(P 231) Investigation of Bone Formation on Collagen-GAG Scaffolds with Varying Composition

• Published in: Tissue engineering. Part A Vol. 14; no. 5; p. 872
• Main Authors: Tierney, C M, Jaasma, MJ, O'Brien, F J
• Format: Journal Article
• Language: English
• Published: 01.05.2008
• ISSN: 1937-3341; 1937-335X

Collagen-glycosaminoglycan (GAG) scaffolds are currently being used in many tissue engineering applications including nerve, cartilage and skin (1). The aim of this study was to investigate the process of in vitro bone formation on a series of collagen-GAG scaffold variants to find an optimal composition for use in bone tissue engineering. Collagen-GAG scaffolds were made as previously described and crosslinked at 105 degree C (2). Scaffolds were divided into two groups: [1] Collagen Variants: 0.5%, 0.75% and 1% Collagen (w/v) and [2] GAG Variants: 0%, 0.022%, 0.044% and 0.088% GAG (w/v). Scaffolds were seeded with 2 x 10 super(6) MC3T3-E1 cells and pre-cultured for 2 days to allow for proliferation before the addition of ascorbic acid and beta -glycerophosphate. Scaffolds were assessed at time points of 1, 3, 7, 14, 21 and 28 days. Cell number, metabolic activity, alkaline phosphatase (pNPP assay) and osteopontin (ELISA) were measured at each time point and scaffolds histologically assessed for mineralisation. Cell number and metabolic activity was highest on the 1% collagen and the 0.088% GAG scaffolds by day 28. Alkaline phosphatase was seen to peak at day 14 although no difference was seen between scaffolds. Nodules of mineralisation within the scaffold stained with alizarin red appeared at day 14 and continued until day 28. The results demonstrate that the 1% collagen and 0.088% GAG scaffolds variants are optimal for bone tissue engineering.