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 decellularised device for use in ACL reconstruction. SFTs were decellularised using a combination of freeze thaw and washes in hypotonic buffer and 0.1% (w/v) SDS in hypotonic buffer plus proteinase inhibitors followed by nuclease treatment and sterilisation using peracetic acid. The decellularised biological scaffold was devoid of cells and cell remnants and contained only 13 ng.mg-1 (dry weight) residual total DNA. Immunohistochemistry showed retention of collagens type I and III and tenascin-C. Quantitative analysis of sulphated sugar and hydroxyproline content revealed a loss of glycosaminoglycans compared to native tissue but no loss of collagen. The decellularised 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 decellularised SFT were not significantly different to native tissue. Decellularised 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 alpha-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 decellularisation. This is discussed in light of the potential role of non-cellular alpha-Gal in the acceleration of wound healing and tissue regeneration in the presence of antibodies to alpha-Gal.