Integration and functional performance of a decellularised porcine superflexor tendon graft in an ovine model of anterior cruciate ligament reconstruction.

The objective was to evaluate the performance of decellularised porcine superflexor tendon (pSFT) as an anterior cruciate ligament (ACL) reconstruction device. The ACL of adult sheep was reconstructed with decellularised pSFT or ovine allograft SFT and animals sacrificed at 4, 12 and 26 weeks (n = 4 per group) for biological evaluation and 26 weeks (n = 6) for biomechanical evaluation of the grafts. Both grafts showed good in vivo performance with no major differences at macroscopic evaluation post euthanasia. Histopathology revealed an inflammatory reaction to both grafts at 4 weeks, which reduced by 26 weeks. There was advanced cellular ingrowth from 12 weeks, ligamentisation of intra-articular grafts, ossification and formation of Sharpey's fibers at the graft/bone junctions. Immunohistochemistry showed that at 4 and 12 weeks, the host response was dominated by CD163+ M2 macrophages and a cell infiltrate comprising α-SMA + myofibroblasts, CD34+ and CD271+ progenitor cells. At 26 weeks the biomechanical properties of decellularised pSFT and oSFT grafts were comparable, with all grafts failing in the intra-articular region. This study provides new insight into constructive remodelling of tendons used for ACL replacement and evidence of integration and functional performance of a decellularised xenogeneic tendon with potential as an alternative for ACL reconstruction.

Decellularisation affects the strain rate dependent and dynamic mechanical properties of a xenogeneic tendon intended for anterior cruciate ligament replacement.

Development of new replacement grafts for anterior cruciate ligament (ACL) repair requires mechanical testing to ensure they can provide joint stability following implantation. A decellularised porcine superflexor tendon (pSFT) has been developed previously as an alternative to current reconstruction methods and subjected to biomechanical analysis. The application of varied strain rates to biological tissues is known to alter their biomechanical properties, however the effects of decellularisation on strain rate dependent and dynamic mechanical behaviour of tissues have not been explored. This study utilised tensile testing to investigate the material properties of native and decellularised pSFTs at three different strain rates (1%.s-1, 10%.s-1 and 100%.s-1). In addition, dynamic mechanical analysis (DMA) was used to ascertain the relative contributions of the solid and fluid phase components of the tissues. Ultimate tensile strength was significantly reduced in decellularised compared with native untreated pSFTs but was unaffected by strain rate. In contrast, toe region moduli increased with increasing strain rate for native tissues, but this effect was not observed in decellularised pSFTs. Linear region moduli were unaffected by strain rate, but were significantly reduced in decellularised pSFT compared with native tissue. Following DMA, significant reductions in dynamic modulus, storage modulus and loss modulus were seen in decellularised compared with native pSFT. Interestingly, the damping ability of the tendons was unaffected by decellularisation, suggesting that solid and fluid phases of the tissue were affected equally. These results, alongside previous studies, suggest that decellularisation affects collagen crimp, tissue swelling and collagen fibre sliding. However, despite these findings, the biomechanical properties of decellularised pSFT remain sufficient to act as an off-the-shelf solution for ACL reconstruction.

Decellularization and Characterization of Porcine Superflexor Tendon: A Potential Anterior Cruciate Ligament Replacement.

The porcine superflexor tendon (SFT) was identified as having appropriate structure and properties for development of a decellularized device for use in anterior cruciate ligament reconstruction. SFTs were decellularized using a combination of freeze-thaw and washes in hypotonic buffer and 0.1% (w/v) sodium dodecyl sulfate in hypotonic buffer plus proteinase inhibitors, followed by nuclease treatment and sterilization using peracetic acid. The decellularized biological scaffold was devoid of cells and cell remnants and contained only 13 ng/mg (dry weight) residual total DNA. Immunohistochemistry showed retention of collagen type I and III and tenascin-C. Quantitative analysis of sulfated sugar and hydroxyproline content revealed a loss of glycosaminoglycans compared with native tissue, but no loss of collagen. The decellularized SFT was biocompatible in vitro and in vivo following implantation in a mouse subcutaneous model for 12 weeks. Uniaxial tensile testing to failure indicated that the gross material properties of decellularized SFTs were not significantly different to native tissue. Decellularized SFTs had an ultimate tensile strength of 61.8 ± 10.3 MPa (±95% confidence limits), a failure strain of 0.29 ± 0.04, and a Young's modulus of the collagen phase of 294.1 ± 61.9 MPa. Analysis of the presence of the α-Gal (galactose-α-1,3-galactose) epitope by immunohistochemistry, lectin binding, and antibody absorption assay indicated that the epitope was reduced, but still present post decellularization. This is discussed in light of the potential role of noncellular α-Gal in the acceleration of wound healing and tissue regeneration in the presence of antibodies to α-Gal.

Vibration stimuli and the differentiation of musculoskeletal progenitor cells: Review of results in vitro and in vivo.

Due to the increasing burden on healthcare budgets of musculoskeletal system disease and injury, there is a growing need for safe, effective and simple therapies. Conditions such as osteoporosis severely impact on quality of life and result in hundreds of hours of hospital time and resources. There is growing interest in the use of low magnitude, high frequency vibration (LMHFV) to improve bone structure and muscle performance in a variety of different patient groups. The technique has shown promise in a number of different diseases, but is poorly understood in terms of the mechanism of action. Scientific papers concerning both the in vivo and in vitro use of LMHFV are growing fast, but they cover a wide range of study types, outcomes measured and regimens tested. This paper aims to provide an overview of some effects of LMHFV found during in vivo studies. Furthermore we will review research concerning the effects of vibration on the cellular responses, in particular for cells within the musculoskeletal system. This includes both osteogenesis and adipogenesis, as well as the interaction between MSCs and other cell types within bone tissue.