Osteoblast-oriented differentiation of BMSCs by co-culturing with composite scaffolds constructed using silicon-substituted calcium phosphate, autogenous fine particulate bone powder and alginate in vitro

Abstract

Autogenous bone graft is the best for spinal fusion in clinics, however, lacking sources, bleeding and infection are limited its practice. Seeking alternative materials are urgent for orthopaedic surgeon. Here, we evaluated osteoblast-oriented differentiation of rabbit BMSCs by co-culturing with composite scaffolds constructed using silicon-substituted-CaP-fine particulate bone powder-alginate. Using CCk8-kit, biocompatibility was evaluated by testing BMSCs proliferation; morphology and survival of osteoblasts within scaffolds were observed using EM and HE staining; growth factors and related genes were detected using RT-PCR. HE staining showed spindle-shaped BMSCs after the 3rd passage; EM data showed that uneven surface and longitudinal section were observed with scattered distribution of 5-100 mm interspaces, which leave enough space for BMSCs adhesion and growth. Interestingly, at 14-day culture with HE staining, osteocytes within the scaffolds grew well with regular shape and integrate structure. RT-PCR results showed that expression levels of BMP2, TGF-b and COL-I, ALP, OPN were increased significantly and time-dependently. Collectively, all mentioned effects were more obvious in co-culture BMSCs with scaffolds than those with other components. Immunohistochemistry showed that positive OPN expression was detected at 7-day co-culturing BMSCs with scaffold, rather than other situations. These results suggest that composite scaffolds constructed with Si-CaP-fine particulate bone powder-alginate have a certain degree of biocompatibility and bioactivity to promote osteoblast-oriented BMSCs differentiation.