Pathak Atul, Kumar Pradeep, Chuttani Krishna, Jain Sanyog, Mishra Anil K, Vyas Suresh P, Gupta Kailash C
Institute of Genomics and Integrative Biology, CSIR, Delhi University Campus, Mall Road, Delhi, India.
ACS Nano. 2009 Jun 23;3(6):1493-505. doi: 10.1021/nn900044f.
Tumor-specific gene delivery constitutes a primary challenge in nonviral mediated gene therapy. In this investigation, branched polyethylenimine (bPEI, 25 kDa) was modified by forming nanoconstructs with a natural polysaccharide, chondroitin sulfate (CS), to impart site-specific property. A library of CS-PEI (CP) nanoconstructs was fabricated by altering the content of CS and evaluated in terms of size, surface charge, morphology, pDNA loading efficiency, pDNA release assay, pDNA protection study, cytotoxicity, and transfection efficiency. In vitro transfection efficiency of CP nanoconstructs was examined in HEK293, HEK293T, HepG2, and HeLa cell lines, while their cytotoxicity was investigated in HepG2 and HeLa cells. DNase I protection assay showed that the plasmid was protected from degradation over a period of time. The CP nanoconstructs possess significantly lower toxicity and enhanced transfection efficiency compared to PEI (25 kDa) and commercial transfection reagents (i.e., superfect, fugene, and GenePORTER 2). Further, the CP nanoconstructs were also found to transfect cells in serum-containing medium. In vivo studies were carried out with pDNA loaded CP-3 nanoconstruct after intravenous (iv) injection in Ehrlich ascites tumor (EAT)-bearing mice. The outcome revealed higher concentration of CP-3 nanoconstruct in tumor mass. These findings demonstrate that CP nanoconstructs could be exploited as carriers for nanomedicine for efficient management of solid tumor.
肿瘤特异性基因递送是非病毒介导的基因治疗中的一项主要挑战。在本研究中，通过与天然多糖硫酸软骨素（CS）形成纳米结构对支化聚乙烯亚胺（bPEI，25 kDa）进行修饰，以赋予其位点特异性特性。通过改变CS的含量制备了一系列CS-PEI（CP）纳米结构，并对其大小、表面电荷、形态、质粒DNA（pDNA）负载效率、pDNA释放测定、pDNA保护研究、细胞毒性和转染效率进行了评估。在HEK293、HEK293T、HepG2和HeLa细胞系中检测了CP纳米结构的体外转染效率，同时在HepG2和HeLa细胞中研究了它们的细胞毒性。DNase I保护试验表明，质粒在一段时间内受到保护未被降解。与PEI（25 kDa）和商业转染试剂（即SuperFect、Fugene和GenePORTER 2）相比，CP纳米结构具有显著更低的毒性和更高的转染效率。此外，还发现CP纳米结构能够在含血清的培养基中转染细胞。对携带艾氏腹水瘤（EAT）的小鼠静脉注射（iv）负载pDNA的CP-3纳米结构后进行了体内研究。结果显示肿瘤块中CP-3纳米结构的浓度更高。这些发现表明，CP纳米结构可作为纳米药物载体用于实体瘤的有效治疗。
