Development and characterisation of a decellularised bovine osteochondral biomaterial for cartilage repair

Abstract

It is proposed that an acellular natural osteochondral scaffold will provide a successful repair material for the early intervention treatment of cartilage lesions, to prevent or slow the progression of cartilage deterioration to osteoarthritis. Here, we investigated the efficacy of methods for the decellularisation of bovine osteochondral plugs. The plugs were subject to four freeze/thaw cycles followed by two cycles of washes in hypotonic solution and low concentration (0.1 % w/v) sodium dodecyl sulphate with protease inhibitors. Plugs were treated with nuclease (DNase and RNase) treatment followed by sterilization in peracetic acid. Full tissue decellularisation was achieved as confirmed by histological analysis and DNA quantification, however the resultant acellular matrix had reduced glycosaminoglycan content which led to an increased percent deformation of cartilage. Furthermore, the acellular scaffold was not reproducibly biocompatible. Additional terminal washes were included in the process to improve biocompatibility, however, this led to visible structural damage to the cartilage. This damage was found to be minimised by reducing the cut edge to cartilage area ratio through decellularisation of larger cuts of osteochondral tissue.

Effects of Chondroitinase ABC-Mediated Proteoglycan Digestion on Decellularization and Recellularization of Articular Cartilage

“…Our decellularization protocol using chABC resulted in a 94% decrease in dsDNA/WW, in keeping with previous studies [ 34 , 35 ]. Furthermore, similar to these previous studies, the final dsDNA content reported here falls below the 50 ng/mg ECM dry weight threshold for prevention of adverse host reactions described by Crapo et al [ 46 ].…”

Biphasic hierarchical extracellular matrix scaffold for osteochondral defect regeneration

“…Thus, preparing an osteochondral ECM scaffold by decellularization can provide a means of recreating osteochondral tissue architecture. Recently, though several studies have reported that osteochondral ECM scaffold was fabricated and applied to osteochondral defect repairing, its evaluation in cell removal effect, stratified structure retention and regeneration mechanism were still uncertain 21,22 .…”

Development and characterisation of a low-concentration sodium dodecyl sulphate decellularised porcine dermis

“…Histological sections from three replicate samples of cellular split-thickness porcine skin, decellularised porcine dermis and decellularised human dermis from each porcine and human donor were stained using 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) for visualisation of nuclear material. 11 A commercially available kit (DNAeasy kit, Qiagen) was used to isolate the DNA from control split-thickness porcine skin, decellularised porcine dermis and decellularised human dermis. Six replicate samples per pig/donor in each group were processed.…”