摘要： 目的：研究富血小板纤维蛋白（plate-rich fibrin, PRF）体外对人牙槽骨成骨细胞(human alveolar osteoblasts, HAOB)增殖分化的影响，探讨HAOB与PRF构建临床组织工程骨的可能性。方法：收集临床拔牙过程中的牙槽骨，采用改良酶消化法体外分离培养HAOB，根据成骨细胞形态学特征及成骨特性对所培养出的细胞进行鉴定，后加入志愿者的PRF分组培养；而后在不同的实验时间点进行细胞增殖CCK-8检测、碱性磷酸酶(ALP)定性检测、钙结节茜素红染色以及成骨相关基因RT-PCR检测。结果：HAOB具有典型的成骨细胞的形态；随时间的延长，细胞数目明显增加（P<0.05），实验组（PRF组）细胞数量明显高于对照组。实验组ALP染色较对照组颜色更深，ALP活性相对较高。经茜素红染色，实验组镜下观察形成的钙结节数量较对照组多。在第7和11天发现实验组成骨相关基因的表达量均高于对照组。结论：采用改良酶消化法分离培养的HAOB具有典型的成骨细胞生物学特性，且成分较为单一；PRF体外具有促进HAOB增殖分化的能力。

关键词: 富血小板纤维蛋白；  , 人牙槽骨成骨细胞；  , 增殖分化；  , 组织工程

Abstract: Objective: The purpose of the study was to evaluate the effects of Choukroun's platelet-rich fibrin (PRF) on proliferation and osteogenic differentiation of  human alveolar osteoblasts (HAOB), and to explore the possibility of bone engineering constructs with PRF and HAOB. Methods: Human alveolar bone was collected during clinical wisdom tooth extraction process. Human alveolar osteoblasts  were isolated and cultured in vitro by improved enzyme digestion. The cultured cells were identified by the morphological characteristics and osteogenic properties of osteoblasts. HAOB was cultured with or without PRF obtained from volunteers. Cell proliferation was measured by CCK-8 method. Qualitative detection of alkaline phosphatase (ALP), alizarin red staining of calcium nodules and RT-PCR of osteogenesis related genes were performed at different experimental time points. Results: The cultured cells were proved to be osteoblasts with a typical morphology and osteogenic properties. As the CCK-8 test results showed, the number of cells were increased significantly (P<0.05) with time and were more higher in the experimental group. ALP activity is relatively high in the experimental group, with a deep color through ALP staining. The number of calcium nodules in the experimental group were more than that of the control group by Alizarin red staining. The relative expression of osteogenesis related genes in the experimentalgroup at 7 and 11 days, were higher than those of the control group. Conclusion: The HAOBs cultured through improved enzyme digestion have typical bionomics of osteoblasts，and their components are single． PRF is demonstrated the ability to promote the proliferation and differentiation of HAOB.

Key words: platelet-rich fibrin(PRF), human alveolar osteoblast, osteogenic differentiation, tissue engineering

中图分类号: 

• R782.2

• Q813