In vitro biocompatibility and osteoblast differentiation of an injectable chitosanano-hydroxyapatite/collagen scaffold

摘要

The purpose of this study was to evaluate the in vitro cell biocompatibility of an in situ forming composite consisting of chitosan (CS), nano-hydroxyapatite and collagen (nHAC), which has a complex hierarchical structure similar to natural bone. MC3T3-E1 mouse calvarial preosteoblasts were cultured on the surface of the injectable CSHAC and CS scaffold. The proliferations of seeded MC3T3-E1 were investigated for 10 days. Cytotoxicity, cell proliferation, and cell expression of osteogenic markers such as alkaline phosphatase (ALP), type 1 collagen (COL-1), RUNX-2, and osteocalcin (OCN) were examined by biochemical assay and reverse transcription polymerase chain reaction. Cell viability and total cellularity (measured by dsDNA) were similar between the two scaffold groups. However, ALP, COL-1, OCN, and RUNX-2 production were significantly greater when osteoblasts were cultured on CSHAC scaffolds. The increase in osteogenic markers production on CSHAC scaffolds indicated that these scaffolds were superior to chitosan-only scaffolds in facilitating osteoblast mineralization. These results demonstrate the potential of the CSHAC scaffolds to be used in bone tissue engineering.