Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative study
Objective: Gene therapy relies on efficient vector for a therapeutic effect. Efficient non-viral vectors are sought as an alternative to viral vectors. Chitosan, a cationic polymer, has been studied for its gene delivery potential. In this work, disulfide bond containing groups were covalently added...
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doaj-5497f00291c048abb3c1084333e931b02020-11-25T00:21:06ZengWolters Kluwer Medknow PublicationsJournal of Pharmacy and Bioallied Sciences0975-74060976-48792013-01-015211111810.4103/0975-7406.111823Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative studyAna V OliveiraAndreia P SilvaDiogo B BitoqueGabriela A SilvaAna M Rosa da CostaObjective: Gene therapy relies on efficient vector for a therapeutic effect. Efficient non-viral vectors are sought as an alternative to viral vectors. Chitosan, a cationic polymer, has been studied for its gene delivery potential. In this work, disulfide bond containing groups were covalently added to chitosan to improve the transfection efficiency. These bonds can be cleaved by cytoplasmic glutathione, thus, releasing the DNA load more efficiently. Materials and Methods: Chitosan and thiolated chitosan nanoparticles (NPs) were prepared in order to obtain a NH3 + :PO4− ratio of 5:1 and characterized for plasmid DNA complexation and release efficiency. Cytotoxicity and gene delivery studies were carried out on retinal pigment epithelial cells. Results: In this work, we show that chitosan was effectively modified to incorporate a disulfide bond. The transfection efficiency of chitosan and thiolated chitosan varied according to the cell line used, however, thiolation did not seem to significantly improve transfection efficiency. Conclusion: The apparent lack of improvement in transfection efficiency of the thiolated chitosan NPs is most likely due to its size increase and charge inversion relatively to chitosan. Therefore, for retinal cells, thiolated chitosan does not seem to constitute an efficient strategy for gene delivery.http://www.jpbsonline.org/article.asp?issn=0975-7406;year=2013;volume=5;issue=2;spage=111;epage=118;aulast=OliveiraChitosangene therapynon-viral vectorsthiolationtransfection efficiency |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ana V Oliveira Andreia P Silva Diogo B Bitoque Gabriela A Silva Ana M Rosa da Costa |
spellingShingle |
Ana V Oliveira Andreia P Silva Diogo B Bitoque Gabriela A Silva Ana M Rosa da Costa Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative study Journal of Pharmacy and Bioallied Sciences Chitosan gene therapy non-viral vectors thiolation transfection efficiency |
author_facet |
Ana V Oliveira Andreia P Silva Diogo B Bitoque Gabriela A Silva Ana M Rosa da Costa |
author_sort |
Ana V Oliveira |
title |
Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative study |
title_short |
Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative study |
title_full |
Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative study |
title_fullStr |
Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative study |
title_full_unstemmed |
Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: A comparative study |
title_sort |
transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: a comparative study |
publisher |
Wolters Kluwer Medknow Publications |
series |
Journal of Pharmacy and Bioallied Sciences |
issn |
0975-7406 0976-4879 |
publishDate |
2013-01-01 |
description |
Objective: Gene therapy relies on efficient vector for a therapeutic effect. Efficient non-viral vectors are sought as an alternative to viral vectors. Chitosan, a cationic polymer, has been studied for its gene delivery potential. In this work, disulfide bond containing groups were covalently added to chitosan to improve the transfection efficiency. These bonds can be cleaved by cytoplasmic glutathione, thus, releasing the DNA load more efficiently. Materials and Methods: Chitosan and thiolated chitosan nanoparticles (NPs) were prepared in order to obtain a NH3 + :PO4− ratio of 5:1 and characterized for plasmid DNA complexation and release efficiency. Cytotoxicity and gene delivery studies were carried out on retinal pigment epithelial cells. Results: In this work, we show that chitosan was effectively modified to incorporate a disulfide bond. The transfection efficiency of chitosan and thiolated chitosan varied according to the cell line used, however, thiolation did not seem to significantly improve transfection efficiency. Conclusion: The apparent lack of improvement in transfection efficiency of the thiolated chitosan NPs is most likely due to its size increase and charge inversion relatively to chitosan. Therefore, for retinal cells, thiolated chitosan does not seem to constitute an efficient strategy for gene delivery. |
topic |
Chitosan gene therapy non-viral vectors thiolation transfection efficiency |
url |
http://www.jpbsonline.org/article.asp?issn=0975-7406;year=2013;volume=5;issue=2;spage=111;epage=118;aulast=Oliveira |
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