A simple and robust method for pre-wetting poly (lactic-co-glycolic) acid microspheres.
A simple and robust method for pre-wetting poly (lactic-co-glycolic) acid microspheres.
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DOI：
10.1177/0885328215577297
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发表时间：
2015-08
影响因子：
2.9
通讯作者：
Day RM
中科院分区：
文献类型：
--
作者：
Wright B;Parmar N;Bozec L;Aguayo SD;Day RM
聚乳酸-羟基乙酸微球适用于多种生物医学程序，以支持细胞、药物、多肽或基因的输送。聚乳酸-乙醇酸的亲水性或润湿性是细胞附着的重要前提条件，可通过暴露在血浆氧或氮气中、用氢氧化钠或氯酸表面水解、浸泡在乙醇和水中或在磷酸盐缓冲盐水或细胞培养液中长时间孵育来实现。本研究的目的是开发一种简单的润湿聚乳酸-羟基乙酸微球的方法，用于细胞递送应用。本文介绍了一种一步乙醇浸泡工艺，即在PLGA微球中加入含血清的培养液和乙醇，浸泡时间为30 min-24 h。该方案提供了一种比传统的两步湿法更有效的方法。人骨骼肌成肌细胞在聚乳酸-羟基乙酸微球上的附着取决于润湿程度、乙醇预润湿引起的表面形貌变化和血清蛋白吸附。乙醇浓度为70%(v/v)和100%时，促进了相似程度的润湿。用35%(v/v)乙醇润湿微球仅在24 h后才能实现。用70%(v/v)乙醇预润湿(3 h以上)微球所允许的血清蛋白吸附显著高于用35%(v/v)乙醇润湿微球(p ≤ 0.01)。在血清蛋白负载的微球上，70%乙醇湿润的构建物上附着的成肌细胞数量比35%(v/v)乙醇部分润湿的构建体上的成肌细胞数量多。用70%(v/v)乙醇处理的微球比用35%(v/v)乙醇处理的微球呈现出更多的皱纹表面，这表明成肌细胞与前者更有效的黏附可能至少部分归因于表面结构的差异。我们得出的结论是，我们的新方案预湿聚(乳酸-羟基乙酸)微球，将生化和结构特征结合到这种生物材料中，可以促进成肌细胞的输送，用于临床环境。
Poly (lactic-co-glycolic) acid microspheres are amenable to a number of biomedical procedures that support delivery of cells, drugs, peptides or genes. Hydrophilisation or wetting of poly (lactic-co-glycolic) acid are an important pre-requisites for attachment of cells and can be achieved via exposure to plasma oxygen or nitrogen, surface hydrolysis with NaOH or chloric acid, immersion in ethanol and water, or prolonged incubation in phosphate buffered saline or cell culture medium. The aim of this study is to develop a simple method for wetting poly (lactic-co-glycolic) acid microspheres for cell delivery applications. A one-step ethanol immersion process that involved addition of serum-supplemented medium and ethanol to PLGA microspheres over 30 min–24 h is described in the present study. This protocol presents a more efficient methodology than conventional two-step wetting procedures. Attachment of human skeletal myoblasts to poly (lactic-co-glycolic) acid microspheres was dependent on extent of wetting, changes in surface topography mediated by ethanol pre-wetting and serum protein adsorption. Ethanol, at 70% (v/v) and 100%, facilitated similar levels of wetting. Wetting with 35% (v/v) ethanol was only achieved after 24 h. Pre-wetting (over 3 h) with 70% (v/v) ethanol allowed significantly greater (p ≤ 0.01) serum protein adsorption to microspheres than wetting with 35% (v/v) ethanol. On serum protein-loaded microspheres, greater numbers of myoblasts attached to constructs wetted with 70% ethanol than those partially wetted with 35% (v/v) ethanol. Microspheres treated with 70% (v/v) ethanol presented a more rugose surface than those treated with 35% (v/v) ethanol, indicating that more efficient myoblast adhesion to the former may be at least partially attributed to differences in surface structure. We conclude that our novel protocol for pre-wetting poly (lactic-co-glycolic) acid microspheres that incorporates biochemical and structural features into this biomaterial can facilitate myoblast delivery for use in clinical settings.