Bioactive silicate materials stimulate angiogenesis in fibroblast and endothelial cell co-culture system through paracrine effect

Angiogenesis is critical in tissue engineering, and bioceramic-induced angiogenesis has been reported. However, the role of other types of cells such as fibroblasts in this bioceramic-induced angiogenesis process has not been reported, and is closer to the in vivo situation of tissue regeneration. In this study, the paracrine effect of silicate bioceramic-induced angiogenesis in the presence of fibroblasts was confirmed by investigating the effects of calcium silicate (CS), one of the simplest silicate bioactive ceramics, on angiogenesis in co-cultures of human dermal fibroblasts (HDF) and human umbilical vein endothelial cells (HUVEC). Results showed that CS extracts stimulated the expression of vascular endothelial growth factor (VEGF) from co-cultured HDF and subsequently enhanced the expression of VEGF receptor 2 on co-cultured HUVEC (co-HUVEC). The endothelial nitric oxide synthase and nitric oxide production in co-HUVEC was then increased to finally initiate the proangiogenesis. During this process, the expression of vascular endothelial cadherin from co-HUVEC was up-regulated, and cadherin proteins were concentrated at the cell junctions to facilitate tube formation. Silicon ions are confirmed to play an important role during silicate bioceramic-inducing angiogenesis, and effective silicon ion concentrations (0.7–1.8 μg ml−1) are proposed.