origin. In Warmblood horses with dorsoproximal P1 fragments, the age (seven years and older) and the presence of more than one fragment in a fetlock significantly increased the risk of lameness. The osteochondral dorsoproximal P1 fragments could be defined as a developmental orthopaedic disease.

  19. Additive manufacturing for in situ repair of osteochondral defects.

    PubMed

    Cohen, Daniel L; Lipton, Jeffrey I; Bonassar, Lawrence J; Lipson, Hod

    2010-09-01

    Tissue engineering holds great promise for injury repair and replacement of defective body parts. While a number of techniques exist for creating living biological constructs in vitro, none have been demonstrated for in situ repair. Using novel geometric feedback-based approaches and through development of appropriate printing-material combinations, we demonstrate the in situ repair of both chondral and osteochondral defects that mimic naturally occurring pathologies. A calf femur was mounted in a custom jig and held within a robocasting-based additive manufacturing (AM) system. Two defects were induced: one a cartilage-only representation of a grade IV chondral lesion and the other a two-material bone and cartilage fracture of the femoral condyle. Alginate hydrogel was used for the repair of cartilage; a novel formulation of demineralized bone matrix was used for bone repair. Repair prints for both defects had mean surface errors less than 0.1 mm. For the chondral defect, 42.8+/-2.6% of the surface points had errors that were within a clinically acceptable error range; however, with 1 mm path planning shift, an estimated approximately 75% of surface points could likely fall within the benchmark envelope. For the osteochondral defect, 83.6+/-2.7% of surface points had errors that were within clinically acceptable limits. In addition to implications for minimally invasive AM-based clinical treatments, these proof-of-concept prints are some of the only in situ demonstrations to-date, wherein the substrate geometry was unknown a priori. The work presented herein demonstrates in situ AM, suggests potential biomedical applications and also explores in situ-specific issues, including geometric feedback, material selection and novel path planning techniques.

  20. Low Grade Juvenile Osteochondritis Dissecans of the Knee

    PubMed Central

    Etcheto, H. Rivarola; Blanchod, C. Collazo; Palanconi, M.; Zordan, J.; Salinas, E. Alvarez; Autorino, C.

    2017-01-01

    Juvenile osteochondritis dissecans (OCD) of the knee is a nosological entity acquired, idiopathic and potentially reversible. Dissects the subchondral bone tissue plane from the underlying bone, making a partial or complete osteochondral detachment, with a loose body. Consensus to treat none surgically poor symptomatic and stable lesions. If the lesion becomes instable or more symptomatic, surgical treatment will be best the option. Recently histological evidence holds is possible find sources of instability in deep layers sub chondral bone, even in patients with ¨stables lesions¨. This condition might be the reason of unfavorable evolution certain cases previously considered as ¨stable or incipient¨, treated with the classic non operative protocols. Objectives: The purpose of the present study consist in present a series of cases of young patients with symptomatic low grade juvenile OCD (grade I-II), treated surgically with subchondral debridement and fixation ¨in situ¨ describing the clinical and imaging findings. Methods: We evaluated 15 cases of symptomatic juvenile OCD of the knee, stables lesion (grade I/ II) according to Di Paola´s classification, who have not responded to conservative therapy for at least 6 months. Results: All patients were treated surgical consecutively with arthroscopically assisted ¨in situ¨ fixation with pins Smart Nail NR, ConMed-Linvatex and for the same group of surgeons. We evaluated the clinical and imagenologic outcomes with MRI for a minimum follow up of six month to one year. No looseness of fastening material or loose bodies in the submitted sample were recorded. The study by MRI imaging techniques using high definition chondral identification evidence allowed the consolidation of the fragment to the 6th month. Conclusion: All patients evolved asymptomatic and returned to the previous activity, with high level of satisfaction.

  1. Volumetric and linear changes at dental implants following grafting with volume-stable three-dimensional collagen matrices or autogenous connective tissue grafts: 6-month data.

    PubMed

    Naenni, Nadja; Bienz, Stefan P; Benic, Goran I; Jung, Ronald E; Hämmerle, Christoph H F; Thoma, Daniel S

    2017-09-18

    The objective of this study was to test whether or not soft tissue augmentation with a volume-stable collagen matrix (VCMX) leads to similar volume gain around dental implants compared to autogenous subepithelial connective tissue graft (SCTG). In 12 adult beagle dogs, immediate implants were placed with simultaneous guided bone regeneration. After 25-45 weeks, soft tissue augmentation was randomly performed using VCMX, SCTG, or a sham-operated control. Impressions were taken pre-op and post-op (tissue augmentation) and again at sacrifice after healing periods of 4, 8, and 24 weeks. They were then digitized to allow for superimposition. Values of linear and volumetric changes were calculated. The median increase (pre-op to post-op) in buccal volume measured 0.92 mm for VCMX, 1.47 mm for SCTG, and 0.24 mm for SH. The values (pre-op to sacrifice) were - 0.25 mm for VCMX, 0.52 mm for SCTG, and - 0.06 mm for group SH. The median ridge width 2 mm below the crest measured - 0.26 mm for VCMX, 0.53 mm for SCTG, and - 0.15 mm for SH (pre-op to sacrifice). Volume augmentation using VCMX and SCTG resulted in an increase in ridge dimension (pre- to post-op). During the follow-up, the volume decreased in all three groups to a level close to the situation prior to surgery. Soft tissue volume augmentation around dental implants is usually performed using the patient's own tissue. This therapy is associated with an increased morbidity due to a second surgical site. Soft tissue volume at implant sites can be augmented using VCMX and SCTG. The gain on top of the ridge appears not to be stable during the follow-up in both groups.

  2. Dysplasia Epiphysealis Hemimelica Treated with Osteochondral Allograft: A Case Report

    PubMed Central

    Anthony, Chris A.; Wolf, Brian R.

    2015-01-01

    Background Dysplasia epiphysealis hemimelica (DEH), or Trevor's disease, is a developmental disorder of the pediatric skeleton characterized by asymmetric osteochondral overgrowth. Methods We present the case of a five year old boy with a two year history of right knee pain and evidence of DEH on imaging who underwent initial arthroscopic resection of his lesion with subsequent recurrence. The patient then underwent osteochondral allograft revision surgery and was asymptomatic at two year follow-up with a congruent joint surface. Results To our knowledge, this is the first reported case of a DEH lesion treated with osteochondral allograft and also the youngest reported case of osteochondral allograft placement in the literature. Conclusions Osteochondral allograft may be a viable option in DEH and other deformities of the pediatric knee. Level of Evidence Level V PMID:26361443

  3. Comparison of three novel biphasic scaffolds for one-stage treatment of osteochondral defects in a sheep model.

    PubMed

    Gervaso, F; Mangiavini, L; Di Giancamillo, A; Boschetti, F; Izzo, D; Zani, D D; Di Giancamillo, M; Tessaro, I; Domenicucci, M; Scalera, F; Domeneghini, C; Crovace, A M; Sannino, A; Peretti, G M

    2016-01-01

    In the last years, several tissue engineering techniques have been applied to develop different kinds of osteochondral substitutes to overcome the scarce reparative properties of this tissue. The aim of this study was to generate and compare three biphasic scaffolds in an osteochondral lesion in a large-animal model. A critical osteochondral defect was generated in the medial femoral condyle of 18 skeletally mature sheep. Three defects were left untreated, the remaining lesions were divided into three groups: 5 lesions were treated with a biphasic scaffold made of collagen type I and small cylinders of Magnesium Hydroxyapatite; 5 lesions were treated with a biphasic substituted formed by collagen type I and Wollastonite, 5 lesions were treated with a scaffold made of collagen type I and small cylinders of Wollastonite/Hydroxyapatite. Animals were sacrificed after 3 months and samples were analyzed by CT and MRI, macroscopic evaluation and histology. Our study demonstrated that one of these novel biphasic scaffolds possesses the potential for being applied for one-stage procedures for osteochondral defects.

  4. Surgical treatment of localized gingival recessions using coronally advanced flaps with or without subepithelial connective tissue graft.

    PubMed

    Bellver-Fernández, Ricardo; Martínez-Rodriguez, Ana-María; Gioia-Palavecino, Claudio; Caffesse, Raul-Guillermo; Peñarrocha, Miguel

    2016-03-01

    A coronally advanced flap with subepithelial connective tissue graft is the gold standard surgical treatment of gingival recessions, since it offers a higher probability of achieving complete root coverage compared with other techniques. However, optimum short- and middle-term clinical results have also been obtained with coronally advanced flaps alone. The aim of the present study was to evaluate the results obtained by the surgical treatment of localized gingival recessions using coronally advanced flaps with or without subepithelial connective tissue graft. The reduction of recession height was assessed, together with the gain in gingival attachment apical to the recession, and total reduction of recession, in a comparative study of two techniques. Twenty-two gingival recessions were operated upon: 13 in the control group (coronally advanced flap) and 9 in the test group (coronally advanced flap associated to subepithelial connective tissue graft). After 18 months, the mean reduction of recession height was 2.2 ± 0.8 mm in the control group and 2.3 ± 0.7 mm in the test group, with a mean gain in gingival attachment of 1.3 ± 0.9 mm and 2.3 ± 1.3 mm, respectively. In percentage terms, the mean reduction of recession height was 84.6 ± 19.6% in the control group and 81.7 ± 17.8% in the test group, with a mean gain in gingival attachment of 20.5 ± 37.4% and 184.4 ± 135.5%, respectively. Significant reduction of gingival recession was achieved with both techniques, though the mean gain in gingival attachment (in mm and as a %) was greater in test group.

  5. Root coverage with a modified laterally positioned flap combined with a subepithelial connective tissue graft in advanced recession.

    PubMed

    Lee, Chun-Teh; Chang, Po-Chun; Touchan, Nawar; Royzman, Daniel

    2014-12-01

    A laterally positioned flap (LPF) combined with a subepithelial connective tissue graft (SCTG) is one of the conventional approaches for resolving gingival recession defects, with the advantages of flap flexibility and extended coverage of the tissue graft. However, thus far, evidence is lacking for the use of this technique for the treatment of advanced gingival recession defects. This report discusses three Miller class III cases with interproximal bone loss and wide and deep defects treated with a combination procedure of a modified laterally positioned flap (mLPF) and SCTG. mLPF combined with SCTG was performed for each case. The defect size and the degree of hypersensitivity at baseline and the final appointment in each case were documented. The three cases had a mean initial defect of 7.7±1.5 mm and a mean residual defect of 1.7±1 mm at the 6-, 3-, and 36-month follow-up, respectively, after the root coverage surgery. The symptom of hypersensitivity was improved, and the patients were satisfied with the clinical outcomes. The results demonstrated that the combination of mLPF with SCTG is promising for treating these advanced cases with respect to obtaining the expected root coverage with the gingival tissue.

  6. Experimental study on the construction of small three-dimensional tissue engineered grafts of electrospun poly-ε-caprolactone.

    PubMed

    Zhu, Guang-Chang; Gu, Yong-Quan; Geng, Xue; Feng, Zeng-Guo; Zhang, Shu-Wen; Ye, Lin; Wang, Zhong-Gao

    2015-02-01

    Studies on three-dimensional tissue engineered graft (3DTEG) have attracted great interest among researchers as they present a means to meet the pressing clinical demand for tissue engineering scaffolds. To explore the feasibility of 3DTEG, high porosity poly-ε-caprolactone (PCL) was obtained via the co-electrospinning of polyethylene glycol and PCL, and used to construct small-diameter poly-ε-caprolactone-lysine (PCL-LYS-H) scaffolds, whereby heparin was anchored to the scaffold surface by lysine groups. A variety of small-diameter 3DTEG models were constructed with different PCL layers and the mechanical properties of the resulting constructs were evaluated in order to select the best model for 3DTEGs. Bone marrow mononuclear cells were induced and differentiated to endothelial cells (ECs) and smooth muscle cells (SMCs). A 3DTEG (labeled '10-4%') was successfully produced by the dynamic co-culture of ECs on the PCL-LYS-H scaffolds and SMCs on PCL. The fluorescently labeled cells on the 3DTEG were subsequently observed by laser confocal microscopy, which showed that the ECs and SMCs were embedded in the 3DTEG. Nitric oxide and endothelial nitric oxide synthase assays showed that the ECs behaved normally in the 3DTEG. This study consequently provides a new thread to produce small-diameter tissue engineered grafts, with excellent mechanical properties, that are perfusable to vasculature and functional cells.

  7. Chemical hydrogels based on a hyaluronic acid-graft-α-elastin derivative as potential scaffolds for tissue engineering.

    PubMed

    Palumbo, Fabio Salvatore; Pitarresi, Giovanna; Fiorica, Calogero; Rigogliuso, Salvatrice; Ghersi, Giulio; Giammona, Gaetano

    2013-07-01

    In this work hyaluronic acid (HA) functionalized with ethylenediamine (EDA) has been employed to graft α-elastin. In particular a HA-EDA derivative bearing 50 mol% of pendant amino groups has been successfully employed to produce the copolymer HA-EDA-g-α-elastin containing 32% w/w of protein. After grafting with α-elastin, remaining free amino groups reacted with ethylene glycol diglycidyl ether (EGDGE) for producing chemical hydrogels, proposed as scaffolds for tissue engineering. Swelling degree, resistance to chemical and enzymatic hydrolysis, as well as preliminary biological properties of HA-EDA-g-α-elastin/EGDGE scaffold have been evaluated and compared with a HA-EDA/EGDGE scaffold. The presence of α-elastin grafted to HA-EDA improves attachment, viability and proliferation of primary rat dermal fibroblasts and human umbilical artery smooth muscle cells. Biological performance of HA-EDA-g-α-elastin/EGDGE scaffold resulted comparable to that of a commercial collagen type I sponge (Antema®), chosen as a positive control.

  8. The effects of early or late treatment of osteochondral defects on joint homoeostasis: an experimental study in rabbits.

    PubMed

    Ozsoy, Mehmet Hakan; Aydogdu, Semih; Taskiran, Dilek; Sezak, Murat; Hayran, Mutlu; Oztop, Fikri; Ozsoy, Arzu

    2009-06-01

    A 3.5 x 4 mm tubular osteochondral defect was created on the right medial femoral condyles of 51 adult rabbits. In the control group (CG), defects were left untreated. In the early-(ETG) and late-(LTG) treatment groups, defects were treated by an osteoperiosteal graft 1 and 12 weeks, respectively, after the index procedure. Synovial fluid (SF) samples were collected regularly and proteoglycan fragments (PF), total collagen (TC) and collagenase (MMP-1) levels were measured. Rabbits were killed at 4 (early period), 12 (intermediate period), or 24 (late period) weeks postoperatively. Histological examination indicated a more successful healing in both grafting groups than in the CG, but without any difference at any time period between the grafting groups. In the CG, PF, and TC levels in SF increased continuously until the late period, indicating an ongoing degenerative activity in the joints. In contrast, SF marker levels in both grafting groups indicated that normalization in joint metabolism could be achieved-at least partially-after treatment. However, PF levels in the SF showed that the treatment of defects in earlier stages might result in better outcomes since the negative effects were more prominent in chronic stages, presumably due to the more prolonged period of disturbed homeostasis. Thus, histological values and SF marker levels indicated that treatment of osteochondral defects at any time of the disease had a positive effect on healing when compared to no treatment. Early treatment might better assist the recovery of joint homeostasis than late treatment.

  9. Transplant of mesenchymal stem cells and hydroxyapatite ceramics to treat severe osteochondral damage after septic arthritis of the knee.

    PubMed

    Adachi, Nobuo; Ochi, Mitsuo; Deie, Masataka; Ito, Yohei

    2005-08-01

    We describe a 21-year-old man with a large osteochondral defect of the knee after septic arthritis, successfully treated by transplant of mesenchymal stem cells (MSC) from bone marrow and a new type of interconnected porous hydroxyapatite ceramic (IP-CHA). We confirmed successful cartilage-like tissue regeneration by a second arthroscopy. Biopsy of the repaired tissue revealed cartilage-like regeneration and bone formation. We were able to regenerate new bone and cartilage-like tissue in a one-stage operation, without sacrificing autologous bone or other tissue. This cultured MSC and IP-CHA hybrid material transplant represents a novel treatment for a severe osteochondral defect after septic arthritis.

  10. Fabrication and development of artificial osteochondral constructs based on cancellous bone/hydrogel hybrid scaffold.

    PubMed

    Song, Kedong; Li, Liying; Yan, Xinyu; Zhang, Yu; Li, Ruipeng; Wang, Yiwei; Wang, Ling; Wang, Hong; Liu, Tianqing

    2016-06-01

    Using tissue engineering techniques, an artificial osteochondral construct was successfully fabricated to treat large osteochondral defects. In this study, porcine cancellous bones and chitosan/gelatin hydrogel scaffolds were used as substitutes to mimic bone and cartilage, respectively. The porosity and distribution of pore size in porcine bone was measured and the degradation ratio and swelling ratio for chitosan/gelatin hydrogel scaffolds was also determined in vitro. Surface morphology was analyzed with the scanning electron microscope (SEM). The physicochemical properties and the composition were tested by using an infrared instrument. A double layer composite scaffold was constructed via seeding adipose-derived stem cells (ADSCs) induced to chondrocytes and osteoblasts, followed by inoculation in cancellous bones and hydrogel scaffolds. Cell proliferation was assessed through Dead/Live staining and cellular activity was analyzed with IpWin5 software. Cell growth, adhesion and formation of extracellular matrix in composite scaffolds blank cancellous bones or hydrogel scaffolds were also analyzed. SEM analysis revealed a super porous internal structure of cancellous bone scaffolds and pore size was measured at an average of 410 ± 59 μm while porosity was recorded at 70.6 ± 1.7 %. In the hydrogel scaffold, the average pore size was measured at 117 ± 21 μm and the porosity and swelling rate were recorded at 83.4 ± 0.8 % and 362.0 ± 2.4 %, respectively. Furthermore, the remaining hydrogel weighed 80.76 ± 1.6 % of the original dry weight after hydration in PBS for 6 weeks. In summary, the cancellous bone and hydrogel composite scaffold is a promising biomaterial which shows an essential physical performance and strength with excellent osteochondral tissue interaction in situ. ADSCs are a suitable cell source for osteochondral composite reconstruction. Moreover, the bi-layered scaffold significantly enhanced cell proliferation compared to the cells seeded on

  11. Optimization of Soft Tissue Management, Spacer Design, and Grafting Strategies for Large Segmental Bone Defects using the Chronic Caprine Tibial Defect Model

    DTIC Science & Technology

    2016-12-01

    AWARD NUMBER: W81XWH-13-2-0087 TITLE: Optimization of Soft Tissue Management, Spacer Design, and Grafting Strategies for Large Segmental Bone...Design, and Grafting Strategies for Large Segmental Bone Defects using the Chronic Caprine Tibial Defect Model 5a. CONTRACT NUMBER 5b. GRANT NUMBER...can be immediately translated into clinical practice. This study highlights the fact that the Chronic Caprine Tibial Defect Model is sufficiently

  12. Osteochondral interface generation by rabbit bone marrow stromal cells and osteoblasts coculture.

    PubMed

    Chen, Kelei; Teh, Thomas Kok Hiong; Ravi, Sujata; Toh, Siew Lok; Goh, James Cho Hong

    2012-09-01

    Physiological osteochondral interface regeneration is a significant challenge. This study aims to investigate the effect of the coculture of chondrogenic rabbit bone marrow stromal cells (rBMSCs) with rabbit osteoblasts in a specially designed two-dimensional (2D)-three-dimensional (3D) co-interface culture to develop the intermediate osteochondral region in vitro. The 2D-3D coculture system was set up by first independently culturing chondrogenic rBMSCs on a scaffold and osteoblasts in cell culture plates, and subsequently placed in contact and cocultured. As control, samples not cocultured with osteoblasts were used. The regulatory effects exerted by osteoblasts on chondrogenic rBMSCs were quantified by real-time polymerase chain reaction. To study the effect of coculture on cells located in different parts of the scaffold, samples were separated into two parts and significantly different gene expression patterns were found between them. In comparison with the control group, a significant moderate downregulation of chondrogenic marker genes, such as Collagen II and Aggrecan was observed. However, the Sox-9 and Collagen I expression increased. More importantly, chondrogenic rBMSCs in the coculture system were shown to form the osteochondral interface layer by expressing calcified cartilage zone specific extracellular matrix marker Collagen X and the hypertrophic chondrocyte marker MMP-13, which were not observed in the control group. Specifically, only the chondrogenic rBMSC layer in contact with the osteoblasts expressed Collagen X and MMP-13, indicating the positive influence of the coculture upon interface formation. Biochemical analyses, histology results, and immunohistochemical staining further supported this observation. In conclusion, this study revealed that specific regulatory stimulations from osteoblasts in the 2D-3D interface coculture system could induce the formation of ostochondral interface for the purpose of osteochondral tissue engineering.

  13. Tissue slice grafts of human renal cell carcinoma: an authentic preclinical model with high engraftment rate and metastatic potential.

    PubMed

    Thong, Alan E; Zhao, Hongjuan; Ingels, Alexandre; Valta, Maija P; Nolley, Rosalie; Santos, Jennifer; Young, Sarah R; Peehl, Donna M

    2014-01-01

    Discovery of curative therapies for renal cell carcinoma (RCC) is hampered by lack of authentic preclinical models. Tumorgrafts, generated by direct implantation of patient-derived tissues into mice, have demonstrated superior ability to predict therapeutic response. We evaluated "tissue slice grafts" (TSGs) as an improved tumorgraft model of RCC. Cores of fresh RCC were precision-cut at 300 µm and implanted under the renal capsule of RAG2(-/-)γC(-/-) mice. Engraftment rate, histology, biomarker expression, genetic fidelity, and metastatic potential were evaluated. Magnetic resonance imaging (MRI) was tested as a noninvasive method to measure tumor volume, and response to a targeted therapy was determined. All 13 cases of RCC engrafted and displayed characteristic histology and biomarkers. TSG volume quantified noninvasively by MRI highly correlated with graft weights, providing a unique tool for monitoring orthotopic growth. Moreover, in 2 cases, cancer cells from TSGs metastasized to clinically relevant sites, including bone. Microarray analysis and DNA sequencing demonstrated a high degree of correlation of global gene expression and von Hippel-Lindau (VHL) status between TSGs and parental tumors. Treatment of TSGs with sunitinib significantly decreased graft weight and mean vessel density compared with controls. The TSG model of RCC faithfully recapitulates tumor pathology, gene expression, genetic mutation, and drug response. The high engraftment rate and metastatic potential of this authentic model, in conjunction with the ability to generate large first-generation animal cohorts and to quantitate tumor volume at the orthotopic site by MRI, proffer significant advantages compared with other preclinical platforms. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. Emerging genetic basis of osteochondritis dissecans.

    PubMed

    Bates, J Tyler; Jacobs, John C; Shea, Kevin G; Oxford, Julia Thom

    2014-04-01

    Genome-wide association studies (GWAS) provide an unbiased approach in the identification of genes that increase the risk for osteochondritis dissecans (OCD). Recent GWAS in humans, horses, and pigs are reviewed and genes identified. The identified genes tended to cluster with respect to function and biologic processes. GWAS in humans are a critical next step in the effort to provide a better understanding of the causes of OCD, which will, in turn, allow preventive strategies for treatment of adolescents and young adults who are at risk for the development of degenerative joint disease due to the effects of OCD.

  15. Three-dimensional osteogenic and chondrogenic systems to model osteochondral physiology and degenerative joint diseases.

    PubMed

    Alexander, Peter G; Gottardi, Riccardo; Lin, Hang; Lozito, Thomas P; Tuan, Rocky S

    2014-09-01

    Tissue engineered constructs have the potential to function as in vitro pre-clinical models of normal tissue function and disease pathogenesis for drug screening and toxicity assessment. Effective high throughput assays demand minimal systems with clearly defined performance parameters. These systems must accurately model the structure and function of the human organs and their physiological response to different stimuli. Musculoskeletal tissues present unique challenges in this respect, as they are load-bearing, matrix-rich tissues whose functionality is intimately connected to the extracellular matrix and its organization. Of particular clinical importance is the osteochondral junction, the target tissue affected in degenerative joint diseases, such as osteoarthritis (OA), which consists of hyaline articular cartilage in close interaction with subchondral bone. In this review, we present an overview of currently available in vitro three-dimensional systems for bone and cartilage tissue engineering that mimic native physiology, and the utility and limitations of these systems. Specifically, we address the need to combine bone, cartilage and other tissues to form an interactive microphysiological system (MPS) to fully capture the biological complexity and mechanical functions of the osteochondral junction of the articular joint. The potential applications of three-dimensional MPSs for musculoskeletal biology and medicine are highlighted.

  16. Silk-Based Electrospun Tubular Scaffolds for Tissue-Engineered Vascular Grafts

    PubMed Central

    Soffer, Leah; Wang, Xianyan; Zhang, Xiaohui; Kluge, Jonathan; Dorfmann, Luis; Kaplan, David L.; Leisk, Gary

    2009-01-01

    Electrospinning was used to fabricate nonwoven nanofibrous tubular structures from Bombyx mori silk fibroin using an all aqueous process. The tubes were prepared for cell studies related to the bioengineering of small diameter vascular grafts. Prior to cell culture, the structures displayed a burst strength of 811±77.2 mmHg, sufficient to withstand arterial pressures. The tensile properties were similar to native vessels, with an ultimate tensile strength of 2.42± 0.48 MPa and a linear modulus of 2.45±0.47 MPa. Human endothelial cells and smooth muscle cells were successfully cultured on the electrospun silk, demonstrating the potential utility of these scaffolds for vascular grafts due to the combination of impressive mechanical properties and biological compatibility. PMID:18419943

  17. Tissue-Engineered Nanofibrous Nerve Grafts for Enhancing the Rate of Nerve Regeneration

    DTIC Science & Technology

    2015-10-01

    examined by staining for neurafilament and observing under a microscope. o Neurite outgrowth stimulating activity: PC-12 cells will be cultured on the...nerve grafts containing NGF and stained for evaluating neurite outgrowth from neurons. Neurite length and percentage of PC-12 cells with neurite...The adult male Sprague-Dawley rats (250-300 grams ) were used. Bone marrow stromal cell isolation and culture: For rat BMSCs isolation and culture

  18. Biocompatible and biodegradable ultrafine fibrillar scaffold materials for tissue engineering by facile grafting of L-lactide onto chitosan.

    PubMed

    Skotak, Maciej; Leonov, Alexei P; Larsen, Gustavo; Noriega, Sandra; Subramanian, Anuradha

    2008-07-01

    A chitosan derivative was prepared with good yields using a "one pot" approach by grafting L-lactide oligomers via ring opening polymerization. Side chains are primarily attached to hydroxyl groups located on carbons 3 and 6 of the glucosamine ring, while the amine group remains nonfunctionalized. By increasing the L-lactide to chitosan ratio, side chain length is controlled. This allows the manipulation of the biodegradation rate and hydrophilicity of the tissue engineering scaffold material. This general synthetic route renders functionalized chitosan soluble in a broad range of organic solvents, facilitating formation of ultrafine fibers via electrospinning. Cytotoxicity tests using fibroblasts (L929 cell line) performed on electrospun L-lactide modified chitosan fibers showed that the specimen with the highest molar ratio of L-lactide (1:24) investigated in this study is the most promising material for tissue engineering purposes, while less stable formulations might still find application in drug delivery vehicles.

  19. Photodynamic damage to cartilage and synovial tissue grafted on a chick's chorioallantoic membrane

    NASA Astrophysics Data System (ADS)

    Fisher, M.; Nahir, A. M.; Kimel, Sol

    1997-09-01

    Rheumatoid arthritis (RA) is a chronic inflammatory disease of the synovial joints causing pain deformities and disability. The highly vascular inflamed synovium has aggressive and destructive characteristics, it invades, erodes and gradually destroys cartilage and underlying bone. Photodynamic therapy (PDT) was performed using the chick chorioallantoic membrane (CAM) model to investigate the vitality of synovium and cartilage implanted on the CAM. Synovium, obtained from human patients, was grafted onto the CAM; gross microscopy and histology proved its vitality 7 days post grafting. Cartilage obtained from rabbit knee joint was also maintained on the CAM for 7 days. Its vitality was demonstrated by histology and by measuring metabolic and enzymatic activity of cartilage cells (chondrocytes) as well as the collagen and proteoglycans content. Selective PDT was performed using aluminum phthalocyanine tetrasulfonate (AlPcS4), a hydrophilic compound, soluble in biological solutions, as a photosensitizer. After irradiation with a diode laser (lambda equals 670 nm, 10 mW) damage was observed in vascularized synovium grafts, whereas avascular cartilage remained intact.

  20. Treatment of Early-Stage Pressure Ulcers by Using Autologous Adipose Tissue Grafts

    PubMed Central

    Marangi, Giovanni Francesco; Pallara, Tiziano; Cagli, Barbara; Schena, Emiliano; Giurazza, Francesco; Faiella, Elio; Zobel, Bruno Beomonte; Persichetti, Paolo

    2014-01-01

    Assessing pressure ulcers (PUs) in early stages allows patients to receive safer treatment. Up to now, in addition to clinical evaluation, ultrasonography seems to be the most suitable technique to achieve this goal. Several treatments are applied to prevent ulcer progression but none of them is totally effective. Furthermore, the in-depth knowledge of fat regenerative properties has led to a wide use of it. With this study the authors aim at introducing a new approach to cure and prevent the worsening of early-stage PUs by using fat grafts. The authors selected 42 patients who showed clinical and ultrasonographic evidence of early-stage PUs. Values of skin thickness, fascial integrity, and subcutaneous vascularity were recorded both on the PU area and the healthy trochanteric one, used as control region. Fat grafting was performed on all patients. At three months, abnormal ultrasonographic findings, such as reduction of cutaneous and subcutaneous thickness, discontinuous fascia, and decrease in subcutaneous vascularity, all were modified with respect to almost all the corresponding parameters of the control region. Results highlight that the use of fat grafts proved to be an effective treatment for early-stage PUs, especially in the care of neurological and chronic bedridden patients. PMID:24818019

  1. Large fresh osteochondral allografts of the knee: a systematic clinical and basic science review of the literature.

    PubMed

    De Caro, Francesca; Bisicchia, Salvatore; Amendola, Annunziato; Ding, Lei

    2015-04-01

    The aim of this study was to conduct an updated review of the literature regarding the clinical and basic science knowledge on osteochondral allograft transplantation in the knee for the treatment of large defects. According to specific criteria, 2 investigators systematically reviewed the literature for clinical and basic science reports regarding osteochondral allograft transplantation; data were independently extracted, pooled, and analyzed. Clinical and functional outcomes, International Knee Documentation Committee and Western Ontario and McMaster Universities Osteoarthritis Index scores, return to sport, quality of life, and survivorship of the grafts were assessed from the clinical articles. Regarding the basic science articles, the effects of allograft storage time, temperature, and different storage media were assessed. Eleven articles reporting on clinical data and 14 articles reporting on basic science data (animal, cell, and biomechanical studies) were selected. The articles included in the review were not homogeneous, and different outcome measures were adopted. Overall excellent results were achieved, with improvement in all objective and subjective clinical scores, a high rate of return to sport, and a survivorship rate of 89% at 5 years. When multiple plugs were implanted, posterior grafts seemed to fail. Only 1 article compared fresh versus frozen grafts, with a greater improvement in scores in the frozen group. Cellular viability and number were reduced during storage, even at low temperatures; polyphenol from green tea and arbutin and higher temperatures favorably influenced cell viability of the cartilage during storage. On the other hand, the structural properties of the extracellular matrix were not influenced by the storage at low temperatures. Integration of the graft to the host was also important, and bony integration was usually achieved; however, on the cartilage side, integration was scant or did not occur, especially in the frozen

  2. Treatment of isolated chondral and osteochondral defects in the knee by autologous matrix-induced chondrogenesis (AMIC).

    PubMed

    Kusano, Taro; Jakob, Roland P; Gautier, Emanuel; Magnussen, Robert A; Hoogewoud, Henri; Jacobi, Matthias

    2012-10-01

    The purpose of this study is to evaluate clinical and radiological outcomes of patients treated with autologous matrix-induced chondrogenesis (AMIC) for full-thickness chondral and osteochondral defects of the femoral condyles and patella. A retrospective evaluation of clinical and radiographic outcomes of patients treated with AMIC for chondral and osteochondral full-thickness cartilage defects of the knee was performed with a mean follow-up of 28.8 ± 1.5 months (range, 13-51 months). Significant improvements in clinical outcome scores (IKDC, Lysholm, Tegner, and VAS pain score) were noted. The largest improvements were seen in the osteochondral subgroup (mean age 25.9 years), whereas patients treated for chondral defects in the patellofemoral joint and on the femoral condyles improved less. Patients in all groups were generally satisfied with their results. MRI evaluation showed that tissue filling was present but generally not complete or homogenous. AMIC is a safe procedure and leads to clinical improvement of symptomatic full-thickness chondral and osteochondral defects and to regenerative defect filling. The value of AMIC relative to other cartilage repair procedures and to the natural course remains undefined. Case series, Level IV.

  3. Focal Defects of the Knee Articular Surface: Evidence of a Regenerative Potential Pattern in Osteochondritis Dissecans and Degenerative Lesions

    PubMed Central

    Gabusi, Elena; Manferdini, Cristina; Paolella, Francesca; Gambari, Laura; Mariani, Erminia

    2017-01-01

    The surgical treatment of knee articular focal lesions may offer heterogeneous clinical results. Osteochondritis dissecans (OCD) lesions showed to heal better than degenerative lesions (DL) but the underlying biological reasons are unknown. We evaluated the basal histological and immunohistochemical characteristics of these lesions analyzing a series of osteochondral fragments from young patients with similar age but presenting different etiology. Osteochondral tissue samples were stained with Safranin O and graded using a histological score. Markers of mesenchymal progenitor cells (CD146), osteoclasts (tartrate-resistant acid phosphatase, TRAP), and vessels (CD34) were evaluated. Histological score showed a higher degeneration of both cartilage and bone compartments in OCD compared to DL fragments. Only CD146-positive cells were found at the same percentage in cartilage compartment of both DL and OCD patients. By contrast, in the bone compartment a significantly higher percentage of CD146, TRAP, and CD34 markers was found in OCD compared to DL patients. These data showed distinct histological characteristics of osteochondral focal lesions located in the same anatomical region but having a different etiology. The higher percentages of these markers in OCD than in DL, mainly associated with a high bone turnover, could help to explain the higher clinical healing potential of OCD patients. PMID:28770227

  4. The effect of interface microstructure on interfacial shear strength for osteochondral scaffolds based on biomimetic design and 3D printing.

    PubMed

    Zhang, Weijie; Lian, Qin; Li, Dichen; Wang, Kunzheng; Hao, Dingjun; Bian, Weiguo; Jin, Zhongmin

    2015-01-01

    Interface integration between chondral phase and osseous phase is crucial in engineered osteochondral scaffolds. However, the integration was poorly understood and commonly failed to meet the need of osteochondral scaffolds. In this paper, a biphasic polyethylene glycol (PEG)/β-tricalcium phosphate (β-TCP) scaffold with enhanced interfacial integration was developed. The chondral phase was a PEG hydrogel. The osseous phase was a β-TCP ceramic scaffold. The PEG hydrogel was directly cured on the ceramic interface layer by layer to fabricate osteochondral scaffolds by 3D printing technology. Meanwhile, a series of interface structure were designed with different interface pore area percentages (0/10/20/30/40/50/60%), and interfacial shear test was applied for interface structure optimization (n=6 samples/group). The interfacial shear strength of 30% pore area group was nearly three folds improved compared with that of 0% pore area percentage group, and more than fifty folds improved compared with that of traditional integration (5.91±0.59 kPa). In conclusion, the biomimetic PEG/β-TCP scaffolds with interface structure enhanced integration show promising potential application for osteochondral tissue engineering. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Osteochondral repair by a novel interconnecting collagen-hydroxyapatite substitute: a large-animal study.

    PubMed

    Sosio, Corrado; Di Giancamillo, Alessia; Deponti, Daniela; Gervaso, Francesca; Scalera, Francesca; Melato, Marco; Campagnol, Marino; Boschetti, Federica; Nonis, Alessandro; Domeneghini, Cinzia; Sannino, Alessandro; Peretti, Giuseppe Michele

    2015-02-01

    A novel three-dimensional bicomponent substitute made of collagen type I and hydroxyapatite was tested for the repair of osteochondral lesions in a swine model. This scaffold was assembled by a newly developed method that guarantees the strict integration between the organic and the inorganic parts, mimicking the biological tissue between the chondral and the osseous phase. Thirty-six osteochondral lesions were created in the trochlea of six pigs; in each pig, two lesions were treated with scaffolds seeded with autologous chondrocytes (cell+group), two lesions were treated with unseeded scaffolds (cell- group), and the two remaining lesions were left untreated (untreated group). After 3 months, the animals were sacrificed and the newly formed tissue was analyzed to evaluate the degree of maturation. The International Cartilage Repair Society (ICRS) macroscopic assessment showed significantly higher scores in the cell- and untreated groups when compared with the cell+ group. Histological evaluation showed the presence of repaired tissue, with fibroblast-like and hyaline-like areas in all groups; however, with respect to the other groups, the cell- group showed significantly higher values in the ICRS II histological scores for "cell morphology" and for the "surface/superficial assessment." While the scaffold seeded with autologous chondrocytes promoted the formation of a reparative tissue with high cellularity but low glycosaminoglycans (GAG) production, on the contrary, the reparative tissue observed with the unseeded scaffold presented lower cellularity but higher and uniform GAG distribution. Finally, in the lesions treated with scaffolds, the immunohistochemical analysis showed the presence of collagen type II in the peripheral part of the defect, indicating tissue maturation due to the migration of local cells from the surroundings. This study showed that the novel osteochondral scaffold was easy to handle for surgical implantation and was stable within the site

  6. Soft tissue grafting to improve the attached mucosa at dental implants: A review of the literature and proposal of a decision tree.

    PubMed

    Bassetti, Mario; Kaufmann, Regula; Salvi, Giovanni E; Sculean, Anton; Bassetti, Renzo

    2015-06-01

    Scientific data and clinical observations appear to indicate that an adequate width of attached mucosa may facilitate oral hygiene procedures thus preventing peri-implant inflammation and tissue breakdown (eg, biologic complications). Consequently, in order to avoid biologic complications and improve long-term prognosis, soft tissue conditions should be carefully evaluated when implant therapy is planned. At present the necessity and time-point for soft tissue grafting (eg, prior to or during implant placement or after healing) is still controversially discussed while clinical recommendations are vague. To provide a review of the literature on the role of attached mucosa to maintain periimplant health, and to propose a decision tree which may help the clinician to select the appropriate surgical technique for increasing the width of attached mucosa. The available data indicate that ideally, soft tissue conditions should be optimized by various grafting procedures either before or during implant placement or as part of stage-two surgery. In cases, where, despite insufficient peri-implant soft tissue condition (ie, lack of attached mucosa or movements caused by buccal frena), implants have been uncovered and/or loaded, or in cases where biologic complications are already present (eg, mucositis, peri-implantitis), the treatment appears to be more difficult and less predictable. Soft tissue grafting may be important to prevent peri-implant tissue breakdown and should be considered when dental implants are placed. The presented decision tree may help the clinician to select the appropriate grafting technique.

  7. On the decellularization of fresh or frozen human umbilical arteries: implications for small-diameter tissue engineered vascular grafts.

    PubMed

    Tuan-Mu, Ho-Yi; Yu, Chen-Hsiang; Hu, Jin-Jia

    2014-06-01

    Most tissues, including those to be decellularized for tissue engineering applications, are frozen for long term preservation. Such conventional cryopreservation has been shown to alter the structure and mechanical properties of tissues. Little is known, however, how freezing affects decellularization of tissues. The purpose of this study was two-fold: to examine the effects of freezing on decellularization of human umbilical arteries (HUAs), which represent a potential scaffolding material for small-diameter tissue-engineered vascular grafts, and to examine how decellularization affects the mechanical properties of frozen HUAs. Among many decellularization methods, hypotonic sodium dodecyl sulfate solution was selected as the decellularizing agent and tested on fresh HUAs to optimize decellularization conditions. The efficiency of decellularization was evaluated by DNA assay and histology every 12 up to 48 h. The optimized decellularization protocol was then performed on frozen HUAs. The stiffness, burst pressure, and suture retention strength of fresh HUAs and frozen HUAs before and after decellularization were also examined. It appeared that freezing decreased the efficiency of decellularization, which may be attributed to the condensed extracellular matrix caused by freezing. While the stiffness of fresh HUAs did not change significantly after decellularization, decellularization reduced the compliance of frozen HUAs. Interestingly, the stiffness of decellularized frozen HUAs was similar to that of decellularized fresh HUAs. Although little difference in stiffness was observed, we suggest avoiding freezing if more efficient and complete decellularization is desired.

  8. Real-time immune cell interactions in target tissue during autoimmune-induced damage and graft tolerance

    PubMed Central

    Miska, Jason; Abdulreda, Midhat H.; Devarajan, Priyadharshini; Lui, Jen Bon; Suzuki, Jun; Pileggi, Antonello; Berggren, Per-Olof

    2014-01-01

    Real-time imaging studies are reshaping immunological paradigms, but a visual framework is lacking for self-antigen-specific T cells at the effector phase in target tissues. To address this issue, we conducted intravital, longitudinal imaging analyses of cellular behavior in nonlymphoid target tissues to illustrate some key aspects of T cell biology. We used mouse models of T cell–mediated damage and protection of pancreatic islet grafts. Both CD4+ and CD8+ effector T (Teff) lymphocytes directly engaged target cells. Strikingly, juxtaposed β cells lacking specific antigens were not subject to bystander destruction but grew substantially in days, likely by replication. In target tissue, Foxp3+ regulatory T (Treg) cells persistently contacted Teff cells with or without involvement of CD11c+ dendritic cells, an observation conciliating with the in vitro “trademark” of Treg function, contact-dependent suppression. This study illustrates tolerance induction by contact-based immune cell interaction in target tissues and highlights potentials of tissue regeneration under antigenic incognito in inflammatory settings. PMID:24567447

  9. A bioreactor test system to mimic the biological and mechanical environment of oral soft tissues and to evaluate substitutes for connective tissue grafts.

    PubMed

    Mathes, Stephanie H; Wohlwend, Lorenz; Uebersax, Lorenz; von Mentlen, Roger; Thoma, Daniel S; Jung, Ronald E; Görlach, Christoph; Graf-Hausner, Ursula

    2010-12-15

    Gingival cells of the oral connective tissue are exposed to complex mechanical forces during mastication, speech, tooth movement and orthodontic treatments. Especially during wound healing following surgical procedures, internal and external forces may occur, creating pressure upon the newly formed tissue. This clinical situation has to be considered when developing biomaterials to augment soft tissue in the oral cavity. In order to pre-evaluate a collagen sponge intended to serve as a substitute for autogenous connective tissue grafts (CTGs), a dynamic bioreactor system was developed. Pressure and shear forces can be applied in this bioreactor in addition to a constant medium perfusion to cell-material constructs. Three-dimensional volume changes and stiffness of the matrices were analyzed. In addition, cell responses such as cell vitality and extracellular matrix (ECM) production were investigated. The number of metabolic active cells constantly increased under fully dynamic culture conditions. The sponges remained elastic even after mechanical forces were applied for 14 days. Analysis of collagen type I and fibronectin revealed a statistically significant accumulation of these ECM molecules (P < 0.05-0.001) when compared to static cultures. An increased expression of tenascin-c, indicating tissue remodeling processes, was observed under dynamic conditions only. The results indicate that the tested in vitro cell culture system was able to mimic both the biological and mechanical environments of the clinical situation in a healing wound. © 2010 Wiley Periodicals, Inc.

  10. Osteochondral Autograft Transplantation Surgery for Metacarpal Head Defects.

    PubMed

    Kitay, Alison; Waters, Peter M; Bae, Donald S

    2016-03-01

    Post-traumatic osteonecrosis of the metacarpal head is a challenging problem, particularly in younger patients in whom arthroplasty may not be a durable option. Although several osteochondral reconstructive options have been proposed, some are associated with considerable donor site morbidity and/or require the use of internal fixation. We present an application of osteochondral autograft transplantation surgery as a treatment option for focal metacarpal head lesions. An osteochondral plug from the non-weight-bearing articular surface of the knee is transferred and press-fit to resurface a focal metacarpal head defect. The technical pearls and pitfalls are reviewed, and an illustrative case is presented.

  11. The effect of storage medium tonicity on osteochondral autograft plug diameter.

    PubMed

    Fening, Stephen D; Mihnovets, Jonathon; Jones, Morgan H; Midura, Ronald J; Miniaci, Anthony

    2011-02-01

    The purpose of this study was to investigate the effect of differing storage medium on osteochondral plug diameter. Four storage conditions were evaluated: air, hypotonic solution (sterile water), isotonic saline solution (0.9% sodium chloride), and hypertonic saline solution (3.0% sodium chloride). Four osteochondral plugs were acquired (4.5-mm harvesting system) from each of 10 fresh calf femurs and randomized to 1 of 4 storage media (N = 40). Micro-computed tomography was used to evaluate the precise diameter of each plug. After a time 0 scan, each plug was placed in a designated storage medium and rescanned at 3 time points over approximately 1 hour. A region of interest was identified from approximately 1 to 6 mm proximal to the tidemark. Custom software automatically calculated the diameter of each plug. The time 0 plug diameter (mean ± 95% confidence interval) for all specimens was 4.66 ± 0.01 mm. There were no significant differences between any of the groups at the baseline scan. There were also no significant differences between the time 0 and subsequent scans of the unsubmerged specimens. However, all of the liquid solutions (hypertonic, isotonic, and hypotonic) resulted in a significant increase in diameter from their baseline scans (P < .05), indicating that a cause may be increased extracellular matrix fluid pressure. Placing an osteochondral plug in a liquid solution increased the diameter of the subchondral bone. Size increase from the storage medium appeared to level off within 14 minutes after placement in solution. Increases in diameter of the plug may alter the ease of insertion of the graft, possibly increasing contact pressure on cartilage during plug implantation. Copyright © 2011 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.

  12. Current management of talar osteochondral lesions

    PubMed Central

    Gianakos, Arianna L; Yasui, Youichi; Hannon, Charles P; Kennedy, John G

    2017-01-01

    Osteochondral lesions of the talus (OLT) occur in up to 70% of acute ankle sprains and fractures. OLT have become increasingly recognized with the advancements in cartilage-sensitive diagnostic imaging modalities. Although OLT may be treated nonoperatively, a number of surgical techniques have been described for patients whom surgery is indicated. Traditionally, treatment of symptomatic OLT have included either reparative procedures, such as bone marrow stimulation (BMS), or replacement procedures, such as autologous osteochondral transplantation (AOT). Reparative procedures are generally indicated for OLT < 150 mm2 in area. Replacement strategies are used for large lesions or after failed primary repair procedures. Although short- and medium-term results have been reported, long-term studies on OLT treatment strategies are lacking. Biological augmentation including platelet-rich plasma and concentrated bone marrow aspirate is becoming increasingly popular for the treatment of OLT to enhance the biological environment during healing. In this review, we describe the most up-to-date clinical evidence of surgical outcomes, as well as both the mechanical and biological concerns associated with BMS and AOT. In addition, we will review the recent evidence for biological adjunct therapies that aim to improve outcomes and longevity of both BMS and AOT procedures. PMID:28144574

  13. Osteochondritis dissecans of the capitellum in adolescents

    PubMed Central

    van Bergen, Christiaan JA; van den Ende, Kimberly IM; ten Brinke, Bart; Eygendaal, Denise

    2016-01-01

    Osteochondritis dissecans (OCD) is a disorder of articular cartilage and subchondral bone. In the elbow, an OCD is localized most commonly at the humeral capitellum. Teenagers engaged in sports that involve repetitive stress on the elbow are at risk. A high index of suspicion is warranted to prevent delay in the diagnosis. Plain radiographs may disclose the lesion but computed tomography and magnetic resonance imaging are more accurate in the detection of OCD. To determine the best treatment option it is important to differentiate between stable and unstable OCD lesions. Stable lesions can be initially treated nonoperatively with elbow rest or activity modification and physical therapy. Unstable lesions and stable lesions not responding to conservative therapy require a surgical approach. Arthroscopic debridement and microfracturing has become the standard initial procedure for treatment of capitellar OCD. Numerous other surgical options have been reported, including internal fixation of large fragments and osteochondral autograft transfer. The aim of this article is to provide a current concepts review of the etiology, clinical presentation, diagnosis, treatment, and outcomes of elbow OCD. PMID:26925381

  14. Immediate placement and provisionalization of maxillary anterior single implant with guided bone regeneration, connective tissue graft, and coronally positioned flap procedures.

    PubMed

    Waki, Tomonori; Kan, Joseph Y K

    2016-01-01

    Immediate implant placement and provisionalization in the esthetic zone have been documented with success. The benefit of immediate implant placement and provisionalization is the preservation of papillary mucosa. However, in cases with osseous defects presenting on the facial bony plate, immediate implant placement procedures have resulted in facial gingival recession. Subepithelial connective tissue grafts for immediate implant placement and provisionalization procedures have been reported with a good esthetic outcome. Biotype conversion around implants with subepithelial connective tissue grafts have been advocated, and the resulting tissues appear to be more resistant to recession. The dimensions of peri-implant mucosa in a thick biotype were significantly greater than in a thin biotype. Connective tissue graft with coronally positioned flap procedures on natural teeth has also been documented with success. This article describes a technique combining immediate implant placement, provisionalization, guided bone regeneration (GBR), connective tissue graft, and a coronally positioned flap in order to achieve more stable peri-implant tissue in facial osseous defect situations.

  15. Surgical Marking Pen Dye Inhibits Saphenous Vein Cell Proliferation and Migration in Saphenous Vein Graft Tissue

    PubMed Central

    Kikuchi, Shinsuke; Kenagy, Richard D; Gao, Lu; Wight, Thomas N; Azuma, Nobuyoshi; Sobel, Michael; Clowes, Alexander W

    2014-01-01

    Objective Markers containing dyes such as crystal violet (CAS 548-62-9) are routinely used on the adventitia of vein bypass grafts to avoid twisting during placement. Since little is known about how these dyes affect vein graft healing and function, we determined the effect of crystal violet on cell migration and proliferation, which are responses to injury after grafting. Methods Fresh human saphenous veins were obtained as residual specimens from leg bypass surgeries. Portions of the vein that had been surgically marked with crystal violet were analyzed separately from those that had no dye marking. In the laboratory, they were split into easily dissected inner and outer layers after removal of endothelium. This f cleavage plane was within the circular muscle layer of the media. Cell migration from explants was measured daily as either 1) % migration positive explants, which exclusively measures migration, or 2) the number of cells on the plastic surrounding each explant, which measures migration plus proliferation. Cell proliferation and apoptosis (Ki67 and TUNEL staining, respectively) were determined in dye-marked and unmarked areas of cultured vein rings. The dose-dependent effects of crystal violet were measured for cell migration from explants as well as proliferation, migration, and death of cultured outer layer cells. Dye was extracted from explants with ethanol and quantified by spectrophotometry. Results There was significantly less cell migration from visibly blue, compared to unstained, outer layer explants by both methods. There was no significant difference in migration from inner layer explants adjacent to blue-stained or unstained sections of vein, because dye did not penetrate to the inner layer. Ki67 staining of vein in organ culture, which is a measure of proliferation, progressively increased up to 6 days in non-blue outer layer and was abolished in the blue outer layer. Evidence of apoptosis (TUNEL staining) was present throughout the wall

  16. The Effect of Pulsatile Loading and Scaffold Structure for the Generation of a Medial Equivalent Tissue Engineered Vascular Graft

    PubMed Central

    Thomas, Lynda V.

    2013-01-01

    Abstract A reliable and cost-effective scaffold for tissue-engineered vascular graft that would not only support cell proliferation and growth but also maintain cell phenotype has been a long-term challenge. In this study, we propose a biodegradable and biomimetic copolymer of gelatin with vinyl acetate synthesized via a graft copolymerization technique to generate tubular scaffolds for vascular tissue engineering. Two fabrication techniques, freeze drying and electrospinning, were used to generate the differing architectures for the scaffolds and characterized. The electrospun scaffolds were found to have a faster rate of mass loss in physiological saline of 81.72% within 4 months compared with 60% mass loss for the freeze-dried samples, though the materials were more crystalline. Vascular (v) smooth muscle cells (SMCs) were seeded on these tubes, which were then subjected to dynamic pulsatile stimulation on a vascular bioreactor for a week. Gross examination of the tissue-engineered constructs revealed that the cells secreted extensive extracellular matrix, with the dynamically conditioned samples exhibiting well-orientated SMCs and collagenous fibers in comparison with growth in static conditions. In addition, the alignment of cells in the direction of strain was greater in the electrospun constructs. The electrospun scaffolds maintained the characteristic contractile phenotype of SMCs, which was confirmed by higher gene expression rates of contractile protein markers like SM22α and calponin. A significant increase in the total matrix components (collagen and elastin) in the electrospun constructs compared with the freeze-dried samples was confirmed by biochemical analysis. The results of this study indicate that a combination approach involving a biomimetic scaffold with the nanofibrillar architecture and good mechanical strength conducive to the growth of SMCs and the use of the pulsatile forces to modulate the cell morphology and phenotypic plasticity of v

  17. S53P4 bioactive glass and fibrin glue for the treatment of osteochondral lesions of the knee - a preliminary in vivo study in rabbits.

    PubMed

    Zazgyva, AncuŢa Marilena; Gurzu, Simona; Jung, Ioan; Nagy, Örs; Mühlfay, Gheorghe; Pop, Tudor Sorin

    2015-01-01

    The role of the subchondral bone and the importance of treating both bone and cartilage in cases of chondral and osteochondral lesions of the knee have been highly emphasized. There are no current studies on the experimental use of bioactive glass S53P4 (BonAlive®) as granules in the treatment of osteochondral lesions of the knee. Our preliminary study was designed to establish an experimental model and assesses the effect of glass granules fixed with fibrin compared to fibrin alone as fillers of the osteochondral defects created in the weight-bearing and partial weight-bearing regions of the distal femur in six adult rabbits. We found that the size of the distal femur in adult domestic rabbits allows the creation of 4 mm diameter and 5 mm deep osteochondral defects on both the medial femoral condyle and the trochlea, bilaterally, without significantly affecting the activity level of the animals. Retention of the glass granules in the defects was achieved successfully using a commercially available fibrin sealant. At five weeks post-implantation, we found macroscopic and microscopic differences between the four types of defects. The use of bioactive glass S53P4 for filling condylar osteochondral defects in rabbit femora led to the initiation of an early bone repair process, observed at five weeks after implantation, while the filling of trochlear defects with fibrin glue resulted in the appearance of cartilaginous tissue characteristic of endochondral ossification.

  18. In vitro generation of whole osteochondral constructs using rabbit bone marrow stromal cells, employing a two-chambered co-culture well design.

    PubMed

    Chen, Kelei; Ng, Kian Siang; Ravi, Sujata; Goh, James C H; Toh, Siew Lok

    2016-04-01

    The regeneration of whole osteochondral constructs with a physiological structure has been a significant issue, both clinically and academically. In this study, we present a method using rabbit bone marrow stromal cells (BMSCs) cultured on a silk-RADA peptide scaffold in a specially designed two-chambered co-culture well for the generation of multilayered osteochondral constructs in vitro. This specially designed two-chambered well can simultaneously provide osteogenic and chondrogenic stimulation to cells located in different regions of the scaffold. We demonstrated that this co-culture approach could successfully provide specific chemical stimulation to BMSCs located on different layers within a single scaffold, resulting in the formation of multilayered osteochondral constructs containing cartilage-like and subchondral bone-like tissue, as well as the intermediate osteochondral interface. The cells in the intermediate region were found to be hypertrophic chondrocytes, embedded in a calcified extracellular matrix containing glycosaminoglycans and collagen types I, II and X. In conclusion, this study provides a single-step approach that highlights the feasibility of rabbit BMSCs as a single-cell source for multilayered osteochondral construct generation in vitro.

  19. Human Bone Marrow Stromal Cells Differentiate Into Corneal Tissue and Prevent Ocular Graft-Versus-Host Disease in Mice.

    PubMed

    Sánchez-Abarca, Luis Ignacio; Hernández-Galilea, Emiliano; Lorenzo, Rebeca; Herrero, Carmen; Velasco, Almudena; Carrancio, Soraya; Caballero-Velázquez, Teresa; Rodríguez-Barbosa, José Ignacio; Parrilla, Marta; Del Cañizo, Consuelo; San Miguel, Jesús; Aijón, José; Pérez-Simón, José Antonio

    2015-01-01

    Clinical trials have assessed the use of human bone marrow stromal cells (hBMSCs) for the treatment of immune-related disorders such as graft-versus-host disease (GVHD). In the current study, we show that GFP(+)-transduced hBMSCs generated from bone marrow migrate and differentiate into corneal tissue after subconjunctival injection in mice. Interestingly, these hBMSCs display morphological features of epithelial, stromal, and endothelial cells and appear at different layers and with different morphologies depending on their position within the epithelium. Furthermore, these cells display ultrastructural properties, such as bundles of intermediate filaments, interdigitations, and desmosomes with GFP(-) cells, which confirms their differentiation into corneal tissues. GFP(+)-transduced hBMSCs were injected at different time points into the right eye of lethally irradiated mice undergoing bone marrow transplantation, which developed ocular GVHD (oGVHD). Remarkably, hBMSCs massively migrate to corneal tissues after subconjunctival injection. Both macroscopic and histopathological examination showed minimal or no evidence of GVHD in the right eye, while the left eye, where no hBMSCs were injected, displayed features of GVHD. Thus, in the current study, we confirm that hBMSCs may induce their therapeutic effect at least in part by differentiation and regeneration of damaged tissues in the host. Our results provide experimental evidence that hBMSCs represent a potential cellular therapy to attenuate oGVHD.

  20. Repair of bone defects in vivo using tissue engineered hypertrophic cartilage grafts produced from nasal chondrocytes.

    PubMed

    Bardsley, Katie; Kwarciak, Agnieska; Freeman, Christine; Brook, Ian; Hatton, Paul; Crawford, Aileen

    2017-01-01

    The regeneration of large bone defects remains clinically challenging. The aim of our study was to use a rat model to use nasal chondrocytes to engineer a hypertrophic cartilage tissue which could be remodelled into bone in vivo by endochondral ossification. Primary adult rat nasal chondrocytes were isolated from the nasal septum, the cell numbers expanded in monolayer culture and the cells cultured in vitro on polyglycolic acid scaffolds in chondrogenic medium for culture periods of 5-10 weeks. Hypertrophic differentiation was assessed by determining the temporal expression of key marker genes and proteins involved in hypertrophic cartilage formation. The temporal changes in the genes measured reflected the temporal changes observed in the growth plate. Collagen II gene expression increased 6 fold by day 7 and was then significantly downregulated from day 14 onwards. Conversely, collagen X gene expression was detectable by day 14 and increased 100-fold by day 35. The temporal increase in collagen X expression was mirrored by increases in alkaline phosphatase gene expression which also was detectable by day 14 with a 30-fold increase in gene expression by day 35. Histological and immunohistochemical analysis of the engineered constructs showed increased chondrocyte cell volume (31-45 μm), deposition of collagen X in the extracellular matrix and expression of alkaline phosphatase activity. However, no cartilage mineralisation was observed in in vitro culture of up to 10 weeks. On subcutaneous implantation of the hypertrophic engineered constructs, the grafts became vascularised, cartilage mineralisation occurred and loss of the proteoglycan in the matrix was observed. Implantation of the hypertrophic engineered constructs into a rat cranial defect resulted in angiogenesis, mineralisation and remodelling of the cartilage tissue into bone. Micro-CT analysis indicated that defects which received the engineered hypertrophic constructs showed 38.48% in bone volume

  1. Evaluation of the stromal vascular fraction of adipose tissue as the basis for a stem cell-based tissue-engineered vascular graft.

    PubMed

    Krawiec, Jeffrey T; Liao, Han-Tsung; Kwan, LaiYee Lily; D'Amore, Antonio; Weinbaum, Justin S; Rubin, J Peter; Wagner, William R; Vorp, David A

    2017-09-01

    One of the rate-limiting barriers within the field of vascular tissue engineering is the lengthy fabrication time associated with expanding appropriate cell types in culture. One particularly attractive cell type for this purpose is the adipose-derived mesenchymal stem cell (AD-MSC), which is abundant and easily harvested from liposuction procedures. Even this cell type has its drawbacks, however, including the required culture period for expansion, which could pose risks of cellular transformation or contamination. Eliminating culture entirely would be ideal to avoid these concerns. In this study, we used the raw population of cells obtained after digestion of human liposuction aspirates, known as the stromal vascular fraction (SVF), as an abundant, culture-free cell source for tissue-engineered vascular grafts (TEVGs). SVF cells and donor-paired cultured AD-MSCs were first assessed for their abilities to differentiate into vascular smooth muscle cells (SMCs) after angiotensin II stimulation and to secrete factors (eg, conditioned media) that promote SMC migration. Next, both cell types were incorporated into TEVG scaffolds, implanted as an aortic graft in a Lewis rat model, and assessed for their patency and composition. In general, the human SVF cells were able to perform the same functions as AD-MSCs isolated from the same donor by culture expansion. Specifically, cells within the SVF performed two important functions; namely, they were able to differentiate into SMCs (SVF calponin expression: 16.4% ± 7.7% vs AD-MSC: 19.9%% ± 1.7%) and could secrete promigratory factors (SVF migration rate relative to control: 3.1 ± 0.3 vs AD-MSC: 2.5 ± 0.5). The SVF cells were also capable of being seeded within biodegradable, elastomeric, porous scaffolds that, when implanted in vivo for 8 weeks, generated patent TEVGs (SVF: 83% patency vs AD-MSC: 100% patency) populated with primary vascular components (eg, SMCs, endothelial cells, collagen, and elastin). Human

  2. Tissue-engineered vascular grafts transform into mature blood vessels via an inflammation-mediated process of vascular remodeling.

    PubMed

    Roh, Jason D; Sawh-Martinez, Rajendra; Brennan, Matthew P; Jay, Steven M; Devine, Lesley; Rao, Deepak A; Yi, Tai; Mirensky, Tamar L; Nalbandian, Ani; Udelsman, Brooks; Hibino, Narutoshi; Shinoka, Toshiharu; Saltzman, W Mark; Snyder, Edward; Kyriakides, Themis R; Pober, Jordan S; Breuer, Christopher K

    2010-03-09

    Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are the earliest tissue-engineered vascular grafts (TEVGs) to be used clinically. These TEVGs transform into living blood vessels in vivo, with an endothelial cell (EC) lining invested by smooth muscle cells (SMCs); however, the process by which this occurs is unclear. To test if the seeded BMCs differentiate into the mature vascular cells of the neovessel, we implanted an immunodeficient mouse recipient with human BMC (hBMC)-seeded scaffolds. As in humans, TEVGs implanted in a mouse host as venous interposition grafts gradually transformed into living blood vessels over a 6-month time course. Seeded hBMCs, however, were no longer detectable within a few days of implantation. Instead, scaffolds were initially repopulated by mouse monocytes and subsequently repopulated by mouse SMCs and ECs. Seeded BMCs secreted significant amounts of monocyte chemoattractant protein-1 and increased early monocyte recruitment. These findings suggest TEVGs transform into functional neovessels via an inflammatory process of vascular remodeling.

  3. Tissue-engineered vascular grafts transform into mature blood vessels via an inflammation-mediated process of vascular remodeling

    PubMed Central

    Roh, Jason D.; Sawh-Martinez, Rajendra; Brennan, Matthew P.; Jay, Steven M.; Devine, Lesley; Rao, Deepak A.; Yi, Tai; Mirensky, Tamar L.; Nalbandian, Ani; Udelsman, Brooks; Hibino, Narutoshi; Shinoka, Toshiharu; Saltzman, W. Mark; Snyder, Edward; Kyriakides, Themis R.; Pober, Jordan S.; Breuer, Christopher K.

    2010-01-01

    Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are the earliest tissue-engineered vascular grafts (TEVGs) to be used clinically. These TEVGs transform into living blood vessels in vivo, with an endothelial cell (EC) lining invested by smooth muscle cells (SMCs); however, the process by which this occurs is unclear. To test if the seeded BMCs differentiate into the mature vascular cells of the neovessel, we implanted an immunodeficient mouse recipient with human BMC (hBMC)-seeded scaffolds. As in humans, TEVGs implanted in a mouse host as venous interposition grafts gradually transformed into living blood vessels over a 6-month time course. Seeded hBMCs, however, were no longer detectable within a few days of implantation. Instead, scaffolds were initially repopulated by mouse monocytes and subsequently repopulated by mouse SMCs and ECs. Seeded BMCs secreted significant amounts of monocyte chemoattractant protein-1 and increased early monocyte recruitment. These findings suggest TEVGs transform into functional neovessels via an inflammatory process of vascular remodeling. PMID:20207947

  4. Current Concepts: Osteochondritis Dissecans of the Capitellum and the Role of Osteochondral Autograft Transplantation.

    PubMed

    Kirsch, Jacob M; Thomas, Jared; Bedi, Asheesh; Lawton, Jeffrey N

    2016-12-01

    Background: Osteochondritis dissecans (OCD) of the capitellum is a painful condition, which often affects young throwing athletes. Our current understanding regarding the etiology, risks factors, diagnosis, and efficacy of the available treatment options has expanded over recent years, however remains suboptimal. Recent data on patient-reported outcomes following osteochondral autograft transplantation (OAT) for the treatment of large osteochondral lesions of the capitellum have been promising but limited. This review seeks to critically analyze and summarize the available literature on the etiology, diagnosis, and reported outcomes associated with OCD of the capitellum and the use of OAT for its treatment. Methods: A comprehensive literature search was conducted. Unique and customized search strategies were formulated in PubMed, Embase, Scopus, Web of Science, and CENTRAL. Combinations of keywords and controlled vocabulary terms were utilized in order to cast a broad net. Relevant clinical, biomechanical, anatomic and imaging studies were reviewed along with recent review articles, and case series. Results: Forty-three articles from our initial literature search were found to be relevant for this review. The majority of these articles were either review articles, clinical studies, anatomic or imaging studies or biomechanical studies. Conclusions: Current evidence suggests that OAT may lead to better and more consistent outcomes than previously described methods for treating large OCD lesions of the capitellum.

  5. Image-Guided Techniques Improve the Short-Term Outcome of Autologous Osteochondral Cartilage Repair Surgeries

    PubMed Central

    Devlin, Steven M.; Hurtig, Mark B.; Waldman, Stephen D.; Rudan, John F.; Bardana, Davide D.; Stewart, A. James

    2013-01-01

    Objective: Autologous osteochondral cartilage repair is a valuable reconstruction option for cartilage defects, but the accuracy to harvest and deliver osteochondral grafts remains problematic. We investigated whether image-guided methods (optically guided and template guided) can improve the outcome of these procedures. Design: Fifteen sheep were operated to create traumatic chondral injuries in each knee. After 4 months, the chondral defect in one knee was repaired using (a) conventional approach, (b) optically guided method, or (c) template-guided method. For both image-guided groups, harvest and delivery sites were preoperatively planned using custom-made software. During optically guided surgery, instrument position and orientation were tracked and superimposed onto the surgical plan. For the template-guided group, plastic templates were manufactured to allow an exact fit between template and the joint anatomy. Cylindrical holes within the template guided surgical tools according to the plan. Three months postsurgery, both knees were harvested and computed tomography scans were used to compare the reconstructed versus the native pre-injury joint surfaces. For each repaired defect, macroscopic (International Cartilage Repair Society [ICRS]) and histological repair (ICRS II) scores were assessed. Results: Three months after repair surgery, both image-guided surgical approaches resulted in significantly better histology scores compared with the conventional approach (improvement by 55%, P < 0.02). Interestingly, there were no significant differences found in cartilage surface reconstruction and macroscopic scores between the image-guided and the conventional surgeries. PMID:26069658

  6. [Osteochondritis dissecans of the acetabulum. Apropos of a case].

    PubMed

    Hardy, P; Hinojosa, J F; Coudane, H; Sommelet, J; Benoit, J

    1992-01-01

    Osteochondritis dissecans acetabuli is a rare affection. The observation presented is one of a 23 year old man, who presented hip blockages in external flexion rotation. X-ray only allowed to find a sequestrum in the acetabulum by tomography-Arthroscanner confirmed the osteochondritis without cartilage rupture. Arthroscopy eliminated a cartilage lesion and directed towards a cavity filled by spongious bone through an extra articular approach.

  7. Juvenile Osteochondritis Dissecans of the Knee. About A Case

    PubMed Central

    Astore, Ignacio; Agotegaray, Juan Ignacio; Comba, Ignacio; Bisiach, Luciana

    2017-01-01

    Introduction: Juvenile osteochondritis dissecans is a pathology that affects the superficial articular cartilage and subchondral bone in patients with open physes. Treatment of this disease is based on patient’s age and the stage of the disease. Methods: 16-year-old patient, athlete, with a history of knee pain on the right side of acute onset, without traumatic history. A physical examination shows pain in the external compartment of the knee. MRI shows a stable lesion that involves the external femoral condyle, over a posterior area of 16 mm by 20mm. Crutches are indicate for walking without body burden. Symptoms continue for six months and there are no changes in MRI. It is decided to do a stabilization with a Herbert type screw. After the surgery, pain persists and in x-ray controls, osteointegration is not observed. Osteosynthesis material is extracted and mosaicplasty is performed. We used Guhl’s intraoperative classification. Results: In this case, for a young patient with Guhl’s lesion type III, the reduction with a Herbert type screw was indicated, as the lesion was stable, of a significant size and congruent. Lesion progressed to type IV in 6 months. Thus, mosaicplasty was performed, obtaining a good functional result according to the physical exam, with a complete range of flexion and extension. A second-look arthroscopic assessment was carried out 2 months after surgery, showing osteointegration and stability of the allogenic graft. Conclusion: The variable of stability of the fragment is very important when determining the treatment. Most of the stable lesions can be successfully treated with a conservative treatment. Also, it has been demonstrated that young patients have a higher rate of healing. Instead, unstable lesions require surgical treatment.

  8. Bridging peripheral nerve defects with a tissue engineered nerve graft composed of an in vitro cultured nerve equivalent and a silk fibroin-based scaffold.

    PubMed

    Tang, Xin; Xue, Chengbin; Wang, Yaxian; Ding, Fei; Yang, Yumin; Gu, Xiaosong

    2012-05-01

    Tissue engineered nerve grafts are considered as a promising alternative to autologous nerve grafts used for peripheral nerve repair. The differences between these two types of nerve grafts are mainly in the regenerative microenvironment established by them. To construct ideal tissue engineered nerve grafts, it is therefore required to develop a better way to introduce biochemical cues into a neural scaffold, as compared to single or combined use of support cells and growth factors. Here, we used a co-culture system of dorsal root ganglia and Schwann cells to create an in vitro formed nerve equivalent, which was introduced into a silk fibroin-based scaffold to furnish a tissue engineered nerve graft (TENG). At 4- and 12- weeks after the TENG was implanted to bridge a 10-mm-long sciatic nerve defect in rats, histological and functional assessments as well as Western blot analysis were performed to evaluate the influences of the TENG on peripheral nerve regeneration. We found that at an early stage of nerve regeneration, the TENG significantly accelerated axonal growth, and up-regulated expressions of N-cadherin and PMP22. Twelve weeks after nerve grafting, the TENG produced a further improved outcome of nerve regeneration and functional recovery, which was more close to that of the autologous nerve graft than that of the silk fibroin-based scaffold. The introduction of an in vitro cultured nerve equivalent into a scaffold might contribute to establishing a native-like microenvironment for nerve regeneration. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. Isolated osteochondral fracture of the patella without patellar dislocation.

    PubMed

    Bhatt, Jay; Montalban, Antonio Santa Cruz; Wang, Kook Hyun; Lee, Hee Du; Nha, Kyung Wook

    2011-01-03

    Chondral fractures of the patella are associated with acute dislocation of the patella. Osteochondral fracture in patellar dislocation is located in the medial facet of the patella. This article presents a case of a 15-year-old female ballerina with isolated displaced osteochondral fracture of the patella without patellar dislocation. She had no history of trauma. A Merchant's view of both knees showed mild subluxation of the patella, a small fragment on the lateral aspect of the knee, and a small defect of the centromedial patella. Axial magnetic resonance imaging (MRI) revealed an osteochondral fragment measuring 13 mm medial to the patella. However, the medial patellofemoral ligament and medial retinaculum were intact. An effusion on the medial side of the patella consistent with hemarthrosis was observed. An absence of a contusion or bone bruise on the lateral femoral condyle was shown. The loose body was removed arthroscopically. Intraoperative findings included a 1.5×2 cm osteochondral fragment. It is unusual that the osteochondral patellar defect site in this patient was in the inferior and central areas of the patella. Patellar chondral fractures without dislocation or patella fracture are rare. Therefore, the possibility of a trivial trauma leading to an osteochondral fracture should be kept in mind in adolescent and young adults who present with knee pain and hemarthrosis. Copyright 2011, SLACK Incorporated.

  10. Development of an Operator-Independent Method for Seeding Tissue-Engineered Vascular Grafts

    PubMed Central

    Udelsman, Brooks; Hibino, Narutoshi; Villalona, Gustavo A.; McGillicuddy, Edward; Nieponice, Alejandro; Sakamoto, Yuki; Matsuda, Shojiro; Vorp, David A.; Shinoka, Toshiharu

    2011-01-01

    The manual seeding of cells onto a biodegradable scaffold by pipetting is an effective method of cell seeding. However, the widespread use and ultimate clinical utility of this technique is limited by operator variability. This study was conducted to evaluate an operator-independent vacuum-seeding method for use in an upcoming clinical trial. Using bone marrow-derived mononuclear cells, we achieved seeding comparable to manually seeded scaffolds in terms of cellular attachment, distribution, and viability in vacuum-seeded grafts at vacuum pressures of −25 to −50 mmHg. In conclusion, we describe an operator-independent seeding method for use in the clinical setting. PMID:21410308

  11. Platelet-Rich Plasma May Improve Osteochondral Donor Site Healing in a Rabbit Model

    PubMed Central

    Smyth, Niall A.; Haleem, Amgad M.; Ross, Keir A.; Hannon, Charles P.; Murawski, Christopher D.; Do, Huong T.; Kennedy, John G.

    2016-01-01

    Purpose The purpose of this study was to assess the effect(s) of platelet-rich plasma (PRP) on osteochondral donor site healing in a rabbit model. Methods Osteochondral donor sites 3 mm in diameter and 5 mm in depth were created bilaterally on the femoral condyles of 12 New Zealand White rabbits. Knees were randomized such that one knee in each rabbit received an intra-articular injection of PRP and the other received saline (placebo). Rabbits were euthanized at 3, 6, and 12 weeks following surgery. Repair tissue was evaluated using the International Cartilage Repair Society (ICRS) macroscopic and histological scores. Results No complications occurred as a result of the interventions. There was no significant difference in macroscopic scores between the 2 groups (5.5 ± 3.8 vs. 3.8 ± 3.5; P = 0.13). Subjective macroscopic assessment determined greater tissue infill with fewer fissures and a more cartilage-like appearance in PRP-treated knees. Overall ICRS histological scores were better in the PRP group compared with the placebo (9.8 ± 2.0 vs. 7.8 ± 1.8; P = 0.04). Histological scores were also higher in the PRP group compared with the placebo group at each time point. Greater glycosaminoglycan and type II collagen content were noted in the repair tissue of the PRP group compared with the placebo group. Conclusion The results of this study indicate that PRP used as an intra-articular injection may improve osteochondral healing in a rabbit model. PMID:26958322

  12. Autologous fat graft in radiated tissue prior to alloplastic reconstruction of the breast: report of two cases.

    PubMed

    Salgarello, Marzia; Visconti, Giuseppe; Farallo, Eugenio

    2010-02-01

    The new indications for radiotherapy in the cure of breast cancer lead to an increasing number of candidates for this adjuvant treatment. However, it raises some concerns with respect to the prosthetic reconstruction of the breast. In fact, the use of implants in the irradiated breast is discouraged by many authors because of the high rate of complications. In these cases autologous tissue reconstruction is being recommended. However, not every patient is a candidate for autologous tissue reconstruction. Furthermore, not all radiotherapies are comparable in dose, timing, and patient tolerance. In this article we present the preliminary results of an alternative surgical management in oncoplastic breast surgery consisting of fat injections before implant placement. Six months after the last radiation treatment, a lumpectomy patient and a modified radical mastectomy patient each underwent two sessions of fat injection prior to implant placement, with a 3-month interval in between sessions. The implants were placed 3 months after the last fat injection. There was a 12-month follow-up for the modified radical mastectomy case and a 18-month follow-up for the lumpectomy case. In both cases we report no postoperative complications, Baker grade 1 capsule contracture, good aesthetic outcome, and high patient satisfaction. The preliminary results show that fat injection in irradiated tissue prior to breast alloplastic reconstruction may reduce the radiation-related complications on implants. Benefits from fat grafting are in keeping with the theoretical basis of this surgery. Larger studies are needed to confirm our observations.

  13. Local application of strontium in a calcium phosphate cement system accelerates healing of soft tissue tendon grafts in anterior cruciate ligament reconstruction: experiment using a rabbit model.

    PubMed

    Kuang, Guan-Ming; Yau, W P; Lu, William W; Chiu, K Y

    2014-12-01

    Healing of soft tissue tendon grafts within the bone tunnel in anterior cruciate ligament (ACL) reconstruction is known to be slower than that of bone-patellar tendon-bone grafts. There are attempts to accelerate healing of the graft within the bone tunnel. One of the methods is the use of strontium-enriched calcium phosphate cement (Sr-CPC). Early results in animal studies have been encouraging, although it is not known whether the accelerated healing was solely caused by the effect of strontium within the cement or by the calcium phosphate cement (CPC) itself. There would be differences between Sr-CPC and conventional CPC in terms of the effect on healing of soft tissue tendon grafts within the bone tunnels in ACL reconstruction. Controlled laboratory study. A total of 30 single-bundle ACL reconstruction procedures were performed in 15 rabbits with the use of an Achilles tendon allograft. The graft on the left limb was coated with Sr-CPC, while that on the right limb was coated with CPC. Three animals each were sacrificed for histological and histomorphometric analyses at 3, 6, 9, 12, and 24 weeks after surgery. In the Sr-CPC group, early formation of Sharpey fibers was present at 6 weeks after surgery, while early remodeling of a graft-fibrocartilage-bone junction was noted at 12 weeks. In the CPC group, early formation of Sharpey fibers was only found at 9 to 12 weeks after surgery. At 24 weeks, a direct enthesis was found in both groups. According to the histomorphometric score, graft healing in the Sr-CPC group took place 3 weeks faster than that in the CPC group at and before 12 weeks; however, there was no difference between the groups at 24 weeks. The local application of strontium in a CPC system leads to accelerated graft healing within the bone tunnels. The use of Sr-CPC to enhance graft-bone healing may improve the clinical results of ACL reconstruction using soft tissue tendon grafts. © 2014 The Author(s).

  14. Tissue-Specific Expression Patterns of MicroRNA during Acute Graft-versus-Host Disease in the Rat

    PubMed Central

    Jalapothu, Dasaradha; Boieri, Margherita; Crossland, Rachel E.; Shah, Pranali; Butt, Isha A.; Norden, Jean; Dressel, Ralf; Dickinson, Anne M.; Inngjerdingen, Marit

    2016-01-01

    MicroRNAs (miRNA) have emerged as central regulators of diverse biological processes and contribute to driving pathology in several diseases. Acute graft-versus-host disease (aGvHD) represents a major complication after allogeneic hematopoietic stem cell transplantation, caused by alloreactive donor T cells attacking host tissues leading to inflammation and tissue destruction. Changes in miRNA expression patterns occur during aGvHD, and we hypothesized that we could identify miRNA signatures in target tissues of aGvHD that may potentially help understand the underlying molecular pathology of the disease. We utilized a rat model of aGvHD with transplantation of fully MHC-mismatched T cell depleted bone marrow, followed by infusion of donor T cells. The expression pattern of 423 rat miRNAs was investigated in skin, gut, and lung tissues and intestinal T cells with the NanoString hybridization platform, in combination with validation by quantitative PCR. MHC-matched transplanted rats were included as controls. In the skin, upregulation of miR-34b and downregulation of miR-326 was observed, while in the intestines, we detected downregulation of miR-743b and a trend toward downregulation of miR-345-5p. Thus, tissue-specific expression patterns of miRNAs were observed. Neither miR-326 nor miR-743b has previously been associated with aGvHD. Moreover, we identified upregulation of miR-146a and miR-155 in skin tissue of rats suffering from aGvHD. Analysis of intestinal T cells indicated 23 miRNAs differentially regulated between aGvHD and controls. Two of these miRNAs were differentially expressed either in skin (miR-326) or in intestinal (miR-345-5p) tissue. Comparison of intestinal and peripheral blood T cells indicated common dysregulated expression of miR-99a, miR-223, miR-326, and miR-345-5p. Analysis of predicted gene targets for these miRNAs indicated potential targeting of an inflammatory network both in skin and in the intestines that may further regulate

  15. Versatility of retroauricular mastoid donor site: a convenient valuable warehouse of various free graft tissues in cosmetic and reconstructive surgery.

    PubMed

    Cho, Jeong Mok; Jeong, Jae Hoon; Woo, Kevin Volt; Lee, Yoon Ho

    2013-01-01

    Soft-tissue deficiency is a critical issue in facial cosmetic and reconstructive surgery. Harvesting autografts from other anatomical sites has been a common practice in overcoming soft-tissue insufficiency for many years. However, donor-site complications and visible scars are of important concerns. Therefore, we would like to introduce an alternative donor site of free-tissue grafts and its inherent advantages: the retroauricular mastoid area located along the mastoid hair line. From August 1991 to June 2011, we performed facial reconstructive surgeries for cosmetic correction of disfigurements from both congenital and complications of previous cosmetic procedures on a total of 213 patients. These patients had undergone either 1 or more facial cosmetic surgeries in the past. In this study, our primary goal focused on revising facial asymmetries or defects from previous surgical scars, tissue contraction, undercorrection, or underdevelopment. For autograft harvesting, we incised an elliptical shape along the retroauricular hairline. We then harvested sufficient amount of skin, dermal fat, fascia, or a piece of the mastoid bone if needed. After harvesting, we closed the incisional area and covered it with a compressive dressing. In evaluation of our results, we compared the preoperative photographs with postoperative and constructed a survey on patient satisfaction. Overall, the patients in this study were greatly satisfied with their surgical results. No major complications were reported. As a result of our long-term study, we believe that the retroauricular mastoid area has been shown to be an indispensable donor site for a variety of autograft tissues in terms of safety, convenience, and versatility of its unique structural composition consisting of skin, dermal fat, fascia, and bone.

  16. Emerging genetic basis of osteochondritis dissecans

    PubMed Central

    Bates, J. Tyler; Jacobs, John C.; Shea, Kevin G.; Oxford, Julia Thom

    2014-01-01

    Genome-wide association studies provide an unbiased approach in the identification of genes that increase the risk for osteochondritis dissecans (OCD). OCD is a disorder of the bone and cartilage that affects humans, horses, pigs, dogs, and other mammals. Recent genome-wide association studies in humans, horses, and pigs are reviewed and genes identified. The identified genes tended to cluster with respect to function and biological processes such as the protein secretion pathway, extracellular matrix molecules, and growth plate maturation. Genome-wide association studies in humans are a critical next step in the effort to provide a better understanding of the causes of OCD, which will, in turn, allow preventive strategies for treatment of adolescent and young adults who are at risk for the development of degenerative joint disease due to the effects of OCD. PMID:24698039

  17. Anatomical Glenoid Reconstruction Using Fresh Osteochondral Distal Tibia Allograft After Failed Latarjet Procedure.

    PubMed

    Sanchez, Anthony; Ferrari, Marcio B; Akamefula, Ramesses A; Frank, Rachel M; Sanchez, George; Provencher, Matthew T

    2017-04-01

    In the treatment of recurrent anterior glenohumeral instability, the Latarjet procedure has been shown to fail. This results in a need for viable revisional procedures for patients who present with this challenging pathology. We report our preferred technique for anatomical glenoid reconstruction using a fresh osteochondral distal tibia allograft after a failed Latarjet procedure. This bony augmentation technique employs a readily available dense, weight-bearing osseous tissue source that has excellent conformity, as well as the added benefit of a cartilaginous surface to correct chondral deficiencies. Given its effectiveness in the Latarjet revision setting and low complication rate, the distal tibia allograft is a reasonable treatment option.

  18. Combination of bone marrow mesenchymal stem cells and cartilage fragments contribute to enhanced repair of osteochondral defects.

    PubMed

    Abbas, Mohammed

    2017-01-01

    Cartilage tissue engineering using stem cells and biomaterials is considered a promising approach despite poor outcomes. We hypothesise that articular cartilage fragments provides native environmental cues to enhance stem cell differentiation. As such we evaluated the chondrogenic differentiation and repair of critical size defect in a human explant osteochondral model (OD) using bone marrow derived mesenchymal stem cells (BM-MSCs) and homogenised cartilage. BM-MSCs were established from the bone-marrow plugs of patients undergoing total knee arthroplasty and characterized. Osteochondral tissue was trimmed and a central drill defect (∼2mm) was made. Chondrogenic repair was evaluated by filling the OD defect area with either BM-MSCs (Group II), homogenized cartilage (Group III) or a combination of both BM-MSCs and homogenized cartilage (Group IV). OD with no added cell or tissue served as control (Group I). Samples were maintained in chondrogenic differentiation medium for 28 days. Microscopic images showed maximal OD closure in Group IV. Partial OD closure was observed in Group II and to a lesser extent in Group III. Haematoxylin-eosin staining revealed immature cartilaginous matrix in Group II and more mature matrix in Group IV. Sircol™ Assay showed increased collagen deposition in both Group II and Group IV. Immunostaining for both groups revealed positive staining for type II collagen. Combining BM-MSCs and homogenised cartilage demonstrated enhanced cartilage formation and defect filling in a human ex-vivo osteochondral model.

  19. Tracheal Replacement Therapy with a Stem Cell-Seeded Graft: Lessons from Compassionate Use Application of a GMP-Compliant Tissue-Engineered Medicine.

    PubMed

    Elliott, Martin J; Butler, Colin R; Varanou-Jenkins, Aikaterini; Partington, Leanne; Carvalho, Carla; Samuel, Edward; Crowley, Claire; Lange, Peggy; Hamilton, Nicholas J; Hynds, Robert E; Ansari, Tahera; Sibbons, Paul; Fierens, Anja; McLaren, Claire; Roebuck, Derek; Wallis, Colin; Muthialu, Nagarajan; Hewitt, Richard; Crabbe, David; Janes, Sam M; De Coppi, Paolo; Lowdell, Mark W; Birchall, Martin A

    2017-06-01

    Tracheal replacement for the treatment of end-stage airway disease remains an elusive goal. The use of tissue-engineered tracheae in compassionate use cases suggests that such an approach is a viable option. Here, a stem cell-seeded, decellularized tissue-engineered tracheal graft was used on a compassionate basis for a girl with critical tracheal stenosis after conventional reconstructive techniques failed. The graft represents the first cell-seeded tracheal graft manufactured to full good manufacturing practice (GMP) standards. We report important preclinical and clinical data from the case, which ended in the death of the recipient. Early results were encouraging, but an acute event, hypothesized to be an intrathoracic bleed, caused sudden airway obstruction 3 weeks post-transplantation, resulting in her death. We detail the clinical events and identify areas of priority to improve future grafts. In particular, we advocate the use of stents during the first few months post-implantation. The negative outcome of this case highlights the inherent difficulties in clinical translation where preclinical in vivo models cannot replicate complex clinical scenarios that are encountered. The practical difficulties in delivering GMP grafts underscore the need to refine protocols for phase I clinical trials. Stem Cells Translational Medicine 2017;6:1458-1464. © 2017 The Authors Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.

  20. Conjugation with RGD Peptides and Incorporation of Vascular Endothelial Growth Factor Are Equally Efficient for Biofunctionalization of Tissue-Engineered Vascular Grafts

    PubMed Central

    Antonova, Larisa V.; Seifalian, Alexander M.; Kutikhin, Anton G.; Sevostyanova, Victoria V.; Matveeva, Vera G.; Velikanova, Elena A.; Mironov, Andrey V.; Shabaev, Amin R.; Glushkova, Tatiana V.; Senokosova, Evgeniya A.; Vasyukov, Georgiy Yu.; Krivkina, Evgeniya O.; Burago, Andrey Yu.; Kudryavtseva, Yuliya A.; Barbarash, Olga L.; Barbarash, Leonid S.

    2016-01-01

    The blend of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) has recently been considered promising for vascular tissue engineering. However, it was shown that PHBV/PCL grafts require biofunctionalization to achieve high primary patency rate. Here we compared immobilization of arginine–glycine–aspartic acid (RGD)-containing peptides and the incorporation of vascular endothelial growth factor (VEGF) as two widely established biofunctionalization approaches. Electrospun PHBV/PCL small-diameter grafts with either RGD peptides or VEGF, as well as unmodified grafts were implanted into rat abdominal aortas for 1, 3, 6, and 12 months following histological and immunofluorescence assessment. We detected CD31+/CD34+/vWF+ cells 1 and 3 months postimplantation at the luminal surface of PHBV/PCL/RGD and PHBV/PCL/VEGF, but not in unmodified grafts, with the further observation of CD31+CD34−vWF+ phenotype. These cells were considered as endothelial and produced a collagen-positive layer resembling a basement membrane. Detection of CD31+/CD34+ cells at the early stages with subsequent loss of CD34 indicated cell adhesion from the bloodstream. Therefore, either conjugation with RGD peptides or the incorporation of VEGF promoted the formation of a functional endothelial cell layer. Furthermore, both modifications increased primary patency rate three-fold. In conclusion, both of these biofunctionalization approaches can be considered as equally efficient for the modification of tissue-engineered vascular grafts. PMID:27854352

  1. Galactose grafting on poly(ε-caprolactone) substrates for tissue engineering: a preliminary study.

    PubMed

    Russo, Laura; Russo, Teresa; Battocchio, Chiara; Taraballi, Francesca; Gloria, Antonio; D'Amora, Ugo; De Santis, Roberto; Polzonetti, Giovanni; Nicotra, Francesco; Ambrosio, Luigi; Cipolla, Laura

    2015-03-20

    The grafting of galactose units onto poly(ε-caprolactone) (PCL) substrates by a wet chemistry two-step procedure is proposed. Even though a reduction of hardness from 0.58-0.31 GPa to 0.12-0.05 GPa is achieved, the chemical functionalization does not negatively affect the tensile modulus (332.2±31.3 MPa and 328.5±34.7 MPa for unmodified and surface-modified PCL, respectively) and strength (15.1±1.3 MPa and 14.8±1.5 MPa as assessed before and after the surface modification, respectively), as well as the mechanical behaviour evaluated through small punch test. XPS and enzyme-linked lectin assay (ELLA) demonstrate the presence, and also the correct exposition of the saccharidic epitope on PCL substrates. The introduction of carbohydrate moieties on the PCL surfaces clearly enhances the hydrophilicity of the substrate, as the water contact angle decreases from 82.1±5.8° to 62.1±4.2°. Furthermore, preliminary biological analysis shows human mesenchymal stem cell viability over time and an improvement of cell adhesion and spreading.

  2. Electrospun collagen-chitosan-TPU nanofibrous scaffolds for tissue engineered tubular grafts.

    PubMed

    Huang, Chen; Chen, Rui; Ke, Qinfei; Morsi, Yosry; Zhang, Kuihua; Mo, Xiumei

    2011-02-01

    The objective of this study is to design a novel kind of scaffolds for blood vessel and nerve repairs. Random and aligned nanofibrous scaffolds based on collagen-chitosan-thermoplastic polyurethane (TPU) blends were electrospun to mimic the componential and structural aspects of the native extracellular matrix, while an optimal proportion was found to keep the balance between biocompatibility and mechanical strength. The scaffolds were crosslinked by glutaraldehyde (GTA) vapor to prevent them from being dissolved in the culture medium. Fiber morphology was characterized using scanning electron microscopy (SEM) and atomic-force microscopy (AFM). Fourier transform infrared spectroscopy (FTIR) showed that the three-material system exhibits no significant differences before and after crosslinking, whereas pore size of crosslinked scaffolds decreased drastically. The mechanical properties of the scaffolds were found to be flexible with a high tensile strength. Cell viability studies with endothelial cells and Schwann cells demonstrated that the blended nanofibrous scaffolds formed by electrospinning process had good biocompatibility and aligned fibers could regulate cell morphology by inducing cell orientation. Vascular grafts and nerve conduits were electrospun or sutured based on the nanofibrous scaffolds and the results indicated that collagen-chitosan-TPU blended nanofibrous scaffolds might be a potential candidate for vascular repair and nerve regeneration. Copyright © 2010 Elsevier B.V. All rights reserved.

  3. Comparison of ADM and Connective Tissue Graft as the Membrane in Class II Furcation Defect Regeneration: A Randomized Clinical Trial

    PubMed Central

    Esfahanian, Vahid; Farhad, Shirin; Sadighi Shamami, Mehrnaz

    2014-01-01

    Background and aims. Furcally-involved teeth present unique challenges to the success of periodontal therapy and influence treatment outcomes. This study aimed to assess to compare use of ADM and connective tissue membrane in class II furcation defect regeneration. Materials and methods. 10 patient with 2 bilaterally class II furcation defects in first and/or second maxilla or man-dibular molar without interproximal furcation involvement, were selected. Four weeks after initial phase of treatment, before and thorough the surgery pocket depth (PD), clinical attachment level to stent (CAL-S), free gingival margin to stent(FGM-S) , crestal bone to stent (Crest-S), horizontal defect depth to stent (HDD-S) and vertical defect depth to stent (VDD-S) and crestal bone to defect depth measured from stent margin. Thereafter, one side randomly treated using connective tissue and DFDBA (study group) and opposite side received ADM and DFDBA (control group). After 6 months, soft and hard tissue parameters measured again in re-entry. Results. Both groups presented improvements after therapies (P & 0.05). No inter-group differences were seen in PD re-duction (P = 0.275), CAL gain (P = 0.156), free gingival margin (P = 0.146), crest of the bone (P = 0.248), reduction in horizontal defects depth (P = 0.139) and reduction in vertical defects depth (P = 0.149). Conclusion. Both treatments modalities have potential of regeneration without any adverse effect on healing process. Connective tissue grafts did not have significant higher bone fill compared to that of ADM. PMID:25093054

  4. Comparison of ADM and Connective Tissue Graft as the Membrane in Class II Furcation Defect Regeneration: A Randomized Clinical Trial.

    PubMed

    Esfahanian, Vahid; Farhad, Shirin; Sadighi Shamami, Mehrnaz

    2014-01-01

    Background and aims. Furcally-involved teeth present unique challenges to the success of periodontal therapy and influence treatment outcomes. This study aimed to assess to compare use of ADM and connective tissue membrane in class II furcation defect regeneration. Materials and methods. 10 patient with 2 bilaterally class II furcation defects in first and/or second maxilla or man-dibular molar without interproximal furcation involvement, were selected. Four weeks after initial phase of treatment, before and thorough the surgery pocket depth (PD), clinical attachment level to stent (CAL-S), free gingival margin to stent(FGM-S) , crestal bone to stent (Crest-S), horizontal defect depth to stent (HDD-S) and vertical defect depth to stent (VDD-S) and crestal bone to defect depth measured from stent margin. Thereafter, one side randomly treated using connective tissue and DFDBA (study group) and opposite side received ADM and DFDBA (control group). After 6 months, soft and hard tissue parameters measured again in re-entry. Results. Both groups presented improvements after therapies (P & 0.05). No inter-group differences were seen in PD re-duction (P = 0.275), CAL gain (P = 0.156), free gingival margin (P = 0.146), crest of the bone (P = 0.248), reduction in horizontal defects depth (P = 0.139) and reduction in vertical defects depth (P = 0.149). Conclusion. Both treatments modalities have potential of regeneration without any adverse effect on healing process. Connective tissue grafts did not have significant higher bone fill compared to that of ADM.

  5. Development of a Sterile Amniotic Membrane Tissue Graft Using Supercritical Carbon Dioxide

    DTIC Science & Technology

    2015-03-04

    electron mi- croscopy (SEM) revealed that the bacteria exposed to SCCO2 remained intact; however, the lipid bilayer and internal cell structures were...tissue to SCCO2 for 10–30 min with the addition of 0– 2 mL of peracetic acid (PAA). Untreated AM tissue inocu- lated with bacteria /spores and the broth...Table 1). SCCO2 alone without the addition of the sterilizing agent PAA was not sufficient to inactivate the S. epidermidis and C. sporogenes bacteria as

  6. Immunoisolation to prevent tissue graft rejection: Current knowledge and future use

    PubMed Central

    David, Anu; Day, James

    2016-01-01

    This review focuses on the concept of immunoisolation and how this method has evolved over the last few decades. The concept of immunoisolation came out of the need to protect allogeneic transplant tissue from the host immune system and avoid systemic side effects of immunosuppression. The latter remains a significant hurdle in clinical translation of using tissue transplants for restoring endocrine function in diabetes, growth hormone deficiency, and other conditions. Herein, we review the most significant works studying the use of hydrogels, specifically alginate and poly (ethylene glycol), and membranes for immunoisolation and discuss how this approach can be applied in reproductive biology. PMID:27188513

  7. Anterior corneal buttons from DSAEK donor tissue can be stored in optisol GS for later use in tectonic lamellar patch grafting.

    PubMed

    Chu, Hsiao-Sang; Hsieh, Min-Chin; Chen, Yan-Ming; Hou, Yu-Chih; Hu, Fung-Rong; Chen, Wei-Li

    2014-06-01

    The aim of this study was to evaluate the feasibility of storing anterior corneal buttons split from donor Descemet stripping automated endothelial keratoplasty (DSAEK) tissue for later use in tectonic lamellar patch grafting. Donor corneas for DSAEK were split into anterior and posterior lamellae using a 350-μm depth microkeratome, and the remaining anterior corneal buttons were stored for up to 4 weeks at 4°C in Optisol GS before use in patch grafting of the cornea and sclera. Optisol GS-preserved anterior lamellar patch grafts successfully restored and maintained corneoscleral integrity after the removal of limbal dermoids (5 cases), after the coverage of the exposed polypropylene suture ends of scleral fixated posterior chamber intraocular lenses (2 cases), corneoscleral melting after pterygium excision (3 cases), sterile corneal ulceration with or without perforation (5 cases), Mooren ulceration (3 cases), and pseudomonas corneal ulceration with impending perforation (1 case). It was also used as a temporary substitute while waiting for a full-thickness corneal graft in a fulminant candida infection after DSAEK (1 case). All grafts remained structurally intact at the last postoperative examination of 19.2 ± 7.6 months after the tectonic surgery. Anterior corneal buttons from DSAEK donor tissue may be stored in Optisol GS for later use in tectonic patch grafting. This method obviates the longer operating time required for the surgeon to manually dissect the donor cornea in the operating room and reduces the wastage of precious donor corneas in countries where there are chronic shortages of eye bank donor tissue.

  8. Insulin-like Growth Factor-I and Slow, Bi-directional Perfusion Enhance the Formation of Tissue-Engineered Cardiac Grafts

    PubMed Central

    Cheng, Mingyu; Moretti, Matteo; Engelmayr, George C.

    2009-01-01

    Biochemical and mechanical signals enabling cardiac regeneration can be elucidated using in vitro tissue-engineering models. We hypothesized that insulin-like growth factor-I (IGF) and slow, bi-directional perfusion could act independently and interactively to enhance the survival, differentiation, and contractile performance of tissue-engineered cardiac grafts. Heart cells were cultured on three-dimensional porous scaffolds in medium with or without supplemental IGF and in the presence or absence of slow, bi-directional perfusion that enhanced transport and provided shear stress. Structural, molecular, and electrophysiologic properties of the resulting grafts were quantified on culture day 8. IGF had independent, beneficial effects on apoptosis (p < 0.01), cellular viability (p < 0.01), contractile amplitude (p < 0.01), and excitation threshold (p < 0.01). Perfusion independently affected the four aforementioned parameters and also increased amounts of cardiac troponin-I (p < 0.01), connexin-43 (p < 0.05), and total protein (p < 0.01) in the grafts. Interactive effects of IGF and perfusion on apoptosis were also present (p < 0.01). Myofibrillogenesis and spontaneous contractility were present only in grafts cultured with perfusion, although contractility was inducible by electrical field stimulation of grafts from all groups. Our findings demonstrate that multi-factorial stimulation of tissue-engineered cardiac grafts using IGF and perfusion resulted in independent and interactive effects on heart cell survival, differentiation, and contractility. PMID:18759675

  9. [Efficacies of treating large area third-degree burns by tangential excision and skin grafting for subcutaneous tissue wounds].

    PubMed

    Song, Guodong; Jia, Jun; Ma, Yindong; Shi, Wen; Wang, Fang; Li, Peilong; Gao, Cong; Zuo, Haibin; Fan, Chunjie; Yang, Tao; Wu, Qiuhe; Shao, Yang

    2014-12-02

    To explore the efficacies of treating patients with large area third-degree burns by tangential excision and skin grafting for subcutaneous tissue wounds. From January 2002 to December 2013, the medical records were retrospectively reviewed for 31 consecutive adult patients with a third-degree burn area exceeding 70% and undergoing tangential excision and skin grafting on subcutaneous tissue wound (TESGSTW) for the first time within 7 days postburn at Burn Center, Affiliated Jinan Central Hospital, Shandong University. For 31 patients, wounds with relative integrity eschar underwent TESGSTW by stages. Tourniquet was not used in some extremities. The relevant clinical data including patient condition on admission, causes of death, blood loss of tangential excision wound, surgical procedures and efficacies in cured group were analyzed. Average age, burn total area and third-degree burn area of 31 patients were (32.4 ± 12.8) years, (89.0 ± 6.2)% and (80.4 ± 7.6)% respectively. There were inhalation injury (n = 25, 80.6%) and early-stage shock before hospitalization (n = 22, 71.0%). Among 18 cured ones (58.1%), 2 patients had a third-degree burn area ≥ 90%. And 13 patients (41.9%) died and 10 of them died at 4 to 19 days postburn. Burn area was a risk factor of burn mortality. Sepsis and multiple organ dysfunction syndrome (MODS) were major mortality causes. Four patients died from early-stage sepsis. Within 14 days postburn, average blood loss volume per 1% tangential excision area in non-tourniquet group was slightly higher than that in the tourniquet group, but the difference was insignificant. For 18 cured patients, TESGSTW were performed 41 times. For 14 patients (77.8%), TESGSTW was performed twice. The average time of the first tangential excision was (4.1 ± 0.6) days postburn, the time interval between the first two tangential excisions was (6.4 ± 2.0) days, the first tangential excision area (33.8 ± 7.6)% and accumulated tangential excision area (58

  10. Enamel matrix derivative (emdogain) or subepithelial connective tissue graft for the treatment of adjacent multiple gingival recessions: a pilot study.

    PubMed

    Alkan, Eylem Ayhan; Parlar, Ates

    2013-01-01

    One treatment approach for achieving healing by regeneration is the application of enamel matrix derivative (EMD) during periodontal surgery. The aim of this randomized clinical study was to compare the efficacy of EMD with a connective tissue graft (CTG) for the treatment of adjacent Miller Class l and II multiple gingival recessions. Twelve systemically healthy subjects with at least two Miller Class I or II multiple gingival recession defects affecting adjacent teeth on both sides of the mouth were enrolled. The surgical protocol was performed for both groups. The 56 recession defects were evaluated for recession depth (RD), recession width (RW), percentage of root coverage (PRC), height of keratinized tissue (HKT), probing depth (PD), and clinical attachment level (CAL). All measurements were repeated at 6 and 12 months. The mean PRC at the final evaluation was 89% ± 17% for the coronally advanced flap (CAF) + EMD group and 93% ± 17% for the CAF + CTG group. Both treatments resulted in statistically significant decreases in RD and RW and increases in HKT at 6 and 12 months. There was also a significant CAL gain for both groups. PD remained shallow over time. The results demonstrated that both procedures were successful in treating Miller Class I and II multiple gingival recessions; however, the greater results of the CAF + CTG group did not reach a statistically significant level.

  11. One-stage Reconstruction of Soft Tissue Defects with the Sandwich Technique: Collagen-elastin Dermal Template and Skin Grafts

    PubMed Central

    Wollina, Uwe

    2011-01-01

    Background: A full-thickness soft tissue defect closure often needs complex procedures. The use of dermal templates can be helpful in improving the outcome. Objective: The objective was to evaluate a sandwich technique combining the dermal collagen–elastin matrix with skin grafts in a one-stage procedure. Materials and Methods: Twenty-three patients with 27 wounds were enrolled in this prospective single-centre observational study. The mean age was 74.8 ± 17.2 years. Included were full-thickness defects with exposed bone, cartilage and/ or tendons. The dermal collagen–elastin matrix was applied onto the wound bed accomplished by skin transplants, i.e. ‘sandwich’ transplantation. In six wounds, the transplants were treated with intermittent negative pressure therapy. Results: The size of defects was ≤875 cm2. The use of the dermal template resulted in a complete and stable granulation in 100% of wounds. Seventeen defects showed a complete closure and 19 achieved a complete granulation with an incomplete closure. There was a marked pain relief. No adverse events were noted due to the dermal template usage. Conclusions: Sandwich transplantation with the collagen–elastin matrix is a useful tool when dealing with full-thickness soft tissue defects with exposed bone, cartilage or tendons. PMID:22279382

  12. The influence of stromal cells on the pigmentation of tissue-engineered dermo-epidermal skin grafts.

    PubMed

    Biedermann, Thomas; Böttcher-Haberzeth, Sophie; Klar, Agnieszka S; Widmer, Daniel S; Pontiggia, Luca; Weber, Andreas D; Weber, Daniel M; Schiestl, Clemens; Meuli, Martin; Reichmann, Ernst

    2015-03-01

    It has been shown in vitro that melanocyte proliferation and function in palmoplantar skin is regulated by mesenchymal factors derived from fibroblasts. In this study, we investigated in vivo the influence of mesenchymal-epithelial interactions in human tissue-engineered skin substitutes reconstructed from palmar- and nonpalmoplantar-derived fibroblasts. Tissue-engineered dermo-epidermal analogs based on collagen type I hydrogels were populated with either human palmar or nonpalmoplantar fibroblasts and seeded with human nonpalmoplantar-derived melanocytes and keratinocytes. These skin substitutes were transplanted onto full-thickness skin wounds of immunoincompetent rats. Four weeks after transplantation the development of skin color was measured and grafts were excised and analyzed with regard to epidermal characteristics, in particular melanocyte number and function. Skin substitutes containing palmar-derived fibroblasts in comparison to nonpalmoplantar-derived fibroblasts showed (a) a significantly lighter pigmentation; (b) a reduced amount of epidermal melanin granules; and (c) a distinct melanosome expression. However, the number of melanocytes in the basal layer remained similar in both transplantation groups. These findings demonstrate that human palmar fibroblasts regulate the function of melanocytes in human pigmented dermo-epidermal skin substitutes after transplantation, whereas the number of melanocytes remains constant. This underscores the influence of site-specific stromal cells and their importance when constructing skin substitutes for clinical application.

  13. The Influence of Stromal Cells on the Pigmentation of Tissue-Engineered Dermo-Epidermal Skin Grafts

    PubMed Central

    Biedermann, Thomas; Böttcher-Haberzeth, Sophie; Klar, Agnieszka S.; Widmer, Daniel S.; Pontiggia, Luca; Weber, Andreas D.; Weber, Daniel M.; Schiestl, Clemens; Meuli, Martin

    2015-01-01

    It has been shown in vitro that melanocyte proliferation and function in palmoplantar skin is regulated by mesenchymal factors derived from fibroblasts. In this study, we investigated in vivo the influence of mesenchymal–epithelial interactions in human tissue-engineered skin substitutes reconstructed from palmar- and nonpalmoplantar-derived fibroblasts. Tissue-engineered dermo-epidermal analogs based on collagen type I hydrogels were populated with either human palmar or nonpalmoplantar fibroblasts and seeded with human nonpalmoplantar-derived melanocytes and keratinocytes. These skin substitutes were transplanted onto full-thickness skin wounds of immunoincompetent rats. Four weeks after transplantation the development of skin color was measured and grafts were excised and analyzed with regard to epidermal characteristics, in particular melanocyte number and function. Skin substitutes containing palmar-derived fibroblasts in comparison to nonpalmoplantar-derived fibroblasts showed (a) a significantly lighter pigmentation; (b) a reduced amount of epidermal melanin granules; and (c) a distinct melanosome expression. However, the number of melanocytes in the basal layer remained similar in both transplantation groups. These findings demonstrate that human palmar fibroblasts regulate the function of melanocytes in human pigmented dermo-epidermal skin substitutes after transplantation, whereas the number of melanocytes remains constant. This underscores the influence of site-specific stromal cells and their importance when constructing skin substitutes for clinical application. PMID:25300246

  14. Laser sintered porous polycaprolacone scaffolds loaded with hyaluronic acid and gelatin-grafted thermoresponsive hydrogel for cartilage tissue engineering.

    PubMed

    Lee, Ming-Yih; Tsai, Wen-Wei; Chen, His-Jung; Chen, Jyh-Ping; Chen, Chih-Hao; Yeh, Wen-Lin; An, Jia

    2013-01-01

    The aim of this study is to evaluate a soft/hard bi-phase scaffold for cartilage tissue engineering. Chondrocyte proliferation, glycoaminoglycan production and total collagen content are compared between laser-sintered porous polycaprolactone (PCL) scaffolds with and without a thermoresponsive hydrogel grafted with hyaluronic acid and gelatin. The in vitro results show that scaffolds loaded with hydrogel have a higher initial chondrocyte attachment than PCL scaffolds. At day 21 and 28, scaffolds loaded with hydrogel have a significantly higher glycosaminoglycan (GAG) production than PCL scaffolds alone, and total collagen content including collagen type II in the hydrogel-loaded group is three times higher than the group without hydrogel. It is concluded that the laser-sintered porous PCL scaffold has good cytocompatibility, and that the hydrogel phase is able to enhance initial chondrocytes attachment as well as GAG and collagen production of chondrocytes. This study suggests that a soft/hard bi-phase scaffold may be used for cartilage tissue engineering to enhance in vitro chondrogenesis.

  15. Characterization of the Natural History of Extracellular Matrix Production in Tissue-Engineered Vascular Grafts during Neovessel Formation

    PubMed Central

    Naito, Yuji; Williams-Fritze, Misty; Duncan, Daniel R.; Church, Spencer N.; Hibino, Narutoshi; Madri, Joseph A.; Humphrey, Jay D.; Shinoka, Toshiharu; Breuer, Christopher K.

    2011-01-01

    Background The extracellular matrix (ECM) is a critical determinant of neovessel integrity. Materials and Methods: Thirty-six (polyglycolic acid + polycaprolactone and poly lactic acid) tissue-engineered vascular grafts seeded with syngeneic bone marrow mononuclear cells were implanted as inferior vena cava interposition grafts in C57BL/6 mice. Specimens were characterized using immunohistochemical staining and qPCR for representative ECM components in addition to matrix metalloproteinases (MMPs). Total collagen, elastin, and glycosaminoglycan (GAG) contents were determined. MMP activity was measured using zymography. Results Collagen production on histology demonstrated an initial increase in type III at 1 week followed by type I production at 2 weeks and type IV at 4 weeks. Gene expression of both type I and type III peaked at 2 weeks, whereas type IV continued to increase over the 4-week period. Histology demonstrated fibrillin-1 deposition at 1 week followed by elastin production at 4 weeks. Elastin gene expression significantly increased at 4 weeks, whereas fibrillin-1 decreased at 4 weeks. GAG demonstrated abundant production at each time point on histology. Gene expression of decorin significantly increased at 4 weeks, whereas versican decreased over time. Biochemical analysis showed that total collagen production was greatest at 2 weeks, and there was a significant increase in elastin and GAG production at 4 weeks. Histological characterization of MMPs showed abundant production of MMP-2 at each time point, while MMP-9 decreased over the 4-week period. Gene expression of MMP-2 significantly increased at 4 weeks, whereas MMP-9 significantly decreased at 4 weeks. Conclusions ECM production during neovessel formation is characterized by early ECM deposition followed by extensive remodeling. PMID:21996715

  16. Fabrication and characterisation of biomimetic, electrospun gelatin fibre scaffolds for tunica media-equivalent, tissue engineered vascular grafts.

    PubMed

    Elsayed, Y; Lekakou, C; Labeed, F; Tomlins, P

    2016-04-01

    It is increasingly recognised that biomimetic, natural polymers mimicking the extracellular matrix (ECM) have low thrombogenicity and functional motifs that regulate cell-matrix interactions, with these factors being critical for tissue engineered vascular grafts especially grafts of small diameter. Gelatin constitutes a low cost substitute of soluble collagen but gelatin scaffolds so far have shown generally low strength and suture retention strength. In this study, we have devised the fabrication of novel, electrospun, multilayer, gelatin fibre scaffolds, with controlled fibre layer orientation, and optimised gelatin crosslinking to achieve not only compliance equivalent to that of coronary artery but also for the first time strength of the wet tubular acellular scaffold (swollen with absorbed water) same as that of the tunica media of coronary artery in both circumferential and axial directions. Most importantly, for the first time for natural scaffolds and in particular gelatin, high suture retention strength was achieved in the range of 1.8-1.94 N for wet acellular scaffolds, same or better than that for fresh saphenous vein. The study presents the investigations to relate the electrospinning process parameters to the microstructural parameters of the scaffold, which are further related to the mechanical performance data of wet, crosslinked, electrospun scaffolds in both circumferential and axial tubular directions. The scaffolds exhibited excellent performance in human smooth muscle cell (SMC) proliferation, with SMCs seeded on the top surface adhering, elongating and aligning along the local fibres, migrating through the scaffold thickness and populating a transverse distance of 186 μm and 240 μm 9 days post-seeding for scaffolds of initial dry porosity of 74 and 83%, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Subchondral pre-solidified chitosan/blood implants elicit reproducible early osteochondral wound-repair responses including neutrophil and stromal cell chemotaxis, bone resorption and repair, enhanced repair tissue integration and delayed matrix deposition

    PubMed Central

    2013-01-01

    Background In this study we evaluated a novel approach to guide the bone marrow-driven articular cartilage repair response in skeletally aged rabbits. We hypothesized that dispersed chitosan particles implanted close to the bone marrow degrade in situ in a molecular mass-dependent manner, and attract more stromal cells to the site in aged rabbits compared to the blood clot in untreated controls. Methods Three microdrill hole defects, 1.4 mm diameter and 2 mm deep, were created in both knee trochlea of 30 month-old New Zealand White rabbits. Each of 3 isotonic chitosan solutions (150, 40, 10 kDa, 80% degree of deaceylation, with fluorescent chitosan tracer) was mixed with autologous rabbit whole blood, clotted with Tissue Factor to form cylindrical implants, and press-fit in drill holes in the left knee while contralateral holes received Tissue Factor or no treatment. At day 1 or day 21 post-operative, defects were analyzed by micro-computed tomography, histomorphometry and stereology for bone and soft tissue repair. Results All 3 implants filled the top of defects at day 1 and were partly degraded in situ at 21 days post-operative. All implants attracted neutrophils, osteoclasts and abundant bone marrow-derived stromal cells, stimulated bone resorption followed by new woven bone repair (bone remodeling) and promoted repair tissue-bone integration. 150 kDa chitosan implant was less degraded, and elicited more apoptotic neutrophils and bone resorption than 10 kDa chitosan implant. Drilled controls elicited a poorly integrated fibrous or fibrocartilaginous tissue. Conclusions Pre-solidified implants elicit stromal cells and vigorous bone plate remodeling through a phase involving neutrophil chemotaxis. Pre-solidified chitosan implants are tunable by molecular mass, and could be beneficial for augmented marrow stimulation therapy if the recruited stromal cells can progress to bone and cartilage repair. PMID:23324433

  18. Tissue reaction of deproteinized bovine bone matrix grafting in ectopic site: histological study on sheep.

    PubMed

    Grossi, João Ricardo Almeida; Bonacin, Rodrigo; Crivelaro, Viviane Rozeira; Giovanini, Allan Fernando; Zielak, João César; Deliberador, Tatiana Miranda

    2016-12-01

    The aim of this paper was to evaluate through histological analysis of the tissue reaction of deproteinized bovine bone matrix (DBBM) when inserted into the site of intramuscular ectopic sheep. In this study, 16 sheep received 3 groups and these were divided into 2 experimental times: Group 1-sham group, Group 2-particulate autogenous bone and Group 3-DBBM granules. All animals underwent surgical procedures for insertion of materials in an ectopic site (muscles of the lower back and after 3 and 6 months postoperatively, the samples were evaluated by histological analysis. The results indicated that the Sham group showed dense collagen fibers and thin, characterizing fibrosis at 3 and 6 months. In the autograft group there was a significant amount of collagen deposition and decreased inflammation at 6 months postoperatively. Group of DBBM, it was noted the presence of dense connective tissue and surrounding remaining particles was observed the formation of with osteoid characteristic tissue. The DBBM demonstrated biocompatibility, osteoconductivity and small osteogenesis capacity on ectopic site.

  19. A rare case of an osteochondral lesion of the tarsal navicular with a subacute stress fracture in a high level athlete.

    PubMed

    Nunag, Perrico; Quah, Colin; Pillai, Anand

    2014-12-01

    In this report, an osteochondral lesion of the tarsal navicular associated with a subacute stress fracture in a professional basketball player surgical treatment is presented. The surgical technique involved extra-articular curettage, bone grafting and plate stabilisation. Postoperative CT scan confirmed that both the osteochondral lesion and the stress fracture healed. The talonavicular joint showed no signs of arthritis on imaging. Clinical foot scores showed marked improvement after surgery. At 6 months patient managed to return to competitive play without pain in the foot and ankle. The outcome of this case indicates that the combination of curettage, bone grating and plate stabilisation works well for this rare and potentially career ending dual lesions. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Initial evaluation of the use of USPIO cell labeling and noninvasive MR monitoring of human tissue-engineered vascular grafts in vivo.

    PubMed

    Nelson, G N; Roh, J D; Mirensky, T L; Wang, Y; Yi, T; Tellides, G; Pober, J S; Shkarin, P; Shapiro, E M; Saltzman, W M; Papademetris, X; Fahmy, T M; Breuer, C K

    2008-11-01

    This pilot study examines noninvasive MR monitoring of tissue-engineered vascular grafts (TEVGs) in vivo using cells labeled with iron oxide nanoparticles. Human aortic smooth muscle cells (hASMCs) were labeled with ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles. The labeled hASMCs, along with human aortic endothelial cells, were incorporated into eight TEVGs and were then surgically implanted as aortic interposition grafts in a C.B-17 SCID/bg mouse host. USPIO-labeled hASMCs persisted in the grafts throughout a 3 wk observation period and allowed noninvasive MR imaging of the human TEVGs for real-time, serial monitoring of hASMC retention. This study demonstrates the feasibility of applying noninvasive imaging techniques for evaluation of in vivo TEVG performance.

  1. The effect of defect localization on spontaneous repair of osteochondral defects in a Gottingen minipig model: a retrospective analysis of the medial patellar groove versus the medial femoral condyle.

    PubMed

    Jung, Martin; Breusch, Steffen; Daecke, Wolfgang; Gotterbarm, Tobias

    2009-04-01

    Various animal models for experimental osteochondral defect healing have been used in orthopaedic research. Two main defect locations were chosen: the patellar groove or the central part of the medial femoral condyles (MFC). To date, it is not clear whether both locations display similar patterns in critical size osteochondral defect healing. We retrospectively analysed both locations in our minipig model hypothesizing that they show similar healing pattern. Thirty-five defects were analysed after three or 12 months. Osteochondral defects were 10 mm deep and 6.3 mm (MFC, n = 19) in diameter or 8 mm and 5.4 mm, respectively (trochlear groove [TG], n = 16). Semi-quantitative histological scoring and histomorphological evaluation were carried out. Both defect locations showed fillings of fibrous and fibrocartilage-like repair tissue. The osseous defect was closed by endochondral bone formation in the MFC. Semi-quantitative scoring did not show differences, whereas qualitative histomorphological analysis more frequently showed cartilaginous repair tissue in MFC defects. There was more frequent subchondral bone cyst formation in MFC location (P = 0.05), TG defects resulted in lower postoperative pain. Both defect localizations are suitable for studies on osteochondral healing. Since regenerating with less hyaline-like repair tissue and less subchondral cyst formation, TG is more favourable for experimental osteochondral defect healing in this model.

  2. In vivo evaluation of hydroxyapatite nanocoating on polyester artificial vascular grafts and possibility as soft-tissue compatible material.

    PubMed

    Kadono, Hiroyuki; Furuzono, Tsutomu; Masuda, Miwa; Okada, Masahiro; Ueki, Mitsuki; Takamizawa, Keiichi; Tanaka, Ryoichi; Miyatake, Kunio; Koyama, Yoshihisa; Takakuda, Kazuo

    2010-01-01

    The efficacy of hydroxyapatite (HAp) nanocoating on polyester vascular grafts was investigated in animal experiments. The HAp nanocrystals were covalently bonded separately between hydroxyl groups on a nanocrystal and alkoxysilyl groups in gamma-methacryloxypropyl triethoxysilane graft polymerized on a polyester substrate. Twelve HAp-coated polyester grafts and 10 control grafts of 20, 30, or 50 mm in length were implanted in canine common carotid arteries. Serious complications or occlusions were not observed in any of the dogs after implantation. A histologic evaluation was conducted by staining with hematoxylin and eosin (HE), the von Willebrand factor (vWf), and alpha-smooth muscle actin (alpha-SMA) around the inner lumen of the grafts. The number of inflammation cells and giant cells in the HAp-coated group was significantly lower than that in the group receiving noncoated grafts (p < 0.05).

  3. Novel Application of Magnetic Resonance Imaging Demonstrates Characteristic Differences in Vasculature at Predilection Sites of Osteochondritis Dissecans

    PubMed Central

    Tóth, Ferenc; Nissi, Mikko J.; Ellermann, Jutta M.; Wang, Luning; Shea, Kevin G.; Polousky, John; Carlson, Cathy S.

    2016-01-01

    Background Understanding the pathogenesis of osteochondrosis/osteochondritis dissecans and other developmental orthopaedic diseases that are thought to occur secondary to defects in vascular supply to growth/epiphyseal cartilage has been hampered by the inability to image the vasculature in this tissue. This is particularly true in human beings due to limitations of current imaging techniques and the lack of availability of appropriate cadaveric samples for histological studies. Hypothesis Susceptibility weighted imaging, an MRI sequence, allows identification of characteristic differences in the vascular architecture in species that are affected by osteochondrosis/osteochondritis dissecans on the femoral condyle (humans and pigs) versus one that is free of the disease (goat). Study design Descriptive laboratory study Materials Distal femora from cadavers of juvenile humans (n=5), pigs (n=3), and goats (n=3) were scanned in a 9.4T MRI scanner using susceptibility weighted imaging. Three-dimensional reconstructions were created and minimum intensity projections were calculated in three planes to enhance visualization of the vascular architecture. Results Susceptibility weighted imaging allowed clear visualization of the epiphyseal vasculature in all species. Vascular architecture, with vessels primarily arising from the perichondrium, was similar in humans and pigs, which are predisposed to osteochondrosis/osteochondritis dissecans, and was starkly different from that present in goats, a species in which there are no reports of osteochondrosis/osteochondritis dissecans. Furthermore, vessels in the distal femoral predilection site disappeared with age in a similar pattern in humans as has been reported previously in pigs. Conclusion Nearly identical vascular architecture at the shared primary predilection site of osteochondrosis/osteochondritis dissecans in the femoral condyles in human beings and pigs suggests that vascular failure, which is known to be central to

  4. Graft laceration and pullout strength of soft-tissue anterior cruciate ligament reconstruction: in vitro study comparing titanium, poly-d,l-lactide, and poly-d,l-lactide-tricalcium phosphate screws.

    PubMed

    Zantop, Thore; Weimann, Andre; Schmidtko, Romana; Herbort, Mirco; Raschke, Michael J; Petersen, Wolf

    2006-11-01

    The aim of part 1 of this study was to compare the structural properties of tibia-graft-interference screw constructs of soft-tissue graft fixation via the anterior cruciate ligament (ACL) by use of different interference screws. In part 2 the influence of graft laceration as a result of 1 single interference screw insertion on the mechanical properties of the graft was evaluated. In part 1 of the study, in a bovine knee model, soft-tissue graft fixation with titanium, biodegradable poly-D,L-lactide (PLDLA), and PLDLA-tricalcium phosphate (TCP) interference screws was performed, and the structural properties were determined by use of a materials testing machine at a rate of 12.5 mm/s. In part 2 the bone was cut after 1 single insertion of titanium, PLDLA, or PLDLA-TCP interference screws, and the mechanical properties of the graft were determined. Soft-tissue graft fixation with a biodegradable PLDLA interference screw showed a statistically significantly higher maximum load when compared with PLDLA-TCP and titanium interference screws. PLDLA-TCP screw fixation revealed a significantly higher ultimate load than titanium screw fixation. The mechanical properties of the grafts after 1 single insertion of PLDLA-TCP interference screws showed a statistically significantly higher stiffness, yield load, and maximum load when compared with the PLDLA and titanium interference screw groups. The results of this study suggest that the use of a titanium screw to fix a soft-tissue graft in cruciate ligament reconstruction surgery may cause damage to the graft. Soft-tissue ACL reconstruction fixed with a PLDLA interference screw resulted in a significantly higher pullout strength than ACL reconstructions fixed with a PLDLA-TCP or titanium interference screw. In contrast to fixation with a biodegradable interference screw, fixation with a titanium interference screw may damage the integrity of a soft-tissue graft in cruciate ligament reconstruction and should therefore be avoided.

  5. Autologous Minced Muscle Grafts: A Tissue Engineering Therapy for the Volumetric Loss of Skeletal Muscle

    DTIC Science & Technology

    2013-07-24

    allotted to one of three time points postinjury: 2, 8, or 16 wk. At these times, tissue was collected for histological and molecular analyses. At 8 and 16...from the CyQUANT kit, and read on a SpectraMax M2 plate reader ( Molecular Devices, Sunnyvale, CA) with excitation at 480 nm and emission at 520 nm. DNA...auto-, homo, and hetero-transplantation]. Biologia (Bratislava) 13: 514–522, 1958. 25. Huijing PA, Jaspers RT. Adaptation of muscle size and myofascial

  6. Use of a titanium mesh "shelter" combined with the soft tissue matrix expansion (tent pole) grafting in the reconstruction of a severely resorbed edentulous mandible. Technical note.

    PubMed

    Sverzut, Cássio Edvard; Trivellato, Alexandre Elias; Sverzut, Alexander Tadeu

    2015-01-01

    Patients wearing complete dentures for a long time usually exhibit lack of bone and keratinized mucosa in the bearing area due to bone resorption. The patients suffering from this phenomenon usually have unstable and non-retentive complete denture, which result in constant trauma to the mucosa, pain, functional limitations and worsening of facial esthetics. An innovative technique has been described in which a novel surgical approach using osseointegrated dental implants as "tent poles" was applied concomitant with particulate autogenous bone graft. The authors claim that the control and maintenance of the surgically expanded soft tissue volume should prevent graft resorption in the long term. Nevertheless, resorption of the bone graft is usually more significant where the bone mass is poorer, in the mandibular body. This paper describes a case of severely resorbed edentulous mandible in which the "tent pole" technique was applied with some modifications. Use of the titanium mesh "shelters" and two additional implants was effective in "protecting" the bone graft in the posterior portion of the mandibular body increasing bone mass volume in this area. Furthermore, we believe that this kind of graft "protection" on the whole residual alveolar ridge can increase the width of bone mass gain optimizing the "tent pole" technique.

  7. The therapeutic effect of low-level laser on repair of osteochondral defects in rabbit knee.

    PubMed

    Kamali, Fahimeh; Bayat, Mohammad; Torkaman, Giti; Ebrahimi, Esmaeel; Salavati, Mahyar

    2007-07-27

    Low level laser therapy (LLLT) has been shown to enhance collagen production and wound healing but its effect on cartilage repair from biomechanical point of view is not known yet. The aim of present study was to evaluate the biomechanical behaviour of repairing osteochondral defect in rabbits which received a pulsed low-level gallium-arsenide (Ga-As) laser irradiation. Osteochondral defects with 5mm diameter and 4mm in depth induced by drilling in right femoral patellar grooves of 41 adolescent male rabbits. They were divided into experimental and control groups. Experimental group received pulsed Ga-As (890nm) laser irradiation with energy density of 4.8J/cm(2). The rabbits in control group received placebo LLLT with shut-down equipment. The control defects were allowed to heal spontaneously. Each group were divided into three subgroups: A, B and C. Subgroups A, B and C were sacrificed on 4, 8, and 16 weeks after surgery. The knee joint were removed, and the defects were examined biomechanically by in situ-indentation method. The thickness, instantaneous and equilibrium indentation stiffness was measured during the test. Data were analysed using ANOVA and independent sample t-test. While no difference was observed in the repaired cartilage biomechanical properties among 4th, 8th, 16th weeks in study groups. The equilibrium indentation stiffness of experimental group was significantly higher in 8th week in comparison with control group. LLLT significantly enhances the stiffness of repairing tissue in the 8th week post injury in osteochondral defects in rabbits.

  8. Fluoroquinolone use in a child associated with development of osteochondritis dissecans

    PubMed Central

    Jacobs, John C; Shea, Kevin G; Oxford, Julia Thom; Carey, James L

    2014-01-01

    Several aetiological theories have been proposed for the development of osteochondritis dissecans. Cartilage toxicity after fluoroquinolone use has been well documented in vitro. We present a case report of a 10-year-old child who underwent a prolonged 18-month course of ciprofloxacin therapy for chronic urinary tract infections. This patient later developed an osteochondritis dissecans lesion of the medial femoral condyle. We hypothesise that the fluoroquinolone therapy disrupted normal endochondral ossification, resulting in development of osteochondritis dissecans. The aetiology of osteochondritis dissecans is still unclear, and this case describes an association between fluoroquinolone use and osteochondritis dissecans development. PMID:25228675

  9. Novel Vanadium-Loaded Ordered Collagen Scaffold Promotes Osteochondral Differentiation of Bone Marrow Progenitor Cells

    PubMed Central

    Cortizo, Ana M.; Ruderman, Graciela; Mazzini, Flavia N.; Molinuevo, M. Silvina; Mogilner, Ines G.

    2016-01-01

    Bone and cartilage regeneration can be improved by designing a functionalized biomaterial that includes bioactive drugs in a biocompatible and biodegradable scaffold. Based on our previous studies, we designed a vanadium-loaded collagen scaffold for osteochondral tissue engineering. Collagen-vanadium loaded scaffolds were characterized by SEM, FTIR, and permeability studies. Rat bone marrow progenitor cells were plated on collagen or vanadium-loaded membranes to evaluate differences in cell attachment, growth and osteogenic or chondrocytic differentiation. The potential cytotoxicity of the scaffolds was assessed by the MTT assay and by evaluation of morphological changes in cultured RAW 264.7 macrophages. Our results show that loading of VOAsc did not alter the grooved ordered structure of the collagen membrane although it increased membrane permeability, suggesting a more open structure. The VOAsc was released to the media, suggesting diffusion-controlled drug release. Vanadium-loaded membranes proved to be a better substratum than C0 for all evaluated aspects of BMPC biocompatibility (adhesion, growth, and osteoblastic and chondrocytic differentiation). In addition, there was no detectable effect of collagen or vanadium-loaded scaffolds on macrophage viability or cytotoxicity. Based on these findings, we have developed a new ordered collagen scaffold loaded with VOAsc that shows potential for osteochondral tissue engineering. PMID:27293438

  10. Joint laminate degradation assessed by reflected ultrasound from the cartilage surface and osteochondral junction

    NASA Astrophysics Data System (ADS)

    Brown, C. P.; Hughes, S. W.; Crawford, R. W.; Oloyede, A.

    2008-08-01

    The ability to quantify and qualify the progression of joint degeneration is becoming increasingly important in surgery. This paper examines the patterns of relative ultrasound reflection from normal, artificially and naturally degraded cartilage-on-bone, particularly investigating the potential of the ratio of reflection coefficients from the surface and osteochondral junction in distinguishing normal from osteoarthritic tissue. To this end, the reflection coefficients from the articular surface and osteochondral junction of normal cartilage-on-bone samples were calculated and compared to samples after the removal of proteoglycans, disruption of the collagen meshwork, delipidization of the articular surface and mechanical abrasion. Our results show that the large variation across normal and degraded joint samples negates the use of an isolated bone reflection measurement and to a lesser extent, an isolated surface reflection. The relative surface to bone reflections, calculated as a ratio of reflection coefficients, provided a more consistent and statistically significant (p < 0.001) method for distinguishing each type of degradation, especially osteoarthritic degradation, and due to the complementary relationship between surface and bone reflections was found to be an effective method for distinguishing degraded from normal tissue in the osteoarthritic joint, independent of the site of initiation of the osteoarthritic process.

  11. [Efficacy of fibrin tissue adhesive in the attachment of autogenous conjuntival graft on primary pterygium surgery].

    PubMed

    Rubin, Michel Risnic; Dantas, Paulo Elias C; Nishiwaki-Dantas, M Cristina; Felberg, Sergio

    2011-01-01

    To compare the efficacy of conjunctival autograft surgery with the attachment to the scleral bed using fibrin tissue adhesive or mononylon 10-0 suture after resection of primary pterygium. A comparative, prospective and randomized clinical trial was performed in 47 eyes of 47 patients with primary medial located pterygium. Group 1 (adhesive) was composed by 21 patients that underwent conjunctival autograft closure with fibrin tissue adhesive (Quixil™) and Group 2 (suture) was composed by 26 patients that underwent pterygium surgery with mononylon 10-0 (Ethicon(®)) suture (suture group) after pterygium excision. All surgeries were performed by the same surgeon. Patients were assessed on the preoperative period and on the 1(st), 14(th) and 21(st) postoperative days. They were followed-up with a questionnaire of ocular discomfort and by the surgical time spent, ocular hyperemia, complications and recurrence signals, being the recurrence also evaluated at the 6th postoperative month. Data were submitted to statistical analysis. A value of p<0.005 was considered statistically significant. The average surgical time was 19.05 ± 6.12 minutes in group 1 (glue) and 48.15 ± 7.13 minutes in the group 2 (suture) (p<0.001). The ocular discomfort scale analysis showed a lower score in the 1(st) (p<0.005), 7(th) (p<0.001) and 21(th) (p<0.001) postoperativ