The effects of methotrexate on the biological properties of dental pulp stem cells

TANG Ying; ZHU Qi; GU Yong-chun; SHEN Yi-fen; FENG Xing-mei

doi:10.15956/j.cnki.chin.j.conserv.dent.2017.02.002

Abstract：AIM:To investigate the effects of methotrexate (MTX) on the biological properties of dental pulp stem cells (hDPSCs).METHODS:hDPSCs were isolated,cultured and identified from dental pulp of young permanent teeth extracted for orthodontic reasons.The hDPSCs were divided into 4 groups,and were cultivated in media containing 0 (control),0.1,1.0 and 10.0 μmol/L MTX respectively.The effects of MTX on the percentage of colony-forming unit (CFU-F),cell proliferation (tested by CCK-8 method),migration (scratch assay),cell circle phase (flow cytometric analysis) and osteogenic potential (Alizarin red staining and Western blot assay of expression of RUNX2,OSX and ALP) were examined.RFSULTS:The capability of CFU-F and migration was significantly reduced by the treatment of MTX concentration in a dose-related manner.The middle and high concentration groups exhibited more inhibition on cell proliferation than the low concentration group (P ＜ 0.05),while there was no statistical difference between 1.0 and 10.0 μmol/L MTX groups (P ＞ 0.05).Flow cytometric analysis showed that MTX treatment inhibited hDPSCs proliferation,and the percentage of S-phase cells increased from 35.4％ to 60.3％-64.5％ (P ＜ 0.05).Whereas there was no statistical difference among the 3 MTX groups (P ＞ 0.05).Alizarin red staining test indicated a decrease of calcified nodules with increase of MTX concentration.However,Western blot analysis showed the expression level of RUNX2,OSX,and ALP had no statistical difference among the 4 groups (P ＞ 0.05).CONCLUSION:MTX has adverse effects on multiple biological properties of hDPSCs,which can be influenced by the drug concentration and time of the treatment.