目的:探索富血小板纤维蛋白(platelet-rich fibrin,PRF)凝胶析出液对人牙髓细胞(human dental pulp cells, hDPCs)体外矿化的影响。方法组织块法培养 hDPCs。采用 Choukroun 一步离心法制备 PRF 凝胶。将新鲜制备的PRF 凝胶浸泡于 DMEM 培养基中,于第7 d 取析出液。用 PRF 凝胶析出液孵育 hDPCs 3 d 后更换矿化诱导液。采用茜素红染色和 RT-PCR 检测人牙髓细胞矿化的潜能。结果矿化诱导21 d 后,茜素红染色观察到实验组有少量钙结节生成,而对照组无钙结节生成;RT-PCR 结果显示,实验组 hDPCs 碱性磷酸酶(ALP)的表达为对照组的1.5倍,差异具有统计学意义(P〈0.05)。结论 PRF 凝胶析出液可促进人牙髓细胞矿化。
Objective This study was designed to investigate the effect of platelet-rich fibrin gel (PRF gel)precipitate liquid on the mineralization of human dental pulp cells (hDPCs)in vitro. Methods The hD-PCs were separated and cultured by using tissue block culture method.PRF gel was prepared by Choukroun's protocols.The newly prepared PRF gel was dipped in DMEM culture media,the precipitate liquid of PRF gel was collected on day 7.hDPCs were treated with mineralization induction solution 3 days after being incubated with the precipitate liquid of PRF gel.The capacity of mineralization was measured by using alizarin red stai-ning and RT-PCR. Results Twenty-one days after mineralization induction,a small amount of mineral-ized nodules on alizarin red staining were observed in experimental group while no mineralized nodule was ob-served in control group;RT-PCR revealed that the expression of alkaline phosphatase (ALP)in experimental group was 1.5 times higher than that in control group,comparison yielded statistical difference (P 〈0.05). Conclusion The precipitate liquid of PRF gel can accelerate the mineralization of hDPCs.