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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 evaluate the effect of newly developed fluorohydroxyapatite (FHA) gel on the proliferation and differentiation of human dental pulp cells (hDPCs). Methods hDPCs were cultured by enzyme digestion. HDPCs were cultured in DMEM medium containing fluorinated hydroxyapatite gel (FHA) extract. Cell proliferation, alkaline phosphatase (ALP) activity assay and real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) were used to detect the expression of dentin sialophosphoprotein ( dentin sialophosphoprotein (DSPP), osteocalcin (OCN) and ALP expression, and evaluate the effect of FHA gel on the proliferation and differentiation of hDPCs. Results There was no significant difference in cell proliferation between FHA group and negative control group at each time point (P> 0.05). The ALP activity of FHA group was lower than that of negative control group on the 3rd and 7th days, while the number of ALP - positive cells was less than that of the negative control group. The results of Real Time PCR showed that the expression of DSPP and OCN in the FHA group was significantly higher than that in the negative control group on the 3rd and 7th day, while the ALP expression was lower than that in the negative control group. Conclusion FHA has no negative effect on the proliferation of human dental pulp cells, but it can promote human dental pulp cells to differentiate into odontoblasts or osteoblasts with biological induction activity.