Cosco Donato, Federico Cinzia, Maiuolo Jessica, Bulotta Stefania, Molinaro Roberto, Paolino Donatella, Tassone Pierfrancesco, Fresta Massimo
Department of Health Sciences, University Magna Græcia of Catanzaro, Catanzaro, Italy ; Interregional Research Center for Food Safety and Health, University of Catanzaro "Magna Græcia", Catanzaro, Italy.
Department of Health Sciences, University Magna Græcia of Catanzaro, Catanzaro, Italy ; Department of Experimental and Clinical Medicine, University Magna Græcia of Catanzaro, Catanzaro, Italy.
Int J Nanomedicine. 2014 May 15;9:2359-72. doi: 10.2147/IJN.S58362. eCollection 2014.
The aim of this study was the evaluation of the effects of two emulsifiers on the physicochemical and technological properties of low molecular weight chitosan/poly (D,L-lactide-co-glycolide) (PLGA) nanoplexes and their transfection efficiency. Nanospheres were prepared using the nanoprecipitation method of the preformed polymer. The mean diameter and surface charge of the nanospheres were investigated by photocorrelation spectroscopy. The degree of binding of the plasmid with the nanoplexes was qualitatively and quantitatively determined. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) testing was performed using HeLa, RPMI8226, and SKMM1 cell lines. Flow cytometry and confocal laser scanning microscopy were used to determine the degree of cellular transfection and internalization of the nanoplexes into cells, respectively. The nanoplexes had a positive zeta potential, and low amounts of PLGA and poloxamer 188 showed a mean colloidal size of ~200 nm with a polydispersity index of ~0.14. The nanoplexes had suitable entrapment efficiency (80%). In vitro experiments showed that the colloidal nanodevices did not induce significant cytotoxicity. The nanoplexes investigated in this study could represent efficient and useful nonviral devices for gene delivery. Use of low amounts of PLGA and poloxamer 188 enabled development of a nanosphere able to transfect cells efficiently. These nanosystems are a helpful platform for delivery of genetic material while preserving therapeutic efficacy.
本研究的目的是评估两种乳化剂对低分子量壳聚糖/聚(D,L-丙交酯-共-乙交酯)(PLGA)纳米复合物的物理化学和技术性质及其转染效率的影响。使用预制聚合物的纳米沉淀法制备纳米球。通过光相关光谱法研究纳米球的平均直径和表面电荷。定性和定量地测定质粒与纳米复合物的结合程度。使用HeLa、RPMI8226和SKMM1细胞系进行MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)测试。分别使用流式细胞术和共聚焦激光扫描显微镜来确定纳米复合物的细胞转染程度和内化到细胞中的情况。纳米复合物具有正的zeta电位,少量的PLGA和泊洛沙姆188显示平均胶体尺寸约为200 nm,多分散指数约为0.14。纳米复合物具有合适的包封效率(80%)。体外实验表明,胶体纳米器件不会诱导明显的细胞毒性。本研究中研究的纳米复合物可以代表用于基因递送的高效且有用的非病毒器件。使用少量的PLGA和泊洛沙姆188能够开发出一种能够有效转染细胞的纳米球。这些纳米系统是在保持治疗效果的同时递送遗传物质的有用平台。