الفهرس | Only 14 pages are availabe for public view |
Abstract Summary Nano-carriers are promising drug carrier systems being developed to deliver the drugs to the target organ. These systems have proven a great success in the treatment of several diseases. Many drug carrier systems have been prepared and successfully utilized in the treatment of skin fungal infections, among which are liposomes, niosomes, transferosomes, ethosomes, solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs). Lipid nanocapsules (LNCs) are lipid nanoparticles with a structure that is a hybrid between polymeric nanoparticles and liposomes. Till current date, only one paper was published on the use of LNCs in treatment of skin fungal infections. Luliconazole is a novel broad spectrum antifungal drug used topically for fungal infections such as Tinea and Onychomycosis. Therefore, the aim of this work was to study the possibilities for using essential oils having antifungal activity (Eucalyptus oil and orange oil) for LNCs preparation instead of using medium chain triglyceride (Miglyol®812) and incorporation of luliconazole in those LNCs in an attempt to modulate its therapeutic efficacy in diseases such as tinea pedis. Summary 192 The work in this thesis was divided into three chapters Chapter I: Preparation and characterization of Lipid nanocapsules using essential oils with antifungal activity The work in this chapter dealt with the formulation, and characterization of essential antifungal oil LNCs. LNCs were prepared by phase inversion method, and characterized for their morphology using transmission electron microscopy, particle size, zeta potential and in vitro antimycotic activity. Stability of eucalyptus oil LNCs and orange oil LNCs stored at 4˚C was tested through monitoring the changes in particle size and zeta potential for six months. The work in this chapter included the following: 1- Preparation of LNCs by phase inversion temperature method. LNCs were prepared using different natural oils with reported antifungal activity (eucalyptus oil and orange oil) instead of using the most commonly used medium chain triglyceride (Miglyol®812). 2- A ternary diagram was used to plot all possible mixtures, composed of distilled water, hydrophilic surfactant (Kolliphor®HS15) and the oil (eucalyptus oil or orange oil) 3- characterization of the prepared LNCs was performed through the following studies: Summary 193 a) Particle size, PDI and zeta potential analysis of freshly prepared LNCs formulations were carried out using zetasizer. b) Samples of LNCs were examined for morphology by transmission electron microscopy. c) Physical stability study on all LNCs formulations was conducted over a storage period of six months at 4° C by the assessment of the effect of storage on the particle size, charge and PDI of the nanoparticles. d) Viscosity determination of selected LNCs (O36 and E35) formulation was carried out using Brookfield rheometer (DV3T). e)The electron micrographs of formulae O36 and E35 showed spherical shaped particles with clear core and shell morphology f) The in-vitro antimycotic activity of selected LNCs formulations (O36), (E35) was evaluated and compared to a standard LNCs formula prepared using Miglyol®812 (M36). The results of this work revealed the following: 1- Successfully prepared LNCs using essential antifungal oils (eucalyptus oil and orange oil). 2- Eucalyptus oil LNCs particle size ranged from 60-402 nm. Summary 194 3- Orange oil LNCs particle size ranged from 26-145 nm. 4- All prepared LNCs were charged, with charges ranging from (-12 to -34 mV). 5- Eucalyptus oil and orange oil LNCs displayed good storage properties as manifested by slight changes in particle size, polydispersity index and zeta potential values. 6- Transmission electron microscopy of LNCs displayed spherical shaped particles with clear core and shell morphology. 7- orange oil based LNCs (O36) was found to follow the typical Newtonian behavior where the viscosity was constant and there was a linear relationship between the shear stress and shear rate 8- Eucalyptus based LNCs (E35) was found to follow a shear thinning or pseudo plastic behavior where a gradual decrease in viscosity occurred upon increasing the shear rate 9- The highest antifungal activity was observed in E35 containing eucalyptus oil (35%w/w) having inhibition zone (3.56 ± 0.12 cm), then O36 containing orange oil (40%w/w) having inhibition zone (2.6 ± 0.1 cm) Summary 195 compared to M36 containing medium chain triglyceride (Miglyol®812) which did not show any antifungal properties. Chapter II: Preparation, characterization, in-vitro and exvivo evaluation of luliconazole loaded lipid nanocapsules (LNCs) The work in this chapter was concerned with the loading luliconazole in the selected eucalyptus oil LNCs and orange oil LNCs. characterization of the prepared formulae was done through the following studies: particle size, zeta potential, invitro release, in-vitro antimycotic activity and skin deposition experiments. Stability study on the LNCs formulations stored at 4˚C was conducted as well. The work in this chapter included the following: 1- Loading luliconazole in the selected eucalyptus oil LNCs. 2- Loading luliconazole in the selected orange oil LNCs. 3- characterization of the prepared LNCs was done through the following studies: a) Particle size and zeta potential analysis of freshly prepared luliconazole LNCs was carried out using Zetasizer. Summary 196 b) Physical stability study on luliconazole based LNCs was conducted over a storage period of four months at 4° C by the assessment of the effect of storage on the particle size, charge and PDI of the nanoparticles. c) Viscosity of luliconazole loaded LNCs were carried out using Brookfield rheometer (DV3T). d) Samples of luliconazole loaded LNCs were examined for morphology by transmission electron microscopy. e) In-vitro release study was performed on the selected luliconazole loaded LNCs formulae. f) Ex-vivo deposition/permeation of the selected luliconazole loaded LNCs formulae was carried out using Franz diffusion apparatus and compared with the oil (eucalyptus oil and orange oil) solution of the drug. g) The in-vitro antimycotic activity of selected LNCs formulations (O36l), (E35l) was evaluated and compared to a standard LNCs formula prepared using Miglyol®812 (M36l). The results of this work revealed the following: 1- Luliconzole orange oil LNCs and luliconazole eucalyptus oil LNCs were successfully prepared using phase inversion temperature method. Summary 197 2- Luliconazole loaded LNCs particle size ranged from 40 - 174 nm. 3- All of the prepared luliconazole loaded LNCs were negatively charged, with zeta potential values ranging from (-14 to -38 mV). 4- Eucalyptus oil luliconazole LNCs and orange oil luliconazole LNCs displayed the best storage properties as manifested by the slight changes in particle size, polydispersity index and zeta potential values. 5- The formulations containing eucalyptus oil as oily phase revealed significant increase in viscosity values (above 20cp) compared to the formulations containing orange oil as oily phase (near to 1cp) (p<0.05). 6- Transmission electron microscopy of luliconazole loaded LNCs displayed spherical shaped particles with clear core and shell morphology. 7- The in-vitro drug release results from eucalyptus oil formulae showed that the highest % luliconazole released after 20 hours was from E26l (75.18±0.76) and the lowest % luliconazole released after 20 hours was from E24l (14.91±1.65). 8- The in-vitro drug release results from orange oil formulae showed that the highest % luliconazole Summary 198 released after 20 hours was from O36l (59.04±3.40) and the lowest % luliconazole released after 20 hours was from O23l (39.29±3.09). 9- Ex vivo skin deposition experiments demonstrated the high potential of the selected LNCs formulae (E35l and O25l) in accumulating the drug into the deeper epidermal and dermal layers of the skin, and hence they were selected for further clinical studies compared to a standard LNCs formula prepared using Miglyol®812 (M36l). 10- The highest antifungal activity was observed in E35l containing eucalyptus oil (35%w/w) having inhibition zone (4.16 ± 0.05 cm), followed by O25l containing orange oil (20%w/w) having inhibition zone (3.4 ± 0.1 cm). The lowest antifungal activity was for M25l having inhibition zone (2.43 ± 0.11 cm). Chapter III: In-vivo and clinical studies of selected luliconazole loaded lipid nanocapsules (LNCs) In this chapter, the therapeutic efficacy of luliconazole eucalyptus oil LNCs and luliconazole orange oil LNCs was done through the following studies: in-vivo animal model using candida albicans for induction of fungal infection and also the selected formulae were clinically tested on patients suffering Summary 199 from tinea pedis, by satisfaction and efficacy report filled by the patients under the doctor supervision. The work in this chapter included the following: 1- In-vivo study was carried on 32 male albino rats to evaluate the antifungal effect of selected luliconazole eucalyptus oil LNCs, luliconazole orange oil LNCs and Miglyol ®812 luliconazole LNCs for 7 days and compared to blank formulae. This study included: a) Fungal burden scoring: 10 blocks from the same animal were implanted onto one plate. Every block was inspected visually for candida albicans growth. The skin blocks yielding fungal growth were regarded as culture positive. Culture negative skin blocks had no growth around any side of the skin block. b) The histopathological study: through staining skin tissue samples with hematoxylin and eosin and Periodic Acid-Schiff. 2- The clinical study was conducted on fourteen patients suffering from tinea pedis fungal lesion. Dermatological examination was performed to detect the type of fungal infections. Fungal scraping using 10% potassium hydroxide (KOH) solution offers an inexpensive, rapid, and useful technique for the daily practice of clinicians and mycologists managing patients with clinically suspected Summary 200 tinea pedis. The patients were photographed before treatment with follow up photographing every week up to 2 weeks or complete improvement. The results of this work revealed the following: 1- Collectively the results of the fungal burden scoring and histopathological study revealed that candida caused skin damage in epidermis and dermis with diffuse inflammatory edema and mononuclear inflammatory cells infiltration. On the topical administration of E35l as antifungal treatment containing luliconazole (antifungal drug) and eucalyptus oil (essential antifungal oil), the results showed that E35l treated candidiasis and its induced inflammation more than E35, O25l, O25, M25l and M25 2- As could be delineated from the results, patients receiving medicated eucalyptus oil LNCs (E35l) displayed significantly better clinical therapeutic outcome compared to plain eucalyptus oil LNCs (E35), medicated orange oil LNCs (O25l) and medicated miglyol® 812 LNCs (M25l). This could be attributed to the synergistic effect of essential antifungal oil (eucalyptus oil) and luliconazole along with the higher skin deposition (no flux rate) potential of the medicated eucalyptus oil LNCs (E35l). |