Department of Periodontology, Faculty of Dentistry, Hacettepe University, Ankara, Turkey.
J Biomed Mater Res B Appl Biomater. 2013 Jan;101(1):28-35. doi: 10.1002/jbm.b.32806. Epub 2012 Sep 15.
The aim of this study is to compare the effects of different platelet-rich plasma (PRP) preparation methods on platelet activity and to investigate the growth factor (GF) release kinetics from PRP-loaded chitosan scaffolds for tissue engineering applications. Flow cytometry analysis showed that centrifugation processes used for PRP preparation did not cause significant effect on platelet activation levels by means of markers investigated. Two different methods were used to prepare PRP-loaded chitosan scaffolds: (i) PRP was added to chitosan gel before freeze-drying to prepare scaffolds called as "GEL" and (ii) PRP was embedded to freeze-dried chitosan scaffolds to prepare scaffolds called as "SPONGE." In addition, nonactivated PRP and PRP activated with type-I collagen were used as control groups. Scanning electron microscopy images demonstrated that, in GEL group, there is no deterioration on the scaffolds porous, 3D, and interconnected structure. GF release kinetics was determined by enzyme-linked immunosorbent assay for platelet-derived GF-BB, transforming GF-β1, and insulin-like GF-1. A sustained release of GFs was achieved in GEL group while a sharp burst release was observed for all the GFs from the SPONGE groups. Moreover, platelet-derived GF-BB, insulin-like GF-1, and transforming GF-β1 releases were prolonged to 20 days in GEL groups, and the biological activities of all GFs released from GEL and SPONGE scaffolds were preserved. This study demonstrated that chitosan scaffold that was called GEL could be an appropriate carrier for PRP applications by providing sustained release of GFs.
本研究旨在比较不同富血小板血浆（PRP）制备方法对血小板活性的影响，并研究 PRP 负载壳聚糖支架的生长因子（GF）释放动力学，以应用于组织工程。流式细胞术分析表明，PRP 制备过程中使用的离心过程不会通过所研究的标志物对血小板激活水平产生显著影响。使用两种不同的方法制备 PRP 负载壳聚糖支架：（i）在冷冻干燥前将 PRP 添加到壳聚糖凝胶中以制备称为“GEL”的支架，和（ii）将 PRP 嵌入冷冻干燥的壳聚糖支架中以制备称为“SPONGE”的支架。此外，还使用非激活的 PRP 和用 I 型胶原激活的 PRP 作为对照组。扫描电子显微镜图像表明，在 GEL 组中，支架的多孔、3D 和相互连接的结构没有恶化。通过酶联免疫吸附试验测定血小板衍生的 GF-BB、转化 GF-β1 和胰岛素样 GF-1 的 GF 释放动力学。在 GEL 组中实现了 GFs 的持续释放，而在所有 GFs 从 SPONGE 组中观察到急剧的爆发释放。此外，血小板衍生的 GF-BB、胰岛素样 GF-1 和转化 GF-β1 的释放在 GEL 组中延长至 20 天，并且从 GEL 和 SPONGE 支架释放的所有 GFs 的生物活性都得到了保留。本研究表明，称为 GEL 的壳聚糖支架可以通过提供 GFs 的持续释放成为 PRP 应用的合适载体。
