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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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Related in: MedlinePlus

(a) FM2-10 recycling rate performed on K562 cells in the presence and absence of Solutol® HS15. Fluorescence intensity was measured every 24 s, over 1 h. (b) FM2-10% endocytosis/min performed on K562 cells, over 60 min and 35–60 min (N = 3, n = 6). *** = <0.001, ** = <0.005, * = < 0.05).
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Fig6: (a) FM2-10 recycling rate performed on K562 cells in the presence and absence of Solutol® HS15. Fluorescence intensity was measured every 24 s, over 1 h. (b) FM2-10% endocytosis/min performed on K562 cells, over 60 min and 35–60 min (N = 3, n = 6). *** = <0.001, ** = <0.005, * = < 0.05).

Mentions: FM2-10 is a membrane dye which is normally used to investigate endocytosis and exocytosis rates, in particular the rate of endocytic vesicle movement through the cell (15, 19). The probe is believed to reversibly bind to the outer leaflet of the cell membrane (and increases in fluorescence intensity) and, during endocytosis, locates within the membrane of endocytic vesicle (19). FM2-10 was employed in this experiment to investigate the effect of Solutol® HS15 on membrane endocytosis rate. The data in Fig. 6a depict the effect of Solutol® HS15 on FM-210 fluorescence intensity changes vs time in a suspension of K562 cells based on the previous demonstration that the increase in fluorescence intensity as a function of time corresponds to the fraction of membrane being internalised via endocytosis per unit of time (15, 20). The profiles obtained illustrate a gradual increase in fluorescence with time for all tested samples, relative to respective control, whereby the effect (slope of the curve) is generally dependent on the concentration of Solutol® HS15 applied to the cells.Fig. 6


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)

(a) FM2-10 recycling rate performed on K562 cells in the presence and absence of Solutol® HS15. Fluorescence intensity was measured every 24 s, over 1 h. (b) FM2-10% endocytosis/min performed on K562 cells, over 60 min and 35–60 min (N = 3, n = 6). *** = <0.001, ** = <0.005, * = < 0.05).
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4300420&req=5

Fig6: (a) FM2-10 recycling rate performed on K562 cells in the presence and absence of Solutol® HS15. Fluorescence intensity was measured every 24 s, over 1 h. (b) FM2-10% endocytosis/min performed on K562 cells, over 60 min and 35–60 min (N = 3, n = 6). *** = <0.001, ** = <0.005, * = < 0.05).
Mentions: FM2-10 is a membrane dye which is normally used to investigate endocytosis and exocytosis rates, in particular the rate of endocytic vesicle movement through the cell (15, 19). The probe is believed to reversibly bind to the outer leaflet of the cell membrane (and increases in fluorescence intensity) and, during endocytosis, locates within the membrane of endocytic vesicle (19). FM2-10 was employed in this experiment to investigate the effect of Solutol® HS15 on membrane endocytosis rate. The data in Fig. 6a depict the effect of Solutol® HS15 on FM-210 fluorescence intensity changes vs time in a suspension of K562 cells based on the previous demonstration that the increase in fluorescence intensity as a function of time corresponds to the fraction of membrane being internalised via endocytosis per unit of time (15, 20). The profiles obtained illustrate a gradual increase in fluorescence with time for all tested samples, relative to respective control, whereby the effect (slope of the curve) is generally dependent on the concentration of Solutol® HS15 applied to the cells.Fig. 6

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
Related in: MedlinePlus