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Mechanism of mucosal permeability enhancement of CriticalSorb® (Solutol® HS15) investigated in vitro in cell cultures.

Shubber S, Vllasaliu D, Rauch C, Jordan F, Illum L, Stolnik S - Pharm. Res. (2014)

Bottom Line: The rate of cell endocytosis was measured in vitro in the presence of Solutol® HS15 using a membrane probe, FM 2-10.This work initially confirms surfactant-like behaviour of Solutol® HS15 in aqueous media, while subsequent experiments demonstrate that the effect of Solutol® HS15 on epithelial tight junctions is different from a 'classical' tight junction opening agent and illustrate the effect of Solutol® HS15 on the cell membrane (endocytosis rate) and F-actin cytoskeleton.This study suggests that its mechanism of action arises primarily from effects on the cell membrane and consequent impacts on the cell cytoskeleton in terms of actin organisation and tight junction opening.

View Article: PubMed Central - PubMed

Affiliation: Division of Drug Delivery and Tissue Engineering, School of Pharmacy Boots Science Building, University of Nottingham, University Park, Nottingham, NG7 2RD, UK.

ABSTRACT

Purpose: CriticalSorb™, with the principal component Solutol® HS15, is a novel mucosal drug delivery system demonstrated to improve the bioavailability of selected biotherapeutics. The intention of this study is to elucidate mechanism(s) responsible for the enhancement of trans-mucosal absorption of biological drugs by Solutol® HS15.

Methods: Micelle size and CMC of Solutol® HS15 were determined in biologically relevant media. Polarised airway Calu-3 cell layers were used to measure the permeability of a panel of biological drugs, and to assess changes in TEER, tight junction and F-actin morphology. The rate of cell endocytosis was measured in vitro in the presence of Solutol® HS15 using a membrane probe, FM 2-10.

Results: This work initially confirms surfactant-like behaviour of Solutol® HS15 in aqueous media, while subsequent experiments demonstrate that the effect of Solutol® HS15 on epithelial tight junctions is different from a 'classical' tight junction opening agent and illustrate the effect of Solutol® HS15 on the cell membrane (endocytosis rate) and F-actin cytoskeleton.

Conclusion: Solutol® HS15 is the principle component of CriticalSorb™ that has shown an enhancement in permeability of medium sized biological drugs across epithelia. This study suggests that its mechanism of action arises primarily from effects on the cell membrane and consequent impacts on the cell cytoskeleton in terms of actin organisation and tight junction opening.

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Confocal micrographs with XZ stack showing distribution of junctional complex protein ZO-1 (left) and F-actin morphology (right) distribution in polarized Calu-3 monolayers following application of Solutol® HS15. White arrows highlight changes in F-actin distribution following Solutol® HS15 application. ZO-1 (green) was immunostained with mouse, anti-human ZO-1 (primary) antibody then with FITC-labelled goat, anti-mouse (secondary) antibody. F-actin was immunostained with Alexafluor 546-labelled phalloidin (red) and nuclei blue. Scale bar represents 25 μm and ‘cross hair’ represents region for Z stack analysis. ImageJ (1.47 K) image software was used to create the overlay images from raw series images.
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Fig5: Confocal micrographs with XZ stack showing distribution of junctional complex protein ZO-1 (left) and F-actin morphology (right) distribution in polarized Calu-3 monolayers following application of Solutol® HS15. White arrows highlight changes in F-actin distribution following Solutol® HS15 application. ZO-1 (green) was immunostained with mouse, anti-human ZO-1 (primary) antibody then with FITC-labelled goat, anti-mouse (secondary) antibody. F-actin was immunostained with Alexafluor 546-labelled phalloidin (red) and nuclei blue. Scale bar represents 25 μm and ‘cross hair’ represents region for Z stack analysis. ImageJ (1.47 K) image software was used to create the overlay images from raw series images.

Mentions: Confocal micrographs of Calu-3 layers following Solutol® HS15 treatment at concentrations below and above the CMC and immunostained for ZO-1 and F-actin distribution are shown in Fig. 5. ZO-1, a junctional protein, normally presents a ‘chicken wire-like’ distribution that resides closer to the apical region of the Calu-3 monolayer. This wire-like structure can be seen in control cells and appears to be retained when Calu-3 cells are treated with solutions of Solutol® HS15 below the CMC (0.01 mM). However, the ZO-1 pattern becomes faintly distorted in places when Calu-3 monolayers are treated Solutol® HS15 above the CMC (1.00 and 5.20 mM). The change in ZO-1 appearance with Solutol® HS15 is different to the dramatic re-distribution that follows from Calu-3 treatment with chitosan (6).Fig. 5


Mechanism of mucosal permeability enhancement of CriticalSorb® (Solutol® HS15) investigated in vitro in cell cultures.

Shubber S, Vllasaliu D, Rauch C, Jordan F, Illum L, Stolnik S - Pharm. Res. (2014)

Confocal micrographs with XZ stack showing distribution of junctional complex protein ZO-1 (left) and F-actin morphology (right) distribution in polarized Calu-3 monolayers following application of Solutol® HS15. White arrows highlight changes in F-actin distribution following Solutol® HS15 application. ZO-1 (green) was immunostained with mouse, anti-human ZO-1 (primary) antibody then with FITC-labelled goat, anti-mouse (secondary) antibody. F-actin was immunostained with Alexafluor 546-labelled phalloidin (red) and nuclei blue. Scale bar represents 25 μm and ‘cross hair’ represents region for Z stack analysis. ImageJ (1.47 K) image software was used to create the overlay images from raw series images.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4300420&req=5

Fig5: Confocal micrographs with XZ stack showing distribution of junctional complex protein ZO-1 (left) and F-actin morphology (right) distribution in polarized Calu-3 monolayers following application of Solutol® HS15. White arrows highlight changes in F-actin distribution following Solutol® HS15 application. ZO-1 (green) was immunostained with mouse, anti-human ZO-1 (primary) antibody then with FITC-labelled goat, anti-mouse (secondary) antibody. F-actin was immunostained with Alexafluor 546-labelled phalloidin (red) and nuclei blue. Scale bar represents 25 μm and ‘cross hair’ represents region for Z stack analysis. ImageJ (1.47 K) image software was used to create the overlay images from raw series images.
Mentions: Confocal micrographs of Calu-3 layers following Solutol® HS15 treatment at concentrations below and above the CMC and immunostained for ZO-1 and F-actin distribution are shown in Fig. 5. ZO-1, a junctional protein, normally presents a ‘chicken wire-like’ distribution that resides closer to the apical region of the Calu-3 monolayer. This wire-like structure can be seen in control cells and appears to be retained when Calu-3 cells are treated with solutions of Solutol® HS15 below the CMC (0.01 mM). However, the ZO-1 pattern becomes faintly distorted in places when Calu-3 monolayers are treated Solutol® HS15 above the CMC (1.00 and 5.20 mM). The change in ZO-1 appearance with Solutol® HS15 is different to the dramatic re-distribution that follows from Calu-3 treatment with chitosan (6).Fig. 5

Bottom Line: The rate of cell endocytosis was measured in vitro in the presence of Solutol® HS15 using a membrane probe, FM 2-10.This work initially confirms surfactant-like behaviour of Solutol® HS15 in aqueous media, while subsequent experiments demonstrate that the effect of Solutol® HS15 on epithelial tight junctions is different from a 'classical' tight junction opening agent and illustrate the effect of Solutol® HS15 on the cell membrane (endocytosis rate) and F-actin cytoskeleton.This study suggests that its mechanism of action arises primarily from effects on the cell membrane and consequent impacts on the cell cytoskeleton in terms of actin organisation and tight junction opening.

View Article: PubMed Central - PubMed

Affiliation: Division of Drug Delivery and Tissue Engineering, School of Pharmacy Boots Science Building, University of Nottingham, University Park, Nottingham, NG7 2RD, UK.

ABSTRACT

Purpose: CriticalSorb™, with the principal component Solutol® HS15, is a novel mucosal drug delivery system demonstrated to improve the bioavailability of selected biotherapeutics. The intention of this study is to elucidate mechanism(s) responsible for the enhancement of trans-mucosal absorption of biological drugs by Solutol® HS15.

Methods: Micelle size and CMC of Solutol® HS15 were determined in biologically relevant media. Polarised airway Calu-3 cell layers were used to measure the permeability of a panel of biological drugs, and to assess changes in TEER, tight junction and F-actin morphology. The rate of cell endocytosis was measured in vitro in the presence of Solutol® HS15 using a membrane probe, FM 2-10.

Results: This work initially confirms surfactant-like behaviour of Solutol® HS15 in aqueous media, while subsequent experiments demonstrate that the effect of Solutol® HS15 on epithelial tight junctions is different from a 'classical' tight junction opening agent and illustrate the effect of Solutol® HS15 on the cell membrane (endocytosis rate) and F-actin cytoskeleton.

Conclusion: Solutol® HS15 is the principle component of CriticalSorb™ that has shown an enhancement in permeability of medium sized biological drugs across epithelia. This study suggests that its mechanism of action arises primarily from effects on the cell membrane and consequent impacts on the cell cytoskeleton in terms of actin organisation and tight junction opening.

Show MeSH