Isolation of mesenchymal stem cells from human alveolar periosteum and effects of vitamin D on osteogenic activity of periosteum-derived cells

Mesenchymal stem cells (MSCs) are present in a variety of tissues and can be differentiated into numerous cell types, including osteoblasts. Among the dental sources of MSCs, the periosteum is an easily accessible tissue, which has been identified to contain MSCs in the cambium layer. However, this source has not yet been widely studied. Vitamin D₃ and 1,25-(OH)₂D₃ have been demonstrated to stimulate in vitro differentiation of MSCs into osteoblasts. In addition, vitamin C facilitates collagen formation and bone cell growth. However, no study has yet investigated the effects of Vitamin D₃ and Vitamin C on MSCs. Here, we present a method of isolating MSCs from human alveolar periosteum and examine the hypothesis that 1,25-(OH)₂D₃ may exert an osteoinductive effect on these cells. We also investigate the presence of MSCs in the human alveolar periosteum and assess stem cell adhesion and proliferation. To assess the ability of vitamin C (as a control) and various concentrations of 1,25-(OH)₂D₃ (10⁻¹⁰, 10⁻⁹, 10⁻⁸, and 10⁻⁷ M) to alter key mRNA biomarkers in isolated MSCs mRNA expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), core binding factor alpha-1 (CBFA1), collagen-1, and osteocalcin (OCN) are measured using real-time polymerase chain reaction (RT-PCR).