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Silk Particle Production Based on Silk/PVA Phase Separation Using a Microfabricated Co-flow Device

机译：基于微细加工同流装置的基于丝绸/ PVA相分离的丝绸颗粒生产

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Polymeric particles are ideal drug delivery systems due to their cellular uptake-relevant size. Microparticles could be developed for direct injection of drug formulations into a diseased site, such as a tumor, allowing for drug retention and slow drug exposure over time through sustained release mechanisms. silk fibroin has shown promise as a biocompatible biomaterial both in research and the clinic. Silk has been previously used to make particles using an emulsion-based method with poly(vinyl alcohol) (PVA). In this study, polydimethylsiloxane-based microfluidic devices were designed, fabricated, and characterized to produce silk particles through self-association of silk when exposed to PVA. Three main variables resulted in differences in particle size and size distribution, or polydispersity index (PDI). Utilizing a co-flow microfluidic device decreased the PDI of the silk particles as compared to an emulsion-based method (0.13 versus 0.65, respectively). With a flow-focusing microfluidics device, lowering the silk flow rate from 0.80 to 0.06 mL/h resulted in a decrease in the median particle size from 6.8 to 3.0 μm and the PDI from 0.12 to 0.05, respectively. Lastly, decreasing the silk concentration from 12% to 2% resulted in a decrease in the median particle size from 5.6 to 2.8 μm and the PDI from 0.81 to 0.25, respectively. Binding and release of doxorubicin, a cytotoxic drug commonly used for cancer treatment, with the fabricated silk particles was evaluated. Doxorubicin loading in the silk particles was approximately 41 µg/mg; sustained doxorubicin release occurred over 23 days. When the cytotoxicity of the released doxorubicin was tested on KELLY neuroblastoma cells, significant cell death was observed. To demonstrate the potential for internalization of the silk particles, both KELLY and THP-1-derived macrophages were exposed to fluorescently labelled silk particles for up to 24 h. With the macrophages, internalization of the silk particles was observed. Additionally, THP-1 derived macrophages exposure to silk particles increased TNF-α secretion. Overall, this microfluidics-based approach for fabricating silk particles utilizing PVA as a means to induce phase separation and silk self-assembly is a promising approach to control particle size and size distribution. These silk particles may be utilized for a variety of biomedical applications including drug delivery to multiple cell types within a tumor microenvironment.
机译：聚合物颗粒由于其与细胞摄取有关的尺寸而成为理想的药物递送系统。可以开发出用于将药物制剂直接注射到患病部位（例如肿瘤）中的微粒，从而通过持续释放机制使药物滞留并随着时间的流逝而缓慢地暴露于药物。丝素蛋白在研究和临床中均显示出作为生物相容性生物材料的前景。以前，丝绸已被用于通过乳液法与聚乙烯醇（PVA）制成颗粒。在这项研究中，基于聚二甲基硅氧烷的微流体装置经过设计，制造和表征，可以在暴露于PVA时通过丝绸的自缔合产生丝绸颗粒。三个主要变量导致了粒度和粒度分布或多分散指数（PDI）的差异。与基于乳液的方法相比，使用并流微流体装置降低了丝绸颗粒的PDI（分别为0.13和0.65）。使用聚流微流控装置，将丝的流速从0.80降低到0.06 mL / h，中值粒径从6.8降低到3.0μm，PDI从0.12降低到0.05。最后，将蚕丝浓度从12％降低到2％导致中值粒径从5.6降低到2.8μm，PDI从0.81降低到0.25。评估了阿霉素（一种通常用于癌症治疗的细胞毒性药物）与制成的蚕丝颗粒的结合和释放。蚕丝颗粒中阿霉素的含量约为41 µg / mg；阿霉素持续释放超过23天。当在KELLY神经母细胞瘤细胞上测试释放的阿霉素的细胞毒性时，观察到明显的细胞死亡。为了证明丝颗粒内部化的潜力，将KELLY和THP-1衍生的巨噬细胞均暴露于荧光标记的丝颗粒中长达24小时。使用巨噬细胞，观察到丝颗粒的内在化。此外，THP-1衍生的巨噬细胞暴露于丝颗粒会增加TNF-α的分泌。总体而言，这种基于微流体的方法利用PVA作为诱导相分离和蚕丝自组装的方法来制备蚕丝颗粒，是一种控制粒度和粒度分布的有前途的方法。这些蚕丝颗粒可用于多种生物医学应用，包括将药物递送至肿瘤微环境内的多种细胞类型。

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