不同培养条件下牙周韧带细胞矿化能力的比较研究  

A comparative study of periodontal ligament cells in their mineral characteristics in vitro

作　　者：曹金芳[1] 李纾[1] 庄昭霞[1] 

机构地区：[1]山东大学口腔医院牙周科,山东济南250012

出　　处：《临床口腔医学杂志》2003年第10期610-612,共3页Journal of Clinical Stomatology

基　　金：山东省自然科学基金资助 (Y98C0 90 38)

摘　　要：目的 :探讨牙周韧带细胞体外常规状态及矿化诱导下钙化特性的差异。方法 :用组织块培养法进行人牙周韧带细胞的原代培养 ,取第 4代细胞用于实验 ,在体外长期培养 ,条件培养组加入矿化诱导液 ,常规培养组不加任何矿化诱导因素 ,倒置显微镜下观察矿化情况 ,茜素红与Von -Kossa染色显示钙盐沉积。结果 :牙周韧带细胞在体外长期培养过程中 ,两组均表现为融合期、复层期、结节期、矿化期 ,形成肉眼可见的白色结节 ,茜素红与Von -Kossa染色均显示结节内有钙盐沉积。但常规培养组矿化所需的时间要比条件培养组长 1周左右。结论 :牙周韧带细胞在体外长期培养过程中 ,无论有无矿化诱导因素存在 ,均具有向矿化组织形成细胞分化的趋势 ,但矿化诱导因素的存在可以促进矿化结节的早期形成。Objective:The purpose of this study was to culture human periodontal ligament cells (PDLC) and to observe the mineraligation with or without dexamethasone (Dex), beta-glycerophosphate (Dex), ascorbic acid (AA) in the culture medium during cultured in vitro. Method:Fibroblasts derived from human periodontal ligament were isolated from periodontal ligament explants of healthy premolars extracted for orthdontic reasons. Only the periodontal ligament attached to the middle third of the root was used. The fourth passages were plated in 12-well plates with 10 mm×10 mm cover-glasses and cultured in complete media consisting of DMEM supplement with or without 10-7M dexamethasone, 10mM beta-glycerophosphate, 50μg/ml ascorbic acid. The cells were incubated for 35 days and then were stained for calcium staining (alizarin red staining and Von Kossa,s silver nitrate staining), respectively.Result:PDLC cultured with or without Dex, GP and AA all underwent differentiation, showing four stages (confluent, multilayer, nodule, and mineralization) of disparate morphological characteristics. The cells were all stained by alizarin red and Von Kossa,s silver nitrate, which indicated that PDLC formed mineralized tissues.Conclusion:Irrespetive of cultured with or without Dex, GP and AA, PDLC all underwent differentiation and formed mineralized tissues in vitro.

关 键 词：牙周韧带细胞 细胞培养 矿化 钙化特性 

分 类 号：R781.4[医药卫生—口腔医学]