Haimanti Mandal1, Sameer S Katiyar1, Rajan Swami1, Varun Kushwah1, Parmeshwar B Katare2, Anand Kumar Meka3, Sanjay K Banerjee2, Amirali Popat4, Sanyog Jain5. 1. Centre for Pharmaceutical Nanotechnology, Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research (NIPER), Sec 67 Mohali, Punjab, India. 2. Drug Discovery Research Center, Translational Health Science and Technology Institute, Faridabad 121001, India. 3. The School of Pharmacy, The University of Queensland, Brisbane, QLD 4072, Australia. 4. The School of Pharmacy, The University of Queensland, Brisbane, QLD 4072, Australia; Inflammatory Disease Biology and Therapeutics Group, Mater Research Institute - The University of Queensland, Translational Research Institute, 37 Kent St, Woolloongabba, QLD 4102, Australia. 5. Centre for Pharmaceutical Nanotechnology, Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research (NIPER), Sec 67 Mohali, Punjab, India. Electronic address: sanyogjain@niper.ac.in.
Abstract
The present work addresses the development and characterization of ε-Poly-l-Lysine/pDNA polyplexes and evaluation for their improved transfection efficacy and safety as compared to polyplexes prepared using Poly-l-Lysine and SuperFect®. Self-assembling polyplexes were prepared by varying the N/P ratio to obtain the optimum size, a net positive zeta potential and gel retardation. The stability in presence of DNase I and serum was assured using gel retardation assay. Their appreciable uptake in MCF-7 and 3.5, 3.79 and 4.79-fold higher transfection compared to PLL/pDNA polyplexes and 1.60, 1.53 and 1.79-fold higher transfection compared to SuperFect®/pDNA polyplexes in MCF-7, HeLa and HEK-293 cell lines respectively, affirmed the enhanced transfection of ε-PLL/pDNA polyplexes which was well supported with in vivo transfection and gene expression studies. The <8% in vitro hemolysis and >98% viability of MCF-7, HeLa and HEK-293 cells in presence of ε-PLL/pDNA polyplexes addressed their safety, which was also ensured using in vivo toxicity studies, where hemocompatibility, unaltered levels of biochemical markers and histology of vital organs confirmed ε-PLL to be an effective and safer alternative for non-viral genetic vectors.
The present work addresses the development and characterization of εpan class="Chemical">-Poly-l-Lysine/pDNA polypn>lexes and evaluation for their impn>roved transfection efficacy and safety as compn>ared to polypn>lexes prepn>ared using n>an class="Chemical">Poly-l-Lysine and SuperFect®. Self-assembling polyplexes were prepared by varying the N/P ratio to obtain the optimum size, a net positive zeta potential and gel retardation. The stability in presence of DNase I and serum was assured using gel retardation assay. Their appreciable uptake in MCF-7 and 3.5, 3.79 and 4.79-fold higher transfection compared to PLL/pDNA polyplexes and 1.60, 1.53 and 1.79-fold higher transfection compared to SuperFect®/pDNA polyplexes in MCF-7, HeLa and HEK-293 cell lines respectively, affirmed the enhanced transfection of ε-PLL/pDNA polyplexes which was well supported with in vivo transfection and gene expression studies. The <8% in vitro hemolysis and >98% viability of MCF-7, HeLa and HEK-293 cells in presence of ε-PLL/pDNA polyplexes addressed their safety, which was also ensured using in vivo toxicity studies, where hemocompatibility, unaltered levels of biochemical markers and histology of vital organs confirmed ε-PLL to be an effective and safer alternative for non-viral genetic vectors.
Authors: Garrett Kelly; Joshua J Milligan; Eric M Mastria; Sarah Kim; Stephanie R Zelenetz; Jarrett Dobbins; Leon Y Cai; Xinghai Li; Smita K Nair; Ashutosh Chilkoti Journal: J Control Release Date: 2022-01-22 Impact factor: 9.776