摘要： 目的：评估牙髓细胞在富血小板纤维蛋白（platelet-rich fibrin, PRF）存在下的体外增殖及成骨分化的能力，为PRF作为支架材料、牙髓细胞作为种子细胞构建组织工程骨，进行前期研究。方法：3月龄比格犬拔除乳磨牙获得乳牙牙髓细胞；恒牙牙髓细胞从16月龄成年比格犬磨牙获得。静脉取血离心10 min获得PRF。实验分4组：对照组（普通培养基不加入PRF）；实验组A（普通培养基加入PRF）；实验组B(成骨诱导培养基不加入PRF)；实验组C（成骨诱导培养基加入PRF）。分别于1、4、7和11d测定细胞数量、MTT值、半定量碱性磷酸酶值、成骨相关基因Q?鄄PCR值，并于21d测定钙结节吸光度值。结果：PRF是一种可以促进牙髓细胞增殖的，无毒性作用的纤维网状支架结构；细胞水平上PRF促进了两种细胞的成骨分化：钙结节以及碱性磷酸酶半定量数值都明显上调（P<0.05）；基因水平上，4个时间点成骨相关基因的表达量都显著增加（P<0.05），且乳牙牙髓细胞的表现均优于恒牙牙髓细胞。结论：可使用PRF和牙髓细胞复合构建组织工程骨。

关键词: 富血小板纤维蛋白；  , 比格犬；  , 牙髓细胞；  , 增殖和成骨分化；  , 组织工程

Abstract: Objective: To evaluate the effects of choukroun's PRF (platelet-rich fibrin) on proliferation and differentiation of dental pulp cells in vitro and making a pre-feasibility assess hunting for the optimum scaffold. Methods: Pulp tissues of deciduous and permanent molars were harvested from 3-month and 16-month old beagle dogs respectively. Dental pulp cells were isolated and cloned. Jugular vein blood were collected from beagle dogs and centrifugation at 3 000 r/min for 10 minutes to obtain PRF. The experiments were divided into four groups: control group (ordinary medium without PRF),  test group A ( ordinary medium with PRF), test group B (osteoinduction medium without PRF), test group C(added PRF in osteoinduction medium ). Cell counts and cytotoxicity tests, semi-quantitative alkaline phosphatase and real time PCR, were performed and assayed at days 1, 4, 7 and 11 of culture. The determination of calcium nodules OD value was in day 21. Results: PRF is a fibrin matrix structure and showed absence of toxic effect on two types of cells. PRF increased a significant proliferation in all cell types. Furthermore, PRF increased a strong differentiation of dental pulp cells including calcium nodules， ALP semi-quantitative（P<0.05） and osteogenesis related genes(P<0.05). Dental pulp cells of exfoliated deciduous teeth showed more obvious than the permanent dental pulp cells.    Conclusion: This in vitro study suggests that PRF scaffolds may be used as a specific regulator of hard-tissue engineering.

Key words: platelet-rich fibrin(PRF), beagle dog, dental pulp cells , cell proliferation and differentiation, tissue engineering

中图分类号: 

• R782.2

• Q813