CAJ | 학술논문

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。
목적：연구부혈소판섬유단백（plate-rich fibrin, PRF）체외대인아조골성골세포(human alveolar osteoblasts, HAOB)증식분화적영향，탐토HAOB여PRF구건림상조직공정골적가능성。방법：수집림상발아과정중적아조골，채용개량매소화법체외분리배양HAOB，근거성골세포형태학특정급성골특성대소배양출적세포진행감정，후가입지원자적PRF분조배양；이후재불동적실험시간점진행세포증식CCK-8검측、감성린산매(ALP)정성검측、개결절천소홍염색이급성골상관기인RT-PCR검측。결과：HAOB구유전형적성골세포적형태；수시간적연장，세포수목명현증가（P<0.05），실험조（PRF조）세포수량명현고우대조조。실험조ALP염색교대조조안색경심，ALP활성상대교고。경천소홍염색，실험조경하관찰형성적개결절수량교대조조다。재제7화11천발현실험조성골상관기인적표체량균고우대조조。결론：채용개량매소화법분리배양적HAOB구유전형적성골세포생물학특성，차성분교위단일；PRF체외구유촉진HAOB증식분화적능력。
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 cul-tured cells were identified by the morphological characteristics and osteogenic properties of osteoblasts. HAOB was cul-tured with or without PRF obtained from volunteers. Cell proliferation was measured by CCK-8 method. Qualitative detec-tion 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 experimental group 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.