Nanoparticles are thought to present a unique hazard to human health. Furthermore, the increasing use of nanomaterials in consumer products has not been accompanied by relevant risk assessments. It is conceivable that skin flexion may assist the translocation of nanoparticles across the stratum corneum. However, current in vitro methodology to study dermal absorption involves the exclusive use of immobile skin within diffusion cells. Therefore, a novel skin-flexing diffusion cell system (“CutaFlex™”) was developed to incorporate reproducible skin flexing (2 flexes min-1; 6 mm maximum amplitude). The initial aims of this Thesis were to characterise the CutaFlex™ system to eliminate the possibility of flexion-induced (experimental) skin damage, demonstrate equivalence with historical permeability data to model compounds and assess the effect of skin flexing on barrier disruption. Subsequent work aimed to investigate the hypothesis that nanoparticles require dermal flexion to penetrate intact skin. In supporting these aims, this Thesis also performed work to assess the correlation between direct measurements of skin barrier function (using tritiated water) and transepidermal water loss (TEWL), the effect of flexing on the performance of topical skin protectants (barrier creams) and to further validate in vitro diffusion cell measurements against in vivo data acquired under identical conditions. The results demonstrated that skin flexing did not alter skin barrier function and that the CutaFlex™ system was in general agreement with historical measurements of skin permeability. Furthermore, controlled chemical or physical damage to the stratum corneum was not exacerbated by skin flexing. Skin flexion did not facilitate the dermal absorption of a range of nanoparticles (quantum dots). However, differences in the partitioning of nanoparticles into the stratum corneum were observed (independent of the degree of flexing), with greater amounts of negatively charged nanoparticles found in the superficial layers of the stratum corneum in comparison with positive or neutral nanoparticles. Flexing had a modest effect on the performance of a skin barrier cream which was limited to low dose applications; an effect tentatively ascribed to flexion-induced movement of cream to previously untreated areas. A poor correlation was found between 3H2O water permeability and TEWL flux. Most importantly, there was excellent agreement between in vitro skin permeability studies and in vivo studies (which used a surrogate measure of skin permeability). To summarise, the data in this Thesis has led to the development and characterisation of a novel diffusion cell (CutaFlex™), capable of simultaneously flexing skin whilst performing dermal absorption measurements comparable with the OECD-compliant models.

In this thesis, a series of in vitro diffusion studies were performed to evaluate the transmembrane permeation of picaridin and oxybenzone across human epidermis and poly(dimethylsiloxane) (PDMS) membrane. Transdermal permeation of picaridin and oxybenzone from four commercially available repellent and sunscreen products was also investigated by using different application concentrations and sequences. The results obtained were then compared to those of the repellent DEET and the sunscreen oxybenzone under identical experimental conditions. Permeation of picaridin and oxybenzone across human epidermis was suppressed when both compounds were used concurrently. Increasing concentration of the test compounds further reduced the permeation percentage of picaridin and oxybenzone. While permeation characteristics were correlative between human epidermis and artificial PDMS membrane, permeability of PDMS membrane was significantly larger than that of human epidermis. This finding was different from concurrent use of DEET and oxybenzone in which a synergistic permeation enhancement was observed between the two substances. Transdermal permeation of picaridin across human epidermis from various commercially available spray preparations was significantly lower than that of DEET from similar spray products, both alone and in combination with sunscreen oxybenzone. Concurrent application of the commercial products resulted in either no change or suppression of permeation of picaridin and oxybenzone. This finding was also different from concurrent application of DEET and oxybenzone using commercial preparations. In addition, permeation of picaridin and oxybenzone across human epidermis was dependent on application concentration, use sequence, and preparation type.It was concluded from this thesis that picaridin would be a better candidate for concurrent application with sunscreen preparations in terms of percutaneous permeation.

In this thesis, a series of in vitro diffusion studies were performed to evaluate the transmembrane permeation of picaridin and oxybenzone across human epidermis and poly(dimethylsiloxane) (PDMS) membrane. Transdermal permeation of picaridin and oxybenzone from four commercially available repellent and sunscreen products was also investigated by using different application concentrations and sequences. The results obtained were then compared to those of the repellent DEET and the sunscreen oxybenzone under identical experimental conditions. Permeation of picaridin and oxybenzone across human epidermis was suppressed when both compounds were used concurrently. Increasing concentration of the test compounds further reduced the permeation percentage of picaridin and oxybenzone. While permeation characteristics were correlative between human epidermis and artificial PDMS membrane, permeability of PDMS membrane was significantly larger than that of human epidermis. This finding was different from concurrent use of DEET and oxybenzone in which a synergistic permeation enhancement was observed between the two substances. Transdermal permeation of picaridin across human epidermis from various commercially available spray preparations was significantly lower than that of DEET from similar spray products, both alone and in combination with sunscreen oxybenzone. Concurrent application of the commercial products resulted in either no change or suppression of permeation of picaridin and oxybenzone. This finding was also different from concurrent application of DEET and oxybenzone using commercial preparations. In addition, permeation of picaridin and oxybenzone across human epidermis was dependent on application concentration, use sequence, and preparation type.It was concluded from this thesis that picaridin would be a better candidate for concurrent application with sunscreen preparations in terms of percutaneous permeation.

Hypothèse : Le nanobroyage d'une suspension de nystatine augmentera son efficacité antifongique in vitro et in vivo. Méthode : Une nanosupension de nystatine a été obtenue en utilisant le broyage humide. Elle a été caractérisée pour sa distribution de taille des particules et pour sa teneur en principe actif. L'activité in vitro a été évaluée contre les souches de C. albicans SC5314 et LAM-1 aux concentrations 12.5 μg/mL jusqu'à 5000 μg/mL. L'efficacité in vivo a été évaluée en utilisant un modèle murin de candidose oropharyngée. Résultats : La taille médiane des particules de la nanosuspension de nystatine a été réduite de 6577 nm à 137 nm. L'analyse CLHP a demontré une teneur de 98.7 ± 0.8%. L'activité in vitro de la nanosuspension était supérieure à la suspension aux concentrations 100 μg/mL à 5000 μg/mL. La charge fongique orale était inférieure dans le groupe traité par la nanosuspension comparativement aux autres groupes. La survie des souris était aussi supérieure. / Hypothesis : Nanomilling of a nystatin suspension will increase its antifungal efficacy in vitro and in vivo. Methods: A nystatin nanosuspension was obtained using wet bead milling. It was characterized for its particle size distribution and for its drug content. In vitro activity was evaluted against C. albicans strains SC5314 and LAM-1 at concentrations of 12.5 μg/mL up to 5000 μg/mL. The in vivo efficacy was evaluated using a murine model of oropharyngeal candidiasis. Results: Median particle size of the nystatin nanosuspension was reduced from 6577 nm to 137 nm. HPLC analysis demonstrated a content assay of 98.7 ± 0.8%. In vitro activity of the nanosuspension was superior to the suspension’s at concentrations ranging from 100 μg/mL to 5000 μg/mL. Oral fungal burdens were inferor in the nanosuspension group compared to the suspension and saline groups. Mice survival was also superior in the nanosuspension group.

Hypothèse : Le nanobroyage d'une suspension de nystatine augmentera son efficacité antifongique in vitro et in vivo. Méthode : Une nanosupension de nystatine a été obtenue en utilisant le broyage humide. Elle a été caractérisée pour sa distribution de taille des particules et pour sa teneur en principe actif. L'activité in vitro a été évaluée contre les souches de C. albicans SC5314 et LAM-1 aux concentrations 12.5 μg/mL jusqu'à 5000 μg/mL. L'efficacité in vivo a été évaluée en utilisant un modèle murin de candidose oropharyngée. Résultats : La taille médiane des particules de la nanosuspension de nystatine a été réduite de 6577 nm à 137 nm. L'analyse CLHP a demontré une teneur de 98.7 ± 0.8%. L'activité in vitro de la nanosuspension était supérieure à la suspension aux concentrations 100 μg/mL à 5000 μg/mL. La charge fongique orale était inférieure dans le groupe traité par la nanosuspension comparativement aux autres groupes. La survie des souris était aussi supérieure. / Hypothesis : Nanomilling of a nystatin suspension will increase its antifungal efficacy in vitro and in vivo. Methods: A nystatin nanosuspension was obtained using wet bead milling. It was characterized for its particle size distribution and for its drug content. In vitro activity was evaluted against C. albicans strains SC5314 and LAM-1 at concentrations of 12.5 μg/mL up to 5000 μg/mL. The in vivo efficacy was evaluated using a murine model of oropharyngeal candidiasis. Results: Median particle size of the nystatin nanosuspension was reduced from 6577 nm to 137 nm. HPLC analysis demonstrated a content assay of 98.7 ± 0.8%. In vitro activity of the nanosuspension was superior to the suspension’s at concentrations ranging from 100 μg/mL to 5000 μg/mL. Oral fungal burdens were inferor in the nanosuspension group compared to the suspension and saline groups. Mice survival was also superior in the nanosuspension group.