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目的:探索动脉血小板浓缩液,在无菌情况下的制备方法。通过免疫组化及物理测量,证实血小板浓缩液中含有多种生长因子,并对比分析其对骨折愈合不同时期的促进作用,为临床应用提供基础资料。方法:选用20只新西兰兔,随机分成2组,分别为空白组及血小板组。制作尺骨中段骨折动物模型,采用指钢板行内固定。在手术之前,先抽取兔股动脉血6ml左右,经枸橼酸钠抗凝后,加低速高速离心,提纯出白色的血小板浓缩液,将其注射回骨折断端。所有操作均为无菌操作。分别在1、2、4及6周处死兔,在骨折处取5mm骨组织标本。切片作PDGF免疫组化染色定性分析及测量骨痂直径定量分析。结果:①6ml的兔动脉血在低速及高速离心后,可分离出0.5ml的血小板浓缩液,浓度在正常兔血小板浓度的3倍以上。②免疫组化染色阳性产物呈棕黄色颗粒状。在空白组术后第1周及第2周PDGF因子为阴性表达,第4周、第6周为弱阳性表达;在血小板浓缩液组,随着处死时间延长,阳性表达率逐渐增高。在相同的处死时间,血小板浓缩液组阳性表达率明显高于空白组。空白组、血小板组兔尺骨骨折标本在1、2、4、6周,均有不同程度骨痂形成,其中以第6周兔骨痂最多,第1周兔以纤维连接为主,仅有少许骨痂形成。空白组与血小板组同期骨痂直径差异有显著性意义。结论:①少量的兔血在严格的实验条件下,能够制备出血小板浓缩液。②兔血小板浓缩液中确含有较高浓度的PDGF生长因子。③血小板浓缩液加速了骨折断端的愈合。
Objective: To explore the preparation method of arterial platelet concentrate under aseptic conditions. Through immunohistochemistry and physical measurements, confirmed platelet concentrates containing a variety of growth factors, and comparative analysis of its promotion of different stages of fracture healing, provide the basic data for clinical applications. Methods: Twenty New Zealand rabbits were randomly divided into 2 groups: blank group and platelet group. Production of the middle of the ulnar fracture animal models, the use of plate fixation. Prior to surgery, rabbit femoral artery blood was drawn to about 6ml, anticoagulated with sodium citrate followed by low-speed, high-speed centrifugation to purify the white platelet concentrate and inject it back to the fracture site. All operations are aseptic. Rabbits were sacrificed at 1, 2, 4, and 6 weeks respectively, and specimens of 5 mm bone were taken at the fracture site. Qualitative analysis of PDGF immunohistochemical staining and quantitative analysis of callus diameter. Results: ① After 6ml rabbit arterial blood was centrifuged at low speed and high speed, 0.5ml platelet concentrate could be isolated at a concentration of more than 3 times of normal rabbit platelet concentration. ② Immunohistochemical positive product was brown granular. The expression of PDGF was negative in the first week and the second week of the blank group, and weakly positive in the fourth week and the sixth week. In the platelet concentrate group, the positive expression rate of PDGF was gradually increased with the prolonging of the execution time. At the same time of sacrifice, the positive rate of platelet concentrate group was significantly higher than that of blank group. In the blank group and the platelet group, ulnar fracture specimens of rabbits were found to have different degrees of callus at 1, 2, 4, and 6 weeks, with the most of the callus in the 6th week. The first week was mainly fibrosis, with only a few Callus formation. There was significant difference in callus diameter between blank group and platelet group over the same period. Conclusion: ① A small amount of rabbit blood can be prepared under strict experimental conditions, platelet concentrates. ② rabbit platelet concentrate does contain higher concentrations of PDGF growth factor. ③ platelet concentrate accelerated fracture healing.