Proliferation and Differentiation of MC3T3-E1 Cells Cultured on Apatite-Coated Honeycomb Collagen

Abstract

The purpose of this study was to develop a new biodegradable bone substitute materials consisting of synthesized hydroxyapatite (HAp) and Type I biodegradable honeycomb collagen sponge (HCS) composites. Apatite-coated Honeycomb collagen composite scaffolds (HAp-HCS) was fabricated and the proliferation and differentiation of MC 3T3-E1 cells were examined for the assessment of their biocompatibility. HAp was combined with Honeycomb Collagen Scaffolds(HCS) using a chemical method. MC 3T3-E1 cells were inoculated into the polystyrene cell culture dishes, HCS and HAp-HCS respectively. The proliferation was assessed using WST-8 assay. The results indicated that MC 3T3-E1 cells grown on the HAp-HCS composite scaffolds showed a higher proliferation rate and spread better than that on the HCS.The differentiation of MC 3T3-E1 cells were characterized by alkaline phosphatase activity. After being cultured in conditioned medium for 14 days, the alkaline phosphatase activities were assessed. The quantitative examination of alkaline phosphatase activity indicated that the cells cultured on the HAp-HCS expressed an activity level about 3 times higher than that on HCS at 14 d. The expression level of characteristic typeⅠcollagen , osteocalcin gene were evaluated using real-time PCR. The typeⅠcollagen gene of cells cultured on HAp-HCS composite scaffolds showed a higher expression level than that on the HCS at 14,21 d . Expression of the osteocalcin mRNA of cells cultured on HAp-HCS showed a higher expression level than that on HCS only at 14 d. The addition of hydroxyapatite in the HCS improved not only the proliferation but also the differentiation of preosteoblast cultured on them. The composite scaffolds showed good biocompatibility and bioactivity. These scaffolds would be promising in bone tissue engineering