Design and Development of Light-Sensitive Chitosan-Based Nanocarriers for Gene Delivery

Abstract:

In this work, we report on the development of a multifunctional and photo-inducible nanoplex made of chitosan (Ch) and hyaluronic acid (HA) for delivery of nucleic acid. Self-assembled Ch/HA nanoparticles were attached to ortho-nitrobenzyl (o-NB) photo-labile molecules (PL)-gold nanoparticles via thiol groups and to QDs-conjugate ssDNA through amide bond linkage to form nanoplexes (Ch:HA:AuPL:QD-DNA). The composition of DNA nanocarriers was validated by nuclear magnetic resonance, transmission electronic microscopy, energy dispersive x-ray spectroscopy, gel electrophoresis and spectrophotometry. The change in zeta potential (34 ± 11 to -26 ± 11 mV) and the loss of the o-NB characteristic peaks in nuclear magnetic resonance spectra, after the exposure of the PL molecule to ultraviolet light, both confirmed the photo-labile properties of the system. The potential of the nanoplexes to induce high cell transfection was assessed by flow cytometry and fluorescent microscopy imaging. Over 30% transfection of HEK-293 was obtained with the nanoplexes after a one-minute exposure of cells to UV light. This corresponds to a 15% increase in the transfection efficiency compared to unexposed Ch:HA:AuPL:QD-DNA nanocarriers. This high transfection efficiency was associated with the unique design of the carrier system and its photo-responsiveness feature for facilitating the DNA release.

• Periodical: Advances in Science and Technology (Volume 86)
• Pages: 75-80
• DOI: https://doi.org/10.4028/www.scientific.net/AST.86.75
• Online since: September 2012
• Authors: Nicolas Duceppe, Maryam Tabrizian
• Keywords: Chitosan, Hyaluronic Acid (HA), Light-Sensitive Non-Viral DNA Nanocarrier, Multifunctional Nanoplex

References

[1] Duceppe N, Tabrizian M. Advances in using chitosan-based nanoparticles for in vitro and in vivo drug and gene delivery. Expert Opin Drug Deliv 2010; 7: 1191-1207.

DOI: 10.1517/17425247.2010.514604

[2] Duceppe N, Tabrizian M. Factors influencing the transfection efficiency of ultra low molecular weight chitosan/hyaluronic acid nanoparticles. Biomaterials 2009; 30: 2625-2631.

DOI: 10.1016/j.biomaterials.2009.01.017

[3] Douglas KL, Piccirillo CA, Tabrizian M. Effects of alginate inclusion on the vector properties of chitosan-based nanoparticles. J Control Release 2006; 115: 354-361.

DOI: 10.1016/j.jconrel.2006.08.021

[4] Douglas KL, Piccirillo CA, Tabrizian M. Cell line-dependent internalization pathways and intracellular trafficking determine transfection efficiency of nanoparticle vectors. Eur J Pharm Biopharm 2008; 68: 676-687.

DOI: 10.1016/j.ejpb.2007.09.002

[5] Sanvicens N, Marco MP. Multifunctional nanoparticles - properties and prospects for their use in human medicine. Trends in Biotechnology 2008; 26: 425-433.

DOI: 10.1016/j.tibtech.2008.04.005

[6] Kim S, Kim JH, Jeon O, Kwon IC, Park K. Engineered polymers for advanced drug delivery. Eur J Pharm Biopharm 2009; 71: 420-430.

DOI: 10.1016/j.ejpb.2008.09.021

[7] Ganta S, Devalapally H, Shahiwala A, Amiji M. A review of stimuli-responsive nanocarriers for drug and gene delivery. J Control Release 2008; 126: 187-204.

DOI: 10.1016/j.jconrel.2007.12.017

[8] Roy D, Cambre JN, Sumerlin BS. Future perspectives and recent advances in stimuli-responsive materials. Prog Polym Sci (Oxford) 2010; 35: 278-301.

DOI: 10.1016/j.progpolymsci.2009.10.008

[9] Wu C, Chen C, Lai J, Chen J, Mu X, Zheng J et al. Molecule-scale controlled-release system based on light-responsive silica nanoparticles. Chem Commun (Camb ) 2008: 2662-2664.

DOI: 10.1039/b804886j

[10] Chandra B, Subramaniam R, Mallik S, Srivastava DK. Formulation of photocleavable liposomes and the mechanism of their content release. Org Biomol Chem 2006; 4: 1730-1740.

DOI: 10.1039/b518359f

[11] Blanc A, Bochet CG. Isotope effects in photochemistry. 1. o-nitrobenzyl alcohol derivatives. J Am Chem Soc 2004; 126: 7174-7175.

DOI: 10.1021/ja049686b

[12] Drake P, Lin YJ. The attachment, modification and release of gold nanoparticles from a photolabile solid phase synthesis resin, a new methodology for nanoparticle surface modification. Thin Solid Films 2006; 515: 245-253.

DOI: 10.1016/j.tsf.2005.12.077

[13] Nagasaki T, Taniguchi A, Tamagaki S. Photoenhancement of transfection efficiency using novel cationic lipids having a photocleavable spacer. Bioconjug Chem 2003; 14: 513-516.

DOI: 10.1021/bc0256603

[14] Kim MS, Diamond SL. Controlled release of DNA/polyamine complex by photoirradiation of a solid phase presenting o-nitrobenzyl ether tethered spermine or polyethyleneimine. Bioorg Med Chem Lett 2006; 16: 5572-5575.

DOI: 10.1016/j.bmcl.2006.08.030

[15] Handwerger RG, Diamond SL. Biotinylated photocleavable polyethylenimine: capture and triggered release of nucleic acids from solid supports. Bioconjug Chem 2007; 18: 717-723.

DOI: 10.1021/bc060280t

[16] Kostiainen MA, Smith DK, Ikkala O. Optically triggered release of DNA from multivalent dendrons by degrading and charge-switching multivalency. Angew Chem Int Ed Engl 2007; 46: 7600-7604.

DOI: 10.1002/anie.200701200

[17] Sandros MG, Behrendt M, Maysinger D, Tabrizian M. InGaP@ZnS-enriched chitosan nanoparticles: A versatile fluorescent probe for deep-tissue imaging. Adv Funct Mater 2007; 17: 3724-3730.

DOI: 10.1002/adfm.200700439

[18] Sandros MG, Shete V, Benson DE. Selective, reversible, reagentless maltose biosensing with core-shell semiconducting nanoparticles. Analyst 2006; 131: 229-235.

DOI: 10.1039/b511591d

[19] Oh E, Hong MY, Lee D, Nam SH, Yoon HC, Kim HS. Inhibition assay of biomolecules based on fluorescence resonance energy transfer (FRET) between quantum dots and gold nanoparticles. J Am Chem Soc 2005; 127: 3270-3271.

DOI: 10.1021/ja0433323

[20] Han G, You CC, Kim BJ, Turingan RS, Forbes NS, Martin CT et al. Light-regulated release of DNA and its delivery to nuclei by means of photolabile gold nanoparticles. Angew Chem Int Ed Engl 2006; 45: 3165-3169.

DOI: 10.1002/anie.200600214

[21] B.K. Pong, B.L. Trout and J.Y. Lee, Modified ligand-exchange for efficient solubilization of CdSe/ZnS quantum dots in water: A procedure guided by computational studies, Langmuir 24 (10) (2008), pp.5270-5276.

DOI: 10.1021/la703431j

[22] Alvarez-Lorenzo C, Concheiro A. Intelligent drug delivery systems: polymeric micelles and hydrogels. Mini Rev Med Chem 2008; 8: 1065-1074.

DOI: 10.2174/138955708785909952

[23] Medintz IL, Uyeda HT, Goldman ER, Mattoussi H. Quantum dot bioconjugates for imaging, labelling and sensing. Nat Mater 2005; 4: 435-446.

DOI: 10.1038/nmat1390