Factors controlling particle size during nebulization of DNA-polycation complexes.

J Lynch, N Behan, Colin Birkinshaw
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引用次数: 11

Abstract

Pulmonary gene therapy has the potential to treat or cure respiratory diseases such as cystic fibrosis. Much work has focused on the delivery of genes to the lung using viral vectors with varying degrees of success. Viral vectors are problematic and undesirable for use in the lung because they can provoke an acute immune response. This study has focused on the characterization of nonviral, polymer-based gene vectors for use with nebulizers. Calf thymus DNA has been used as a model, and was complexed with each of the three polycations; 22 kDa linear polyethyleneimine, 25 kDa branched polyethyleneimine, and 29.5 kDa polylysine using water, glucose solution, and phosphate-buffered saline (PBS) as carrier liquids. Fourier transform infrared spectroscopy has shown that the DNA retains the B form during the complex formation. The complexes prepared at N:P ratios of 10, have been nebulized using a vibrating plate nebulizer and the particle size and Zeta potentials measured before and after nebulization. The particle size distributions of the DNA complexes prepared in water and glucose solution were unimodal before and after nebulization with a small increase in particle size following nebulization. Choice of complexing polymer is shown to have only a small effect on particle size with the dominant effect coming from the ionic character of the dispersion fluid. Complexes prepared in PBS, although originally unimodal, showed pronounced agglomeration on nebulization. With all polymers in water or glucose solution, the Zeta potential increases after nebulization, but with PBS as the carrier liquid the potential falls and is clearly associated with the observed agglomeration. Gel electrophoresis shows that the complexing polymers protect the DNA through the nebulization process in all cases.

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dna -多阳离子配合物雾化过程中粒径的控制因素。
肺基因疗法有可能治疗或治愈呼吸系统疾病,如囊性纤维化。许多工作都集中在利用病毒载体将基因输送到肺部上,并取得了不同程度的成功。病毒载体是有问题的,不希望在肺中使用,因为它们可以引起急性免疫反应。本研究的重点是非病毒性的,基于聚合物的基因载体的特征,用于雾化器。小牛胸腺DNA被用作模型,并与三种聚合阳离子中的每一种都进行了复合体;22 kDa线性聚乙烯亚胺,25 kDa支化聚乙烯亚胺,29.5 kDa聚赖氨酸,以水、葡萄糖溶液和磷酸盐缓冲盐水(PBS)为载体液体。傅里叶变换红外光谱表明,DNA在络合物形成过程中保持B型。在N:P比为10的条件下,用振动板喷雾器雾化制备了配合物,并测量了雾化前后的粒径和Zeta电位。在水和葡萄糖溶液中制备的DNA复合物在雾化前后的粒径分布呈单峰分布,雾化后粒径略有增加。络合聚合物的选择对粒径的影响很小,主要影响来自分散液的离子特性。在PBS中制备的配合物,虽然最初是单峰的,但在雾化过程中表现出明显的团聚。所有聚合物在水或葡萄糖溶液中雾化后,Zeta电位增加,但以PBS为载体液时电位下降,并且与观察到的团聚明显相关。凝胶电泳表明,在所有情况下,络合聚合物通过雾化过程保护DNA。
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