Insight into Polycation Chain Length Affecting Transfection Efficiency by O-methyl-free N,N,N-trimethyl Chitosans As Gene Carriers

Overview

Journal Pharm Res

Specialties Pharmacology
Pharmacy

Date 2013 Oct 9

PMID 24100950

Citations 1

Authors

Tao Xu

Suhang Wang

Zhengzhong Shao

Affiliations

Soon will be listed here.

Abstract

Purpose: The structure-function relationship and mechanism of polycations as gene carriers have attracted considerable research interest in recent years. The present study was to investigate the relationship between polycation chain length and transfection efficiency (RCL-TE), and the corresponding mechanism by O-methyl-free N,N,N-trimethyl chitosans (TMCs) as gene carriers.

Methods: Four TMCs with various chain lengths were synthesized and used to evaluate the RCL-TE. To investigate the details of RCL-TE, a number of factors such as cytotoxicity, cellular uptake efficiency, cellular uptake pathway and intracellular trafficking, were evaluated.

Results: In comparison to short chain length TMCs (S-TMCs), long chain length ones (L-TMCs) mediated higher gene expression. The polyplexes formed by L-TMCs and pDNA showed higher stability. The cellular uptake pathway and intracellular trafficking of these TMC/pDNA polyplexes were different. These above factors are probably the key ones in RCL-TE rather than polycation-DNA binding affinity, polyplex particle size in water, zeta potential, serum, cytotoxicity, and cellular uptake efficiency.

Conclusions: For rational design of chitosan-based polycations as gene carriers, polycations with relative long chain lengths are more favorable and more attention should be paid to polyplex stability, function of uncomplexed polycation chains, cellular uptake pathway, and intracellular trafficking.

Citing Articles

Elucidating the role of free polycationic chains in polycation gene carriers by free chains of polyethylenimine or N,N,N-trimethyl chitosan plus a certain polyplex.

Xu T, Liu W, Wang S, Shao Z Int J Nanomedicine. 2014; 9:3231-45.

PMID: 25061299 PMC: 4086671. DOI: 10.2147/IJN.S64308.

References

Yue Y, Jin F, Deng R, Cai J, Chen Y, Lin M . Revisit complexation between DNA and polyethylenimine - Effect of uncomplexed chains free in the solution mixture on gene transfection. J Control Release. 2010; 155(1):67-76. DOI: 10.1016/j.jconrel.2010.10.028. View

Song Y, Wang H, Zeng X, Sun Y, Zhang X, Zhou J . Effect of molecular weight and degree of substitution of quaternized cellulose on the efficiency of gene transfection. Bioconjug Chem. 2010; 21(7):1271-9. DOI: 10.1021/bc100068f. View

Thomas J, Rekha M, Sharma C . Unraveling the intracellular efficacy of dextran-histidine polycation as an efficient nonviral gene delivery system. Mol Pharm. 2011; 9(1):121-34. DOI: 10.1021/mp200485b. View

Ma P, Buschmann M, Winnik F . One-step analysis of DNA/chitosan complexes by field-flow fractionation reveals particle size and free chitosan content. Biomacromolecules. 2010; 11(3):549-54. DOI: 10.1021/bm901345q. View

Koping-Hoggard M, Varum K, Issa M, Danielsen S, Christensen B, Stokke B . Improved chitosan-mediated gene delivery based on easily dissociated chitosan polyplexes of highly defined chitosan oligomers. Gene Ther. 2004; 11(19):1441-52. DOI: 10.1038/sj.gt.3302312. View

Jiang H, Kim Y, Arote R, Nah J, Cho M, Choi Y . Chitosan-graft-polyethylenimine as a gene carrier. J Control Release. 2006; 117(2):273-80. DOI: 10.1016/j.jconrel.2006.10.025. View

Rejman J, Bragonzi A, Conese M . Role of clathrin- and caveolae-mediated endocytosis in gene transfer mediated by lipo- and polyplexes. Mol Ther. 2005; 12(3):468-74. DOI: 10.1016/j.ymthe.2005.03.038. View

van der Aa M, Huth U, Hafele S, Schubert R, Oosting R, Mastrobattista E . Cellular uptake of cationic polymer-DNA complexes via caveolae plays a pivotal role in gene transfection in COS-7 cells. Pharm Res. 2007; 24(8):1590-8. PMC: 1915651. DOI: 10.1007/s11095-007-9287-3. View

Godbey W, Wu K, Mikos A . Poly(ethylenimine) and its role in gene delivery. J Control Release. 1999; 60(2-3):149-60. DOI: 10.1016/s0168-3659(99)00090-5. View

10.

Kulkarni R, Wu D, Davis M, Fraser S . Quantitating intracellular transport of polyplexes by spatio-temporal image correlation spectroscopy. Proc Natl Acad Sci U S A. 2005; 102(21):7523-8. PMC: 1140437. DOI: 10.1073/pnas.0501950102. View

11.

Germershaus O, Mao S, Sitterberg J, Bakowsky U, Kissel T . Gene delivery using chitosan, trimethyl chitosan or polyethylenglycol-graft-trimethyl chitosan block copolymers: establishment of structure-activity relationships in vitro. J Control Release. 2007; 125(2):145-54. DOI: 10.1016/j.jconrel.2007.10.013. View

12.

Kean T, Roth S, Thanou M . Trimethylated chitosans as non-viral gene delivery vectors: cytotoxicity and transfection efficiency. J Control Release. 2005; 103(3):643-53. DOI: 10.1016/j.jconrel.2005.01.001. View

13.

Thanou M, Florea B, Geldof M, Junginger H, Borchard G . Quaternized chitosan oligomers as novel gene delivery vectors in epithelial cell lines. Biomaterials. 2002; 23(1):153-9. DOI: 10.1016/s0142-9612(01)00090-4. View

14.

Deng R, Yue Y, Jin F, Chen Y, Kung H, Lin M . Revisit the complexation of PEI and DNA - how to make low cytotoxic and highly efficient PEI gene transfection non-viral vectors with a controllable chain length and structure?. J Control Release. 2009; 140(1):40-6. DOI: 10.1016/j.jconrel.2009.07.009. View

15.

Jintapattanakit A, Mao S, Kissel T, Junyaprasert V . Physicochemical properties and biocompatibility of N-trimethyl chitosan: effect of quaternization and dimethylation. Eur J Pharm Biopharm. 2008; 70(2):563-71. DOI: 10.1016/j.ejpb.2008.06.002. View

16.

Forrest M, Pack D . On the kinetics of polyplex endocytic trafficking: implications for gene delivery vector design. Mol Ther. 2002; 6(1):57-66. DOI: 10.1006/mthe.2002.0631. View

17.

Gebhart C, Kabanov A . Evaluation of polyplexes as gene transfer agents. J Control Release. 2001; 73(2-3):401-16. DOI: 10.1016/s0168-3659(01)00357-1. View

18.

Eltoukhy A, Siegwart D, Alabi C, Rajan J, Langer R, Anderson D . Effect of molecular weight of amine end-modified poly(β-amino ester)s on gene delivery efficiency and toxicity. Biomaterials. 2012; 33(13):3594-603. PMC: 3326358. DOI: 10.1016/j.biomaterials.2012.01.046. View

19.

Reilly M, Larsen J, Sullivan M . Polyplexes traffic through caveolae to the Golgi and endoplasmic reticulum en route to the nucleus. Mol Pharm. 2012; 9(5):1280-90. DOI: 10.1021/mp200583d. View

20.

von Gersdorff K, Sanders N, Vandenbroucke R, De Smedt S, Wagner E, Ogris M . The internalization route resulting in successful gene expression depends on both cell line and polyethylenimine polyplex type. Mol Ther. 2006; 14(5):745-53. DOI: 10.1016/j.ymthe.2006.07.006. View