血清浓度及培养时间对人牙周膜细胞生物学活性的影响

The effect of serum concentration and culture time on the biologic activity of human periodontal ligament cells

目的:探讨血清浓度及培养时间对人牙周膜细胞(HPDLCs)生物学活性的影响。方法:用四唑盐比色法(MTT),BCA蛋白浓度、碱性磷酸酶及羟脯氨酸试剂盒,罗氏诊断cobas誖钙浓度测定试剂盒,放射免疫法检测10mL/L及50mL/L2种浓度的新生牛血清对HPDLCs增殖(1、4、7、10、14、21d)、蛋白质合成、碱性磷酸酶活性、胶原合成、钙吸收及骨钙素分泌(4、7d)的影响。采用SPSS13.0软件包中的双因素方差分析及t检验对数据进行分析。结果:含50mL/L新生牛血清培养液在各时间点均显著促进第4代(P4代)HPDLCs增殖、蛋白合成,显著抑制P4代HPDLCs碱性磷酸酶活性、钙吸收及培养7d时骨钙素分泌,但对细胞胶原合成无影响。在P4代HPDLCs培养过程中,研究选用的2种血清浓度与培养时间仅对细胞增殖、碱性磷酸酶活性及骨钙素分泌有交互效应。结论:血清浓度及培养时间能够影响HPDLCs的生物学活性。

PURPOSE： To investigate the effect of serum concentration and culture time on the biologic activity of human periodontal ligament cells （HPDLCs）. METHODS： The proliferation （1,4,7,10,14,21d）, the protein synthesis, the ALP activity, the collagen synthesis, the calcium intake and the osteocalcin secretion（4,7d） of the fourth passage （P4） HPDLCs treated with 10mL/L or 50mL/L neonatal bovine serum （NBS） were examined using MTT method, BCA protein concentration,ALP activity,hydroxyproline and Roche Diagnostics cobas~ calcium content reagent kits and radio-immunity assay. The data were subjected to two-way analysis of variance and t test using SPSS13.0 software package. RESULTS： 50mL/L NBS could significantly increase the proliferation（1,4,7,10,14,21d） and the protein synthesis（4,7d） of the P4 HPDLCs. 50mL/L NBS could significantly inhibit their ALP activity, the calcium intake （4,7d） and the osteocalcin secretion （7d）. But it had no effect on the collagen synthesis. During cell culture, the two serum concentrations and the culture time only had interaction on the proliferation, ALP activity and osteocalcin secretion of the P4 HPDLCs. CONCLUSIONS： The serum concentration and culture time could affect the biologic activity of HPDLCs. Supported by National Natural Science Foundation of China（Grant No.30973354）.