富血小板血浆在犬骨缺损修复中的应用研究

Study on the application of platelet-rich plasma in canine bone defect repair

为了初步探讨富血小板血浆(platelet-rich plasma, PRP)在犬大段骨缺损修复中发挥的作用,试验采用二次离心法处理抗凝血,制备PRP;采用酶联免疫吸附试验(ELISA)法测定被氯化钙与牛凝血酶激活后的PRP组和血清对照组中血小板衍生生长因子(PDGF)和转化生长因子β1(TGF-β1)的浓度;在3只比格犬的双侧股骨上分别构建3个直径为10 mm的圆形穿透性皮质骨缺损模型,随机分成对照组、自体骨移植组和自体骨+PRP移植组进行骨缺损修复的动物试验,4,8周时采样制作骨组织病理切片,检测组织新生毛细血管和新生骨小梁情况。结果表明:利用二次离心法可以分离制备出PRP,其富集的倍数平均为3.64倍;激活后的PRP释放出来的PDGF浓度为(435.00±49.41)pg/mL,TGF-β1浓度为(44.47±3.81)pg/mL,与血清对照组相比差异极显著(P<0.01);自体骨+PRP移植组的骨缺损修复程度较好,其骨组织新生毛细血管计数和新生骨小梁面积百分比极显著高于其他两组(P<0.01)。说明PRP可促进犬大段骨缺损修复,可在临床上治疗犬大段骨缺损。

In order to preliminarily investigate the role of platelet-rich plasma(PRP) in canine large segmental bone defect repair, PRP was prepared by double centrifugation treatment of anticoagulated blood;enzyme-linked immunosorbent assay(ELISA) was used to determine the concentrations of platelet-derived growth factor(PDGF) and transforming factor-β1(TGF-β1) in PRP activated by calcium chloride and bovine thrombin. Three 10 mm-diameter circular penetrating cortical bone defect models were constructed on the bilateral femur of three Beagle dogs and randomly divided into control group, autologous bone graft group and autologous bone+PRP graft group for animal experiment on bone defect repair. At 4 and 8 weeks, bone histopathological sections were sampled for detection. The results showed that PRP could be isolated and prepared by double centrifugation method, and the average multiplicity of enrichment was 3.64;the average concentration of PDGF released from activated PRP was(435.00±49.41) pg/mL, the mean concentration of TGF-β1 was(44.47±3.81) pg/mL, and the diffrence was extremely highly significant compared with the serum control group(P<0.01). Bone pathological tissue sections showed better bone defect repair in the autologous bone+PRP graft group, which had a significantly higher bone tissue neocapillary count and percentage of new bone trabecular area than the other two groups(P<0.01). The results indicated that PRP had a positive promoting effect on canine large segmental bone defect repair, which provided a new idea for the clinical treatment of canine large segmental bone defect.