Low-dose β-glycerophosphate induced differentiation of dental pulp stem cells into odontoblasts and expressions of relevant factors / 中国组织工程研究

ABSTRACT

BACKGROUND:The induced concentration for osteoblasts is often introduced as reference to induce odontoblast differentiation. However, there are no reports on other concentrations. OBJECTIVE: To observe the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein during low-dose β-glycerophosphate-induced differentiation of dental pulp stem cels into odontoblasts. METHODS:Human dental pulp stem cels were isolated and cultured, and then induced by different concentrations of inducing solution to differentiate into adipocytes and osteoblasts, which could verify the multi-directional differentiation ability of human dental pulp stem cels. Under 5 mmol/L β-glycerophosphate, dental pulp stem cels differentiated into odontoblasts. At 7, 14, 21, 28 days of culture, RNA samples were extracted from dental pulp stem cels in each group, and reverse-transcription PCR was used to detect the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein. Mineralized nodules were detected by alizarin red S staining to show the successfuly osteogenesis induction. RESULTS AND CONCLUSION: Human dental pulp stem cels could be induced to adipocytes and osteoblasts. The results of reverse-transcription PCR showed that the dental pulp stem cels under 5 mmol/L β-glycerophosphate could increase the expression of dentin matrix protein-1 and dentin sialoprotein, but downregulate the expression of matrix extracelular phosphoglycoprotein at 7, 14, 21 days. At 28 days of culture, dental pulp stem cels were al successfuly mineralized detected by alizarin red S. There were some red mineralized nodules. These findings indicate that the 5 mmol/L β-glycerophosphate can induce the differentiation of dental pulp stem cels into odontoblasts successfuly, up-regulate the mRNA expression of dentin sialoprotein and dentin matrix protein-1, and meanwhile down-regulate the mRNA expression of matrix extracelular phosphoglycoprotein.