Jantakee Kanyaluck, Prapan Ausanai, Chaiwaree Saranya, Suwannasom Nittiya, Kaewprayoon Waraporn, Georgieva Radostina, Tragoolpua Yingmanee, Bäumler Hans
Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand.
The Graduate School, Chiang Mai University, Chiang Mai 50200, Thailand.
Polymers (Basel). 2021 Jan 21;13(3):334. doi: 10.3390/polym13030334.
Non-sericin (NS) extract was produced from the ethanolic extract of silk cocoons. This extract is composed of both carotenoids and flavonoids. Many of these compounds are composed of substances of poor aqueous solubility. Thus, this study focused on the development of a carrier system created from biocompatible and biodegradable materials to improve the biological activity of NS extracts. Accordingly, NS was incorporated into human serum albumin template particles with MnCO (NS-HSA MPs) by loading NS into the preformed HAS-MnCO microparticles using the coprecipitation crosslinking dissolution technique (CCD-technique). After crosslinking and template dissolution steps, the NS loaded HSA particles are negatively charged, have a size ranging from 0.8 to 0.9 µm, and are peanut shaped. The degree of encapsulation efficiency ranged from 7% to 57% depending on the initial NS concentration and the steps of adsorption. In addition, NS-HSA MPs were taken up by human lung adenocarcinoma (A549 cell) for 24 h. The promotion of cellular uptake was evaluated by flow cytometry and the results produced 99% fluorescent stained cells. Moreover, the results from CLSM and 3D fluorescence imaging confirmed particle localization in the cells. Interestingly, NS-HSA MPs could not induce inflammation through nitric oxide production from macrophage RAW264.7 cells. This is the first study involving the loading of non-sericin extracts into HSA MPs by CCD technique to enhance the bioavailability and biological effects of NS. Therefore, HSA MPs could be utilized as a carrier system for hydrophobic substances targeting cells with albumin receptors.
非丝胶蛋白（NS）提取物由蚕茧的乙醇提取物制备而成。该提取物含有类胡萝卜素和黄酮类化合物。这些化合物中的许多都由水溶性差的物质组成。因此，本研究着重于开发一种由生物相容性和可生物降解材料制成的载体系统，以提高NS提取物的生物活性。相应地，通过共沉淀交联溶解技术（CCD技术）将NS加载到预先形成的HAS-MnCO微粒中，从而将NS掺入含碳酸锰的人血清白蛋白模板颗粒（NS-HSA MPs）中。经过交联和模板溶解步骤后，负载NS的HSA颗粒带负电荷，尺寸范围为0.8至0.9μm，呈花生形状。包封效率取决于初始NS浓度和吸附步骤，范围为7%至57%。此外，NS-HSA MPs被人肺腺癌（A549细胞）摄取24小时。通过流式细胞术评估细胞摄取的促进情况，结果显示99%的细胞呈荧光染色。此外，共聚焦激光扫描显微镜（CLSM）和三维荧光成像的结果证实了颗粒在细胞中的定位。有趣的是，NS-HSA MPs不会通过巨噬细胞RAW264.7细胞产生一氧化氮来诱导炎症。这是第一项通过CCD技术将非丝胶蛋白提取物加载到HSA MPs中以提高NS生物利用度和生物学效应的研究。因此，HSA MPs可作为一种载体系统，用于将疏水性物质靶向具有白蛋白受体的细胞。
