Post-Graduation Program in Dentistry, Fluminense Federal University, Niterói, Rio de Janeiro, Brazil.
Laboratory of Ultrastructure and Cellular Biology Hertha Meyer, Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
J Biomed Mater Res A. 2018 May;106(5):1373-1380. doi: 10.1002/jbm.a.36346. Epub 2018 Jan 31.
Platelet-rich fibrin membranes are biomaterials widely used for therapeutic purposes, and canonically produced through the processing of peripheral blood with fixed-angle rotor centrifuges. In this work, we evaluate the in vitro stability and release of cytokines and growth factors when these biomaterials are produced with a horizontal swing-out clinical centrifuge. Membranes produced from the blood of 14 donors were morphologically evaluated by scanning electron microscopy and fluorescence microscopy, and their stability was assessed by photographic recording after incubation in culture medium for up to 28 days. The release of 27 cytokines and growth factors was monitored for three weeks through a multiparametric immunoassay. The fibrin membranes presented complex three-dimensional structure with a high density of nucleated cells. A large release of growth factors [platelet derived growth factor, fibroblastic growth factor (bFGF), and vascular endothelial growth factor] was detected in the first 24 h, followed by time-dependent decay, maintaining significant concentrations after three weeks. Both anti-inflammatory and pro-inflammatory cytokines presented different release peaks, maintaining high rates of elution for up to 21 days. Chemokines of relevance in tissue repair [RANTES, granulocyte colony-stimulating factor (G-CSF)] were also produced in large quantities throughout the experimental period. The present results demonstrate that blood-derived fibrin membranes with high structural stability and cell content can be generated by horizontal centrifugation, being able of a prolonged production/release of growth factors and pro- and anti-inflammatory cytokines. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1373-1380, 2018.
富血小板纤维蛋白膜是一种广泛用于治疗目的的生物材料，通常通过使用固定角度转子离心机处理外周血来制备。在这项工作中，我们评估了使用水平摆动式临床离心机制备这些生物材料时细胞因子和生长因子的体外稳定性和释放。通过扫描电子显微镜和荧光显微镜对 14 位供体的血液制备的膜进行形态学评估，并通过在培养物中孵育长达 28 天的摄影记录评估其稳定性。通过多参数免疫测定监测 27 种细胞因子和生长因子的释放长达三周。纤维蛋白膜具有复杂的三维结构，有大量有核细胞。在最初的 24 小时内检测到大量生长因子（血小板衍生生长因子、成纤维细胞生长因子（bFGF）和血管内皮生长因子）的释放，随后呈时间依赖性衰减，在三周后仍保持显著浓度。抗炎和促炎细胞因子均表现出不同的释放峰值，在长达 21 天的时间内保持高洗脱率。组织修复中相关的趋化因子[RANTES、粒细胞集落刺激因子（G-CSF）]也在整个实验期间大量产生。这些结果表明，通过水平离心可以产生具有高结构稳定性和细胞含量的血液衍生纤维蛋白膜，能够长时间持续释放生长因子和促炎及抗炎细胞因子。© 2018 Wiley Periodicals, Inc. J 生物材料 Res 部分 A：106A：1373-1380，2018 年。
