Number Of Dopa-positive Melanocytes Research Articles

Protective effects of adipose-derived stem cells against UVB-induced skin pigmentation

Background: Hyperpigmentation, mainly following UV-irradiation, can cause major cosmetic concerns. Human adipose tissue-derived stem cells (ASCs) have been reported to serve as whitening agents through a paracrine effect. However, there have been few reports on the direct effects of ASCs on skin pigmentation following UVB-irradiation.Methods: To evaluate the effect of ASCs on UVB-irradiated mouse skin, UVB-irradiation alone was applied to one side of the backs of mice (melanin-processing hairless mouse, HRM-2) as a control, and UVB-irradiation plus injection of ASCs was applied to the contralateral side. Skin pigmentation and histology were evaluated and the number of DOPA-positive melanocytes in the mouse skin was counted. The absolute value of ΔL* via a colorimeter was measured to evaluate the degree of skin pigmentation. The effects of ASCs on the melanogenic activities of mouse skin were examined by measuring the tyrosinase activity and the melanin contents in the epidermis of the mouse skin.Results: Skin pigmentation was suppressed in the ASC-injected side. Moreover, the change in skin thickness following UVB irradiation was reduced in the ASC-injected side. The number of DOPA-positive melanocytes in the ASC-injected side (139 ± 18 cells/mm2) was significantly lower than that in the control side (239 ± 48 cells/mm2). The tyrosinase activity (67.4 ± 9.8% of that of the control side) and melanin content (63.4 ± 5.7% of that of the control side) of the ASC-injected side were also significantly reduced.Conclusions: Collectively, these results suggest that ASCs injected subcutaneously into the backs of mice can attenuate tanning following UVB-irradiation, through suppression of tyrosinase activity.

Hairless Descendants of Mexican Hairless Dogs: An Experimental Model for Studying Hypertrophic Scars

In previous studies, the author noticed hypertrophic scar formation in hairless dogs. The purpose of this study was to evaluate both grossly and histopathologically hypertrophic scars in hairless dogs to explore any similarities with hypertrophic scars in humans and to introduce these dogs as an appropriate model for further investigation on hypertrophic scarring. Full-thickness wounds were made on the dorsolumbar skin of hairless dogs. Hypertrophic scarring was examined with three methods: clinical observations, dihydroxyphenylalanine (DOPA)-positive melanocytes, and skin histopathology. Hairless dogs clinically developed the formation of hyperpigmented and hypertrophic scars that did not extend beyond the original wound margins. In hypertrophic scars of hairless dogs, the split epidermal sheet showed an increased number of DOPA-positive melanocytes with well-developed dendrites exhibiting activated melanocytes. There were very few DOPA-positive melanocytes in the repaired skin of haired dogs. Histopathologic examinations demonstrated that hypertrophic scars were fully reepithelialized and granulation tissue formation was accompanied by inflammatory cell infiltration. There was remodeling of thick collagens and fine elastic fibers in the course of hypertrophic scar formation. Experimental hypertrophic scars produced in hairless dogs have morphologic properties similar to those of human hypertrophic scars.

Chimeric Human Epidermal Reconstructs to Study the Role of Melanocytes and Keratinocytes in Pigmentation and Photoprotection

Chimeric epidermal reconstructs made with Negroid melanocytes and Caucasoid keratinocytes (or vice versa) were studied before and after UVB irradiation to understand the respective roles of these cells in tanning and photoprotection, especially lipoperoxidation and enzymatic defences against free radicals. Using this approach, we have confirmed overall the theory of the epidermal melanin unit. We have also shown that melanocytes of poorly tanning Caucasoids, which have a comparatively higher content of unsaturated fatty acids in their cell membrane, are more prone to the peroxidative effects of UV light, and that keratinocytes participate in photoprotection via phototype-dependent antioxidant enzyme activities, especially for catalase.

Regulatory Effects of Heat on Normal Human Melanocyte Growth and Melanogenesis: Comparative Study with UVB

Although energy-rich ultraviolet B (UVB) is considered to be primarily responsible for most of the effects associated with solar radiation, small energy recorded as heat appears to contribute to the biologic effects of solar radiation on the skin. We compared the effects of heat and UVB on normal human melanocyte functions. In monolayer culture the following was found. (i) Heat-treated melanocytes showed an increased dendricity and exhibited a larger cell body compared with nontreated melanocytes. (ii) After multiple treatments with UVB (20 mJ per cm2, 312 nm) or heat (42 degrees C for 1 h) for 3 d, melanocytes had a lower survival than nontreated melanocytes, but they resumed proliferation within 6 d in the same manner as seen in control. (iii) The expression levels of cell cycle regulators, p53 and p21 proteins, were increased after multiple treatments with UVB or heat. (iv) The tyrosinase (dopa-oxidase) activity per cell was increased after the multiple treatments with UVB or heat. (v) The number of dopa-positive melanocytes in coculture with keratinocytes in epithelial sheets was greatly increased by UVB or heat treatments. (vi) Similarly, the increased number of tyrosinase-related protein 1 positive melanocytes was seen in skin equivalents after UVB (100 mJ per cm2) or heat (42 degrees C for 1 h) treatments for 7 d. These results suggest that heat shares significant biologic activities with UVB in melanocyte functions. These results could be considered as one of the protective or adaptive responses of the skin pigmentary system to the environment.

Age-related changes in skin color and histologic features of hairless descendants of Mexican Hairless dogs

Summary Age-related changes in skin color and histologic features of hairless descendants of Mexican Hairless dogs were investigated and compared with those of haired descendants of Mexican Hairless dogs and Beagles. According to age, dogs studied were allotted to 4 groups: 0 to 2 weeks, 4 to 5 months, 1 to 1.5 years, and 3 to 4 years old. Skin color, histologic features, and numbers of dihydroxyphenylalanine (dopa)-positive melanocytes were examined. The luminance values measured, using a spectrophotometer, decreased with advancing age up to 1.5 years, but they increased again at 3 to 4 years in hairless dogs. The number of dopa-positive melanocytes in hairless dogs decreased with advancing age, whereas there were no dopa-positive melanocytes in haired dogs and Beagles. Histologically, the epidermis of newborn hairless pups was thick. The border between the epidermis and dermis was wavy, and epidermal ingrowths were found projecting into the dermis. As hairless dogs grew older, the epidermis became thinner and flatter. Although numbers of hair follicles and sebaceous and apocrine sweat glands were apparently fewer in hairless dogs than in haired dogs and Beagles, these structures were detected at least up to 4 years of age. On the other hand, haired dogs and Beagles had a thin epidermis at birth and aging had little effect on their epidermal structures. The dermis of hairless dogs contained fewer mast cells than did that of haired dogs and Beagles.

Histological, Biochemical, and Ultrastructural Studies on Hyperpigmented Human Skin Xenografts

The mechanisms for hyperpigmentation observed in human cutaneous xenografts placed on athymic nude mice was investigated. Histologic, biochemical, histochemical, and ultrastructural examinations were performed on human skin prior to grafting and at various times ranging from 2 weeks to 30 weeks post-grafting (PG). Hyperpigmentation was macroscopically visible on the graft as early as 4-6 weeks. The number of Dopa-positive melanocytes per unit area was increased at 2 weeks PG and remained elevated until 20 weeks PG. The surface area of the melanocytes, a measure of the activity of the cells, also increased significantly and remained above the pre-grafting size throughout the study. Western blot analysis using tyrosinase specific antibody (alpha Ty-SP) revealed the presence of tyrosinase exclusively in the grafted skin from 2 weeks to 12 weeks PG tested. Histological and ultrastructural observations revealed the presence of numerous dendritic melanocytes, indeterminant clear cells suggestive of Langerhans cells, and dermal melanophages. The results of this study suggest that the observed hyperpigmentation in grafted tissue is caused by an increase in the number of Dopa-positive melanocytes and probably from enhanced melanin production. Extracts of proteins from the xenografts exhibited prominent differences in low and high molecular proteins between pre- and post-grafted skin. Among them, the exclusive appearance of a protein doublet with apparent mw approximately 14 kDa was found in grafted skin, and subsequent studies indicate it has potent effects on melanocyte function.

Treatment of idiopathic guttate hypomelanosis with liquid nitrogen: Light and electron microscopic studies

Idiopathic guttate hypomelanosis is a common skin disorder of unknown cause. Our studies have shown that significantly fewer dopa-positive melanocytes are in the white macules of idiopathic guttate hypomelanosis than in normal skin. By electron microscopy we observed that the melanocytes in the lesional skin were round and less dendritic with fewer melanosomes than in normal pigment cells. Lesions gently frozen with liquid nitrogen repigmented in 6 to 8 weeks. The number of dopa-positive melanocytes was significantly greater in the repigmented areas than in untreated lesions but less than in normal skin.

Tyrosinase Activity in the First Coat of Agouti and Black Mice

Tyrosinase activity was compared in the skin and hair bulbs of young black and agouti mice between 4 and 12 days old. Differences in activity were found to be maximal in both the hair and skin at the time of yellow pigment synthesis in agouti mice. Histological examination suggested that the number of dopa-positive melanocytes is similar in the hair bulbs of agouti and black mice. The level of SH-compounds in the hair bulb was examined and found to be elevated in agouti tissue at the time of phaeomelanin formation. It was shown that sulphydryl compounds such as cysteine and glutathione have an inhibitory effect on tyrosinase, and it is possible that the elevated levels of SH-compounds are responsible for a reduction in tyrosinase activity in agouti mice. In agouti hair bulbs, this effect can be reversed in vitro by addition of copper.

Differential analysis of experimental hypermelanosis induced by UVB, PUVA, and allergic contact dermatitis using a brownish guinea pig model.

In moderately colored guinea-pig skin, UVB, PUVA, and allergic contact dermatitis were shown to induce hyperpigmentation that resembled the pigmentary changes observed in mongoloid human skin. Using this model, we examined the effects of chemical agents, including tyrosinase inhibitors and sunscreen agents, on the color changes induced by UV irradiation. The daily exposure of brownish guinea-pig skin to UVB irradiation at a variety of energies for 3 successive days induced clearly visible black pigmentation on the irradiated rectangular areas of the flank within a few days of irradiation, the maximum being reached about 1 week after irradiation, i.e., similar to the changes that occur in pigmented human skin. Split epidermal sheets prepared from untreated pigmented guinea pigs exhibited 200-400 melanocytes/mm2; 1 week after UV irradiation, the applied areas show an increased number of strongly dopa-positive melanocytes with stout dendrites (800-1,000 cells/mm2). UVA irradiation following an intraperitoneal injection of 8-methoxypsoralen (8-MOP) also produced black pigmentation 1 week after irradiation, and this was paralleled by a marked increase in the number of dopa-positive melanocytes in dopa-reacted split epidermal sheets. Allergic contact dermatitis produced by the application of 1-phenylazo-2-naphthol induced hyperpigmentation after an interval of about 14 days in 10 of the 21 allergy-acquiring animals examined. This induced pigmentation was accompanied by an increase in the number of dopa-positive melanocytes as compared to the number seen in controls. In contrast, allergic contact dermatitis produced by the application of dinitrochlorobenzene failed to induce such a high ratio of postpigmentation, with only 3 of the 21 allergy-acquiring animals showing hyperpigmentation and 5 showing depigmentation; in the latter, there was a slight decrease in the number of dopa-positive melanocytes. To study the preventive effect of tyrosine inhibitors on UVB-induced pigmentation, daily topical applications of these compounds were performed after three daily UVB irradiations. Treatment with 10% hydroquinone for 10 days interrupted UVB-induced pigmentation and resulted in a marked reduction in the number of epidermal melanocytes as compared to the number found in UVB-irradiated, untreated control skin.

Idiopathic Guttate Hypomelanosis

Idiopathic guattate hypomelanosis (IGH) has been studied histochemically (split-dopa) and ultrastructurally in three patients. Affected skin was compared to normal surrounding skin. Argentic stains revealed a decrease in the melanin content of affected epidemis and pigment granules were irregularly distributed. Split-dopa preparations showed a decrease in the the number of dopa-positive melanocytes in hypomelanotic macules. Most of the melanocytes were rounded in from and either lacked dendrites or had fragmented ones. Ultrastructural study confirmed the progressive loss of epidermal melanocytes in skin affected by IGH and revealed two types of melanocytes, healthy melanocytes with normal melanogenic activity and melanocytes containing few or no immature melanosomes but showing no cellular alterations. Depigmentation in IGH seems to occur in two stages, loss of melanogenic activity in certain malanocytes and elimination of inactive melanocytes. The basic defect in IGH melanocytes may reflect the aging process.

The Effects of UVB and 7, 12, Dimethylbenz(α)anthracene (DMBA) on Epidermal Melanocytes of the Tail in C57BL Mice

The morphological and numerical changes in the epidermal melanocyte system of the tail of C57BL mice were studied after exposure to 7, 12 dimethylbenz(alpha)anthracene (DMBA) followed by UVB irradiation. The dorsal aspect of the tail was exposed for 5 days per week, for a total duration of 10 weeks, to a daily dosage of 0.1 J/cm2 UVB (peak 310 nm). Once a week, 0.2 ml of 0.15% DMBA in acetone was locally applied to the irradiated areas. Biopsies were studied by the combined skin-splitting DOPA and electron microscopic techniques. After 10 weeks of DMBA treatment the following changes were observed: DMBA treatment the following changes were observed: the original brick-like arrangement of melanocytes became confluent, melanocytes were irregularly shaped, dendrites shortened and clumped together, and the outer root sheaths of the hair follicles became covered with melanocytes. There was a significant increase in the number of DOPA positive melanocytes at the end of the first week in DMBA and DMBA + UVB treated skin. Ultrastructurally, an increase in melanosome formation in melanocytes and transfer into keratinocytes was found, as well as redistribution of melanosomes from singlets and doublets into larger groups. Damage of melanocytes by the DMBA treatment was seen, but no inflammation or tumor formation was observed.

The role of the epidermis in the histopathogenesis of lichen amyloidosus. Histochemical correlations.

Summary.— Biopsies from 10 patients with lichen amyloidosus were investigated histochemically with special reference to the epidermal changes. The number of dopa-positive melanocytes was markedly decreased in the epidermis overlying the amyloid deposits, indicating that melanocytes are also involved and probably destroyed by the disease process. No increase in lysosomal enzyme activity could be detected in epidermal cells undergoing degeneration or in the epidermis immediately above the amyloid deposits. Respiratory enzyme activity was moderately reduced in the epidermis overlying the amyloid deposits. These histochcmical findings confirm that the epidermis is likely to be involved in the histopathogenesis of lichen amyloidosus. It is suggested that, prior to the deposition of amyloid, a specific cellular tolerance may develop to the presence of necrotic epidermal cells in the region of the dermo-epidermal junction.

Melanocytes of Nail Matrix and Nail Pigmentation

Histologic studies of melanocytes of nail matrices with or without nail pigmentation were made in 15 specimens taken from 12 Japanese subjects. The following results were obtained: (1) Melanin granules were present in the normal nail matrix, (2) dopa-positive melanocytes were also present in the lower two to four layers of normal nail matrix, but few or none in the lowest layer, (3) histological findings of the nail matrix, corresponding to pigmented bands, revealed an increase in activity and number of dopa-positive melanocytes, (4) of five patients with pigmented band on nail, none was found to be nevoid in character.