氟羟基磷灰石凝胶对人牙髓细胞增殖和分化的影响  

Effects of fluorohydroxyapatite gel on proliferation and differentiation of human dental pulp cells

作　　者：王倩倩[1] 邓旭亮[1] 陈海峰[2] 高学军[1] 

机构地区：[1]北京大学口腔医学院.口腔医院,100081 [2]北京大学工学院生物工程系

出　　处：《现代口腔医学杂志》2014年第2期90-94,共5页Journal of Modern Stomatology

基　　金：科技部国际重大科学研究计划项目(2012CB933903);北京大学口腔医学院青年科研基金项目(PKUSS20130103)

摘　　要：目的评价新研制的氟羟基磷灰石(fluorohydroxyapatite,FHA)凝胶对人牙髓细胞(human dental pulp cells,hDPCs)增殖和分化的影响。方法采用酶消化法培养hDPCs。用含氟羟基磷灰石凝胶(FHA)浸提液的DMEM培养液培养hDPCs。分别于1,3,5,7天通过CCK-8实验检测细胞增殖、碱性磷酸酶(alkaline phosphatase,ALP)活性的检测以及实时定量逆转录聚合酶链反应检测分化基因牙本质涎磷蛋白(dentin sialophosphoprotein,DSPP)、骨钙蛋白(osteocalcin,OCN)和ALP表达,评估FHA凝胶对hDPCs增殖和分化的影响。结果 FHA组各时间点细胞增殖与阴性对照组无明显差异(P>0.05),第3,7天的ALP活性低于阴性对照组,ALP染色阳性细胞少于阴性对照组。Real TimePCR结果显示FHA组细胞第3,7天DSPP和OCN表达显著高于阴性对照组,而ALP表达低于阴性对照组。结论 FHA对人牙髓细胞的增殖无负面作用,但是可以促进人牙髓细胞向成牙或成骨向分化,有生物诱导活性。Objective To investigate the effects of Fluorohydroxyapatite gel on proliferation and differentiation of human dental pulp cells. Methods HDPCs were harvested from impacted third molars of healthy adult individuals using enzyme digestion, expanded and cultured. The hDPCs were cultured in DMEM medium containing Fluorohydroxyapatite gel （FHA） eluate for 7 days. Cell proliferation was assessed by CCK-8 cell proliferation kit on days 1, 3, 5 and 7. Alkaline phosphatase activity, real-time reverse transcription polymerase chain reaction （Real Time-PCR） were used to examine the differentiation of hDPCs on day 1,3,7. Results The proliferation of hDPCs in FHA gel eluate was higher than negative control group（P〉0.05）. On day 3 and 7, the ALP activity of hDPCs was lower than negative control group, and the positive staining cells were also less than control group on day 7. The expression of DSPP and OCN genes were significantly up-regulated compared with negative control group, while ALP gene expression was down-regulated compared with control group. Conclusion FHA gel has good biocompatibility to hDPCs, but can promote the odonto and osteogenic differentiaion of hDPCs.

关 键 词：氟羟基磷灰石 人牙髓细胞 细胞增殖 细胞分化 盖髓剂 

分 类 号：R780.2[医药卫生—口腔医学]