Enhanced Osteogenic and Angiogenic-Related Gene Expression of Human Dental Stem Cells on Biphasic Calcium Phosphate Scaffold Treated with Vascular Endothelial Growth Factor: Part I

Abstract

To study the effectiveness of vascular endothelial growth factor (VEGF) added with biphasic calcium phosphate (BCP) on the expression of osteogenesis and angiogenesis-related gene in dental stem cells (DSCs). The cells were treated with three different modalities; BCP group, VEGF group, and VEGF-added-BCP. The optimal BCP and VEGF concentrations were determined. The cells were harvested at four different time intervals (day 3, day 7, day 10 and day 14) and were subjected to RNA isolation. Osteogenesis and angiogenesis-regulated genes was amplified using reverse transcriptase-PCR (RT-PCR). The RT-PCR products were then electrophoresed. The gel images were captured using Image Analyser AlphaEaseFC™. Angiogenesis and osteogenesis genes were clearly expressed in DSCs in response to treatments with 75mg/ml BCP and 5ng/ml VEGF. Angiogenesis gene VEGF was highly expressed by VEGF treatment group but showed some changes when added with BCP. Osteogenesis genes (BMP-2 and OPN) were positively affected by both BCP and VEGF. Some genes were expressed at an earlier time interval compared to the other genes depending on the type of treatments. BCP treatment induced high expression of initial-regulated osteogenesis genes (BMP-2 and OPN). Combination of BCP and VEGF modality on DSCs was suggested to initiate osteogenesis and angiogenesis-related gene expressions earlier than the other modalities. In our design, uncontrolled release of VEGF protein showed inhibition of BMP-2 at mRNA level in VEGF-BCP combined group.