MDP⁃钙盐对牙髓干细胞增殖和成牙本质分化的影响

Effects of MDP⁃calcium salts on the proliferation and odontogenic differentiation of dental pulp stem cells

目的:探索甲基丙烯酰氧基癸基磷酸二氢酯(MDP)⁃钙盐对牙髓干细胞(DPSCs)成牙本质分化的影响。方法:使用不同比例的10⁃MDP与CaCl2合成三种MDP⁃钙盐;提取人DPSCs,通过CCK⁃8实验研究不同浓度MDP⁃钙盐对细胞增殖的影响,碱性磷酸酶(ALP)活力测试选取最适浓度,茜素红染色和实时荧光定量PCR研究MDP⁃钙盐对DPSCs成牙本质分化的影响。结果:一定浓度范围MDP⁃钙盐对DPSCs增殖无抑制作用,MDP⁃钙盐促进DPSCs的ALP活力的最佳浓度为0.2 mg/mL(P<0.05),0.2 mg/mL MDP⁃2钙盐促进DPSCs钙结节形成(P<0.05),0.2 mg/mL MDP⁃1钙盐和MDP⁃2钙盐对DPSCs中牙本质基质蛋白⁃1(DMP⁃1)的分泌有促进作用(P<0.05),0.2 mg/mL MDP⁃0.5钙盐和MDP⁃1钙盐对DPSCs中牙本质涎磷蛋白(DSPP)的分泌有促进作用(P<0.05),0.2 mg/mL MDP⁃0.5钙盐和MDP⁃2钙盐对DPSCs中骨钙素(OCN)的分泌有促进作用(P<0.05),0.2 mg/mL MDP⁃0.5钙盐、MDP⁃1钙盐和MDP⁃2钙盐对DPSCs中Runt相关转录因子2(RUNX2)的分泌有促进作用(P<0.05)。结论:MDP⁃钙盐在一定浓度范围内对DPSCs增殖无抑制作用,0.2 mg/mL MDP⁃钙盐对DPSCs成牙本质分化有促进作用。

Objective:To investigate the effects of methacryloyloxydecyl dihydrogen phosphate(MDP)⁃calcium salts on the prolif⁃eration and odontogenic differentiation of dental pulp stem cells(DPSCs).Methods:Three types of MDP⁃calcium salts were synthe⁃sized using different ratios of 10⁃MDP and CaCl2,and Human DPSCs were harvested and cultured in medium.The effect of different concentrations of MDP⁃calcium salts on cell proliferation was determined by CCK⁃8 assay.Alkaline phosphatase(ALP)activity was measured to screen the best concentration for subsequent experiments.Additionally,alizarin red staining,and real⁃time quantitative polymerase chain reaction(qRT⁃PCR)were employed to study the influence of a specific concentration of MDP⁃calcium salts on the odontogenic differentiation of DPSCs.Results:A certain concentration range of MDP⁃calcium salts had no inhibitory effect on the pro⁃liferation of DPSCs.Compared with the control group,0.2 mg/mL was the optimal concentration of MDP⁃calcium salts in inducing ALP activity of DPSCs(P<0.05).Alizarin red staining results showed that 0.2 mg/mL MDP⁃2 calcium salts promoted the formation of calci⁃um nodules in DPSC 0.2 mg/mL MDP⁃1 calcium salts and 0.2 mg/mL MDP⁃2 calcium salts promoted the secretion of odontogenic dif⁃ferentiation marker dental matrix protein⁃1(DMP⁃1)(P<0.05).Additionally,0.2 mg/mL MDP⁃0.5 calcium salts and 0.2 mg/mL MDP⁃1 calcium saltsenhance the secretion of dentin sialophosphoprotein(DSPP)(P<0.05).Furthermore,0.2 mg/mL MDP⁃0.5 calci⁃um salts and 0.2 mg/mL MDP⁃2 calcium salts also enhanced the secretion of osteocalcin(OCN)(P<0.05).Lastly,0.2 mg/mL MDP⁃0.5 calcium salts,0.2 mg/mL MDP⁃1calcium salts and 0.2 mg/mL MDP⁃2 calcium salts stimulated the secretion of runt⁃related tran⁃scription factor 2(RUNX2)(P<0.05).Conclusions:MDP⁃calcium salts can not inhibit the proliferation of DPSCs within a certain concentration range.0.2 mg/mL MDP⁃calcium salts can promote the odontogenic differentiation of DPSCs.