Skip to main content
SpringerLink
Log in

Molecular Architecture Governs Cytotoxicity and Gene Transfection Efficacy of Polyethylenimine Based Nanoplexes in Mammalian Cell Lines

  • Published:
Journal of Inorganic and Organometallic Polymers and Materials Aims and scope Submit manuscript

Abstract

Polyethylenimine (pEI) is a potent cationic transfection agent that is commonly used in conjunction with other inorganic and organometallics formulations. However, the toxicity that is intrinsic to pEI constitutes a major obstacle for both in vitro and in vivo applications. In this study, the roles of pEI molecular architectures and effects of nitrogen/phosphate (N/P) molar ratio on the cytotoxic potential and in vitro transfection performance were elucidated utilizing linear (L-pEI) and branched (B-pEI) 25 kDa pEI. Both L-pEI and B-pEI were able to self-assemble with the plasmid DNA to form positively charged nanoplexes having an average hydrodynamic diameter of 100–150 nm. In vitro cytotoxicity and transfection performance were assessed with three model mammalian cell lines namely, L929, COS-7 and HeLa cells. Transfection profiles over a wide range of N/P ratio exemplify a similar trend across three different cell lines, with B-pEI nanoplexes consistently showing maximum transgene expression of enhanced green fluorescent protein at an earlier N/P ratio compared to L-pEI nanoplexes. However, high transfection activity was maintained over a wider N/P ratio for L-pEI nanoplexes. While the absolute size of the nanoplexes exhibited no discernible effect on the transfection performance, cytotoxicity of free un-bound B-pEI was identified as the dictating factor responsible for the observed global cellular response. Collectively, our findings provided critical insights into the role of pEI molecular architecture in cellular transfection and are expected to lay the foundation for the rational design and development of advanced pEI-based inorganic and organometallic gene delivery systems.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. U. Griesenbach, S. Ferrari, D.M. Geddes, E.W. Alton, Gene Ther. 9, 1344 (2002)

    Article  CAS  Google Scholar 

  2. M. Fiandaca, J. Forsayeth, K. Bankiewicz, Exp. Neurol. 209, 51 (2008)

    Article  CAS  Google Scholar 

  3. G. Hsich, M. Sena-Esteves, X.O. Breakefield, Hum. Gene Ther. 13, 579 (2002)

    Article  CAS  Google Scholar 

  4. J.M. Isner, Nature 415, 234 (2002)

    Article  CAS  Google Scholar 

  5. I.M. Verma, N. Somia, Nature 389, 239 (1997)

    Article  CAS  Google Scholar 

  6. M.D. Brown, A.G. Schatzlein, I.F. Uchegbu, Int. J. Pharm. 229, 1 (2001)

    Article  CAS  Google Scholar 

  7. D.W. Pack, A.S. Hoffman, S. Pun, P.S. Stayton, Nat. Rev. Drug Discov. 4, 581 (2005)

    Article  CAS  Google Scholar 

  8. O. Boussif, F. Lezoualc’h, M.A. Zanta, M.D. Mergny, D. Scherman, B. Demeneix, J.P. Behr, Proc. Natl. Acad. Sci. USA 92, 7297 (1995)

    Article  CAS  Google Scholar 

  9. M. Neu, D. Fischer, T. Kissel, J. Gene Med. 7, 992 (2005)

    Article  CAS  Google Scholar 

  10. V. Cebrian, F. Martin-Saavedra, C. Yague, M. Arruebo, J. Santamaria, N. Vilaboa, Acta Biomater. 7, 3645 (2011)

    Article  CAS  Google Scholar 

  11. Y.K. Buchman, E. Lellouche, S. Zigdon, M. Bechor, S. Michaeli, J.P. Lellouche, Bioconjug. Chem. 24, 2076 (2013)

    Article  CAS  Google Scholar 

  12. Y. Manoharan, Q. Ji, T. Yamazaki, S. Chinnathambi, S. Chen, S. Ganesan, J.P. Hill, K. Ariga, N. Hanagata, Int. J. Nanomed. 7, 3625 (2012)

    CAS  Google Scholar 

  13. D. Ang, C.Y. Tay, L.P. Tan, P.R. Preiser, R.V. Ramanujan, Mater. Sci. Eng. C 31, 1445 (2011)

    Article  CAS  Google Scholar 

  14. W. Wei, C. Xu, H. Wu, J. Huazhong, Univ. Sci. Technol. Med. Sci. 24, 618 (2004)

    CAS  Google Scholar 

  15. S. Xenariou, U. Griesenbach, S. Ferrari, P. Dean, R.K. Scheule, S.H. Cheng, D.M. Geddes, C. Plank, E.W. Alton, Gene Ther. 13, 1545 (2006)

    Article  CAS  Google Scholar 

  16. Y. Liu, D.C. Wu, W.D. Zhang, X. Jiang, C.B. He, T.S. Chung, S.H. Goh, K.W. Leong, Angew. Chem. Int. Ed. Engl. 44, 478 (2005)

    Google Scholar 

  17. W. Amer, L. Wang, A. Amin, L. Ma, H. Yu, J. Inorg. Organomet. Polym. 20, 605 (2010)

    Article  CAS  Google Scholar 

  18. A. Beyerle, M. Irmler, J. Beckers, T. Kissel, T. Stoeger, Mol. Pharm. 7, 727 (2010)

    Article  CAS  Google Scholar 

  19. M. Giovanni, C.Y. Tay, M.I. Setyawati, J.P. Xie, C.N. Ong, R. Fan, J. Yue, L. Zhang, D.T. Leong, Environ. Toxicol. (2014). doi:10.1002/tox.22015

    Google Scholar 

  20. M.I. Setyawati, C.Y. Tay, S.L. Chia, S.L. Goh, W. Fang, M.J. Neo, H.C. Chong, S.M. Tan, S.C.J. Loo, K.W. Ng, J.P. Xie, C.N. Ong, N.S. Tan, D.T. Leong, Nat. Commun. 4, 1673 (2013)

    Article  CAS  Google Scholar 

  21. C.Y. Tay, W. Fanru, M.I. Setyawati, S.L. Chia, K.S. Tan, C.H.L. Hong, D.T. Leong, ACS Appl. Mater. Interfaces 6, 6248 (2014)

    Article  CAS  Google Scholar 

  22. C.Y. Tay, M.I. Setyawati, J.P. Xie, W.J. Parak, D.T. Leong, Adv. Funct. Mater. 24, 5936 (2014)

    Article  CAS  Google Scholar 

  23. L. Wightman, R. Kircheis, V. Rossler, S. Carotta, R. Ruzicka, M. Kursa, E. Wagner, J. Gene Med. 3, 362 (2001)

    Article  CAS  Google Scholar 

  24. J.W. Wiseman, C.A. Goddard, D. McLelland, W.H. Colledge, Gene Ther. 10, 1654 (2003)

    Article  CAS  Google Scholar 

  25. J. Intra, A.K. Salem, J. Control. Release 130, 129 (2008)

    Article  CAS  Google Scholar 

  26. S. Choosakoonkriang, B.A. Lobo, G.S. Koe, J.G. Koe, C.R. Middaugh, J. Pharm. Sci. 92, 1710 (2003)

    Article  CAS  Google Scholar 

  27. C. Zhang, P. Yadava, J. Hughes, Methods 33, 144 (2004)

    Article  CAS  Google Scholar 

  28. K. Morimoto, M. Nishikawa, S. Kawakami, T. Nakano, Y. Hattori, S. Fumoto, F. Yamashita, M. Hashida, Mol. Ther. 7, 254 (2003)

    Article  CAS  Google Scholar 

  29. Q.-Q. Zhao, J.L. Chen, T.F. Lv, C.X. He, G.P. Tang, W.Q. Liang, Y. Tabata, J.Q. Gao, Biol. Pharm. Bull. 32, 706 (2009)

    Article  CAS  Google Scholar 

  30. S. Kawakami, Y. Ito, P. Charoensit, F. Yamashita, M. Hashida, J. Pharmacol. Exp. Ther. 317, 1382 (2006)

    Article  CAS  Google Scholar 

  31. U. Lungwitz, M. Breunig, T. Blunk, A. GopFerich, Eur. J. Pharm. Biopharm. 60, 247 (2005)

    Article  CAS  Google Scholar 

  32. S. De Smedt, J. Demeester, W. Hennink, Pharm. Res. 17, 113 (2000)

    Article  Google Scholar 

  33. K. Ariga, Q. Ji, J.P. Hill, N. Kawazoe, G. Chen, Expert Opin. Biol. Ther. 9, 307 (2009)

    Article  CAS  Google Scholar 

  34. B.P. Bastakoti, M. Inoue, S. Yusa, S.H. Liao, K.C. Wu, K. Nakashima, Y. Yamauchi, Chem. Commun. (Camb) 48, 6532 (2012)

    Article  CAS  Google Scholar 

  35. B.P. Bastakoti, S. Ishihara, S.Y. Leo, K. Ariga, K.C. Wu, Y. Yamauchi, Langmuir 30, 651 (2014)

    Article  CAS  Google Scholar 

  36. B.P. Bastakoti, K.C. Wu, M. Inoue, S. Yusa, K. Nakashima, Y. Yamauchi, Chemistry 19, 4812 (2013)

    Article  CAS  Google Scholar 

  37. M. Ramanathan, K. Hong, Q. Ji, Y. Yonamine, J.P. Hill, K. Ariga, J. Nanosci. Nanotechnol. 14, 390 (2014)

    Article  CAS  Google Scholar 

  38. E. Ruiz-Hitzky, M. Darder, P. Aranda, K. Ariga, Adv. Mater. 22, 323 (2010)

    Article  CAS  Google Scholar 

  39. J. Suh, Y. Lee, S. Han, Bioorg. Med. Chem. Lett. 8, 1331 (1998)

    Article  CAS  Google Scholar 

  40. M.L. Forrest, G.E. Meister, J.T. Koerber, D.W. Pack, Pharm. Res. 21, 365 (2004)

    Article  CAS  Google Scholar 

  41. N.P. Gabrielson, D.W. Pack, Biomacromolecules 7, 2427 (2006)

    Article  CAS  Google Scholar 

  42. E. Frohlich, Int. J. Nanomed. 7, 5577 (2012)

    Article  Google Scholar 

  43. E. Wagner, M. Cotton, R. Foisner, M.L. Birnstiel, Proc. Natl. Acad. Sci. USA 88, 4255 (1991)

    Article  CAS  Google Scholar 

  44. M. Ogris, P. Steinlein, M. Kursa, K. Mechtler, R. Kircheis, E. Wagner, Gene Ther. 5, 1425 (1998)

    Article  CAS  Google Scholar 

  45. Z. Dai, C. Wu, Macromolecules 45, 4346 (2012)

    Article  CAS  Google Scholar 

  46. K. Kunath, A. von Harpe, H. Petersen, D. Fischer, K. Voigt, T. Kissel, U. Bickel, Pharm. Res. 19, 810 (2002)

    Article  CAS  Google Scholar 

  47. D. Fischer, Y. Li, B. Ahlemeyer, J. Krieglstein, T. Kissel, Biomaterials 24, 1121 (2003)

    Article  CAS  Google Scholar 

  48. S.M. Moghimi, P. Symonds, J.C. Murray, A.C. Hunter, G. Debska, A. Szewczyk, Mol. Ther. 11, 990 (2005)

    Article  CAS  Google Scholar 

  49. J. Sun, F. Zeng, H. Jian, S. Wu, Polym. Chem. 4, 5810 (2013)

    Article  CAS  Google Scholar 

  50. S. Boeckle, K. von Gersdorff, S. van der Piepen, C. Culmsee, E. Wagner, M. Ogris, J. Gene Med. 6, 1102 (2004)

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to acknowledge financial support from National University of Singapore and Nanyang Technological University. C.Y.T gratefully acknowledges support from the Lee Kuan Yew Postdoctoral Fellowship.

Author information

Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore, 117585, Singapore

    Chor Yong Tay, Nandita Menon & David Tai Leong

  2. School of Materials Science and Engineering, Nanyang Technological University, Nanyang Avenue, Singapore, 639798, Singapore

    Lay Poh Tan

Authors
  1. Chor Yong Tay
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nandita Menon
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. David Tai Leong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lay Poh Tan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Chor Yong Tay or Lay Poh Tan.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tay, C.Y., Menon, N., Leong, D.T. et al. Molecular Architecture Governs Cytotoxicity and Gene Transfection Efficacy of Polyethylenimine Based Nanoplexes in Mammalian Cell Lines. J Inorg Organomet Polym 25, 301–311 (2015). https://doi.org/10.1007/s10904-014-0135-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10904-014-0135-8

Keywords

Search

Navigation