青霉素和链霉素对炎症牙髓干细胞的增殖、成牙成骨分化能力及TNF-α表达的影响

Effect of Penicillin-Streptomycin on the proliferation,odonto-and osteogenic differentiation potential and the expression of the inflammatory cytokines TNF-α of the inflamed human dental pulp stem cells

目的:探讨青霉素和链霉素对炎症牙髓干细胞的增殖、成牙成骨分化能力及肿瘤坏死因子α(tumor necrosis factorα,TNF-α)表达的影响。方法:采用酶消化法分离培养炎症牙髓干细胞,采用甲基噻唑基四唑(MTT)比色法、碱性磷酸酶活性检测、Western blot及实时定量RT-PCR等方法分析青霉素和链霉素作用于炎症牙髓干细胞后,其增殖和成牙成骨分化指标(核心结合因子、牙本质涎蛋白/牙本质涎磷蛋白、骨钙素)及TNF-α表达的变化。结果:青霉素和链霉素作用于炎症牙髓干细胞后,与未加抗生素的培养组细胞的增殖能力及成骨/成牙相关蛋白、基因的表达无明显差异(P>0.05),而TNF-α的表达则低于普通培养组(P<0.01)。结论:青霉素和链霉素降低炎症牙髓干细胞的炎性因子TNF-α表达,对细胞的增殖及成牙/成骨分化能力无明显影响。

Objective：To observe the effects of Penicillin-Streptomycin on the proliferation , odonto-and osteogenic differentiation and the expression of the inflammatory cytokines of inflamed human dental pulp stem cells （iDPSCs） in vitro.Methods：iDPSCs were isolated from inflamed human dental pulp and cultured respectively in alpha minimum essential medium （α-MEM） or containing 100 U/mL Penicillin-Streptomycin.The proliferation ability of iDPSCs was evaluated by MTT colorimetric assay .Alkaline phosphotase （ALP） activity, real-time RT-PCR and Western blot were used to examine the odonto /osteogenic potential-the the expression of RUNX2/RUNX2,DSP/DSPP,OCN/OCN and the expression of TNF-αof iDPSCs.Results：MTT assay showed that 100U/ml Penicillin-Strepto-mycin had no effect on the proliferation of iDPSCs , ALP activity assay , real-time RT-PCR and Western blot showed that there was no difference between Penicillin-Streptomycin group and α-MEM group （ P＆gt;0.05）.The expression of TNF-αdecreased in Penicillin-Streptomycin group （P＆lt;0.01）.Conclusions：Penicillin-Streptomycin significantly suppressed the expression of inflammatory cytokines in inflammatory dental pulp stem cells , and has no noticeable effect on the proliferation and differentiation of the cells .