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Formulation and Storage of Platelet-Rich Plasma Homemade Product

机译：富含血小板的血浆自制产品的配方和储存

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The platelet-rich plasma (PRP) is an autologous biotherapy based on platelet-healing properties. Here, we developed a simple and reproducible PRP purification protocol based on two successive centrifugations. We evaluated different centrifugation speeds and time-storage durations on the platelet quantity and quality. Sterility and stability of our PRP homemade product were also performed. We prepared PRP from 54 healthy volunteers. We tested activation state, reactivity, and stability of platelets by flow cytometry using basal and adenosine diphosphate (ADP)-induced P-selectin expression markers; growth factor release after platelet activation by an enzyme-linked immunosorbent assay (ELISA); platelet aggregation capacity by aggregrometry assays; clot formation and retraction by thromboelastography; and platelet morphology by ultrastructural analysis. About 130 and 250 g successive speed centrifugations further concentrated platelets while preserving their bioactivity during 6?h (after that, platelet functions were significantly altered). In these conditions, we obtained a highly concentrated pure PRP product (with a low leukocyte count) suitable to study platelet properties. To avoid the loss of efficacy, we recommend injecting PRP under 3?h after preparation.
机译：富血小板血浆（PRP）是基于血小板修复特性的自体生物疗法。在这里，我们基于两次连续离心，开发了一种简单且可重现的PRP纯化方案。我们根据血小板的数量和质量评估了不同的离心速度和时间存储时间。我们的PRP自制产品也具有无菌性和稳定性。我们从54名健康志愿者那里准备了PRP。我们使用基础和二磷酸腺苷（ADP）诱导的P-选择素表达标记通过流式细胞仪测试了血小板的活化状态，反应性和稳定性。通过酶联免疫吸附测定（ELISA）激活血小板后生长因子释放;通过凝集测定法测定血小板的聚集能力;血栓弹力图显示血凝块的形成和收缩;和血小板形态的超微结构分析。连续约130和250 g离心进一步浓缩血小板，同时在6？h内保持其生物活性（此后，血小板功能发生了显着变化）。在这些条件下，我们获得了适合研究血小板特性的高浓度纯PRP产品（白细胞计数低）。为避免药效下降，我们建议在准备后3小时内注射PRP。

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