重组全长人釉原蛋白对人成纤维细胞黏附、增殖、迁移的影响  

Effects of rhAm on attachment,proliferation and immigration of human fibroblasts

作　　者：林智恺[1] 束蓉[1] 程岚[1] 宋忠臣[1] 郭秋曼[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·口腔医学院牙周科,上海200011 [2]上海市口腔医学研究所、上海市口腔医学重点实验室,上海200011

出　　处：《上海口腔医学》2013年第1期7-12,共6页Shanghai Journal of Stomatology

基　　金：国家自然科学基金(81070838,81271156);上海交通大学医工交叉研究基金(YG2011MS31))~~

摘　　要：目的：比较重组25kDa全长人釉原蛋白（recombinanthumanamelogenin，rhAm）和提取的猪釉基质蛋白（enamelmatrixproteins，EMPs）对人牙周膜成纤维细胞（humanperiodontalligamentfibroblasts．HPDLF）和人包皮成纤维细胞（foreskinfibroblasts，HFF）生物学特性的影响，初步探讨rhAm促进牙周组织再生的可能机制。方法：选择BL21／pET28a—His—SUMO—rhAm表达系统，在适宜的条件下，经诱导表达和酶切纯化．得到25kDa全长rhAm．采用SDS—PAGE和Western印迹鉴定，乙酸法提纯EMPs。体外培养HPDLF和HFF．采用不同浓度的rhAm和EMPs分别刺激HPDLF、HFF，观察并比较2种蛋白对细胞黏附、增殖和迁移能力的影响。采用SAS5．0软件包对数据进行统计学分析。结果：10—20μg／mLrhAm能显著促进人牙周膜成纤维细胞和人包皮成纤维细胞的黏附、增殖和迁移（P〈0．05），其作用效果与200μg／mLEMPs相似（P〉0．05）。结论：25kDarhAm和EMPs对成纤维细胞具有相似的生物学作用。rhAm和EMPs可通过增强牙周膜成纤维细胞的生物学活性．从而促进牙周组织再生。PURPOSE： To compare the effects of 25 kDa full-length rhAm and porcine EMPs on cell behaviors of human periodontal ligament fibroblasts （HPDLF） and foreskin fibroblasts（HFF）. METHODS： rhAm was induced by BL21/ pET28a-His-SUMO-rhAm express system, and 25 kDa full-length rhAm was analyzed by SDS-PAGE and Western blot. EMPs were extracted by acetic acid method. HPDLF and HFF were cultured in vitro. The cells were treated with rbAm and EMPs at different concentrations. The cell adhesion, proliferation and migration assays were qualitatively analyzed. The data was statistically analyzed with SAS 5.0 software package. RESULTS： 10-20 Ixg/mL rhAm significantly promoted the adhesion, proliferation and migration of HPDLF and HFF （P〈0.05）, but no significant difference between two proteins was found （P〉0.05）. CONCLUSIONS： 25 kDa rhAm and EMPs shows similar biological effects on fibroblast, which indicates that rhAm may play an important role in the periodontal regeneration through the activation of fibroblasts. Supported by National Natural Science Foundation of China （81070838, 81271156） and Biomedical Engineering Cross Research Foundation of Shanghai Jiao Tong University （YG2011MS31）.

关 键 词：重组人釉原蛋白 猪釉基质蛋白 成纤维细胞 增殖 黏附 迁移 

分 类 号：R780.2[医药卫生—口腔医学]