caa a22 4500 477126189 CHVBK 20180405111650.0 cr unu---uuuuu 170330e19970901xx s 000 0 eng 10.1023/A:1012106907708 doi (NATIONALLICENCE)springer-10.1023/A:1012106907708 N-Trimethyl Chitosan Chloride as a Potential Absorption Enhancer Across Mucosal Surfaces: In Vitro Evaluation in Intestinal Epithelial Cells (Caco-2) [Elektronische Daten] [Awie Kotzé, Henrik Lueβen, Bas de Leeuw, (A)Bert de Boer, J. Verhoef, Hans Junginger] Purpose. Previous studies have established that chitosan hydrochloride and glutamate are potent absorption enhancers for large hydrophilic compounds across mucosal surfaces. However, these compounds lack solubility at neutral pH values. A partially quaternized and well-soluble derivative of chitosan, N-trimethyl chitosan chloride, was synthesized and the effects of this polymer on the transepithelial electrical resistance and permeability of intestinal epithelial cells were investigated in vitro. Methods. N-trimethyl chitosan chloride was synthesized by reductive methylation and characterized with NMR. The effect of this polymer (1.0−2.5% w/v) on the transepithelial electrical resistance of intestinal epithelial cells, using Caco-2 cell monolayers, was investigated. Permeation of the hydrophilic model compounds [l4C]-mannitol (MW 182.2), FITC-Dextran (MW 4400) and the peptide drug buserelin (MW 1299.5), in the presence of N-trimethyl chitosan chloride (1.5−2.5% w/v), was followed for 3 hours. The transport process of the fluorescent marker, FITC-Dextran 4400, across the cell monolayers was visualised with confocal laser scanning microscopy. Viability of the cells was checked with the trypan blue exclusion technique. Results. N-trimethyl chitosan chloride was found to be a perfectly water-soluble, partially quaternized (about 12%) derivative of chitosan. This polymer (1.5−2.5% w/v) caused a pronounced and immediate reduction (25−85%) in the transepithelial electrical resistance of Caco-2 cells. Large increases in the transport rate of [!4C]-mannitol (32−60 fold), FITC-Dextran 4400 (167−373 fold) and buserelin (28−73 fold) were demonstrated. Confocal laser scanning microscopy confirmed that N-trimethyl chitosan chloride opens the tight junctions of intestinal epithelial cells to allow increased transport of hydrophilic compounds through the paracellular transport pathway. No deleterious effects to the cells could be demonstrated with trypan blue. Conclusions. The potential use of N-trimethyl chitosan chloride as an absorption enhancer across mucosal surfaces could be an important contribution towards the development of effective delivery systems for hydrophilic drugs. Plenum Publishing Corporation, 1997 N-trimethyl chitosan chloride nationallicence absorption enhancer nationallicence paracellular transport nationallicence transepithelial electrical resistance nationallicence Caco-2 cell monolayers nationallicence Kotzé Awie Department of Pharmaceutics, Potchefstroom University for Christian Higher Education, 2520, Potchefstroom, Republic of South Africa aut Lueβen Henrik Department of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300, RA Leiden, The Netherlands aut de Leeuw Bas Department of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300, RA Leiden, The Netherlands aut de Boer (A)Bert A Department of Pharmacology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300 RA, Leiden, The Netherlands aut Verhoef J. Department of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300, RA Leiden, The Netherlands aut Junginger Hans Department of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300, RA Leiden, The Netherlands aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/9(1997-09-01), 1197-1202 0724-8741 14:9<1197 1997 14 11095 https://doi.org/10.1023/A:1012106907708 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012106907708 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kotzé Awie Department of Pharmaceutics, Potchefstroom University for Christian Higher Education, 2520, Potchefstroom, Republic of South Africa aut NATIONALLICENCE 700 1- Lueβen Henrik Department of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300, RA Leiden, The Netherlands aut NATIONALLICENCE 700 1- de Leeuw Bas Department of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300, RA Leiden, The Netherlands aut NATIONALLICENCE 700 1- de Boer (A)Bert A Department of Pharmacology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300 RA, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Verhoef J. Department of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300, RA Leiden, The Netherlands aut NATIONALLICENCE 700 1- Junginger Hans Department of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, 2300, RA Leiden, The Netherlands aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/9(1997-09-01), 1197-1202 0724-8741 14:9<1197 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer