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Synthesis and application of photodegradable microspheres for spatiotemporal control of protein delivery

机译:合成和可光降解的微球体的应用蛋白质递送的时空控制

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摘要

Here, we present a photodegradable microparticle system that can be employed to entrap and deliver bioactive proteins to cells during culture. By using a photosensitive delivery system, experimenters can achieve a wide variety of spatiotemporally regulated release profiles with a single microparticle formulation, thereby enabling one to probe many questions as to how protein presentation can be manipulated to regulate cell function. Photodegradable microparticles were synthesized via inverse suspension polymerization with a mean diameter of 22 μm, and degradation was demonstrated upon exposure to several irradiation conditions. The protein-loaded depots were incorporated into cell cultures and release of bioactive protein was quantified during the photodegradation process. This phototriggered release allowed for the delivery of TGF-β1 to stimulate PE25 cells and for the delivery of fluorescently labeled Annexin V to assay apoptotic 3T3 fibroblasts during culture. By incorporating these photoresponsive protein delivery depots into cell culture, new types of experiments are now possible to test hypotheses about how individual or multiple soluble factors might affect cell function when presented in a uniform, temporally varying, or gradient manner.
机译:这里,我们介绍可光降解的微粒系统,可用于在培养过程中诱捕和将生物活性蛋白传递给细胞。通过使用光敏递送系统,实验者可以实现具有单个微粒制剂的各种时尚调节的释放曲线,从而能够探讨如何操纵蛋白质呈现以调节细胞功能的许多问题。通过平均直径为22μm的逆悬浮聚合来合成可光降解的微粒,并且在暴露于几种照射条件下证明降解。将蛋白质加载的储物掺入细胞培养物中,在光降解过程中定量了生物活性蛋白的释放。这种光电触发释放允许递送TGF-β1以刺激PE25细胞,并在培养过程中递送荧光标记的膜蛋白V以测定凋亡3T3成纤维细胞。通过将这些光响应蛋白质递送贮存掺入细胞培养物中,现在可以在呈现均匀,时间变化或梯度方式时,测试新类型的实验可以测试关于个体或多种可溶性因子的假设可能影响细胞功能。

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