In-vitro cytotoxicity of Polyethyleneimine on HeLa and Vero Cells

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Date

2014

Journal Title

Journal ISSN

Volume Title

Type

Article

Publisher

International Journal of Innovation and Applied Studies

Series Info

International Journal of Innovation and Applied Studies;Vol. 5 No. 3 Mar. 2014, pp. 192-199

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Scientific Journal Rankings

Abstract

The success of gene therapy depends on the choice of a suitable vector that is biocompatible and efficient in delivering therapeutic DNA into disease cells. After more than two decades, such an ideal vector is still a wish. Viral vectors though naturally evolved to transfect cells are immunogenic. As alternatives, non-viral vectors such as polyethyleneimine have been exploited. We decided to investigate the in-vitro cytotoxicity of branched polyethyleneimine 800D, 25kD and linear 20kD on HeLa and Vero cells. At exponential phase, cells were exposed to polymers at concentration range of 0.5 to 1000mg/ml. Cells were MTT assayed after 24, 48 and 72hours for viability (IC50). Linear PEI was less toxic than the branched PEI in both cells. The IC50 (mg/ml) values (Mean±SEM, n=6) post 72hours of PEI800D, 25kD and PEI20kD on HeLa cells were 2.42±0.22, 2.92±0.59, and 3.03±0.11 and for Vero cells, 7.42±0.29, 7.26±0.12, and 6.89±0.53 respectively. Two tailed t-test (P<0.05) of each polymer on both cells 72hours post dosing gave a significant P value of <0.0001. The results indicate that branched PEI800D, PEI25kD and linear PEI20kD are differently apoptotic to HeLa and Vero cells. The toxicity also time, cell line and concentration dependent. More research aimed at improving biocompatibility and transfection efficiency is needed.

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Keywords

HeLa, Vero, Polyethyleneimine, Poly L-lysine, Cytotoxicity, Dextran

Citation

[1] S. N. Uddin, Cationic lipids in non-viral gene delivery systems. Biotechnology and Molecular Biology Review, 2(3): 058- 067, 2007. [2] Panno, J. (2010). Viruses: Cornerstones of Gene Therapy. Gene Therapy, In: Gene Therapy Treatments and Cures for Genetic Disease. Rev. Ed. Bang Printing USA. pp. 55-71, 2010. [3] T. W. Writh and Y. H. Seppo, History of gene therapy. Journal of Gene, 525(2):162-169, 2013. [4] M. Morille, C. Passirani, A, Vonarbourg, A. Clavreul and J. P. Benoit, Progress in developing cationic vectors for non-viral system gene therapy against cancer, Biomaterials, 29:3477-3496, 2008. [5] F. W. Huang, H. Y. Wang, C. Li, H. F., Wang, Y. X. Sun and J. Feng, PEGylated PEI-based biodegradable polymers as nonviral gene vector, Acta Biomaterialia, 6:4285-4295, 2010. [6] H. Lv, S. Zhang, B. Wang, S. Cui, and J, Yan, Toxicity of cationic lipids and cationic polymers in gene delivery, Journal of Control Release, 114:100-109, 2006. [7] Gascón, Pozo-Rodríguez and Solinís,2013 Non-Viral Delivery Systems in Gene Therapy, Gene Therapy - Tools and Potential Applications, F. Martin (Ed.), ISBN: 978-953-51-1014-9, InTech, DOI: 10.5772/52704. Available: http://www.intechopen.com/books/gene-therapy-tools-and-potential-applications/non-viral-delivery-systems-in-genetherapy, (July 7, 2013). [8] C. Tros de Ilarduya, Y. Sun, and N. Duzgunes, Gene delivery by lipoplexes and polyplexes. European Journal of Pharmaceutical Sciences. 40:159-170. [9] K. Kunath, D. von Harpe, H. Fisher, U. Petersen, K. Bickel, T. Voight and T. Kissel, Low molecular weight polyethyleneimine as a non-viral vector for DNA delivery: comparison of physicochemical properties, transfection efficiency and in vivo distribution with high-molecular polyethyleneimine, Journal of Control Release, 89:113-125, 2003. [10] M. Lungwitz, M. Breunig, T. Blunk, and A. Go-pferich, Polyethylene mine based non-viral gene delivery systems, Eur. J. Pharm. Bio pharm, 60:247-266, 2005. [11] Y. H. Kim, J. H. Park, M. Lee, Y. H. Kim, T. G. Park, and S. W. Kim, Polyethyleneimine with acid-labile linkages as a biodegradable gene carries, Journal of Control Release, 103:209-219, 2005. [12] M. Breunig, U. Lungwitz, R. Liebl, C. Fontanari, J. Klar, and Kurtz A, et al,. Gene delivery with low molecular weight linear polyethyleneimine, J. Gene Med, 7(10):1287-98, 2005. [13] W. T. Godbey, K. K. Wu, A.G. Mikos and G. J. Hirasaki, Improved packing of polyethyleneimine/DNA complexes increases transfection efficiency, Gene Therapy, 6:1380-1388, 1999. [14] W. T. Godbey, K. K. Wu and A.G. Mikos, Size matter: molecular weight affects the efficiency of polyethyleneimine as a gene delivery vehicle, J. Biomed. Mater.Res, 45:268-275, 2001 [15] M. R. Park, K. O. Han, I. K. Han, M .H. Cho, J. W. Nah, Y. J. Choi, and C. S Cho, Degradable polyethyleneimine-alt-poly (ethylene glycol) copolymers as novel gene carriers, Journal of Controlled Release, 105:367-380, 2005. [16] A. F. Vargas, The Human Genome Project and its importance in clinical medicine, International Congress Series, 1237: 3 –13, 2002. [17] K. Lundstrom and T. Boulikas, Viral and Non-viral Vectors in Gene Therapy: Technology Development and Clinical Trials, Technology in Cancer Research and Treatment, 2(5): 471-485, 2003. [18] O. Kafil and Y. Omidi, Cytotoxic Impact of Linear and Branched Polyethleneimine Nanostructures in A431 cells, Bioimpacts, 1(1):23-30, 2011. [19] J. R. Goss, W. F. Goins, D. Lacomis, M. Mata, J. C. Gloriso, and D. J. Fink, Herpes simplex-mediated gene transfer of the nerve growth factor protects against peripheral neuropathy in streptozotocin-induced diabetes in the mouse, Diabetes, 51:2227-2232, 2002. [20] M. Giacca and S. Zacchigna, Virus-mediated gene delivery for human gene therapy, Journal of Controlled Release, 161:377-388, 2012. [21] S. Yla-Herttuala, Endgame: Glybera finally recommended for approval as the first therapy drug in the European Union, Molecular Therapy, 20: 1831-1832, 2012 [22] S. C.W. Richardosn, N. G. Pattrick, N. Lavignac, P, Ferruti and R. Ducan, Intracellular fate of bioresponsive poly(amidoamine) in vitro and in vivo, Journal of Controlled Release, 142:78-88, 2010. [23] Y. J. Choi, S. J. Kang, Y. J. Kim, Y.B. Lim and H.W. Chung, Comparative studies on the genotoxicity and cytotoxicity of polymeric gene carriers polyethyleneimine (PEI) and polyamidoamine (PAMAM) dendrimers in Jurkat T-cells, Drug and Chemical Toxicology, 33(4):357-366, 2010. [24] B.I. Florea, C. Meaney, H. E. Junginger, and G. Borchard, Transfection efficiency and toxicity of polyethyleneimine in differentiated Calu-3 and non-differentiated COS-1 cell culture, AAPS Pharm. Sci., 4(3):E12, 2002. [25] M. Turk, S. Dincer, I.G. Yulug and E. Piskin, In vitro transfection of HeLa cells with temperature sensitive polycationic copolymers, Journal of Controlled Release, 96:325-340, 2004. [26] J. H. Jeong, S. H. Song, D.W. Lim, H. Lee, and T.G. Park, DNA transfection using linear poly(ethyleneimine) prepared by concentration acid hydrolysis of poly(2-ethyl-2-oxazoline), Journal of controlled Release, 73:391-399, 2001. [27] M.P. Xiong, M.L. Forrest, G. Ton, A. Zhao, N.M. Davies and G.S, Kwon, ,Poly(aspartate-g-PEI800), a polyethyleneimine analogue of low toxicity and high transfection efficiency for gene delivery, Biomaterials, 28(32): 4889-4990,2007 [28] D. Fischer, T. Bieber, Y. Li, and H.P. Elasser, (1999). A novel non-viral vector for DNA delivery for DNA delivery based on low molecular weight, branched polyethyleneimine: effect of molecular weight on transfection efficiency and cytotoxicity, Pharma. Res., 16:1273-1279, 1999 [29] A.C. Hunter, Molecular hurdles in polyfectin design and mechanistic background to polycation induced cytotoxicity. Adv. Drug Del. Rev., 58:1523-1531, 2006

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