Abstract
The aim of this study was to investigate the effects of a type of hydrophobic moiety, extent of N-substitution (ES), and degree of quaternization (DQ) of chitosan (CS) on the transepithelial electrical resistance and permeability of Caco-2 cells monolayer, using fluorescein isothiocyanate dextran 4,400 (FD-4) as the model compound for paracellular tight junction transport. CS was substituted with hydrophobic moiety, an aliphatic aldehyde (n-octyl) or aromatic aldehyde (benzyl), for the improved hydrophobic interaction with cell membrane, and they were quaternized with Quat-188 to render CS soluble. The factors affecting the epithelial permeability have been evaluated in the intestinal cell monolayers, Caco-2 cells. Cytotoxicity was evaluated by using the trypan blue and MTT viability assay. The results revealed that at pH 7.4 CSQ appeared to increase cell permeability in dose-dependent manner, and this effect was relatively reversible at the lower doses of 0.05–1.25 mM. The higher DQ and ES caused the higher permeability of FD-4. Cytotoxicity of CSQ was concentration, %DQ, and %ES dependent. Substitution with hydrophobic moiety caused decreasing in permeability of FD-4 and cytotoxicity by benzyl group had more effect than octyl group. These studies demonstrated that these novel modified chitosan derivatives had potential for using as absorption enhancers.
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Abbreviations
- BzCSQ:
-
N-Benzyl chitosan Quat-188
- CS:
-
Chitosan
- CSA:
-
Chitosan acetate
- CSQ:
-
Chitosan Quat-188
- DQ:
-
Degree of Quat-188
- ES:
-
The extent of N-substitution
- FD-4:
-
Fluorescein isothiocyanate dextran 4,400
- MTT:
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide
- OctCSQ:
-
N-Octyl chitosan Quat-188
- TEER:
-
The transepithelial electrical resistance
- TM-Bz-CS:
-
Methylated N-(4-N,N-dimethylaminobenzyl) chitosan
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Acknowledgements
The authors wish to thank Commission of Higher Education (Thailand), The Thailand Research Funds through the Golden Jubilee Ph.D. Program (Grant No. PHD/0114/2550), The National Research Council of Thailand, and Silpakorn University Research and Development Institute for financial support.
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Kowapradit, J., Opanasopit, P., Ngawhirunpat, T. et al. In vitro Permeability Enhancement in Intestinal Epithelial Cells (Caco-2) Monolayer of Water Soluble Quaternary Ammonium Chitosan Derivatives. AAPS PharmSciTech 11, 497–508 (2010). https://doi.org/10.1208/s12249-010-9399-7
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DOI: https://doi.org/10.1208/s12249-010-9399-7