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-189- Hair Biology(1) 9/5 Room-D
CELL KINETICS IN HAIR CYCLE OF C3H MICE.
Norimitsu Saitoh, Yukinori Ohta, Minako Tatsuta, Kensei Katsuoka.
Department of Dermatology, Kitasato University School of Medicine, Sagamihara, Japan.
Recent reports suggested the follicular stem cell in bulge region is sufficient in hair growth. Regarding the hair cycle, however, little is known about the central molecular event inducing anagen phase. C3H mice, representing anagen phase induced by hair-plucking, could be an useful mice model for hair research. In this study, we evaluated the sequential change in the distribution of proliferating cell by using anti-PCNA (proliferating cell nuclear antigen) antibody immunohistochemically, and attempted to elucidate growth/differentiation-related gene expression in hair cycle by RT-PCR. PCNA was strongly expressed both in the mid-portion and matrix of hair follicule at 7 days after plucking, whereas at 10-14 days (mid-anagen) PCNA positive cells were located only in matrix. C-myc, proto-oncogene linked with epithelial cell growth/apoptosis, was overexpressed in pre-anagen phase. These suggested that cells in mid-portion of hair follicule proliferate transiently before plucking-induced anagen phase. Moreover, c-myc overexpression might be related to the entry of hair regrowth stage.
-190- Hair Biology(1) 9/5 Room-D
HAIR CYCLE OF MOUSE VIBRISSA FOLLICLES.
Chika Hanzawa Hamada, Akihiro Ishino, Masahiro Tajima.
Shiseido Reserch Center, Yokohama, Japan.
The follicle-organ culture is an important tool for the study of hair growth mechanism. Previously, we reported the effect of growth factors in organ culture of mouse vibrissa follicle(1). In this study, we characterized the synchronization of growth of vibrissa follicles and discussed the relationships between the diameter or the length of hair shaft and the growth ability on organ culture. The growth ability decreased in proportion to the length of shaft. The follicles with less than 25 ¦Ìm-diameter shaft (low-diameter follicles) elongated more than 2 mm by 3 days. In from 20 to 35 days-old-mice, almost follicles classified the low-diameter follicles. In from 35 to 50 days-old-mice, the low-diameter follicles were not observed. In from 50 to 70 days-old-mice, approximately half of follicles classified the low-diameter follicles. It was suggested that the hair cycles of mice vibrissa follicles synchronized in young, but not in aging. (1)Uzuka M. & Hanzawa C., Jap. J. Dermatol. :104,979-987(1994) (in Japanese)
-191- Hair Biology(1) 9/5 Room-D
INDUCTION OF APOPTOSIS ON MURINE HAIR FOLLICLE CELLS.
Noriaki Nakagawa, Kazuto Hamada.
Cosmetics Laboratory, Kanebo Ltd, Kanagawa, Japan.
We have previously reported that apoptosis occurred from the anagen phase to the catagen phase and, in the catagen phase, apoptotic cells increased, compared with those in the anagen phase. But the mechanism of induction of apoptosis is still unclear. In this study, to induce apoptosis on the murine hair follicle cells ( MHC ), we have examined how Ca concentration added to culture media influenced the attachment of the MHC to culture dish and moreover, whether apoptosis occurred on the MHC by disruption of attachment to the culture dish. The MHC were cultured on type I collagen coated dish with different Ca concentrations and the cells floating and attaching to the dish were measured. Then fragmentated DNA was also analyzed. Moreover, the MHC were cultured on agarose coated dish for complete suspension culture and fragmentated DNA was analyzed. As the results, the MHC on the culture dish were increased and fragmentated DNA was reduced according to higher Ca concentration. Moreover, fragmentated DNA from the MHC cultured on the agarose coated dish was increased, compared with that on type I collagen coated dish. All these results suggest induction of apoptosis on MHC by disruption of attachment of to culture dish is one method to elucidate the mechanism of induction of apoptosis during hair cycle.
-192- Hair Biology(1) 9/5 Room-D
ANALYSIS OF APOPTIC CELL DEATH IN HUMAN HAIR FOLLICLE.
Tsutomu Soma, Masashi Ogo, Jun Suzuki, Tadahito Takahashi, Toshihiko Hibino, Yasuhisa Nakayama.
Life Science Laboratories, Shiseido Research Center, Yokohama, Japan.
It has long been suggested that apoptosis plays an important role for the transition from the growth phase (anagen) to the regression phase (catagen) in hair cycle. We analyzed the apoptic cell death in human hair follicles in vivo and in vitro. In anagen hair follicles, positive stainings for TUNEL reaction were observed at the keratogenous zone of hair bulb and the inner most layer of the outer root sheath (ORS). While, in early catagen hair follicles, the lower bulb cells around dermal papilla and the cells in outer layer of the ORS became strongly positive, suggesting that the apoptosis in this area is related to the process of the catagen phase. We also detected the mRNA expression of four ICE family proteases (ICE, ICErel II, CPP-32 and Mch3) in anagen hair follicles by in situ hybridization. Moreover, when human anagen hair follicles were isolated and cultured in the presence of TGF-b, similar catagen-like morphology including cell death pattern were induced as seen in early catagen. Our data showed that apoptosis in catagen hair occurs in a distinct pattern and multiple forms of ICE family proteases would be involved in the regulation of hair cycle. Furthermore, TGF-b may be related to the induction of the catagen phase.
-193- Hair Biology(1) 9/5 Room-D
TOPICAL LIPOSOME TARGETING OF MELANIN AND DYES TO THE HAIR FOLLICLE IN HUMAN STUDIES.
L. Li E. Baranov, R.M. Hoffman.
AntiCancer, Inc., San Diego, USA.
We have developed a method of hair-follicle-selective delivery based on topical application of phosphatidylcholine liposomes. Liposome-entrapped melanins, proteins, and genes have been selectively targeted to the hair follicle of mice by topical application. New results from initial human studies which have shown that topical application of a] liposomes labeled with Dilc (a lipophilic fluorescent dye), b] liposome-entrapped red-fluorescent dye rhodamine, and c] liposome-entrapped melanin conjugated with the fluorescent dye Texas red cadaverine can effectively target hair follicles below the epidermis, including the hair bulb. The pattern of dye distribution in hair follicles is time-dependent, dose-dependent, and hair-cycle dependent. In contrast, topical application of free rhodamine and free fluorescent-dye-conjugated melanin do not result in significant accumulation of these substances to the hair follicle below the epidermis. Thus, the results have shown that liposomes are highly effective for delivery of small and large compounds to hair follicles by topical application in human studies. Fluorescent-dye-conjugated melanin is an easily traced marker to observe the efficiency of melanin delivery to the hair follicles by topical application in liposomes. Further studies will be done with the topical delivery of melanin as well as with the other beneficial compounds including genes, all entrapped in liposomes, for the purpose of hair coloring and regulating hair growth.