Study Of Compound Minoxidil On The Mouse Hair Growth

AIM To investigate the best concentration of the combination of ginseng extract and minoxidil by the culture of mouse follicle in vitro. We employ the mouse skin in vitro and use transdermal ffusion assembly to inspect the best concentration of azone as the penetration enhancer of the combination of ginseng extract and minoxidil. We study the best combination to promote the hair growth by whole animal experiments. We study the security of the combination of ginseng extract and minoxidil with acute toxicity trial, stimulation trial and hypersensitivity trial on animal skins which provide the foundation for clinical application. METHODS1 The routine culture of hair follicle: The whole hair follicles of C57BL/6J mice in anagen are picked up under the anatomic microscope. Then put them in Willimas E culture medium containing four different concentrations of minoxidil and six different concentrations of ginseng extrac and nine different concentrations of combination minoxidil with ginseng extract. Incubate them 7 days in hatch box which is 37Cand contains 5% CO2. The length of hair shaft is measured once every 24 hours.2 The 3H-TdR culture of hair follicle: The methods are the same as the above. After culturing 24 hours, 3H-TdR 50uL (0.5uCi) is added to every aperture. After another twenty hours, the follicles are picked up from every aperture and washed three times with PBS (phosphate-buffered saline)(about thirty minutes). The hair follicles are fixed twice (about twenty minutes) with 10% trichloroacetic acid under ice bath and fixed once with ice-water (about ten minutes). They are digested to lysis completely with 0.3 mol/L NaOH in water bath with 37C(about thirty hours). The supernatant is collected to 999 type fibrous membrane with ZT -III type cell collecting device and is dried by ultra-red rays. The flicker liquid is added and LS-6500 counting equipment is used to measure it.3 The transdermal absorption study of compound minoxidil: By using transdermal ffusion assembly, the back skin of mouse in vitro is fixed to receiving pool with the internal skin directed to receiving pool. There is saline in the receiving pool and combinatinal agent with different concentration of azone in the releasing pool. Put the transdermal ffusion assembly in the water bath shake with homoiothermy. The temperature is controlled between 36~37C. We sample 0.5mL each time at the second fourth eighth 10th 12th and 24th hour and complement the same amount of saline simultaneously. The receiving fluid is centrifugated 10 minutes with 7000g/min and 20uL supernatant is measured by HPLC.4 The study of pharmacodynamics: The hair of mouse back is taken off with 8% sodium sulfide 24 hours before administration. The compound agents with different concentrations are applied to the skin of mouse twice a day for 2 weeks. Each time, 0.1 mL agent is spreaded to skin of back steady. The results are observed 2 weeks later and the condition of hair growth is observed in hairless region.5 Reliability experiment:5.1 Acute toxicity trial of skin: Take off the hair of rabbit backs with 8% sodiumsulfide 24 hours before applying agents. Damaged skin is made through scarification with bullet. 2mL of mixture agent and 2mL of the control are applied to the backs of the rabbits respectively. The backs of the rabbits are washed with warm water 24 hours later. After taking off the agents, the topical and total body conditions are observed at the 1 st, 24th, 48th, 72th hour and on the seventh day.5.2 Cutaneous stimulation trial: The method of taking off hair is the same to the above. Utilizing the skins of both sides as control, we apply the 1 mL trial drug and 1 ml control drug on the backs of rabbits. 24 later, the backs of the rabbits with remaining drug are washed with warm water and the potential erythema and edema are observed. The drugs are used once per day and lasted for one week. After taking off the trial drugs at last time, the possible erythema and edema of topical skin are observed successively after the removal of the...