PURPOSE: To verify or refute the mechanism of permeation enhancement with thiolated polymers via GSH by the use of NaFlu as marker for the paracellular permeation. METHODS: The capability of 0.5% polycarbophil cysteine conjugate (PCP-Cys) to reduce 0.02% oxidized glutathione (GSSG) was evaluated via iodometric titration in aqueous solution. Glutathione in its reduced form (GSH; 0.1%-0.4%) and in combination with 0.5% PCP-Cys were tested for their permeation enhancement of sodium fluorescein (NaFlu) and fluorescence labeled bacitracin (bac-FITC) used as paracellular markers. Permeation studies across guinea pig duodenum were carried out in Ussing-type chambers. Opening of the tight junctions was additionally monitored by transepithelial electrical resistance (TEER) measurements. RESULTS: PCP-Cys (0.5%) was shown to reduce 22.0%+/-8.2% of GSSG (0.02%) to GSH in aqueous solution at pH 7.0 and 37 degrees C within 3 h. Permeation of NaFlu was shown to depend on the concentration of GSH. The apparent permeability coefficient (Papp) of NaFlu in buffer only was 4.98+/-0.5*10(-6), while in the presence of 0.4% GSH a Papp of 9.31+/-0.92*10(-6) was achieved, representing an enhancement ratio (R = Papp enhancer system/Papp control) of 1.86. The combination of GSH (0.4%) with PCP-Cys (0.5%) led to a significant (p < 0.001) improvement of R for NaFlu up to 2.93 accompanied by a decrease in TEER of 20.3%+/-1.4%. Incubation of bac-FITC with the same GSH/PCP-Cys combination led to an enhancement ratio of 2.06 within 3 h. CONCLUSION: GSH plays an important role in the opening of tight junctions of intestinal epithelia. It would appear that PCP-Cys is able to reduce GSSG, prolonging the concentration of GSH at the apical membrane, resulting in significantly enhanced paracellular transport.
PURPOSE: To verify or refute the mechanism of permeation enhancement with thiolated polymers via GSH by the use of NaFlu as marker for the paracellular permeation. METHODS: The capability of 0.5% polycarbophil cysteine conjugate (PCP-Cys) to reduce 0.02% oxidized glutathione (GSSG) was evaluated via iodometric titration in aqueous solution. Glutathione in its reduced form (GSH; 0.1%-0.4%) and in combination with 0.5% PCP-Cys were tested for their permeation enhancement of sodium fluorescein (NaFlu) and fluorescence labeled bacitracin (bac-FITC) used as paracellular markers. Permeation studies across guinea pig duodenum were carried out in Ussing-type chambers. Opening of the tight junctions was additionally monitored by transepithelial electrical resistance (TEER) measurements. RESULTS:PCP-Cys (0.5%) was shown to reduce 22.0%+/-8.2% of GSSG (0.02%) to GSH in aqueous solution at pH 7.0 and 37 degrees C within 3 h. Permeation of NaFlu was shown to depend on the concentration of GSH. The apparent permeability coefficient (Papp) of NaFlu in buffer only was 4.98+/-0.5*10(-6), while in the presence of 0.4% GSH a Papp of 9.31+/-0.92*10(-6) was achieved, representing an enhancement ratio (R = Papp enhancer system/Papp control) of 1.86. The combination of GSH (0.4%) with PCP-Cys (0.5%) led to a significant (p < 0.001) improvement of R for NaFlu up to 2.93 accompanied by a decrease in TEER of 20.3%+/-1.4%. Incubation of bac-FITC with the same GSH/PCP-Cys combination led to an enhancement ratio of 2.06 within 3 h. CONCLUSION:GSH plays an important role in the opening of tight junctions of intestinal epithelia. It would appear that PCP-Cys is able to reduce GSSG, prolonging the concentration of GSH at the apical membrane, resulting in significantly enhanced paracellular transport.
Authors: W C Barrett; J P DeGnore; S König; H M Fales; Y F Keng; Z Y Zhang; M B Yim; P B Chock Journal: Biochemistry Date: 1999-05-18 Impact factor: 3.162
Authors: T Uchiyama; A Kotani; T Kishida; H Tatsumi; A Okamoto; T Fujita; M Murakami; S Muranishi; A Yamamoto Journal: J Pharm Sci Date: 1998-04 Impact factor: 3.534
Authors: K Gradauer; S Dünnhaupt; C Vonach; H Szöllösi; I Pali-Schöll; H Mangge; E Jensen-Jarolim; A Bernkop-Schnürch; R Prassl Journal: J Control Release Date: 2012-12-08 Impact factor: 9.776
Authors: K Gradauer; J Barthelmes; C Vonach; G Almer; H Mangge; B Teubl; E Roblegg; S Dünnhaupt; E Fröhlich; A Bernkop-Schnürch; R Prassl Journal: J Control Release Date: 2013-10-16 Impact factor: 9.776