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.
1 Institute of Medical and Biological Engineering, School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds , Leeds, United Kingdom .
Gemma
G
Jones
2 Institute of Medical and Biological Engineering, School of Mechanical Engineering, University of Leeds , Leeds, United Kingdom .
Anthony
A
Herbert
3 Tissue Regenix Group, The Biocentre Innovation Way , York, United Kingdom .
Helen
H
Berry
1 Institute of Medical and Biological Engineering, School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds , Leeds, United Kingdom .
Jennifer Helen
JH
Edwards
2 Institute of Medical and Biological Engineering, School of Mechanical Engineering, University of Leeds , Leeds, United Kingdom .
John
J
Fisher
1 Institute of Medical and Biological Engineering, School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds , Leeds, United Kingdom .
Eileen
E
Ingham
D000818:Animals; D016118:Anterior Cruciate Ligament; D005109:Extracellular Matrix / Q000737:chemistry* / Q000637:transplantation*; D013552:Swine; D013708:Tendon Injuries / Q000378:metabolism* / Q000473:pathology / Q000601:surgery*; D013710:Tendons / Q000737:chemistry*
anterior cruciate ligament; decellularization; tendon