Abstract
Ketoconazole (KTZ) is a potential antifungal drug to control resistant and recurring opportunistic fungal infections. The study aimed to prepare KTZ loaded solid lipid nanoparticles (KTZ-SLNs) by high pressure homogenization method followed by optimization using Design Expert software and characterization for particle size, polydispersity index (PDI), zeta potential, morphology, in-vitro antifungal activity (susceptible and resistant strains), in-vitro cellular uptake (L929 and J774A.1 cell lines) and in-vitro hemolysis. Cellular uptake studies using dermal fibroblasts and Candida albicans were carried out with rhodamine B (RhB) and fluorescein isothiocyanate (FITC) labelled SLNs. Finally, acute skin irritation (EpiDerm (TM) and rabbit skin) and histological assessments were performed to confirm safety concern. Results showed that the optimized spherical KTZ-SLN4 exhibited mean size of 292 +/- 6.3 nm, optimal zeta potential (-24.39) and SPAN value of 2.0. In-vitro antifungal efficacy of KTZ-SLN4 exhibited 75-95% and 50-75% reduction in MIC (minimum inhibitory concentration) value as compared to the free drug suspension (KTZ-SUS) and marketed product (KTZ-MKT), respectively. In-vitro hemolysis confirmed the biocompatibility at explored concentration. Developed SLNs exhibited significant (p < 0.05) cellular internalization both by dermal fibroblasts (site of infection) and Candida albicans. Furthermore, rhodamine probed SLNs (RhB-SLN4) showed 12.6 fold higher penetration up to dermal region of skin as evidenced with confocal laser scanning microscopy (CLSM). Thus, KTZ-SLN4 demonstrated hemocompatibility, substantial penetration into rat skin, and improved efficacy against fungal strains. Conclusively, KTZ-SLNs can be a promising alternative to conventional formulations with significant clinical impact to control menace of fungal resistance and recurrence.