Abstract
Porous β-CaSiO3/β-Ca3(PO4)2 (β-CS/β-TCP) composite scaffolds have been previously shown to promote bone formation in vivo. However, the mechanisms underlying such beneficial effects remain unclear. In this study, we recreated an extracellular environment using the extracts of β-CS/β-TCP composites developed in our previous in vivo study, and investigated the effects of the extracts on osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBMSCs) and its related mechanisms. The angiogenic potential of the extracts was also evaluated using human umbilical vein endothelial cells (HUVECs). In the absence of osteogenic supplements, the osteogenic differentiation of rBMSCs was detected by alkaline phosphatase (ALP) activity assay and the messenger RNA expression of a panel of osteoblast markers. The results showed that the soluble ions of porous β-CS/β-TCP composites were capable of promoting cell viability, directly inducing cell differentiation. The increase in phosphorylation of AMP-activated protein kinase (AMPK) and ERK1/2 were observed in rBMSCs cultured in β-CS/β-TCP composite extracts. The ALP expression, calcium deposition, and ERK1/2 phosphorylation of rBMSCs, which was promoted by ions released from β-CS/β-TCP composites, were blocked by an AMPK inhibitor, Compound C. These results indicate that bioactive ions extracted from β-CS/β-TCP composites could stimulate the osteogenic differentiation of rBMSCs via the AMPK-Erk1/2 pathway. Interestingly, the secretion of vascular endothelial growth factor and the viability of HUVECs were shown to be enhanced in the presence of extracts from the β-CS/β-TCP composite scaffolds. Our findings suggest that 50 or 80% wt. CS could promote bone regeneration by stimulating osteogenesis and angiogenesis.
Published Version
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