DEVELOPMENT OF NANOSTRUCTURED LIPID CARRIER CINCHONINE DELIVERY SYSTEM FOR ACCELERATING HAIR GROWTH

• Main Author: Hariyanti
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/75709
• Institution: Institut Teknologi Bandung

Background: Androgenetic alopecia (androgenic alopecia) is a disturbance of the hair growth and regeneration cycle due to the sensitivity of the hair follicle and dermal papilla to high levels of dihydrotestosterone. Cinchonine has activity to accelerate hair growth through telangiectasia mechanisms and activation of the wnt/?-catenin gene. In order to optimize its activity, cinchonine must be able to penetrate and reach the site of action of the hair follicle and dermal papilla. Cinchonine is practically insoluble in water, resulting in low penetration into the hair follicle and dermal papilla. The low cinchonine penetration is exacerbated by the presence of a hair follicle barrier. To overcome the characteristics of cinchonine and hair follicle barrier, we need a delivery system that can increase the penetration of cinchonine, namely Nanostructured Lipid Carriers (NLC). Objective: To develop an NLC delivery system to increase cinchonine penetration into hair follicles and dermal papillae. Methodology: The study began with the confirmation and validation of the HPLC method for determining cinchonine levels in the NLC delivery system development process, which included maximum wavelength, linearity (r2 ), sensitivity (LoD and LoQ), precision (intraday and inter days), accuracy, and selectivity. Formula development begins with lipid screening, optimizing the ratio of lipids, surfactants and cosurfactants. NLC was prepared using microemulsification and ultrasonification methods with an effective dose of 0.18% cinchonine. Characterization of optimum NLC-CN was carried out by determining particle size, polydispersity index (PDI), zeta potential using PSA, particle morphology using TEM, melting point analysis using DSC, entrapment efficiency and cinchonine release test using HPLC. The main parameter of the optimum NLC-cinchonine (NLC-CN) has a particle size in the range of 300 – 640 nm with the smallest PDI. After that, it was continued with optimization of preservatives for the manufacture of NLC-CN serum. NLC-CN serum were analyzed for particle size, PDI, viscosity, pH, and entrapment efficiency. After that, it was continued with stability tests at 3 storage conditions (40oC, room temperature and 4oC). In vitro analysis was carried out by determining localization of NLC with the nile red marker compound and male Swiss Webster mice using a Confocal Laser Scanning Microscope and diffusion using Franz Diffusion Cell with the skin of male Swiss Webster mice in active and inactive follicle conditions. In vivo analysis was carried out by determining skin and eye irritation tests using male New Zealand rabbits. Parameters observed in the dermal irritation test were erythema and edema indices, while parameters observed in the eye irritation test were the degree of irritation/corrosion of the conjunctiva, cornea, and iris. The activity test for accelerating hair growth was carried out using male Swiss Webster mice strain (androgenetic alopecia model was induced using testosterone, subcutaneously injected at a dose of 10 mg/kg BW/day for 21 days) with parameters observing baldness (excessive loss), hair texture become shorter and thinner. The data obtained were analyzed using statistics including diffusion data and the effect of NLC-CN serum on hair length. Results: Cinchonine has a maximum wavelength of 289 nm; with the validation parameters of the analytical method including linearity with a correlation coefficient (r2 ) of 0.9998; LoD of 1.2016 µg/mL; LoQ of 4.0054 µg/mL; high precision with SD and KV values of < 2%; high accuracy with percent recovery of 99 – 101% and high selectivity with RF of 4±0.3 minutes and TF of 1. Optimum NLC-CN formula with composition (in % w/v) cinchonine 0.18; stearic acid 1,8; oleic acid 0.2; tween 80 3.5; glycerin 2.5; aquadion up to 100. The optimum NLC-CN has characteristics including a particle size of 567.6 ± 15.4 nm; PDI of 0.343 ± 0.045; zeta potential of -36.70 mV; the morphology of the NLC-CN particles is spherical with a size of 500 nm; melting point of 61.50oC; entrapment efficiency of 94.85 ± 1.92%; with a release of 10.20 ± 0.49% at 180 minutes. Optimum NLC-CN serum was obtained by adding 0.2% DMDM hydantoin. The optimum NLC-CN serum has characteristics including a particle size of 575.6 ± 23.36 nm; PDI of 0.289 ± 0.011; viscosity of 1.22 ± 0.04 cP; and entrapment efficiency of 92.55 ± 6.19%. NLC-CN serum is stable under storage conditions in refrigerator (4 oC) for 12 months. NLC-CN serum was able to localize in hair follicles and was able to increase cinchonine penetration (p<0.05) from the diffusion data of serum NLC-CN opened hair follicles at 8 hours (17.64 ± 1.35 mg/cm2 ) higher than serum follicular NLC-CN was closed hair follicles (11.33 ± 0.29 mg/cm2 ) and follicular CN solution was opened hair follicles (4.81 ± 1.18 mg/cm2 ). In vivo analysis for skin and eye irritation tests showed that NLC-CN serum was non-irritant with a primary irritation index of 0.0. The hair growth accelerating activity test showed a significantly different increase in hair length (p<0.5) on day 21 in the NLC-CN serum test group of 0.69 ± 0.03 cm compared to the positive control of 0.34 ± 0 .01 cm (increased by 106.75%). Skin histology showed that at each point of organoleptic observation there was an increase in the size and number of hair follicles, and also dermal papillae. Conclusion: Cinchonine can be formulated in NLC delivery system. The system is able to accommodate an effective dose of cinchonine (0.18%), increase the stability, skin penetration, and hair growth accelerating activity with a good safety profile.