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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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Effect of Solutol® HS15 on Calu-3, Caco-2 and A549 cells assessed by metabolic activity (a) and LDH assays (b). Solutol® HS15 was used at concentrations above and below the Critical Micelle Concentration (Fig. 2). Data expressed as relative metabolic activity and presented as the mean ± SD with N = 3 and n = 4. Data to summarize Solutol® HS15 concentrations that cause 50% reduction in cell viability (EC50, mM) and LDH leakage in tested cell lines shown in table (c). Statistical analysis: MTS assay, EC50P values: Calu-3 to Caco-2 < 0.05, Caco-2 to A549 > 0.05 and Calu-3 to A459 > 0.05. LDH assay EC50P values; Calu-3 to Caco-2 < 0.05, Caco-2 to A549 > 0.05 and Calu-3 to A459 < 0.05 conducted using t-test. Overall statistical difference between each test conducted for MTS and LDH P value < 0.05, conducted using one-way ANOVA.
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Fig3: Effect of Solutol® HS15 on Calu-3, Caco-2 and A549 cells assessed by metabolic activity (a) and LDH assays (b). Solutol® HS15 was used at concentrations above and below the Critical Micelle Concentration (Fig. 2). Data expressed as relative metabolic activity and presented as the mean ± SD with N = 3 and n = 4. Data to summarize Solutol® HS15 concentrations that cause 50% reduction in cell viability (EC50, mM) and LDH leakage in tested cell lines shown in table (c). Statistical analysis: MTS assay, EC50P values: Calu-3 to Caco-2 < 0.05, Caco-2 to A549 > 0.05 and Calu-3 to A459 > 0.05. LDH assay EC50P values; Calu-3 to Caco-2 < 0.05, Caco-2 to A549 > 0.05 and Calu-3 to A459 < 0.05 conducted using t-test. Overall statistical difference between each test conducted for MTS and LDH P value < 0.05, conducted using one-way ANOVA.

Mentions: Figure 3a summarises data from the MTS assay, employed to assess the effect of Solutol® HS15 on the viability of lung derived Calu-3 and A549, as well as intestinal Caco-2 cells. A general decline in cell metabolic activity is seen for all three tested cell lines as the concentration of Solutol® HS15 solution is increased. A concentration-dependent effect is also seen for LDH release from the cells (Fig. 3b).Fig. 3


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)

Effect of Solutol® HS15 on Calu-3, Caco-2 and A549 cells assessed by metabolic activity (a) and LDH assays (b). Solutol® HS15 was used at concentrations above and below the Critical Micelle Concentration (Fig. 2). Data expressed as relative metabolic activity and presented as the mean ± SD with N = 3 and n = 4. Data to summarize Solutol® HS15 concentrations that cause 50% reduction in cell viability (EC50, mM) and LDH leakage in tested cell lines shown in table (c). Statistical analysis: MTS assay, EC50P values: Calu-3 to Caco-2 < 0.05, Caco-2 to A549 > 0.05 and Calu-3 to A459 > 0.05. LDH assay EC50P values; Calu-3 to Caco-2 < 0.05, Caco-2 to A549 > 0.05 and Calu-3 to A459 < 0.05 conducted using t-test. Overall statistical difference between each test conducted for MTS and LDH P value < 0.05, conducted using one-way ANOVA.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig3: Effect of Solutol® HS15 on Calu-3, Caco-2 and A549 cells assessed by metabolic activity (a) and LDH assays (b). Solutol® HS15 was used at concentrations above and below the Critical Micelle Concentration (Fig. 2). Data expressed as relative metabolic activity and presented as the mean ± SD with N = 3 and n = 4. Data to summarize Solutol® HS15 concentrations that cause 50% reduction in cell viability (EC50, mM) and LDH leakage in tested cell lines shown in table (c). Statistical analysis: MTS assay, EC50P values: Calu-3 to Caco-2 < 0.05, Caco-2 to A549 > 0.05 and Calu-3 to A459 > 0.05. LDH assay EC50P values; Calu-3 to Caco-2 < 0.05, Caco-2 to A549 > 0.05 and Calu-3 to A459 < 0.05 conducted using t-test. Overall statistical difference between each test conducted for MTS and LDH P value < 0.05, conducted using one-way ANOVA.
Mentions: Figure 3a summarises data from the MTS assay, employed to assess the effect of Solutol® HS15 on the viability of lung derived Calu-3 and A549, as well as intestinal Caco-2 cells. A general decline in cell metabolic activity is seen for all three tested cell lines as the concentration of Solutol® HS15 solution is increased. A concentration-dependent effect is also seen for LDH release from the cells (Fig. 3b).Fig. 3

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