Department of Experimental Surgery, Centro Médico Nacional "20 de Noviembre," ISSSTE, Mexico City, Mexico.
Departamento de Cirugía, Facultad de Medicina, UNAM, Mexico City, Mexico.
J Tissue Eng Regen Med. 2020 Apr;14(4):600-608. doi: 10.1002/term.3024. Epub 2020 Mar 9.
Peripheral blood mononuclear cells (PBMCs) contain a cell fraction of mononuclear progenitor cells (MPCs), which own significant angiogenic potential. Autologous transplant of PBMC and/or platelet-rich plasma (PRP) promotes endothelial cells differentiation in experimental lower limb ischemia, which is considered a safe and effective strategy to support revascularization, either in animal models or clinical trials. In addition, thrombin has been proposed to enrich biological scaffolds, hence increasing MPC viability after intramuscular administration, whereas proangiogenic mediators such as vascular endothelial growth factor (VEGF), tumor necrosis factor alpha (TNF-α), inhibitor of the plasminogen activator-1 (PAI-1), and chemokine (CXCL1; GRO-α) participate in the endothelial response to ischemia, through their proangiogenic effects over endothelial cells proliferation, survival, migration, endothelial integrity maintenance, and physiologic vascular response to injury. In the present study, we describe the effect of autologous PBMCs transplant and PRP, either with or without thrombin, over proangiogenic mediators (measured by enzyme-linked immunosorbent assay) and revascularization response (angiographic vascular pattern at 30 days after vascular occlusion) in a rat model of lower limb ischemia. The group treated with PBMC + PRP significantly induced PAI-1, an effect that was prevented by the addition of thrombin. Furthermore, treatment with PBMC + PRP + thrombin resulted in the induction of VEGF. GRO-α showed a sensitive induction of all proangiogenic mediators. All treatments significantly stimulated revascularization, according to angiographic assessment, whereas higher effect was observed with PBMC + PRP treatment (p < .0001). In conclusion, autologous PBMC transplant stimulates revascularization during experimental ischemia of the lower limb, whereas particular effects over proangiogenic and fibrinolytic mediators may be attributed to PBMCs and its combination with PRP and thrombin.
外周血单个核细胞（PBMCs）包含单核祖细胞（MPCs）的细胞部分，其具有显著的血管生成潜力。自体 PBMC 和/或富含血小板的血浆（PRP）的移植促进了实验性下肢缺血中的内皮细胞分化，这被认为是一种安全有效的策略，可以支持再血管化，无论是在动物模型还是临床试验中。此外，已经提出了凝血酶来丰富生物支架，从而增加肌肉内给药后 MPC 的存活率，而血管内皮生长因子（VEGF）、肿瘤坏死因子 alpha（TNF-α）、纤溶酶原激活物-1（PAI-1）抑制剂和趋化因子（CXCL1；GRO-α）等促血管生成介质通过其对内皮细胞增殖、存活、迁移、内皮完整性维持和对损伤的生理性血管反应的促血管生成作用参与到对缺血的内皮反应中。在本研究中，我们描述了自体 PBMC 移植和 PRP，无论是否添加凝血酶，对促血管生成介质（通过酶联免疫吸附试验测量）和下肢缺血大鼠模型中再血管化反应（血管闭塞后 30 天的血管造影血管模式）的影响。用 PBMC + PRP 治疗的组显著诱导了 PAI-1，这种作用可以通过添加凝血酶来预防。此外，用 PBMC + PRP + 凝血酶治疗导致了 VEGF 的诱导。GRO-α对所有促血管生成介质都有敏感的诱导作用。所有治疗均根据血管造影评估显著刺激了再血管化，而用 PBMC + PRP 治疗则观察到更高的效果（p<0.0001）。总之，自体 PBMC 移植在实验性下肢缺血中刺激再血管化，而对促血管生成和纤溶介质的特殊作用可能归因于 PBMC 及其与 PRP 和凝血酶的组合。
