摘要
目的:体外培养长结合上皮(long junctional epithelium,LJE)细胞,研究其生物学特性并与结合上皮细胞(junctional epithelium,JE)进行比较。方法:LJE取自慢性牙周炎患者翻瓣手术切除的标本,无菌剪刀剪去沟内上皮,采用酶消化法和无血清的角化上皮细胞培养液培养。JE取自健康正畸牙或阻生牙,同法培养。采用细胞分析仪、流式细胞仪和激光共聚焦显微镜比较2种细胞在形态、大小、细胞活力、分子表达及增殖凋亡上的差异。采用SPSS11.5软件包对数据进行独立样本t检验。结果:LJE和JE细胞形态相似,LJE活力低于JE细胞但不存在统计差异(P>0.05)。LJE细胞直径为(19.03±0.19)μm,体积为(4.29±0.19)×103fL;JE细胞直径为(17.16±0.95)μm,体积为(3.20±0.52)×103fL,2种细胞大小有显著差异(P<0.05)。LJE细胞的倍增时间比JE细胞长。CK19在LJE中表达比JE细胞弱,而CK6、CK13在2种细胞中的表达相似。与对照组相比,LJE细胞的早期、晚期凋亡率分别是(4.62±2.16)%和(9.46±1.84)%,而JE细胞分别是(0.47±0.33)%和(3.84±0.98)%,2组细胞的凋亡率有显著差异(P<0.05)。结论:LJE和JE细胞在大小、CK19表达、增殖、凋亡上存在差异,对进一步了解LJE的生物学特性和牙周炎损伤、修复过程有一定意义。
RPOSE：To culture long junctional epithelium（LJE） cells in vitro and compare its biological characteristics with junctional epithelium（JE） cells.METHODS：LJE samples were collected from the patients with chronic periodontitis during flap surgery.Sulcular parts of tissues were removed by sterile scissor then enzymatic method and keratinocyte-serum free medium（KSFM） were used for culturing.JE samples were obtained from healthy bicuspid premolars or third morlars for orthodontic or impaction reasons and the cells were cultured in the same way as LJE.The two types of cells were compared in cell orphology,dimension,viability,proliferation,apoptosis and cytokeratin expression by means of cytoanalyzer,confocal laser scanning microscope and flowcytometry.Statistical analysis was performed using SPSS11.5 software package for independent-samples t test.RESULTS：LJE and JE cells exhibited similar cell morphology.Although the viability of LJE cells were lower than JE cells,there were no statistical significance（P〉0.05）.The mean diameter and volume of LJE cells were（19.03±0.19） μm and（4.29±0.19）×103 fL versus（17.16±0.95） μm and（3.20± 0.52）×103fL of JE cells（P〈0.05）.The doubling time of LJE cells,was longer than that of JE cells.CK19 expression of LJE cells was weaker than of JE cells and there were no difference in the expression pattern of CK6 and CK13.Compared with control cells,the early-and late-stage apoptosis ratio of LJE cells were（4.62±2.16）% and（9.46±1.84）%,respectively which were all higher than JE cells which were（0.47 ±0.63）% and（3.84±0.98）%（P〈0.05）.CONCLUSION：There is difference between LJE and JE cells in dimension,CK19 expression,proliferation time and apoptosis.This study provides some clinical significance and aid in understanding the biological characteristics of LJE and the healing process of periodontitis.
出处
《上海口腔医学》
CAS
CSCD
2009年第4期386-391,共6页
Shanghai Journal of Stomatology
基金
国家自然科学基金(30672314)
上海市科学技术委员会资助项目(08D22271100)
上海市重点学科建设项目(S30206)~~
关键词
长结合上皮
结合上皮合
细胞培养
角蛋白
增殖
凋亡
Long junctional epithelium
Junctional epithelium
Cell culture
Cytokeratin
Proliferation
Apoptosis
