活血化瘀补肾壮骨中药促进BMP基因克隆转染犬牙髓细胞增殖的实验研究

THE EXPERIMENTAL STUDY ON THE PROMOTIVE EFFECT OF CHINESE TRADITION MEDICINE ON PROLIFERATION OF DOG DENTAL PULP CELLS TRANSFECTED BY BMP2 GENE

目的观察活血化瘀补肾壮骨含药血清对骨形成蛋白2(bone morphogenetic proteins 2,BMP2)绿色荧光融合蛋白(green-fluorescent protein,GFP)真核表达质粒(pEGFP-N1-BMP2)在体外转染的犬牙髓细胞(dogdental pulp cells,DDPCs)增殖情况的影响,探讨活血化瘀补肾壮骨中药在牙本质改建形成中的作用机制。方法将BMP2-DDPCs分别于活血化瘀补肾壮骨含药血清组(A组)、无药血清组(B组)和胎牛血清组(C组)中培养。分别于24、48、72h用溴化-3-(4,5-二甲基噻唑基-2)-2,5二苯基四氮唑[3-(4,5-dimethythiazol-2-y1)-2,5-diphenyl tetrazolium bromide,MTT]法检测BMP2-IDDPCs增殖活性。结果 BMP2-DDPCs分别在A、B、C 3组中培养后,细胞的形态相同。3组细胞随着培养时间的延长,细胞数量逐渐增加,A组增加更为显著,与其他2组差异有统计学意义(P<0.05)。结论活血化瘀补肾壮骨中药含药血清作用于BMP2-DDPCs,促进了该细胞增殖活性,血化瘀补肾壮骨中药含药血清大大增加了组织工程重要环节的种子细胞的数量。

Objective To observe the effect of Chinese tradition medicine serum on the proliferation of dog dental pulp cells（DDPCs） transfected with pEGFP- N1 -BMP2 eukaryotic expression plasmid in vitro. To investigate the mechanism of Chinese tradition medicine on formation of dentin. Methods BMP2 -DDPCs were cultivated respectively in Chinese tradition medicine serum group（group A） ,non- medicine serum group （group B ） and fetal bovine serum group （group C ）. The proliferation activity of BMP2 -DDPCs was detected respectively by the 3 - （4,5 -dimethythiazol- 2 -yl） -2,5 - diphenyl tetrazolium bromide（MTI？） after 24 hours ,48 hours and 72hours. Results After BMP2 - DDPCs were cultivated in three kinds of serum, there were no differences in the cells morphology, but there were differences in the number of the cells. With the prolongation of culture time, the number of the cells gradually increased in the 3 groups, among which medicine serum group increased more prominently. There were significant differences between Chinese traditional medicine serum group and the other two groups （P 〈 0. 05 ）. Conclusion The Chinese tradition medicine could promote proliferation of BMP2 -DDPCs and obviously increase the quantity of seed cells in dentin tissue engineering.