Abrica-González Paulina, Zamora-Justo José Alberto, Sotelo-López Antonio, Vázquez-Martínez Guillermo Rocael, Balderas-López José Abraham, Muñoz-Diosdado Alejandro, Ibáñez-Hernández Miguel
Instituto Politécnico Nacional, Basic Sciences Department, Unidad Profesional Interdisciplinaria de Biotecnología, 07340, Mexico City, Mexico.
Instituto Politécnico Nacional, Biochemistry Department, Escuela Nacional de Ciencias Biológicas, 11340, Mexico City, Mexico.
Nanoscale Res Lett. 2019 Jul 30;14(1):258. doi: 10.1186/s11671-019-3083-y.
Currently, gold nanoparticles have found applications in engineering and medical sciences, taking advantage from their properties and characteristics. Surface plasmon resonance, for instance, is one of the main features for optical applications and other physical properties, like high density, that represents the key for cellular uptake. Among other applications, in the medical field, some diseases may be treated by using gene therapy, including monogenetic or polygenetic disorders and infections. Gene adding, suppression, or substitution is one of the many options for genetic manipulation. This work explores an alternative non-viral method for gene transfer by using gold nanoparticles functionalized with organic polymers; two routes of synthesis were used: one of them with sodium borohydride as reducing agent and the other one with chitosan oligosaccharide as reducing and stabilizing agent. Gold nanoparticles conjugated with chitosan, acylated chitosan and chitosan oligosaccharide, were used to evaluate transfection efficiency of plasmid DNA into cell culture (HEK-293). Physical and chemical properties of gold nanocomposites were characterized by using UV-Vis Spectroscopy, ξ-potential, and transmission electron microscopy. Furthermore, the interaction between gold nanoparticles and plasmid DNA was demonstrated by using agarose gel electrophoresis. Transfection tests were performed and evaluated by β-galactosidase activity and green fluorescence protein expression. The percentage of transfection obtained with chitosan, acylated chitosan, and chitosan oligosaccharide were of 27%, 33%, and 60% respectively.
目前，金纳米颗粒凭借其性质和特点已在工程学和医学领域得到应用。例如，表面等离子体共振是光学应用的主要特性之一，而诸如高密度等其他物理性质则是细胞摄取的关键。在医学领域的其他应用中，一些疾病可以通过基因疗法进行治疗，包括单基因或多基因疾病以及感染。基因添加、抑制或替换是基因操作的众多选择之一。这项工作探索了一种通过使用有机聚合物功能化的金纳米颗粒进行基因转移的非病毒替代方法；采用了两种合成路线：一种以硼氢化钠作为还原剂，另一种以壳寡糖作为还原剂和稳定剂。将与壳聚糖、酰化壳聚糖和壳寡糖共轭的金纳米颗粒用于评估质粒DNA导入细胞培养物（HEK-293）的转染效率。通过紫外可见光谱、ξ电位和透射电子显微镜对金纳米复合材料的物理和化学性质进行了表征。此外，通过琼脂糖凝胶电泳证明了金纳米颗粒与质粒DNA之间的相互作用。通过β-半乳糖苷酶活性和绿色荧光蛋白表达进行转染测试并评估。壳聚糖、酰化壳聚糖和壳寡糖获得的转染百分比分别为27%、33%和60%。
