两种离心程序制备的富血小板纤维蛋白结构分析

Structure analysis of platelet-rich fibrin derived from two centrifugation procedures

背景:富血小板纤维蛋白因其特性已在临床中广泛应用,但不同离心程序制造出的纤维蛋白结构及成分会有不同,进而影响临床效果。目的:比较两种特定离心程序制备的富血小板纤维蛋白的组织特点。方法:采集40例自愿捐献者的静脉血,随机分2组,每组20例,一组通过低速离心(1500 r/min离心14 min)程序获得改良富血小板纤维蛋白,另一组通过中速离心(3000 r/min离心10 min)程序获得传统白细胞富血小板纤维蛋白。通过苏木精-伊红染色、免疫组化技术、扫描电镜及透射电镜观察两组纤维蛋白中的白细胞、纤维蛋白基质分布情况及差异。结果与结论:①苏木精-伊红染色:两组纤维蛋白均含有较多白细胞,多集中在中间层并偏向于红细胞端层,凝胶层大部分为纤维蛋白基质,其中改良富血小板纤维蛋白中的白细胞在凝胶层有一些散在均匀分布;②免疫组化:两组纤维蛋白在中间层区域含有大量白细胞,包括单核细胞和中性粒细胞,改良富血小板纤维蛋白中的中性粒细胞比例大于白细胞富血小板纤维蛋白(P﹤0.05),两组单核细胞比例比较差异无显著性意义(P>0.05);③扫描电镜:两组纤维蛋白的白细胞分布广泛,其中改良富血小板纤维蛋白中纤维蛋白基质空间内白细胞和血小板数量稍多,纤维基质条索排列略紧密;④透射电镜:两组纤维蛋白中间层聚集较多的中性粒细胞、单核细胞,血小板密集并含量较多,纤维蛋白基质排列致密,其中改良富血小板纤维蛋白中的纤维基质排列略密集,白细胞含量及血小板稍多;⑤结果表明:因含有的白细胞数量差别,改良富血小板纤维蛋白可能比白细胞富血小板纤维蛋白在促进组织愈合中发挥更积极的作用。

BACKGROUND:Platelet-rich fibrin has been widely used in clinic because of its characteristics,but the structure and composition of fibrin produced by different centrifugation procedures will be different,which will affect the clinical effect.OBJECTIVE:To compare the histological characteristics of platelet-rich fibrin prepared by the different centrifugation procedures.METHODS:Venous blood samples from 40 voluntary patients were divided into two groups(n=20).In group A,modified platelet-rich fibrin was obtained by low-speed centrifugation(1500 r/min centrifugation for 14 minutes).In group B,traditional platelet-rich fibrin was obtained by medium-speed centrifugation(3000 r/min for 10 minutes).Hematoxylin-eosin staining,immunohistochemistry,scanning electron microscopy,and transmission electron microscope were used to analyze the difference of leukocytes and fibrin matrix in two groups of platelet-rich fibrin.RESULTS AND CONCLUSION:(1)Hematoxylin-eosin staining:Both groups contained more leucocytes,mostly concentrated in the middle layer and biased towards the end layer of erythrocytes.Gel layer was mainly composed of fibrin matrix,and the leucocytes were coagulated in the advanced platelet-rich fibrin.The glue layer was scattered and evenly distributed.(2)Immunohistochemistry:The two groups contained a large number of leucocytes in the middle layer,including monocytes and neutrophils.The proportion of neutrophils in the modified platelet-rich fibrin was greater than that of the leukocyte-rich platelet fibers(P<0.05).There was no significant difference in the ratio of monocytes between the two groups(P>0.05).(3)Scanning electron microscopy:The leucocytes of the two groups were distributed slightly widely.The numbers of leucocytes and platelet in the advanced platelet-rich fibrin were slightly higher than those in leucocyte-and platelet-rich fibrin.The fiber matrix cords were slightly tightly arranged in the modified platelet-rich fibrin.(4)Transmission electron microscopy:The two groups of intermediate layers gathered a large number of platelets and many neutrophils and monocytes.The protein matrix was densely arranged,among which the fiber matrix in the modified platelet-rich fibrin was slightly denser than that in leucocyte-and platelet-rich fibrin.(5)The results showed that the modified platelet-rich fibrin may be better than leucocyte-and platelet-rich fibrin due to the difference in the number of leucocytes.The advanced platelet-rich fibrin may play a more active role in promoting tissue healing.