The Effect of Medications Used in Mesotherapy on Hair Organ Culture and Culture of Dermal Papilla Cells / 대한피부과학회지

BACKGROUND: Hair loss including androgenetic alopecia and chronic telogen effluvium is recognized increasingly as a physically and psychologically harmful medical condition. Mesotherapy is considered as a new therapeutic modality for hair loss. OBJECTIVE: We studied to determine the effect of medications used in mesotherapy on hair organ culture and culture of dermal papilla cells. METHODS: First, occipital hair follicles were collected from patients with androgentic alopecia and separated into single hair follicles. The single hair follicles were cultured in William E media mixed with mesotherapy medications such as lidocaine, placental extract, Pondil(R), CRP-1000(R), and mixture of all these medications at different concentrations (1, 10, 50 microliter). On the 8th day, the cultured single hairs were stained with H&E and the length of those was measured under a microscope to compare with control group. Immunofluorescent study was performed to check expression of Ki-67, Bcl-2 and Bax on the hairs. Second, dermal papilla cells were isolated from occipital anagen hairs of patients with androgenetic alopecia and cultured in Dulbeco's modified Eagle's medium (DMEM). The mesotherapy medicines were added to the medium with one and two thousand dermal papilla cells, respectively. At the 3rd day, survival of the cells was evaluated with ELISA method comparing with control group. RESULTS: There were no statistical differences of the length of the hairs and the survival of the dermal papilla cells between experimental and control groups. With Bcl-2, we couldn't see any differences between experimental and control groups. With Ki-67, experimental groups showed less expression than control group. On the contrary, experimental groups showed more expression than control group in case of Bax. CONCLUSION: We can conclude from the results that the four medications used in mesotherapy are not effective for growth of cultured hair follicles and survival of cultured dermal papilla cells. However, more study would be needed for the establishment of objective and scientific evidences supporting mesotherapy and we should be in search for new medications for mesotherapy.

Effects of Substance P on the Hair Growth in Human Hair Follicle Organ Culture / 대한피부과학회지

BACKGROUND: In vitro, some neuropeptides, including substance P(SP), act as a growth factor. The cyclic growth of the richly innervated hair follicle offers a model for probing such functions in a complex, developmentally regulated tissue interaction system under the physiologic condition. Dissecting the role of neuropeptides in this system may also reveal as yet obscure neural mechanisms of hair growth control. OBJECTIVE: The purpose of this study was to investigate the effect of SP on human hair growth using a recently described model in which isolated hair follicles are grown in vitro. METHODS: After the healthy human hair follicles without any visible damage were collected, they were cultured in DMEM with several combination of supplements including insulin, hydrocortisone, sodium selenite, human transferrin, fetal bovine serum at 37 degrees C in an atmosphere of 5% CO2 /95% air incubator, and SP was added to the media. The culture media were supplemented with final concentration of 10(-6),10(-7),10(-8) M SP dissolved in DMEM. The results were evaluated by measuring linear hair fiber growth and hair follicle morphology on light microscopy and electron microscopy and by measuring radioisotope uptake of [methyl-3H] thymidine and [U-14C] leucine of hair follicle. RESULTS: The following results were obtained from this study. 1. SP did not have an statistically significant effect on the rate of linear hair growth in cultured hair follicles. However, it prolonged the anagen stage of hair cycle. 2. We could not find morphological differences of hair follicles cultured in SP groups compared with those cultured in control group. 3. DNA and protein synthesis in hair follicles increased steadily for 5 days of culture. CONCLUSION: From the results, we can conclude that SP has growth-stimulatory effect and especially prolongs the duration of anagen phase without affecting the rate of linear hair growth.

Growth Factors Influencing the Morphology and Growth Rate of Hair Follicles in a Human Hair Organ Culture / 대한피부과학회지

BACKGROUND: In order to study hair biology, a hair organ culture system is necessary. However satisfactory hair culture systems have not been established. OBJECTIVE: The purpose of this study was to examine the effectiveness of growth factors and to establish a hair organ culture system for studying hair biology and to evaluate the effectiveness of growth factors. METHOD: After the healthy human anagen hair follicles were collected without any visible damage, they were cultured in William E medium with several combinations of growth factors including insulin, hydrocortisone, sodium selenite, human transfemn, fetal calf serum and epidermal growth factor at 37C in an atmosphere of 5% CO2/air incubation. The culture medium was changed every 3 days. The results were evaluated by measuring hair growth and hair follicle morphology. RESULTS: The results of this study are summarized as follows; 1) In the medium composed of insulin, hydrocortisone,sodium selenite and human transferrin, the human hair follicles continued to grow at an in vivo rate of 0.3mm in a day over 10 days without change of gross and microscopic morphology. 2) In the medium containing insulin and/or hydrocortisone the growing rate of the human hair follicles was similar to that in vivo, but the follicles revealed premature entry into catagen at 2-6 days in the culture macroscopically and microscopically. 3) Adding fetal calf serum to the above medium made the hair follicles retain the freshly isolated hair follicles morphology for 10 days in culture, even though they grew somewhat slower than the in vivo rate from 6 days in culture. 4) The effectiveness of EGF mimics the in vivo depilation of EGF in sheep. CONCLUSION: To supplement insulin, hydrocortisone, sodium selenite, transferrin as growth factors, William E medium was necessary for maintenance of an in vivo growth rate and the morphology the anagen hair follicles. This culture system is not enough, but it might be useful for investigation of the physiology, biology of hair follicles as well as pharmacology and toxicology in hair.