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阳离子聚乳酸-羟基乙酸共聚物纳米球作为DNA载体的制备与表征

Preparation and characterization of cationic PLGA nanospheres as DNA carriers.

作者信息

Ravi Kumar M N V, Bakowsky U, Lehr C M

机构信息

Department of Biopharmaceutics and Pharmaceutical Technology, Saarland University, D 66123, Saarbrücken, Germany.

出版信息

Biomaterials. 2004 May;25(10):1771-7. doi: 10.1016/j.biomaterials.2003.08.069.

Abstract

Nanoparticles formulated from biodegradable polymers such as poly(lactic acid) (PLA) and poly(lactide-co-glycolide) (PLGA) are being extensively investigated as non-viral gene delivery systems due to their controlled release characteristics and biocompatibility. PLGA nanoparticles for DNA delivery are mainly formulated by an emulsion-solvent evaporation technique using PVA as a stabilizer generating negatively charged particles and heterogeneous size distribution. The objective of the present study was to formulate cationically modified PLGA nanoparticles with defined size and shape that can efficiently bind DNA. An Emulsion-diffusion-evaporation technique to make cationic nanospheres composed of biodegradable and biocompatible co-polyester PLGA has been developed. PVA-chitosan blend was used to stabilize the PLGA nanospheres. The nanospheres were characterized by atomic force microscopy (AFM), photon-correlation spectroscopy (PCS), and Fourier transform infrared spectroscopy (FTIR). Zeta potential and gel electrophoresis studies were also performed to understand the surface properties of nanospheres and their ability to condense negatively charged DNA. The designed nanospheres have a zeta potential of 10mV at pH 7.4 and size under 200nm. From the gel electrophoresis studies we found that the charge on the nanospheres is sufficient to efficiently bind the negatively charged DNA electrostatically. These cationic PLGA nanospheres could serve as potential alternatives of the existing negatively charged nanoparticles.

摘要

由可生物降解聚合物如聚乳酸(PLA)和聚丙交酯-乙交酯共聚物(PLGA)制备的纳米颗粒,因其控释特性和生物相容性,正作为非病毒基因递送系统被广泛研究。用于DNA递送的PLGA纳米颗粒主要通过乳液-溶剂蒸发技术制备,使用聚乙烯醇(PVA)作为稳定剂,产生带负电荷的颗粒且粒径分布不均一。本研究的目的是制备具有特定尺寸和形状、能有效结合DNA的阳离子修饰PLGA纳米颗粒。已开发出一种乳液-扩散-蒸发技术来制备由可生物降解且生物相容的共聚酯PLGA组成的阳离子纳米球。使用PVA-壳聚糖共混物来稳定PLGA纳米球。通过原子力显微镜(AFM)、光子相关光谱法(PCS)和傅里叶变换红外光谱法(FTIR)对纳米球进行表征。还进行了zeta电位和凝胶电泳研究,以了解纳米球的表面性质及其凝聚带负电荷DNA的能力。所设计的纳米球在pH 7.4时zeta电位为10mV,粒径小于200nm。从凝胶电泳研究中我们发现,纳米球上的电荷足以通过静电作用有效结合带负电荷的DNA。这些阳离子PLGA纳米球可作为现有带负电荷纳米颗粒的潜在替代品。

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