Abstract: PURPOSE: To investigate the effects of different hypoxic concentrations on biological characteristics of human dental pulp stem cells in vitro. METHODS: Impacted mandibular third molars were extracted from healthy individuals, and the dental pulp stem cells were cultured by tissue block enzyme digestion. Cells cultured under the conditions of 3%, 5% and 21% oxygen concentration for 7 days were set as 3% hypoxia group, 5% hypoxia group, and 21% nomoxia group, respectively. Flow cytometry was used to detect cell surface markers, cell cycle and apoptosis. Cell proliferation was measured by CCK-8 method. Transwell chamber assay was used to detect migration ability. Statistical analysis was completed by SPSS 20.0 software package. RESULTS: The expression rates of CD44, CD29 and D73 of the subculture cells were 97.25%, 99.36% and 99.60%, respectively. The proliferation ability of dental pulp stem cells was the strongest in 5% hypoxia group, and weakest in 3% hypoxia group, with significant difference(P<0.05). The apoptosis rate had no significant difference among various concentrations of oxygen(P>0.05). Compared with 21% nomoxia group, the proportion of dental pulp stem cells in G1 phase was significantly lower than that in 3% hypoxia group and 5% hypoxia group(P<0.05), and cell in S phase was significantly higher than that in 3% hypoxia group and 5% hypoxia group(P<0.05). The migration ability was the strongest in 3% hypoxia group, and weakest in 21% nomoxia group, with significant difference(P<0.05). CONCLUSIONS: Different concentrations of hypoxia have great influence on the morphology, proliferation, migration and cell cycle of human dental pulp stem cells in vitro with little impact on cell apoptosis.

Key words: Hypoxic concentration, In vitro culture, Human dental pulp stem cells, Biological characteristics