title: Decellularized cartilage may be a chondroinductive material for osteochondral tissue engineering.
abstract: Extracellular matrix (ECM)-based materials are attractive for regenerative medicine in their ability to potentially aid in stem cell recruitment, infiltration, and differentiation without added biological factors. In musculoskeletal tissue engineering, demineralized bone matrix is widely used, but recently cartilage matrix has been attracting attention as a potentially chondroinductive material. The aim of this study was thus to establish a chemical decellularization method for use with articular cartilage to quantify removal of cells and analyze the cartilage biochemical content at various stages during the decellularization process, which included a physically devitalization step. To study the cellular response to the cartilage matrix, rat bone marrow-derived mesenchymal stem cells (rBMSCs) were cultured in cell pellets containing cells only (control), chondrogenic differentiation medium (TGF-β), chemically decellularized cartilage particles (DCC), or physically devitalized cartilage particles (DVC). The chemical decellularization process removed the vast majority of DNA and about half of the glycosaminoglycans (GAG) within the matrix, but had no significant effect on the amount of hydroxyproline. Most notably, the DCC group significantly outperformed TGF-β in chondroinduction of rBMSCs, with collagen II gene expression an order of magnitude or more higher. While DVC did not exhibit a chondrogenic response to the extent that DCC did, DVC had a greater down regulation of collagen I, collagen X and Runx2. A new protocol has been introduced for cartilage devitalization and decellularization in the current study, with evidence of chondroinductivity. Such bioactivity along with providing the 'raw material' building blocks of regenerating cartilage may suggest a promising role for DCC in biomaterials that rely on recruiting endogenous cell recruitment and differentiation for cartilage regeneration.
authors.affiliation: University of Kansas Medical Center, Kansas City, Kansas, United States of America.
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Sutherland
authors.affiliation: Bioengineering Graduate Program, University of Kansas, Lawrence, Kansas, United States of America.
authors.forename: Emily C
authors.initials: EC
authors.lastname: Beck
authors.affiliation: Orbis Biosciences, Kansas City, Kansas, United States of America.
authors.forename: S Connor
authors.initials: SC
authors.lastname: Dennis
authors.affiliation: Children's Mercy Hospital, Cardiac Surgery Research Lab, Ward Family Center for Congenital Heart Disease, Kansas City, Missouri, United States of America.
authors.forename: Gabriel L
authors.initials: GL
authors.lastname: Converse
authors.affiliation: Children's Mercy Hospital, Cardiac Surgery Research Lab, Ward Family Center for Congenital Heart Disease, Kansas City, Missouri, United States of America.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Hopkins
authors.affiliation: Department of Chemical and Petroleum Engineering, University of Kansas, Lawrence, Kansas, United States of America; Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, United States of America.
authors.forename: Cory J
authors.initials: CJ
authors.lastname: Berkland
authors.affiliation: Bioengineering Graduate Program, University of Kansas, Lawrence, Kansas, United States of America; Department of Chemical and Petroleum Engineering, University of Kansas, Lawrence, Kansas, United States of America.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Detamore
mesh_terms: D000818:Animals; D002358:Cartilage, Articular / Q000737:chemistry* / Q000166:cytology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D020219:Chondrogenesis; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D005109:Extracellular Matrix / Q000737:chemistry*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013552:Swine; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; C493717:Runx2 protein, rat; D003094:Collagen