Comparative Study of Diethylaminoethyl-Chitosan and Methylglycol-Chitosan as Potential Non-Viral Vectors for Gene Therapy

Sergei V. Raik, Stanislav Andranovitš, Valentina A. Petrova, Yingying Xu, Jenny Ka-Wing Lam, Gordon Morris, Alexandra V. Brodskaia, Luca Casettari, Andreii S. Kritchenkov, Yury A. Skorik

Research output: Contribution to journalArticle

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Abstract

In this paper, we compared the transfection efficiency and cytotoxicity of methylglycol-chitosan (MG-CS) and diethylaminoethyl-chitosan (DEAE-CSI and DEAE-CSII with degrees of substitution of 1.2 and 0.57, respectively) to that of Lipofectamine (used as a reference transfection vector). MG-CS contains quaternary amines to improve DNA binding, whereas the DEAE-CS exhibits pH buffering capability that would ostensibly enhance transfection efficiency by promoting endosomal escape. Gel retardation assays showed that both DEAE-CS and MG-CS bound to DNA at a polysaccharide:DNA mass ratio of 2:1. In Calu-3 cells, the DNA transfection activity was significantly better with MG-CS than with DEAE-CS, and the efficiency improved with increasing polysaccharide:DNA ratios. By contrast, the efficiency of DEAE-CSI and DEAE-CSII was independent of the polysaccharide:DNA ratio. Conversely, in the transfection-recalcitrant JAWSII cells, both Lipofectamine and MG-CS showed significantly lower DNA transfection activity than in Calu-3 cells, whereas the efficiency of DEAE-CSI and DEAE-CSII was similar in both cell lines. The toxicity of DEAE-CS increased with increasing concentrations of the polymer and its degree of substitution, whereas MG-CS demonstrated negligible cytotoxicity, even at the highest concentration studied. Overall, MG-CS proved to be a more efficient and less toxic transfection agent when compared to DEAE-CS.
Original languageEnglish
Article number442
Number of pages15
JournalPolymers
Volume10
Issue number4
DOIs
Publication statusPublished - 14 Apr 2018

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Gene therapy
Chitosan
DNA
Polysaccharides
Cytotoxicity
Substitution reactions
2-diethylaminoethanol
methylglycol chitosan
Poisons
Toxicity
Amines
Assays
Gels
Cells
Polymers

Cite this

Raik, Sergei V. ; Andranovitš, Stanislav ; Petrova, Valentina A. ; Xu, Yingying ; Lam, Jenny Ka-Wing ; Morris, Gordon ; Brodskaia, Alexandra V. ; Casettari, Luca ; Kritchenkov, Andreii S. ; Skorik, Yury A. / Comparative Study of Diethylaminoethyl-Chitosan and Methylglycol-Chitosan as Potential Non-Viral Vectors for Gene Therapy. In: Polymers. 2018 ; Vol. 10, No. 4.
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title = "Comparative Study of Diethylaminoethyl-Chitosan and Methylglycol-Chitosan as Potential Non-Viral Vectors for Gene Therapy",
abstract = "In this paper, we compared the transfection efficiency and cytotoxicity of methylglycol-chitosan (MG-CS) and diethylaminoethyl-chitosan (DEAE-CSI and DEAE-CSII with degrees of substitution of 1.2 and 0.57, respectively) to that of Lipofectamine (used as a reference transfection vector). MG-CS contains quaternary amines to improve DNA binding, whereas the DEAE-CS exhibits pH buffering capability that would ostensibly enhance transfection efficiency by promoting endosomal escape. Gel retardation assays showed that both DEAE-CS and MG-CS bound to DNA at a polysaccharide:DNA mass ratio of 2:1. In Calu-3 cells, the DNA transfection activity was significantly better with MG-CS than with DEAE-CS, and the efficiency improved with increasing polysaccharide:DNA ratios. By contrast, the efficiency of DEAE-CSI and DEAE-CSII was independent of the polysaccharide:DNA ratio. Conversely, in the transfection-recalcitrant JAWSII cells, both Lipofectamine and MG-CS showed significantly lower DNA transfection activity than in Calu-3 cells, whereas the efficiency of DEAE-CSI and DEAE-CSII was similar in both cell lines. The toxicity of DEAE-CS increased with increasing concentrations of the polymer and its degree of substitution, whereas MG-CS demonstrated negligible cytotoxicity, even at the highest concentration studied. Overall, MG-CS proved to be a more efficient and less toxic transfection agent when compared to DEAE-CS.",
keywords = "diethylaminoethyl-chitosan, gene delivery, cell transfection, methylglycol-chitosan, polyplex",
author = "Raik, {Sergei V.} and Stanislav Andranovitš and Petrova, {Valentina A.} and Yingying Xu and Lam, {Jenny Ka-Wing} and Gordon Morris and Brodskaia, {Alexandra V.} and Luca Casettari and Kritchenkov, {Andreii S.} and Skorik, {Yury A.}",
year = "2018",
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Raik, SV, Andranovitš, S, Petrova, VA, Xu, Y, Lam, JK-W, Morris, G, Brodskaia, AV, Casettari, L, Kritchenkov, AS & Skorik, YA 2018, 'Comparative Study of Diethylaminoethyl-Chitosan and Methylglycol-Chitosan as Potential Non-Viral Vectors for Gene Therapy', Polymers, vol. 10, no. 4, 442. https://doi.org/10.3390/polym10040442

Comparative Study of Diethylaminoethyl-Chitosan and Methylglycol-Chitosan as Potential Non-Viral Vectors for Gene Therapy. / Raik, Sergei V.; Andranovitš, Stanislav; Petrova, Valentina A.; Xu, Yingying; Lam, Jenny Ka-Wing; Morris, Gordon; Brodskaia, Alexandra V.; Casettari, Luca; Kritchenkov, Andreii S.; Skorik, Yury A.

In: Polymers, Vol. 10, No. 4, 442, 14.04.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Comparative Study of Diethylaminoethyl-Chitosan and Methylglycol-Chitosan as Potential Non-Viral Vectors for Gene Therapy

AU - Raik, Sergei V.

AU - Andranovitš, Stanislav

AU - Petrova, Valentina A.

AU - Xu, Yingying

AU - Lam, Jenny Ka-Wing

AU - Morris, Gordon

AU - Brodskaia, Alexandra V.

AU - Casettari, Luca

AU - Kritchenkov, Andreii S.

AU - Skorik, Yury A.

PY - 2018/4/14

Y1 - 2018/4/14

N2 - In this paper, we compared the transfection efficiency and cytotoxicity of methylglycol-chitosan (MG-CS) and diethylaminoethyl-chitosan (DEAE-CSI and DEAE-CSII with degrees of substitution of 1.2 and 0.57, respectively) to that of Lipofectamine (used as a reference transfection vector). MG-CS contains quaternary amines to improve DNA binding, whereas the DEAE-CS exhibits pH buffering capability that would ostensibly enhance transfection efficiency by promoting endosomal escape. Gel retardation assays showed that both DEAE-CS and MG-CS bound to DNA at a polysaccharide:DNA mass ratio of 2:1. In Calu-3 cells, the DNA transfection activity was significantly better with MG-CS than with DEAE-CS, and the efficiency improved with increasing polysaccharide:DNA ratios. By contrast, the efficiency of DEAE-CSI and DEAE-CSII was independent of the polysaccharide:DNA ratio. Conversely, in the transfection-recalcitrant JAWSII cells, both Lipofectamine and MG-CS showed significantly lower DNA transfection activity than in Calu-3 cells, whereas the efficiency of DEAE-CSI and DEAE-CSII was similar in both cell lines. The toxicity of DEAE-CS increased with increasing concentrations of the polymer and its degree of substitution, whereas MG-CS demonstrated negligible cytotoxicity, even at the highest concentration studied. Overall, MG-CS proved to be a more efficient and less toxic transfection agent when compared to DEAE-CS.

AB - In this paper, we compared the transfection efficiency and cytotoxicity of methylglycol-chitosan (MG-CS) and diethylaminoethyl-chitosan (DEAE-CSI and DEAE-CSII with degrees of substitution of 1.2 and 0.57, respectively) to that of Lipofectamine (used as a reference transfection vector). MG-CS contains quaternary amines to improve DNA binding, whereas the DEAE-CS exhibits pH buffering capability that would ostensibly enhance transfection efficiency by promoting endosomal escape. Gel retardation assays showed that both DEAE-CS and MG-CS bound to DNA at a polysaccharide:DNA mass ratio of 2:1. In Calu-3 cells, the DNA transfection activity was significantly better with MG-CS than with DEAE-CS, and the efficiency improved with increasing polysaccharide:DNA ratios. By contrast, the efficiency of DEAE-CSI and DEAE-CSII was independent of the polysaccharide:DNA ratio. Conversely, in the transfection-recalcitrant JAWSII cells, both Lipofectamine and MG-CS showed significantly lower DNA transfection activity than in Calu-3 cells, whereas the efficiency of DEAE-CSI and DEAE-CSII was similar in both cell lines. The toxicity of DEAE-CS increased with increasing concentrations of the polymer and its degree of substitution, whereas MG-CS demonstrated negligible cytotoxicity, even at the highest concentration studied. Overall, MG-CS proved to be a more efficient and less toxic transfection agent when compared to DEAE-CS.

KW - diethylaminoethyl-chitosan

KW - gene delivery

KW - cell transfection

KW - methylglycol-chitosan

KW - polyplex

U2 - 10.3390/polym10040442

DO - 10.3390/polym10040442

M3 - Article

VL - 10

JO - Polymers

JF - Polymers

SN - 2073-4360

IS - 4

M1 - 442

ER -