The purpose of this study was to investigate the effect of PPRP (pure PRP) and LPRP (PRP with leukocytes) on recovery from limb ischemia and on expression of growth factors involved in angiogenesis, myogenesis and fibrogenesis. PPRP and LPRP prepared by centrifugation were added to cultures of C2C12 and NIH3T3 cells (1 or 10% PRPs) to evaluate alterations in cell metabolism and expression of growth factors by MTT, ELISA and RT-qPCR, respectively. To evaluate in vivo regenerative effects, PRPs were injected into the ischemic limbs of BALB/c mice and muscle mass/strength and histomorphometry were evaluated after 30 days. Mice treated with PRPs after limb ischemia showed an increase in the size of myofibers and muscle strength, reduced fibrosis and adipocytes, and decreased capillary density and necrosis scores compared to untreated mice. In cell culture, serum deprivation reduced the viability of C2C12 and NIH3T3 cells to about 50%, but the addition of 1% PRPs completely recovered this loss. Both PRPs, downregulated most of the tested genes; however, angiogenic gene in C2C12 and the fibrogenic genes and in NIH3T3 cells were upregulated by LPRP. PPRP and LPRP had similar effects in regulation of genes involved in angiogenesis, myogenesis and fibrogenesis. However, the presence of leucocytes did not significantly affect regenerative activities of PRP in the ischemic limb.

中文翻译：

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富血小板血浆对小鼠缺血肢体的再生作用


本研究的目的是研究 PPRP（纯 PRP）和 LPRP（含白细胞的 PRP）对肢体缺血恢复以及对参与血管生成、肌生成和纤维生成的生长因子表达的影响。将离心制备的 PPRP 和 LPRP 添加到 C2C12 和 NIH3T3 细胞培养物（1 或 10% PRP）中，分别通过 MTT、ELISA 和 RT-qPCR 评估细胞代谢和生长因子表达的变化。为了评估体内再生效果，将 PRP 注射到 BALB/c 小鼠的缺血肢体中，并在 30 天后评估肌肉质量/强度和组织形态计量学。与未治疗的小鼠相比，肢体缺血后接受 PRP 治疗的小鼠的肌纤维尺寸和肌肉强度有所增加，纤维化和脂肪细胞减少，毛细血管密度和坏死评分降低。在细胞培养中，血清剥夺使 C2C12 和 NIH3T3 细胞的活力降低至约 50%，但添加 1% PRP 完全恢复了这种损失。两种 PRP 均下调了大多数测试基因；然而，LPRP 上调了 C2C12 中的血管生成基因以及 NIH3T3 细胞中的纤维生成基因。 PPRP 和 LPRP 在调节涉及血管生成、肌生成和纤维生成的基因方面具有相似的作用。然而，白细胞的存在并没有显着影响缺血肢体中 PRP 的再生活性。