Journal
BIOMATERIALS
Volume 27, Issue 7, Pages 1043-1053Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2005.07.032
Keywords
chondrocyte; gene expression; surface modification; PCR; fibronectin; vitronectin; cartilage tissue engineering
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Optimizing re-differentiation of clinically relevant cell sources on biomaterial substrates in serum containing (S+) and serum-free (SF) media is a key consideration in scaffold-based articular cartilage repair strategies. We investigated whether the adhesion and post-expansion re-differentiation of human chondrocytes could be regulated by controlled changes in substrate surface chemistry and composition in S+ and SF media following gas plasma (GP) treatment. Expanded human nasal chondrocytes were plated on gas plasma treated (GP+) or untreated (GP-) poly(ethylene glycol)-terephthalate-poly(butylene terephthalate) (PEGT/PBT) block co-polymer films with two compositions (low or high PEG content). Total cellularity, cell morphology and immunofluorescent staining of vitronectin (VN) and fibronectin (FN) integrin receptors were evaluated, while post-expansion chondrogenic phenotype was assessed by collagen types I and II mRNA expression. We observed a direct relationship between cellularity, cell morphology and re-differentiation potential. Substrates supporting high cell adhesion and a spread morphology (i.e. GP+ and low PEG content films), resulted in a significantly greater number of cells expressing alpha(5)beta(1) FN to alpha(V)beta(3) VN integrin receptors, concomitant with reduced collagen type II/I mRNA gene expression. Substrates supporting low cell adhesion and a spherical morphology (GP- and high PEG content films) promoted chondrocyte redifferentiation indicated by high collagen type II/I gene expression and a low percentage of alpha(5)beta(1) FN integrin expressing cells. This study demonstrates that cell-substrate interactions via alpha(5)beta(1) FN integrin mediated receptors negatively impacts expanded human nasal chondrocyte re-differentiation capacity. GP treatment promotes cell adhesion in S + media but reverses the ability of low PEG content PEGT/PBT substrates to maintain chondrocyte phenotype. We suggest alternative cell immobilization techniques to GP are necessary for clinical application in articular cartilage repair. (c) 2005 Elsevier Ltd. All rights reserved.
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