Intervention with platelet-rich fibrin in rabbit models after microvascular anastomosis surgery of the femoral artery: changes in vascular structure and soft tissue

doi:10.12307/2023.182

Abstract

Abstract: BACKGROUND: Recently, numerous basic and clinical data have shown the superiority of platelet-rich fibrin in the regeneration and repair of soft and hard tissues, while there are few relevant reports in the field of microvascular anastomosis.
OBJECTIVE: To evaluate the efficacy of platelet-rich fibrin applied to the femoral artery after microvascular anastomosis surgery in a rabbit model.
METHODS: Thirty-six adult New Zealand white rabbits of either sex were randomly divided into experimental group and control group (n=18 per group) after establishing femoral artery transection-reanastomosis models. In the experimental group, autologous ear venous blood were extracted and centrifuged to prepare platelet-rich fibrin gel, and the prepared gel was immediately applied to the anastomotic site of the femoral artery. The control group was given the same volume of normal saline. Incision infection, healing, and adhesion were evaluated at 3, 7, 14 days after surgery. Vascular structure of the femoral artery at the anastomotic site was observed using hematoxylin-eosin staining. Immunohistochemistry detection was performed to analyze the expression levels of vascular endothelial growth factor, platelet-derived growth factor, and transforming growth factor β1.
RESULTS AND CONCLUSION: The overall number of incision infections in the experimental group was lower than that in the control group animals (P < 0.05), and the number of incision infections showed no significant changes in each group (P > 0.05). The overall incision healing level in the experimental group was better than that in the control group (P < 0.05), and the incision healing showed no significant changes in the experimental group (P > 0.05) but changed significantly in the control group at different time points (P < 0.05). The degree of femoral artery adhesion in the operation area was generally lighter in the experimental group than the control group (P < 0.05). The degree of adhesion had insignificant changes in the experimental group at different time points (P > 0.05) but gradually increased in the control group after operation (P < 0.05). The results of hematoxylin-eosin staining showed better repair of vascular endothelial cells and inner elastic lamina in the experimental group than the control group. Compared with the control group, the reconstruction of the intima was more complete in the experimental group, with less necrosis, proliferation, and migration of vascular smooth muscle cells in the media and traumatic hyperplasia of the extima. The overall healing condition of the vascular wall was better in the experimental group than the control group. Immunohistochemical results revealed that the expression levels of vascular endothelial growth factor, platelet-derived growth factor, and transforming growth factor β1 in the experimental group were higher than those in the control group (P < 0.05). To conclude, platelet-rich fibrin can promote the growth of vascular endothelial cells, contribute to the in situ reconstruction of various layers of vascular tissue, reduce postoperative complications, such as infection and adhesion, in the surgical area, and shorten the repair and healing time after vascular injury. This novel treatment method is expected to improve the therapeutic effect of arterial anastomosis in microsurgery.

Key words: blood concentrate, platelet-rich fibrin, growth factor, microsurgery, microvascular anastomosis, vascular endothelial cell injury, thrombus, revascularization

CLC Number:

Lu Yangyang, Yan Ruihong, Qiu Zhongpeng, Dai Yi, Wang Zixin, Wang Weishan, Shi Chenhui, Du Xinhui, Li Gang. Intervention with platelet-rich fibrin in rabbit models after microvascular anastomosis surgery of the femoral artery: changes in vascular structure and soft tissue[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(11): 1659-1668.

Figures/Tables 13

术后7 d时，对照组兔股动脉吻合口区域断裂的血管开始修复，可见部分血管内皮细胞生长，内弹力膜层部分恢复，血管内膜层大部分中断，血管中膜平滑肌细胞坏死开始增殖，纤维结缔组织和血管外膜层结缔组织生长明显。而实验组兔股动脉吻合口区域断裂的血管基本修复，血管内皮细胞生长活跃，内弹力膜层大部分恢复，血管内膜层基本连续，仅少部分中断，血管中膜平滑肌细胞逐渐修复，血管中膜、外膜结构恢复良好，界线基本清晰，见图4。术后14 d时，对照组兔股动脉吻合口区域断裂的血管修复基本完整，血管腔内可见血栓形成，血管腔较为狭窄，血管内皮细胞生长活跃，内弹力膜层大部分恢复，血管内膜层部分是由平滑肌细胞增殖迁移构成，血管中膜平滑肌细胞过度增殖，纤维结缔组织和血管外膜层结缔组织生长明显，血管内膜、中膜、外膜结构紊乱，界线不清。实验组中观察到兔股动脉吻合口区域断裂的血管修复完整，无原有结构异位出现的现象，血管腔内未见血栓形成，无明显狭窄，血管内皮细胞生长良好，血管内弹力膜层基本恢复正常，血管内膜层连续且修复完整，血管中膜平滑肌细胞无过度增殖现象，血管中膜层及外膜层结构原位修复完好且界线清晰，见图4。总之，苏木精-伊红染色结果显示：对照组中血管内皮细胞开始生长的时间为术后第7天，而实验组中术后第3天就观察到了血管内皮细胞生长现象；同样内弹力膜层开始出现的时间在对照组和实验组中分别为术后第7天和第3天；实验组中血管内皮细胞整体生长情况较对照组更为活跃，实验组血管内膜层的重建更为完善，对照组血管中膜平滑肌细胞坏死、迁移及创伤性过度增殖情况比实验组严重，实验组术后3，7，14 d时血管组织各层结构的恢复情况均优于对照组。 2.4 两组兔术区股动脉组织免疫组织化学染色结果富血小板纤维蛋白成分中的VEGF、PDGF及转化生长因子β1在组织损伤的修复过程中起着重要作用。实验组与对照组在术后3，7，14 d分别进行VEGF、PDGF及转化生长因子β1的免疫组化实验，结果发现实验组的血管组织管腔结构大体上更加完整，于3个时间节点上均比对照组修复得完好。同时，实验组中3种生长因子所对应的棕色颗粒数量及表达面积均多于对照组，见图5-7。 "

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