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Background The objective of this study was to investigate the humoral immune response to xenogeneic antigens administered during the fetal state utilizing a baboon-to-pig model. Methods Nine fetuses from an alpha-1,3-galactosyltransferase gene knockout (GalT- KO) MGH-miniature swine sow underwent transuterine ultrasound-guided intraportal injection of T-cell depleted baboon bone marrow (B- BM) at mid-gestation. Two juvenile GalT-KO swine undergoing direct B- BM intraportal injection were used as controls. Results Postnatal humoral tolerance was induced in the long-term surviving piglets as demonstrated by the absence of any antibody response to baboon donor cells. In addition, a second intraportal B- BM administration at 2.5 months post-birth led to no antibody formation despite re-exposure to xenogeneic antigens. This B-cell unresponsiveness was abrogated only when the animal was exposed subcutaneously to third-party xenogeneic and allogeneic antigens, suggesting that the previously achieved humoral non-responsiveness was donor specific. In comparison, the two juvenile GalT-KO control swine demonstrated increasing anti-baboon IgM and IgG levels following intraportal injection. Conclusions In summary, xenogeneic B-cell tolerance was induced through in utero intraportal exposure to donor cells and this tolerance persisted following postnatal rechallenge with donor B- BM, but was lost on exposure to third-party antigen, possibly as a result of cross-reactive antibody formation. [ABSTRACT FROM AUTHOR]

The objective of this study was to assess the ability of tissue engineered cartilage to adhere to and integrate with porous polyethylene (PPE) in vivo and to evaluate the biomechanical integrity of the bond formed at the interface. Porcine auricular, articular, and costal chondrocytes were suspended in fibrin gel polymer and placed between discs of PPE to form tri-layer constructs. Controls consisted of fibroblasts suspended in gel or gel alone between the discs. Constructs were implanted into nude mice for 6, 12, and 18 weeks. Upon harvest, specimens were evaluated for neocartilage formation and integration into the PPE, using histological, dimensional (mass, thickness, diameter), and biomechanical (adhesion strength, interfacial stiffness, failure energy and failure strain) analyses. Neotissue was formed in all experimental constructs, consisting mostly of neocartilage integrating with discs of PPE. Control samples contained only fibrous tissue. Biomechanical analyses demonstrated that adhesion strength, interfacial stiffness, and failure energy were all significantly higher in the chondrocyte-seeded samples than in fibroblast-seeded controls, with the exception of costal constructs at 12 weeks, which were not significantly greater than controls. In general, failure strains did not vary between groups. In conclusion, porous polyethylene supported the growth of neocartilage that formed mechanically functional bonds with the PPE. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 103B: 983-991, 2015. [ABSTRACT FROM AUTHOR]

This study investigated a dual crosslinking paradigm, consisting of (a) photocrosslinking with Rose Bengal (RB) and green light followed by (b) chemical crosslinking with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), and N-hydroxysuccinimide (NHS) to enhance collagen gel stiffness. In group 1, 50 μL collagen constructs of 2% (w/v) type I collagen containing 10 μ M RB were allowed to gel spontaneously at 37°C. In group 2, the spontaneous gels were exposed to green light (532 nm). In group 3, the photochemically crosslinked gels were subsequently treated with a 1-h exposure to 33 m M EDC/6 m M NHS. Samples ( n = 18) were subjected to 0.08% (w/v) collagenase digestion, and the storage modulus of samples was measured by rheometry. Viability of encapsulated chondrocytes was measured by live/dead assay. Chondrocytes were ≥95% viable in all constructs at 10 days in vitro. Resistance to collagenase digestion increased as; spontaneous gels (2 h) < photochemical gels (3-4 h) < dual crosslinked gels (>24 h). The storage modulus of dual-crosslinked constructs was increased 5-fold over both photocrosslinked and spontaneous gels. As the dual crosslinking paradigm did not reduce encapsulated chondrocyte viability, these crosslinked collagen hydrogels could be a useful tool for the practical delivery of encapsulated chondrocytes to articular defects. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 1332-1338, 2015. [ABSTRACT FROM AUTHOR]

An approximate solution for the effects of high strain rates, and gradual strength degradation, on the penetration resistance of penetrometers can be obtained by combining the strain-path method with the classical upper bound theorem. The stream path calculations require the integration of the material constitutive equation along the streamlines. Unless the geometry is simple so that the integration can be evaluated analytically, numerical procedures are required to backtrack streamlines. The strain at any location is calculated by finding the streamline that passes through the given point and integrating the strain rate along that streamline from its inlet boundary. Thus, the calculations can be complicated, and errors can be accumulated during the calculation procedure. This paper presents an efficient approach for evaluating cumulative strains around penetrometers without the need to backtrack individual streamlines. In this approach, the strain components are treated as field variables. The global solution is obtained using the streamline upwind Petrov-Galerkin method. The new method together with an Eulerian-based finite element formulation was used to study the cone penetration test and evaluate the effect of strain softening on the cone resistance. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Healing articular cartilage defects remains a significant clinical challenge because of its limited capacity for self-repair. While delivery of autologous chondrocytes to cartilage defects has received growing interest, combining cell-based therapies with growth factor delivery that can locally signal cells and promote their function is often advantageous. We have previously shown that PEG thiol-ene hydrogels permit covalent attachment of growth factors. However, it is not well known if embedded chondrocytes respond to tethered signals over a long period. Here, chondrocytes were encapsulated in PEG hydrogels functionalized with transforming growth factor-beta 1 (TGF-β1) with the goal of increasing proliferation and matrix production. Tethered TGF-β1 was found to be distributed homogenously throughout the gel, and its bioactivity was confirmed with a TGF-β1 responsive reporter cell line. Relative to solubly delivered TGF-β1, chondrocytes presented with immobilized TGF-β1 showed significantly increased DNA content, and GAG and collagen production over 28 days, while maintaining markers of articular cartilage. These results indicate the potential of thiol-ene chemistry to covalently conjugate TGF-β1 to PEG to locally influence chondrocyte function over 4 weeks. Scaffolds with other or multiple tethered growth factors may prove broadly useful in the design of chondrocyte delivery vehicles for cartilage tissue engineering applications. © 2014 The Authors. Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc.: 102A: 4464-4472, 2014. [ABSTRACT FROM AUTHOR]