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

摘要

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>0.05).The expression of TNF-αdecreased in Penicillin-Streptomycin group (P<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 .%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 factor α， 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。