Rosell-Valle Cristina, Martín-López María, Campos Fernando, Chato-Astrain Jesús, Campos-Cuerva Rafael, Alaminos Miguel, Santos González Mónica
Unidad de Producción y Reprogramación Celular de Sevilla (UPRC), Red Andaluza de Diseño y Traslación de Terapias Avanzadas (RADyTTA), Seville, Spain.
Escuela Internacional de Doctorado Universidad de Sevilla, Seville, Spain.
Front Bioeng Biotechnol. 2022 Aug 23;10:908250. doi: 10.3389/fbioe.2022.908250. eCollection 2022.
Fibrin is widely used for tissue engineering applications. The use of blood derivatives, however, carries a high risk of transmission of infectious agents, necessitating the application of pathogen reduction technology (PRT). The impact of this process on the structural and biomechanical properties of the final products is unknown. We used normal plasma (PLc) and plasma inactivated by riboflavin and ultraviolet light exposure (PLi) to manufacture nanostructured cellularized fibrin-agarose hydrogels (NFAHs), and then compared their structural and biomechanical properties. We also measured functional protein C, prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and coagulation factors [fibrinogen, Factor (F) V, FVIII, FX, FXI, FXIII] in plasma samples before and after inactivation. The use of PLi to manufacture cellularized NFAHs increased the interfibrillar spacing and modified their biomechanical properties as compared with cellularized NFAH manufactured with PLc. PLi was also associated with a significant reduction in functional protein C, FV, FX, and FXI, and an increase in the international normalized ratio (derived from the PT), APTT, and TT. Our findings demonstrate that the use of PRT for fibrin-agarose bioartificial tissue manufacturing does not adequately preserve the structural and biomechanical properties of the product. Further investigations into PRT-induced changes are warranted to determine the applications of NFAH manufactured with inactivated plasma as a medicinal product.
纤维蛋白广泛应用于组织工程领域。然而，使用血液衍生物存在较高的感染因子传播风险，因此需要应用病原体灭活技术（PRT）。该过程对最终产品的结构和生物力学性能的影响尚不清楚。我们使用正常血浆（PLc）和经核黄素及紫外线照射灭活的血浆（PLi）制备了纳米结构的细胞化纤维蛋白-琼脂糖水凝胶（NFAH），然后比较了它们的结构和生物力学性能。我们还测量了灭活前后血浆样本中的功能性蛋白C、凝血酶原时间（PT）、活化部分凝血活酶时间（APTT）、凝血酶时间（TT）以及凝血因子[纤维蛋白原、因子（F）V、FVIII、FX、FXI、FXIII]。与用PLc制备的细胞化NFAH相比，使用PLi制备细胞化NFAH增加了纤维间间距并改变了其生物力学性能。PLi还与功能性蛋白C、FV、FX和FXI的显著降低以及国际标准化比值（由PT得出）、APTT和TT的增加有关。我们的研究结果表明，在纤维蛋白-琼脂糖生物人工组织制造中使用PRT并不能充分保留产品的结构和生物力学性能。有必要对PRT引起的变化进行进一步研究，以确定用灭活血浆制备的NFAH作为医药产品的应用。
