Development and characterization of an acellular porcine medial meniscus for use in tissue engineering.
The objectives of this study were to characterize fresh porcine menisci and develop a decellularization protocol with a view to the generation of a biocompatible and biomechanically functional scaffold for use in tissue engineering/regeneration of the meniscus. Menisci were decellularized by exposing the tissue to freeze-thaw cycles, incubation in hypotonic tris buffer, 0.1% (w/v) sodium dodecyl sulfate in hypotonic buffer plus protease inhibitors, nucleases, hypertonic buffer followed by disinfection using 0.1% (v/v) peracetic acid and final washing in phosphate-buffered saline. Histological, immunohistochemical, and biochemical analyses of the decellularized tissue confirmed the retention of the major structural proteins. There was, however, a 59.4% loss of glycosaminoglycans. The histoarchitecture was unchanged, and there was no evidence of the expression of the major xenogeneic epitope, galactose-alpha-1,3-galactose. Biocompatibility of the acellular scaffold was determined by using contact cytotoxicity and extract cytotoxicity tests. Decellularized tissue and extracts were not cytotoxic to cells. Biomechanical properties were determined by indentation and tensile tests, which confirmed the retention of biomechanical properties following decellularization. In conclusion, this study has generated data on the production of a biocompatible, biomechanically functional scaffold for use in meniscal repair.
Stapleton
Thomas W
TW
Institute of Molecular and Cellular Biology, The University of Leeds, Leeds, United Kingdom. t.w.stapleton01@leeds.ac.uk
Ingram
Joanne
J
Katta
Jaynath
J
Knight
Richard
R
Korossis
Sotirios
S
Fisher
John
J
Ingham
Eileen
E
D000818:Animals; D001672:Biocompatible Materials; D001696:Biomechanical Phenomena; D002356:Cartilage; D002474:Cell-Free System; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D005260:Female; D006025:Glycosaminoglycans; D006245:Hardness Tests; D006909:Hydroxyproline; D008592:Menisci, Tibial / Q000033:anatomy & histology / Q000166:cytology* / Q000648:ultrastructure; D034421:Sus scrofa; D013718:Tensile Strength; D023822:Tissue Engineering / Q000379:methods*; D016707:Tissue Fixation; D054457:Tissue Scaffolds; D016474:Weight-Bearing; D000519:alpha-Galactosidase
D001672:Biocompatible Materials; D006025:Glycosaminoglycans; D000519:alpha-Galactosidase; D006909:Hydroxyproline