β-磷酸三钙复合改良型富血小板纤维蛋白对兔成骨细胞增殖与分化的影响  

Effect of combination of advanced platelet-rich fibrin and β-tricalcium phosphate on the proliferation and differentiation of rabbit osteoblasts

作　　者：高也 芦帅 孙勇 GAO Ye;LU Shuai;SUN Yong(School of Stomatology,Southwest Medical University,Luzhou,Sichuang 646000,China;Department of Stomatology,Pujiang People’s Hospital,Chengdu,Sichuang 611630,China;Department of Stomatology,General Hospital of Western Theater,Chengdu,Sichuang 610083,China)

机构地区：[1]西南医科大学口腔医学院,四川泸州646000 [2]蒲江县人民医院口腔科,成都611630 [3]西部战区总医院口腔科,成都610083

出　　处：《重庆医学》2023年第24期3686-3692,共7页Chongqing medicine

基　　金：军事口腔医学国家重点实验室开放课题(2016KB01)。

摘　　要：目的 探讨β-磷酸三钙(β-TCP)复合改良型富血小板纤维蛋白(A-PRF)对兔成骨细胞增殖和分化的影响。方法 选取新西兰乳兔取颅顶骨制备成骨细胞,成年雄性新西兰大白兔取耳中央动脉血制备兔A-PRF。实验分为4组:A-PRF(A-PRF组)、β-TCP(β-TCP组)及β-TCP+A-PRF复合物(复合组)分别与成骨细胞共培养,空白组无材料仅培养成骨细胞。扫描电镜观察成骨细胞在不同材料上的形态结构,黏附、生长及增殖情况;细胞计数试剂盒8(CCK-8)定量分析4组成骨细胞增殖情况;ELISA检测Ⅰ型胶原蛋白(Col-Ⅰ)、碱性磷酸酶(ALP)和骨钙素(OCN)分泌情况;茜素红染色检测矿化结节的形成情况。结果 成骨细胞与材料复合培养后,电镜下见细胞在β-TCP表面和内部空隙中,以及A-PRF材料表面生长增殖。CCK-8结果显示,β-TCP组培养前5 d,细胞生长增殖受到抑制,其余3组细胞未见明显异常;随培养时间延长细胞增殖明显,培养过程中复合组和A-PRF组细胞数量明显高于β-TCP组(P<0.05)。成骨相关蛋白检测结果显示:Col-Ⅰ、ALP和OCN的分泌均随时间延长而增加,同一时间点Col-Ⅰ、ALP、OCN分泌量β-TCP组最高,复合组和A-PRF组次之,空白组最低(P<0.05)。培养4周后检测矿化结节,复合组生成最多,β-TCP组和A-PRF组次之,空白组最低(P<0.05)。结论 β-TCP和A-PRF复合物对成骨细胞增殖具有明显促进作用,且能有效促进其成骨相关蛋白分泌和矿化结节的沉积,具有良好的体外成骨效应,是理想的骨移植材料。Objective To investigatethe effects of combination of advanced platelet-rich fibrin(A-PRF) and β-tricalcium phosphate(β-TCP) on the proliferation and differentiation of rabbit osteoblasts.Methods The cranial parietal bone was extracted from neonatal New Zealand rabbits to prepare osteoblasts,and the central arterial blood of ear was extracted from adult male New Zealand rabbits to prepare rabbit A-PRF.The experiment rabbits were divided into four groups:rabbit A-PRF(the A-PRF group),β-TCP(the β-TCP group) and β-TCP+rabbit A-PRF complex(the composite group) were cultured with osteoblasts,respectively,and in the blank group osteoblasts were cultured without materials.The morphology,adhesion,growth and proliferation of osteoblasts on different materials were observed by scanning electron microscope.The proliferation of osteoblasts in the four groups was quantitatively analyzed by cell counting kit 8(CCK-8) staining.ELASA was used to detect the expression of collagen-Ⅰ(Col-Ⅰ),alkaline phosphatase(ALP) and osteocalcin(OCN).Alizarin red staining was used to detect the formation of mineralized nodules.Results After the composite culture of osteoblasts with the materials,the growth and proliferation of the osteoblasts were observed on the surface of β-TCP and the inner space,as well as on the surface of rabbit A-PRF under electron microscope.CCK-8 results showed that the proliferation of osteoblasts was inhibited at the first five days in the β-TCP group,and the osteoblasts showed no significant abnormalities in the other three groups.The cell proliferation was obvious with the prolongation of culture time,and the number of osteoblasts in the composite group and the A-PRF group was significantly higher than that in the β-TCP group(P<0.05).Osteogenic related protein detection showed that the secretion of Col-Ⅰ,ALP and OCN increased with time,the β-TCP group showed the most significant increase,followed by the composite group and the A-PRF group,and the blank group had the lowest increase(P<0.05).After four

关 键 词：Β-磷酸三钙 改良富血小板纤维蛋白 成骨细胞 细胞增殖 骨相关蛋白 骨再生 

分 类 号：R318.08[医药卫生—生物医学工程]