Rodrigues Susana, Cardoso Lurdes, da Costa Ana M Rosa, Grenha Ana
CBMR-Centre for Biomedical Research, University of Algarve, Faculty of Sciences and Technology, Campus de Gambelas, Faro 8005-139, Portugal.
CIQA-Algarve Chemistry Research Centre and Department of Chemistry and Pharmacy, Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, Faro 8005-139, Portugal.
Materials (Basel). 2015 Aug 28;8(9):5647-5670. doi: 10.3390/ma8095268.
Chitosan (CS) and chondroitin sulfate (CHS) are natural polymers with demonstrated applicability in drug delivery, while nanoparticles are one of the most explored carriers for transmucosal delivery of biopharmaceuticals. In this work we have prepared CS/CHS nanoparticles and associated for the first time the therapeutic protein insulin. Fluorescein isothiocyanate bovine serum albumin (FITC-BSA) was also used to enable comparison of behaviors regarding differences in molecular weight (5.7 kDa 67 kDa). Nanoparticles of approximately 200 nm and positive zeta potential around +20 mV were obtained. These parameters remained stable for up to 1 month at 4 °C. Proteins were associated with efficiencies of more than 50%. The release of FITC-BSA in PBS pH 7.4 was more sustained (50% in 24 h) than that of insulin (85% in 24 h). The biocompatibility of nanoparticles was tested in Calu-3 and A549 cells by means of three different assays. The metabolic assay MTT, the determination of lactate dehydrogenase release, and the quantification of the inflammatory response generated by cell exposure to nanoparticles have indicated an absence of overt toxicity. Overall, the results suggest good indications on the application of CS/CHS nanoparticles in respiratory transmucosal protein delivery, but the set of assays should be widened to clarify obtained results.
壳聚糖（CS）和硫酸软骨素（CHS）是天然聚合物，在药物递送方面已显示出适用性，而纳米颗粒是生物药物经粘膜递送研究最多的载体之一。在这项工作中，我们制备了CS/CHS纳米颗粒，并首次将治疗性蛋白质胰岛素与之结合。还使用了异硫氰酸荧光素牛血清白蛋白（FITC-BSA）来比较分子量差异（5.7 kDa对67 kDa）下的行为。获得了直径约200 nm、zeta电位为+20 mV左右的纳米颗粒。这些参数在4°C下可稳定长达1个月。蛋白质结合效率超过50%。FITC-BSA在pH 7.4的磷酸盐缓冲盐溶液（PBS）中的释放比胰岛素更持久（24小时内释放50%，而胰岛素为85%）。通过三种不同的测定方法在Calu-3和A549细胞中测试了纳米颗粒的生物相容性。代谢测定MTT、乳酸脱氢酶释放的测定以及细胞暴露于纳米颗粒产生的炎症反应的定量表明没有明显毒性。总体而言，结果表明CS/CHS纳米颗粒在呼吸道经粘膜蛋白质递送中的应用有良好前景，但应扩大测定方法的范围以阐明所得结果。
