无机三氧化物聚合物对胎鼠颅骨细胞细胞周期及增殖影响研究

Influence of MTA culture with fetal rat osteoblasts on cell cycle and proliferation

目的探讨与胎鼠颅骨细胞共培养时,无机三氧化物聚合物(MTA)对其细胞周期及增殖的影响。方法研究于2012年1月至2013年1月在湖北省十堰市人民医院临床研究所进行。用胰蛋白酶和Ⅱ型胶原酶混合多次消化获得原代胎鼠颅骨细胞,将细胞与MTA共培养(MTA组)4、7、14及21 d,细胞培养加入聚氯乙烯作为对照组。采用流式细胞仪(PI染色)及细胞增殖-毒性试剂盒法(CCK-8法)检测细胞周期中G0/G1、S和G2/M期的细胞数变化及细胞增殖情况。结果 G0/G1期第21天时MTA组细胞明显多于对照组(P<0.05);S期细胞数量在第4、14、21天时MTA组明显少于对照组(P<0.05);G2/M期细胞数量在第4、14天时MTA组明显多于对照组(P<0.05),在第7、21天时MTA组明显低于对照组(P<0.05)。细胞增殖指数(PI):MTA组在第4、7天时均略高于对照组,在第14天时开始降低,第21天时明显低于对照组(P<0.05)。CCK-8检测结果显示实验各组细胞均随培养时间延长而增殖,第14天达到峰值,后开始略有减弱。与对照组比较,MTA组在第4、7天细胞吸光度值略高,随着培养时间继续增加,MTA组吸光度值略低于对照组,但差异均无统计学意义(P>0.05)。结论胎鼠颅骨细胞可以在MTA表面很好地增殖,并且MTA可能通过调节细胞周期来决定细胞的增殖能力,并在7 d后降低细胞增殖速度,这可能对细胞的成骨分化存在一定推动作用。

Objective To research the influence of MTA on cell cycle and proliferation when co-cultured with fetal rat osteoblasts.Methods The structure and proliferation of osteoblasts that were co-cultured on the surface of MTA plates were observed with CCK-8 method respectively. Otherwise,the cell cycles of the osteoblasts were also investigated. The results were analyzed with SPSS 10.0 software.Results The flow cytometer results showed that in the G0/G1 stage cellsproportion of MTA groups were more than control groups（P〉0.05）significantly on the 21 th day（P〈0.05）.The cells incubated with the MTA in the S stage were significantly lower compared with those of control group on the 4th,7th and 21 th day. In the G2/M stage,the MTA group cells were lower than control group（P〈0.05）. Otherwise,in MTA group,the cells on 4th and 14 th were significantly more than control group（P〈0.05）.The PI value increased on the 4th and 7th day and started to reduce on the 14 th day. The results of CCK-8 showed that the cells proliferated at the peak value after being incubated for 14 days. There was no significant difference between the experiment and control group.Conclusion The osteoblasts could not only adhere and proliferate properly on the surface of MTA plates,but also display developing proficiency. The influence on the local microenvironment of absorbable plates may be one of the factors that induce postoperative complications.