注射型富血小板纤维蛋白对成骨细胞影响的研究

The effect of injection platelet-rich fibrin on rabbit osteoblasts

目的观察和探讨注射型富血小板纤维蛋白(injection platelet-rich fibrin,i-PRF)对兔成骨细胞增殖、分化及细胞骨架的影响。方法取新西兰乳兔颅顶骨通过改良胶原酶和胰酶分段消化结合法获取兔成骨细胞,并通过原代及传代培养后进行细胞鉴定。将实验分为实验组及对照组分别培养兔成骨细胞。CCK-8法检测培养1、3、5、7 d细胞增殖的OD值;碱性磷酸酶(ALP)法检测培养5、7、9 d的细胞分化情况并进行半定量分析,罗丹明-鬼笔环肽染色观察培养12、24、48 h的细胞骨架形态。结果CCK-8检测示,随培养时间延长,细胞数量增加(P<0.05),在各时间节点实验组A值均大于对照组;ALP染色显示随培养时间延长,在各个时间节点,细胞碱性磷酸酶蓝染颗粒表达情况均表现为实验组优于对照组;细胞骨架观察显示在各时间节点,实验组细胞骨架较对照组排列更加均匀,形态更加伸展。结论i-PRF可以促进成骨细胞增殖、分化,对细胞骨架的排列和伸展亦有促进作用。

Objective To explore the effect of injection platelet-rich fibrin(i-PRF)on the proliferation,differentiation and cytoskeleton of rabbit osteoblasts.Methods Osteoblasts were obtained by modified collagenase and trypsin digestion combined with tissue block culture from the cranial bone of New Zealand milk rabbits,and the cells were identified after primary culture and subculture.The cultures were divided into the experimental group and the control group.Cell proliferation on the 1st,the 3rd,the 5th and the 7th day was detected by CCK-8 method.The differentiation of osteoblasts on the 5th,the 7thand the 9thday was detected by ALP method(BCIP/NBT method).The cytoskeleton on 12 h,24 h and 48 h was observed by using rhodamine-phalloidin dye.Results CCK-8 test showed that the number of cells was increased with the extension of culture time(P<0.05),and the OD value in the experimental group was greater than that in the control group(P<0.05).ALP test showed that the expression of alkaline phosphatase blue dye granules at each time point in the experimental group was better than that in the control group.Cytoskeleton observation showed that the cytoskeleton of the experimental group was more arranged and extended than that in the control group at each time point.Conclusion i-PRF can promote the proliferation and differentiation of osteoblasts and promote the arrangement and extension of cytoskeleton.