Preparation, characterization and transfection efficiency of nanoparticles composed of alkane-modified polyallylamine

Document Type : Research Paper


1 Neurogenic Inflammation Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Medical Biotechnology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

4 Targeted Drug Delivery Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

5 Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran


Although viral vectors are considered efficient gene transfer agents, their board application has been limited by toxicity, immunogenicity, mutagenicity and small gene carrying capacity. Non-viral vectors are safe but they suffer from low transfection efficiency. In the present study, polyallylamine (PAA) in two molecular weights (15 and 65 kDa) was modified by alkane derivatives in order to increase transfection activity and to decrease cytotoxicity.
Materials and Methods:
Modified PAA was synthesized using three alkane derivatives (1-bromobutane, 1-bromohexane and 1-bromodecane) in different grafting percentages (10, 30 and 50). The condensation ability of modified PAA was determined by ethidium bromide test. The prepared polyplexes, complexes of modified PAA and DNA, were characterized by size and zeta potential. Transfection activity of polyplexes was checked in Neuro2A cells. The cytotoxicity of vector was examined in the same cell line.
DNA condensation ability of PAA was decreased after modification but modified polymer could still condense DNA at moderate and high carrier to plasmid (C/P) ratios. Most of polyplexes composed of modified polymer had mean size less than 350 nm. They showed a positive zeta potential, but some vectors with high percentage of grafting had negative surface charge. Transfection efficiency was increased by modification of PAA by 1-bromodecane in grafting percentages of 30 and 50%. Modification of polymer reduced polymer cytotoxicity especially in C/P ratio of 2.
Results of the present study indicated that modification of PAA with alkane derivatives can help to prepare gene carriers with better transfection activity and less cytotoxicity.


1. Thomas M, Klibanov AM. Non-viral gene therapy: polycation-mediated DNA delivery. Appl Microbiol Biotechnol. 2003; 62: 27-34.
2. Verma IM, Somia N. Gene therapy - promises, problems and prospects. Nature. 1997; 389: 239-242.
3. Kullberg M, McCarthy R, Anchordoquy TJ. Systemic tumor-specific gene delivery. J Control Release. 2013; 172: 730-736.
4. Nayerossadat N, Maedeh T, Ali PA. Viral and nonviral delivery systems for gene delivery. Adv Biomed Res. 2012; 1:27.
5. Wang T, Upponi JR, Torchilin VP. Design of multifunctional non-viral gene vectors to overcome physiological barriers: Dilemmas and strategies. Int J Pharm. 2012; 427: 3-20.
6. Yin H, Kanasty RL, Eltoukhy AA, Vegas AJ, Dorkin JR, Anderson DG. Non-viral vectors for gene-based therapy. Nat Rev Genet. 2014; 15: 541-555.
7. Oskuee RK, Mohtashami E, Golami L, Malaekeh-Nikouei B, Cationic liposomes-polyallylamine-plasmid nanocomplexes for gene delivery. J Exp Nanosci. 2014; 9: 1026-1034.
8. Nimesh S, Kumar R, Chandra R. Novel polyallylamine-dextran sulfate-DNA nanoplexes: highly efficient non-viral vector for gene delivery. Int J Pharm. 2006; 320: 143-149.
9. Pathak A, Aggarwal A, Kurupati RK, Patnaik S, Swami A, Singh Y, et al. Engineered polyallylamine nanoparticles for efficient in vitro transfection. Pharm Res. 2007; 24: 1427-1440.
10. Pisskin E, Dincer S, Turk M. Gene delivery: intelligent but just at the beginning. J Biomater Sci Polym Ed. 2004; 15: 1181-1202.
11. Somia N, Verma IM. Gen therapy: trials and tribulations. Nat. Rev. Genet. 2000; 1: 91-99.
12. Boussifi O, Delair T, Brua C, Veron L, Pavirani A, Kolbe HV. Synthesis of polyallylamine derivatives and their use as gene transfer vectors in vitro. Bioconjug Chem. 1999; 10: 877-883.
13. Neu M, Fischer D, Kissel T. Recent advances in rational gene transfer vector design based on poly(ethylene imine) and its derivatives. J Gene Med. 2005; 7: 992-1009.
14. Gabrielson NP, Pack DW. Acetylation of polyethylenimine enhances gene delivery via weakened polymer/DNA interactions. Biomacromolecules. 2006; 7: 2427-2435.
15. Mahato M, Rana G, Kumar P, Sharma AK. Tetramethy lguanidiniumolyallyl-amine (Tmg‐PA): A new class of nonviral vector for efficient gene transfection.  J Polym Sci. 2012; 50: 2344-2355.
16. Zanta MA, Boussifi O, Adib A, Behr J.-P. In vitro gene delivery to hepatocytes with galactosylated polyethylenimine. Biocon-jug. Chem. 1997;  8: 839-844.
17. Tros de Ilarduya C., Sun Y., Duzgunes N., Gene delivery by lipoplexes and polyplexes. Eur J Pharm Sci. 2010; 40: 159-170.
18. Kuhn PS, Levin Y, Barbosa MC. Charge inversion in DNA–amphiphile complexes: possible application to gene therapy. Physica A. 1999; 274: 8–18.
19. Dehshahri A, Oskuee RK, Shier WT, Hatefi A, Ramezani M. Gene transfer efficiency of high primary amine content, hydrophobic, alkyl-oligoamine derivatives of polyethylenimine. Biomaterials. 2009; 30: 4187-4194.
20. Oskuee RK, Dehshahri A, Shier WT, Ramezani M. Modified polyethylene-imine: self-assembled nanoparticle forming ploymer for pDNA delivery. Iran J Basic Med Sci. 2008; 11: 33-40.