Preparation and Characterization of Chitosan/β-cyclodextrin Nanoparticles Containing Plasmid DNA Encoding Interleukin-12

Abstract

Background:

Interleukin-12 (IL-12) as a cytokine has been proved to possess antitumor effects via stimulating the immune system. Non-viral gene delivery systems offer several advantages, including easiness in production, low cost, safety; low immunogenicity and can carry higher amounts of genetic material without limitation on their sizes.

Methods:

pUMVC3-hIL12 loaded Low Molecular Weight chitosan/β-cyclodextrin (LMW CS/CD) nanoparticles were prepared using ionotropic gelation method and characterized in terms of size, zeta potential, polydispersity index, morphology, loading efficiency and cytotoxicity against the CT-26 colon carcinoma cell line.

Results:

All prepared particles were spherical in shape and nano-sized (171.3±2.165 nm, PdI: 0.231±0.014) and exhibited a positive zeta potential (34.3±1.55). The nanoparticles demonstrated good DNA encapsulation efficiencies (83.315%±2.067). Prepared pUMVC3-hIL12 loaded LMW CS/CD nanoparticles showed no cell toxicity in murine CT-26 colon carcinoma cells. At the concentration of 0.1 µg/ml of nanoparticles, the transfection ability was obviously higher than that of the naked DNA.

Conclusion:

LMW CS/CD-plasmid DNA nanoparticles encoding IL-12 prepared using ionotropic gelation method with no toxic effect on the tested cells can be considered as a basis for further gene delivery studies both in vitro and in vivo to enhance the expression of IL-12.

Key words

chitosan - β-cyclodextrin - plasmid - nanoparticle - interleukin-12

• References

• 1 Hallaj-Nezhadi S, Valizadeh H, Dastmalchi S et al. Preparation of chitosan-plasmid DNA nanoparticles encoding interleukin-12 and their expression in CT-26 colon carcinoma cells. J Pharm Pharm Sci 2011; 14: 181-195
• 2 George JA. Gene therapy progress and prospects: Adenoviral vectors. Gene ther 2003; 10: 1135-1141
• 3 Schmidt-Wolf GD, Schumidt-Wolf IGH. Non-viral and hybrid vectors in human gene therapy: An update. Trends Mol Med 2003; 9: 67-72
• 4 Davis ME. Non-viral gene delivery systems. Curr Opin Biotech 2002; 13: 128-131
• 5 Al-Dosari MS, Gao X. Nonviral gene delivery: Principle, limitations and recent progress. AAPS 2009; 11: 671-681
• 6 Morille M, Passirani C, Vonarbourg A et al. Progress in developing cationic vectors for non-viral systemic gene therapy against cancer. Biomaterials 2008; 29: 3477-3496
• 7 Teijeiro-Osorio D, Remunan-Lopez C, Alonso MJ. Chitosan/cyclodextrin nanoparticles can efficiently transfect the airway epithelium in vitro. Eur J Pharm Biopharm 2009; 71 (02) 257-263
• 8 Arun R, Ashok KCK, Sravanti VVNSS. Cuclodextrins as drug carrier molecule: A review. Scientia Pharmaceutica 2008; 76: 567-598
• 9 Zhang JT, Chua LS, Lynn DM. Multilayer thin films that sustain the release of functional DNA under physiological conditions. Langmuir 2004; 20 (19) 8015-8021
• 10 Jessel N, Oulad-Abdelghani M, Meyer F et al. Multiple and time-scheduled in situ DNA delivery mediated be beta-cyclodextrin embedded in a polyelectolyte multilayer. Proc Nat Acad Sci USA 2006; 103 (23) 8618-8621
• 11 Irie T, Otagiri M, Sunada M. et al. Cyclodextrin-induced hemolysis and shape changes of human erythrocytes in vitro. J Pharmacobiodyn 1982; 5 (09) 741-744
• 12 Kilsdonk EPC, Yancey PG, Stoudt GW et al. Cellular cholesterol efflux mediated by cyclodextrins. J Biol Chem 1995; 270 (29) 17250-17256
• 13 Cryan SA, Holohan A, Donohue R et al. Cell transfection with polycationic cyclodextrine vectors. Eur J Pharm Sci 2004; 21 (05) 625-633
• 14 Mao S, Shuai X, Unger F et al. The depolymerization of chitosan: Effects on physicochemical and biological properties. Int J Pharm 2004; 281 (1–2) 45-54
• 15 Wang W, Bo S, Li S et al. Determination of the Mark-Houwink equation for chitosans with different degrees of deacetylation. Int J Biol Macromol 1991; 13 (05) 281-285
• 16 Mao HQ, Roya K, Troung-Le VL et al. Chitosan-DNA nanoparticles as gene carrier: Synthesis, characterization and transfection efficiency. J Controlled Release 2001; 70 (03) 399-421
• 17 Tan J, Newton CA, Djeu JY et al. Unjection of complementary DNA encoding interleukin-12 inhibits tumor establishment at a distant site in a murine renal carcinoma model. P Am Assoc Canc Res 1996; 56: 3399-3403
• 18 Diez S, Migueliz I, Ilarduya CT. Targeted cationic poly (D,L-lactic-co-glycolic acid) nanoparticles for gene delivery to cultured cells. Cell Mol Biol Lett 2009; 14 (02) 347-362
• 19 Ranaldi G, Mariglianoa I, Vespignani I et al. The effect of chitosan and other polycations on tight junction permeability in the human intestinal Caco-2 cell line. J Nutr Biochem 2002; 13 (03) 157-167
• 20 Bowman K, Leong KW. Chitosan nanoparticles for oral drug and gene delivery. Int J Nanomedicine 2006; 1 (02) 117-128
• 21 Csaba N, Kping-Hggard M, Alonso MJ. Ionically crosslinked chitosan/tripolyphosphate nanoparticles for oligonucleotide and plasmid DNA delivery. Int J Pharm 2009; 382: 205-214
• 22 Li XW, Lee DKL, Chan ASC et al. Sustained expression in mammalian cells with DNA complexed with chitosan nanoparticles. Biochim Biophys Acta 2003; 1630 (01) 7-18
• 23 Maeda Y, Kimura Y. Antitumor effects of various low-molecular-weight chitosans are due to increased natural killer activity of intestinal intraepithelial lymphocytes in sarcoma 180-bearing mice. J Nutr 2004; 134 (04) 945-950
• 24 Dhiman HK, Ray AR, Panda AK. Three-dimensional chitosan scaffold-based MCF-7 cell culture for the determination of the cytotoxicity of tamoxifen. Biomaterials 2005; 26 (09) 979-986
• 25 Qi LF, Xu ZR, Li Y et al. In vitro effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 cells. World J Gastroentero 2005; 11 (33) 5136-5141
• 26 Nge KL, Nitar N, Chandrkrachang S et al. Chitosan as a growth stimulator in orchid tissue culture. Plant Sci 2006; 170: 1185-1190