BACKGROUND: Stratum corneum lipids serve as a water retainer as well as permeability barrier by forming a multi-lamellae structure in the stratum corneum. The major constituent of these lipids, ceramides, have been shown to be predominantly associated with both functions. OBJECTIVE: Exposure of human epidermis to ultraviolet(UV) irradiation leads to changes in the physiologic and biochemical features of the skin. In order to investigate the effect of UVB irradiation on the skin barrier function in hairless mice, transepidermal water loss (TEWL) and lipid composition of stratum corneum were evaluated in hairless mice. METHODS: Hairless mice were irradiated 3 times weekly for 3 weeks with suberythemal dose (0.6MED, Group I) and minimal erythemal dose(MED), Group II) of UB. The mice of Group III received high dose of UVB(3MED) on the back in a single exposure. The control was Group IV. TEWL measured by evaporimeter and lipid composition of stratum corneum appraised by high performance thin layer chromatography(HPTLC) were evaluated weekly for 3 weeks. RESULTS: 1. Each time it was measured, the values of TEWL in group I were lower than group IV, but the difference was not significant. The peak value of TEWL in group II was 8.2+/-1.56 g/cm/h on the 7th day. The increase in TEWL was markedly significant at this point(P<0.001). Although the values of TEWL on the 14th and 21th day in group E increased compared with those of the control group, the significance of the values decreased (P<0.05). 2. The peak value of TEWL in group III was 9.88+/-1.13 g/cm/h on the 2nd day, showed a markedly significant increase compared with that of the control group(P<0.001). The values of TEWL decreased to the level of the control group on the 14th day. 3. The lipid(cholesterol sulfate, ceramide and neutral lipid) and total lipid mass in group 1 were insignificantly larger than that of the group IV measured each time. On the 7th and 14th day, the amount of each three lipid and total lipid mass significantly increased (P<0.05). On the 21th day, the amount of ceramide and neutral lipid showed a significant increase(P<0.05), furthermore the total lipid mass increased pronouncedly(P<0.01) in group II. 4. The amount of the 3 kinds of lipid and total lipid mass in group III significantly increased compared with those of the control group on the 2nd day(P<0.05). After the 7th day, no significant difference of the lipid mass except neutral lipid compared with that of the control group was seen. Comparing the 2nd and 14th day, there was a significant decrease in the amount of ceramide and total lipid mass(P<0.05) CONCLUSION: These results results suggest that considerable amount of UVB irradiation given in single or repeated exposure causes the disruption of skin barrier function, but a compensatory increase of skin lipid, especially ceramide, protests it from further damage and also improves skin barrier function.
Animals ; Ceramides ; Epidermis ; Humans ; Mice ; Mice, Hairless* ; Permeability ; Skin*

Chinese herbal medicine ultrafine powder has become a research hotspot for the addition of cosmetic raw materials. Dendrobium candidum is a traditional Chinese herbal medicine. Its extract and stem extract are already cosmetic raw materials and its water extract has the effect of preventing photoaging,but D. candidum ultrafine powder has not been accepted as a raw material for cosmetics,and no relevant research on photoaging prevention has been reported. In this experiment,the ultra-fine powder and fine powder of D. candidum to prevent photoaging were observed and compared,and its mechanism of action was discussed to provide a basis for the prevention of skin photoaging products. Seventy-two female ICR mice were randomly divided into normal group,model group,solvent group,titanium dioxide(Ti O2) group,isooctyl salicylate(2-ES) group,D. candidum ultrafine powder 1(DP1),ultrafine powder 2(DP2) and fine powder(DP3) groups. The photoaging model was established by ultraviolet irradiation for 8 weeks,and the model was intervened while modeling. The skin wrinkle grade,elastic parameters,skin microcirculation blood flow,skin structure and pathological changes(skin thickness,skin collagen fiber,elastic fiber) were observed,the skin transforming growth factor-β1(TGF-β1),Smad3 levels were determined,and the type Ⅰ and type Ⅲ collagen,matrix metalloproteinase-1(MMP-1),activated protein-1(AP-1),VEGF expression were detected. The results showed that ultrafine powder(DP1,DP2) significantly reduced the wrinkle level and skin blood flow of the model mice(P<0. 05,P<0. 01); DP1,DP2 and DP3 could significantly reduce the thickness of the epidermis(P<0. 001),improve collagen fiber,elastic fiber hyperplasia,and distortion and decrease VEGF expression,and DP1 is better than DP2 and DP3; each group could up-regulate type Ⅰ collagen,down-regulate type Ⅲ collagen,AP-1,MMP-1 protein expression,and DP1 improvement optimal. However,it has no obvious effect on TGF-β1 and Smad3. The ultrafine powder and fine powder of D. candidum have certain preventive effect on photoaging,and the effect of ultrafine powder is better than that of fine powder. Ultrafine powder may down-regulate the expression of type Ⅲ collagen,AP-1 and MMP-1 by up-regulating type Ⅰ collagen. Inhibition of collagen degradation plays a role in preventing photoaging.
Animals ; Dendrobium ; Female ; Mice ; Mice, Hairless ; Mice, Inbred ICR ; Skin ; Skin Aging ; Ultraviolet Rays

BACKGROUND: Moisturizers induce skin hydration and then increase flexibility and elasticity, making the skin soft and smooth, and protecting it against environmental stimuli. OBJECTIVE: The aim of this work was to study the role of intercorneocyte lipid layers and desmosomes in the mechanism of moisturization. METHODS: Transepidermal water loss (TEWL) and capacitance were measured and the morphologic changes of the intercorneocyte lipid layers and desmosomes with electron microscopy, using ruthenium tetroxide (RuO4) postfixation, following the application of glycerin, propylene glycol, and a mixture of glycerin and propylene glycol for a 2 hour period to the epidermis of hairless mice were measured. RESULTS: 1. The TEWL was significantly increased in all three groups; glycerin, propylene glycol, and mixture of glycerin and propylene glycol. The increase of TEWL after the application of glycerin was maintained from the second to the forth hour after application which was statistically significant, after the application of propylene glycol it was maintained for 5 hours, and after the application of a mixture of glycerin and propylene glycol, for 6 hours. 2. The capacitance also was increased in all three experimental groups, compared to the control group. However there was no statistical significance. 3. Light microscopic findings showed no specific changes in all three groups, compared to the control group. 4. Ultrastructural observation by electron microscope, using RuO4 postfixation, showed widening of the intercorneocyte lipid layers in all three groups. In contrast to glycerin in which the results showed detachment of the desmosomes without changes in the intercorneocyte lipid layers, propylene glycol showed interruption and undulation of the intercorneocyte lipid layers and expansion of the lacunae spaces. A mixture of glycerin and propylene glycol showed interruption and undulation of the intercorneocyte lipid layers, detachment of the desmosomes, and, partial, formation of lacunae. CONCLUSION: These results suggest that the moisturizing effects of glycerin result from an increased detachment of the desmosomes and widening of the intercorneocyte lipid layers and then an increase in the water holding capacity of the stratum corneum. Propylene glycol, a chemical penetration enhancer, induce widening, interruption, and undulation of the lipid layers and expansion of the lacunae space. In the mixture of glycerin and propylene glycol, propylene glycol potentiate and continue the moisturizing effects of the glycerin.
Animals ; Desmosomes* ; Elasticity ; Epidermis ; Glycerol* ; Mice ; Mice, Hairless ; Microscopy, Electron ; Pliability ; Propylene Glycol* ; Ruthenium ; Skin

BACKGROUND: Topical irritants disrupt the cutaneous permeability barrier through the removal of stratum comeum lipids. This perturbation of barrier integrity stimulates a variety of homeostatic repair responses that ultimately result in the normalization of bamer function. Object To measure the effect of desoxymethasone ointment, vaseline and hydrobase on the barrier recovery of benzalkonium chloride (BKC) imtated skin. MATERIALS AND METHODS: The left flank skin of 2-3 monthold hairless mice was treated with BKC and then desoxymethasone ointment, vaseline and hydrobase were applied. Transepidermal water loss (TEWL) was checked after 0, 3, 6, 9, 12, 15, 18, 21 and 24 hours. Electron microscopic examination was performed after 3 and 24 hours after desoxymethasone, vaseline and hydrobase had been applied. RESULTS: The recovery of TEWL was most prominantly observed in the desoxymethasone ointment treated group followed by vaseline and hydrobase. Electron microscopic examination using ruthenium tetroxide fixation revealed that secretion and numbers of lamellar bodies and complete formatice of lipid bilayers were most prominent at desoxymethasone ointment and vaseline treated group. CONCLUSION: Desoxymethasone ointment, vaseline and hydrobase can be good agents in improving bamer recovery after exposure to irritant material.
Animals ; Benzalkonium Compounds* ; Desoximetasone* ; Irritants ; Lipid Bilayers ; Mice ; Mice, Hairless* ; Permeability ; Petrolatum* ; Ruthenium ; Skin*

Ultraviolet (UV) irradiation is an environmental factor that causes skin aging, and is also a major factor leading to cumulative alterations of skin structure, function and appearance. To investigate the effects of Selenium (Sel) on UV-induced skin aging, hairless mice were treated for 4 weeks with UV irradiation and topical application of Sel. Then, the effects of Sel were measured in the skin of these mice via histological analysis and Western blotting. According to the results of wrinkle formation analysis, the topical application of Sel induced a reduction in wrinkling formation in the damaged skin of the UV-irradiated mice. Additionally, our histological analysis demonstrated that the skin thickness in the Sel-treated group was less than in the UV-irradiated group. Furthermore, in an effort to investigate the mechanisms underlying the effects of Sel, the expression levels of matrix-metalloproteinase (MMP) and MAPK protein were assessed in both groups. The application of Sel induced a reduction in MMP-1 expression levels to the levels observed in the non-irradiated group. However, the expression level of MMP-9 was increased slightly in the Sel application group as compared with the vehicle application group. Additionally, the levels of ERK phosphorylation were increased by the application of Sel, but the levels of JNK and p38 were not altered by Sel treatment. These results suggest the possibility that Sel should be considered as a skin aging-protective and therapeutic drug candidate, which functions via the regulation of MMP expression levels.
Animals ; Blotting, Western ; Mice ; Mice, Hairless ; Phosphorylation ; Selenium ; Skin ; Skin Aging

BACKGROUND: Acetone disrupts the cutaneous permeability barrier through the removal of stratum corneum lipids. This pertubation of barrier integrity stimulates a variety of homeostatic repair that ultimately results in the normalization of barrier function. OBJECT: To measure the effect of steroid on the barrier recovery of acetone applied skin. MATERIAL AND METHODS: The flank skin of 8~10 week old hairless mice was treated with acetone and then topical and systemic steroids were applied. Transepidermal water loss(TEWL) was checked after 0, 3, 6, 12 and 24 hours. Electron and light microscopic examination and ion capture cytochemistry were performed after 3, 6, 12 and 24 hours after systemic and topical steroids had been applied. RESULTS: The results were as follows ; 1) During 3~6 hours after experiment, the recovery rate of TEWL was most prominent in the group of acetone applied animal than other groups. 2) After 12 hours after acetone applied, formation of new stratum corneum was found in the groups of acetone applied or acetone applied skin with topical steroid application. But loss of stratum corneum was observed in the groups of high or low dose steroid injection. 3) Ruthenium tetroxide staining of acetone alone or topical steroid treated specimens after 12 hours experiment revealed that the lipid bilayer was partly impaired and fragmented. Intercellular spaces were widening and the lipid bilayer disappeared or was damaged in the groups of high or low dose steroid injection. 4) Six hours after acetone application, pattern of calcium distribution had been partially reestabilished in the group of acetone alone or topical steroid treated animals. But calcium content was still sparse and decreased from the stratum granulosum to basale in the groups of high or low dose steroid injection. CONCLUSION: In summary the present study demonstrates that steroid treatment acutely delays recovery rate of TEWL, inhibits normalization of calcium gradient or epidermal lipid synthesis that leads to abnormalities in permeability barrier homeostasis.
Acetone ; Animals ; Calcium ; Extracellular Space ; Histocytochemistry ; Homeostasis ; Lipid Bilayers ; Mice ; Mice, Hairless* ; Permeability ; Ruthenium ; Skin ; Steroids

S-methyl-(L)-methionine (SMM), also known as vitamin U, is commercially available as skin care cosmetic products for its wound healing and photoprotective effects. However, the low skin permeation expected of SMM due to its hydrophilic nature with a log P value of −3.3, has not been thoroughly addressed. The purpose of this study thus was to evaluate the effect of skin permeation enhancers on the skin permeation/deposition of SMM. Among the enhancers tested for the in vitro skin permeation and deposition of SMM, oleic acid showed the most significant enhancing effect. Moreover, the combination of oleic acid and ethanol further enhanced in vitro permeation and deposition of SMM through hairless mouse skin. Furthermore, the combination of oleic acid and ethanol significantly increased the in vivo deposition of SMM in the epidermis/dermis for 12 hr, which was high enough to exert a therapeutic effect. Therefore, based on the in vitro and in vivo studies, the combination of oleic acid and ethanol was shown to be effective in improving the topical skin delivery of SMM, which may be applied in the cosmetic production process for SMM.
Animals ; Ethanol ; In Vitro Techniques* ; Mice ; Mice, Hairless ; Oleic Acid ; Skin Care ; Skin* ; Vitamin U ; Wound Healing

PURPOSE: The objective of this study is to evaluate the heparin effect for a viability of random-pattern dorsal flap in hairless mouse. METHODS: A caudally-based random dorsal flap, measuring 1.5 x 5 m, was designed and heparin was applied topically after microneedling. Twenty five male hairless mice were randomly divided into five groups: control (Group 1, n=5); received only microneedling (Group 2, n=5), only heparin (Group 3, n=5), microneedling with saline (Group 4, n=5), and microneedling with heparin group (group 5, n=5) to the flap during 7 days. The number of the capillaries were compared between the experimental groups and control group with respect to neovascularization after heparin application using imaging analysis program under hematoxylin-eosin stain. The capillary blood flow was measured by laser Doppler flowmetry. After seven days each animal was evaluatedfor the percentage area of the flap survival. Mann- Whitnety U test and Kruskal-Wallis statistical analysis of survival relationships was performed. RESULTS: It can be observed increased number of the blood vessels in the experimental groups however it was not statistically significant. Blood flow of the haparin with microneedling group maintained higher than other experimental groups. Treated microneedling and heparin mice were significantly better flap viability than in controls (flap survival 67% and 54.4 mm2 respectively; p< 0.01). Positive correlation was shown between flap survival rate and laser Doppler flux value only at first day after surgery. CONCLUSION: Heparin has a beneficial effect on capillary flow and improve peripheral circulatory disturbances in random pattern flaps.
Animals ; Blood Vessels ; Capillaries ; Heparin ; Humans ; Laser-Doppler Flowmetry ; Male ; Mice ; Mice, Hairless ; Skin ; Survival Rate

BACKGROUND: The cornified cell envelope(CE) which is formed during the terminal differentiation of keratinocytes, is a specialized structure which forms a structurally and functionally complete permeability barrier. OBJECTIVE: The purpose of our study is to investigate the effects of changes in the calcium ions on keratinocyte differentiation, especially in the expression of CE protein. METHODS:The permeability barrier of hairless mice was disrupted by tape-stripping and then exposed to the air or occluded with a water-vapor impermeable membrane, and iontophoresis was done without permeability barrier perturbation. Skin specimens were prepared for ion capture cytochemistry and immunohistochemistry with anti-K5, anti-K10, anti-K6, anti-involucrin and anti-loricrin. RESULTS: The calcium gradient which disappeared after tape-stripping was restored at 36 h after tape-stripping with air exposure and at 60 h after tape-stripping with occlusion. The change in calcium ions produced by both positive and negative iontophoresis showed recovery at 6 h. Expression of basal K5 showed a slight decrease and expression of suprabasal K10 showed an increase at 12 h with air exposure after tape-stripping, tape stripping with occlusion, and iontophoresis. Expression of K6 appeared at 12 h after tape-stripping and then in the whole epidermis at 36 h with air exposure after tape-stripping and tape stripping with occlusion and focally appeared in the stratum granulosum and stratum spinosum after iontophoresis. Expression of involucrin was increased at 12 h with air exposure after tape-stripping and iontophoresis and was extended to the lower spinous layers in tape-stripping with occlusion. Expression of loricrin in air exposure after tape-stripping, tape-stripping with occlusion and iontophoresis was similar to that of normal skin. CONCLUSION: The changes in calcium ions without permeability barrier perturbation are related to the expression of CE protein. It is thought that calcium ions in the epidermis have an important role in the terminal differentiation of keratinocytes.
Animals ; Calcium* ; Epidermis ; Histocytochemistry ; Immunohistochemistry ; Ions ; Iontophoresis ; Keratinocytes* ; Membranes ; Mice ; Mice, Hairless ; Permeability ; Skin

BACKGROUND: The cornified cell envelope(CE) which is formed during the terminal differentiation of keratinocytes, is a specialized structure which forms a structurally and functionally complete permeability barrier. OBJECTIVE: The purpose of our study is to investigate the effects of changes in the calcium ions on keratinocyte differentiation, especially in the expression of CE protein. METHODS:The permeability barrier of hairless mice was disrupted by tape-stripping and then exposed to the air or occluded with a water-vapor impermeable membrane, and iontophoresis was done without permeability barrier perturbation. Skin specimens were prepared for ion capture cytochemistry and immunohistochemistry with anti-K5, anti-K10, anti-K6, anti-involucrin and anti-loricrin. RESULTS: The calcium gradient which disappeared after tape-stripping was restored at 36 h after tape-stripping with air exposure and at 60 h after tape-stripping with occlusion. The change in calcium ions produced by both positive and negative iontophoresis showed recovery at 6 h. Expression of basal K5 showed a slight decrease and expression of suprabasal K10 showed an increase at 12 h with air exposure after tape-stripping, tape stripping with occlusion, and iontophoresis. Expression of K6 appeared at 12 h after tape-stripping and then in the whole epidermis at 36 h with air exposure after tape-stripping and tape stripping with occlusion and focally appeared in the stratum granulosum and stratum spinosum after iontophoresis. Expression of involucrin was increased at 12 h with air exposure after tape-stripping and iontophoresis and was extended to the lower spinous layers in tape-stripping with occlusion. Expression of loricrin in air exposure after tape-stripping, tape-stripping with occlusion and iontophoresis was similar to that of normal skin. CONCLUSION: The changes in calcium ions without permeability barrier perturbation are related to the expression of CE protein. It is thought that calcium ions in the epidermis have an important role in the terminal differentiation of keratinocytes.
Animals ; Calcium* ; Epidermis ; Histocytochemistry ; Immunohistochemistry ; Ions ; Iontophoresis ; Keratinocytes* ; Membranes ; Mice ; Mice, Hairless ; Permeability ; Skin