Biodegradable insulin-loaded PLGA microspheres fabricated by three different emulsification techniques: Investigation for cartilage tissue engineering
Biodegradable insulin-loaded PLGA microspheres fabricated by three different emulsification techniques: Investigation for cartilage tissue engineering
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DOI：
10.1016/j.actbio.2010.12.014
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发表时间：
2011-04-01
影响因子：
9.7
通讯作者：
Ringe, Jochen
中科院分区：
文献类型：
--
作者：
Andreas, Kristin;Zehbe, Rolf;Ringe, Jochen
生长、分化和迁移因子促进组织的工程化，但需要在延长的时间段内以限定的梯度施用。在这项研究中，胰岛素作为软骨组织工程的生长因子和可生物降解的PLGA输送装置。目的是比较研究三种不同的微胶囊化技术，固体-油-水（s/o/w），水-油-水（w/o/w）和油-油-水（o/o/w），用于制备负载胰岛素的PLGA微球的蛋白负载效率，释放和降解动力学，释放的蛋白的生物活性和微球的吞噬功能。三种乳化技术制备的胰岛素PLGA微球表面光滑、球形，Zeta电位为负。制备技术不影响颗粒降解，也不诱导人白细胞的吞噬作用。使用圆二色光谱和基于MCF 7细胞的增殖测定显示从微球递送结构完整和生物活性的胰岛素。然而，胰岛素负载效率（w/o/w约80%，s/o/w 60%，和o/o/w 25%）和胰岛素释放动力学受微囊化技术的影响。结果表明，w/o/w微球是最合适的，提供了高的包封率和低的初始突释，因此这些最终用于软骨组织工程。胰岛素释放的w/o/w PLGA微球刺激软骨形成相当大的软骨细胞高密度小球培养，如所确定的蛋白多糖和II型胶原蛋白的分泌增加。我们的研究结果应该鼓励进一步的研究应用蛋白质负载PLGA微球结合细胞移植或无细胞原位组织工程植入物再生软骨。(C)2010 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
Growth, differentiation and migration factors facilitate the engineering of tissues but need to be administered with defined gradients over a prolonged period of time. In this study insulin as a growth factor for cartilage tissue engineering and a biodegradable PLGA delivery device were used. The aim was to investigate comparatively three different microencapsulation techniques, solid-in-oil-in-water (s/o/w), water-in-oil-in-water (w/o/w) and oil-in-oil-in-water (o/o/w), for the fabrication of insulin-loaded PLGA microspheres with regard to protein loading efficiency, release and degradation kinetics, biological activity of the released protein and phagocytosis of the microspheres. Insulin-loaded PLGA microspheres prepared by all three emulsification techniques had smooth and spherical surfaces with a negative zeta potential. The preparation technique did not affect particle degradation nor induce phagocytosis by human leukocytes. The delivery of structurally intact and biologically active insulin from the microspheres was shown using circular dichroism spectroscopy and a MCF7 cell-based proliferation assay. However, the insulin loading efficiency (w/o/w about 80%, s/o/w 60%, and o/o/w 25%) and the insulin release kinetics were influenced by the microencapsulation technique. The results demonstrate that the w/o/w microspheres are most appropriate, providing a high encapsulation efficiency and low initial burst release, and thus these were finally used for cartilage tissue engineering. Insulin released from w/o/w PLGA microspheres stimulated the formation of cartilage considerably in chondrocyte high density pellet cultures, as determined by increased secretion of proteoglycans and collagen type II. Our results should encourage further studies applying protein-loaded PLGA microspheres in combination with cell transplants or cell-free in situ tissue engineering implants to regenerate cartilage. (C) 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.