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基于具有正细胞靶向能力的 halloysite 纳米管-碳点杂化多功能纳米载体作为一种有潜力的非病毒基因载体的口服基因治疗。

Halloysite nanotubes-carbon dots hybrids multifunctional nanocarrier with positive cell target ability as a potential non-viral vector for oral gene therapy.

机构信息

Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), University of Palermo, Viale delle Scienze, Ed. 17, 90128 Palermo, Italy.

Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche (STEBICEF), University of Palermo, Viale delle Scienze, Ed. 17, 90128 Palermo, Italy; Institute of Biomedicine and Molecular Immunology, CNR, IBIM, Via Ugo La Malfa, 153, 90146 Palermo, Italy.

出版信息

J Colloid Interface Sci. 2019 Sep 15;552:236-246. doi: 10.1016/j.jcis.2019.05.062. Epub 2019 May 20.

Abstract

HYPOTHESIS

The use of non-viral vectors for gene therapy is hindered by their lower transfection efficiency and their lacking of self-track ability.

EXPERIMENTS

This study aims to investigate the biological properties of halloysite nanotubes-carbon dots hybrid and its potential use as non-viral vector for oral gene therapy. The morphology and the chemical composition of the halloysite hybrid were investigated by means of high angle annular dark field scanning TEM and electron energy loss spectroscopy techniques, respectively. The cytotoxicity and the antioxidant activity were investigated by standard methods (MTS, DPPH and HO, respectively) using human cervical cancer HeLa cells as model. Studies of cellular uptake were carried out by fluorescence microscopy. Finally, we investigated the loading and release ability of the hybrid versus calf thymus DNA by fluorescence microscopy, circular dichroism, dynamic light scattering and ζ-potential measurements.

FINDINGS

All investigations performed confirmed the existence of strong electrostatic interactions between the DNA and the halloysite hybrid, so it shows promise as a multi-functional cationic non-viral vector that has also possesses intracellular tracking capability and promising in vitro antioxidant potential.

摘要

假设

非病毒载体在基因治疗中的应用受到其转染效率较低和缺乏自我跟踪能力的限制。

实验

本研究旨在探讨纳米管-碳点杂化材料的生物学特性及其作为口服基因治疗非病毒载体的潜在用途。采用高角环形暗场扫描透射电子显微镜和电子能量损失谱技术分别对纳米管-碳点杂化材料的形貌和化学组成进行了研究。采用 MTS、DPPH 和 HO 分别作为标准方法,以人宫颈癌 HeLa 细胞为模型,对细胞毒性和抗氧化活性进行了研究。通过荧光显微镜研究了细胞摄取情况。最后,通过荧光显微镜、圆二色性、动态光散射和 ζ-电位测量研究了杂化材料对小牛胸腺 DNA 的负载和释放能力。

发现

所有的研究都证实了 DNA 与纳米管-碳点杂化材料之间存在强烈的静电相互作用,因此它有望成为一种多功能阳离子非病毒载体,具有细胞内跟踪能力和体外抗氧化潜力。

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