title: Effects of Chondroitinase ABC-Mediated Proteoglycan Digestion on Decellularization and Recellularization of Articular Cartilage.
abstract: Articular cartilage has a limited capacity to heal itself and thus focal defects often result in the development of osteoarthritis. Current cartilage tissue engineering strategies seek to regenerate injured tissue by creating scaffolds that aim to mimic the unique structure and composition of native articular cartilage. Decellularization is a novel strategy that aims to preserve the bioactive factors and 3D biophysical environment of the native extracellular matrix while removing potentially immunogenic factors. The purpose of this study was to develop a procedure that can enable decellularization and recellularization of intact articular cartilage matrix. Full-thickness porcine articular cartilage plugs were decellularized with a series of freeze-thaw cycles and 0.1% (w/v) sodium dodecyl sulfate detergent cycles. Chondroitinase ABC (ChABC) was applied before the detergent cycles to digest glycosaminoglycans in order to enhance donor chondrocyte removal and seeded cell migration. Porcine synovium-derived mesenchymal stem cells were seeded onto the decellularized cartilage scaffolds and cultured for up to 28 days. The optimized decellularization protocol removed 94% of native DNA per sample wet weight, while collagen content and alignment were preserved. Glycosaminoglycan depletion prior to the detergent cycles increased removal of nuclear material. Seeded cells infiltrated up to 100 μm into the cartilage deep zone after 28 days in culture. ChABC treatment enhances decellularization of the relatively dense, impermeable articular cartilage by reducing glycosaminoglycan content. ChABC treatment did not appear to affect cell migration during recellularization under static, in vitro culture, highlighting the need for more dynamic seeding methods.
authors.affiliation: Center for Biomedical Engineering, Brown University, Providence, Rhode Island, United States of America.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Bautista
authors.affiliation: Division of Biology and Medicine, Brown University, Providence, Rhode Island, United States of America.
authors.forename: Hee Jun
authors.initials: HJ
authors.lastname: Park
authors.affiliation: Center for Biomedical Engineering, Brown University, Providence, Rhode Island, United States of America.
authors.forename: Courtney M
authors.initials: CM
authors.lastname: Mazur
authors.affiliation: Department of Orthopaedics, Warren Alpert Brown Medical School of Brown University/Rhode Island Hospital, Providence, Rhode Island, United States of America.
authors.forename: Roy K
authors.initials: RK
authors.lastname: Aaron
authors.affiliation: Department of Orthopaedics, Warren Alpert Brown Medical School of Brown University/Rhode Island Hospital, Providence, Rhode Island, United States of America.|Providence VA Medical Center, Providence, Rhode Island, United States of America.
authors.forename: Bahar
authors.identifier: http://orcid.org/0000-0002-7395-2315
authors.initials: B
authors.lastname: Bilgen
mesh_terms: D000818:Animals; D002358:Cartilage, Articular / Q000737:chemistry*; D018929:Cell Culture Techniques; D002465:Cell Movement; D002478:Cells, Cultured; D019765:Chondroitin ABC Lyase / Q000737:chemistry*; D005109:Extracellular Matrix / Q000737:chemistry*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D011509:Proteoglycans / Q000737:chemistry*; D013552:Swine; D013583:Synovial Membrane / Q000166:cytology* / Q000378:metabolism; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D011509:Proteoglycans; D019765:Chondroitin ABC Lyase