Instituto de Biología Molecular en Medicina y Terapia Génica, Centro Universitario de Ciencias de la Salud, Universidad de Guadalajara, Jalisco, Mexico
Tecnologico de Monterrey, Escuela de Medicina y Ciencias de la Salud, Jalisco, Mexico
Turk J Haematol. 2023 Feb 28;40(1):43-49. doi: 10.4274/tjh.galenos.2023.2022.0445. Epub 2023 Jan 31.
Platelet-rich plasma (PRP) is used in multiple coagulation disorders. Its therapeutic effectiveness relies on technical procedures related to PRP procurement and preservation because free radicals induce platelet activation and aging. This work aims to elucidate the oxidative mechanisms involved in activation of platelets obtained from PRP during storage.
One hundred ten PRP batches were obtained from healthy donors and kept under stirring at a temperature of 20-24 °C. Protein extraction was performed from platelet homogenates and plasma at different times of storage from day 1 to 20. The activities of antioxidant markers such as catalase (CAT), superoxide dismutase, and ceruloplasmin, as well as fibrinolytic protein activity metalloproteases 2 and 3, plasmin, and urokinase plasminogen activator, were analyzed by zymography assays. Oxidized proteins were also determined.
Significant activity of antioxidant enzymes and fibrinolytic molecules was observed on day 5 of storage in PRP homogenates, which increased over time and was concomitantly correlated with oxidized protein levels. Reverse enzymatic activity patterns were observed in plasma, except for CAT, which remained unchanged.
Storage conditions of platelets from PRP for up to 5 days induced in vitro platelet activation by oxidative damage and proteolysis. This finding confirms the need for proper management of these blood products to preserve their viability and functionality.
富含血小板的血浆 (PRP) 被用于多种凝血障碍。其治疗效果依赖于与 PRP 采集和保存相关的技术程序,因为自由基会诱导血小板的激活和老化。这项工作旨在阐明在储存过程中从 PRP 获得的血小板激活所涉及的氧化机制。
从健康供体中获得了 110 份 PRP 批次,并在 20-24°C 的温度下进行搅拌保存。在从第 1 天到第 20 天的不同储存时间,从血小板匀浆和血浆中提取蛋白质。通过酶谱分析测定抗氧化标记物(如过氧化氢酶 (CAT)、超氧化物歧化酶和铜蓝蛋白)以及纤维蛋白溶解蛋白活性金属蛋白酶 2 和 3、纤溶酶和尿激酶纤溶酶原激活物的活性。还测定了氧化蛋白。
在 PRP 匀浆中储存的第 5 天观察到抗氧化酶和纤维蛋白溶解分子的显著活性,随着时间的推移,其活性增加,并与氧化蛋白水平呈正相关。在血浆中观察到相反的酶活性模式,除了 CAT 保持不变。
PRP 血小板储存 5 天以内的条件会导致体外血小板通过氧化损伤和蛋白水解而激活。这一发现证实了需要对这些血液制品进行适当的管理,以保持其活力和功能。