釉基质蛋白对猪骨髓基质细胞增殖和根面附着生长的影响  

Effects of enamel matrix proteins on the proliferation and attachment of porcine bone marrow stromal cells on autogeneous root surface in vitro

作　　者：宋爱梅[1] 束蓉[1] 谢玉峰[1] 吴安平[1] 刘晓峰[1] 张秀丽[2] 

机构地区：[1]上海交通大学医学院附属第九人民医院口腔内科,200011 [2]上海交通大学医学院附属第九人民医院

出　　处：《实用口腔医学杂志》2006年第5期679-683,共5页Journal of Practical Stomatology

基　　金：国家863计划资助课题(2002AA205011);上海市教育发展基金会资助课题(2002-02)

摘　　要：目的:研究釉基质蛋白(enamel matrix proteins,EMPs)对体外培养的猪骨髓基质细胞(bone marrowstrom alcells,BMSCs)增殖和在根面附着生长的影响,为EMPs联合应用BMSCs修复牙周组织缺损提供理论依据。方法:抽取猪髂骨骨髓,全血培养法获得骨髓基质细胞。培养液中EMPs的浓度分别为25、50、100、200μg/ml,以不加EMPs为对照。MTT法测定各组细胞的增殖活性。对实验数据行单因素方差分析和SNK法组间比较,检验水准为α=0.05。制备猪自体牙根片,以200μg/mlEMPs处理组为实验组,对照组不用EMPs处理。接种BMSCs后培养7d,HE染色和扫描电镜观察。选取800倍下标准视野,计数每个视野中的细胞数,取4个视野均值。采用配对t检验法作统计学分析,检验水准为α=0.05。结果:猪BMSCs在含有EMPs的培养液中生长良好。EMPs对BMSCs的促增殖作用呈浓度和时间依赖性,200μg/ml浓度的EMPs从实验的第3天开始显著促进猪BMSCs增殖。HE染色显示BMSCs在根片表面附着良好。扫描电镜观察表明实验组附着生长的BMSCs数量较多,与对照组相比具有显著差异。结论:EMPs在200μg/ml浓度时可显著促进猪BMSCs增殖,并促进其在根面的附着生长,提示EMPs和BMSCs可以联合应用以修复牙周组织缺损。Objective：To study the effects of enamel matrix proteins（EMPs） on the proliferation of porcine bone marrow stromal cells （BMSCs） and on the adhesion of BMSCs cultured on autogeneous root surface. Methods： BMSCs were obtained from porcine bone marrow aspiration and cultured in DMEM medium with 100 ml/L FBS. The cells of third passage were exposed to EMPs at 25,50,100 and 200 μg/ml respectively for 9 days. Controls were BMSCs cultured in DMEM medium without EMPs. BMSCs proliferation rate was examined by MTT assay. Root dentine specimens were prepared from extracted teeth of three porcines and distributed into 2 groups. In the control group, BMSCs were directly cultured on the specimen surface. BMSCs of the experiment group were cultured on the specimen surface pretreated with EMPs at a concentration of 200 μg/ml. After 7 d incubation, the medium was removed, specimens were fixed, processed for HE staining and SEM examination. BMSCs adherence was measured by counting the number of cells within a standard test area. The data were analysed by paired t test. Results ： BMSCs could well attached and spread on EMPs-coated substrata. The proliferation of BMSCs was significantly stimulated by EMPs in a dose- and time-dependent manner, and EMPs at a concentration of 200 μg/ml significantly enhanced BMSCs proliferation from day 3 over the experiment. A continuous line of BMSCs along the root surface was observed by HE staining, and SEM observation demonstrated a significant increase in number of adherent BMSCs treated by EMPs when compared with the controls（ P 〈 0.05 ）. Conclusions： EMPs at the concentration of 200 μg/ml can stimulate the proliferation of BMSCs and enhance the BMSCs attachment and growth on the root surface.

关 键 词：釉基质蛋白 骨髓基质细胞 增殖 根面 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]