Excellent color-matched repigmentation of human vitiligo can be obtained by mini-punch grafting using a machine in combination with ultraviolet therapy

A new method is reported here for treatment of vitiligo by using mini-punch grafting using a machine (mMG) in combination with ultraviolet light B (UVB) therapy. After UVB had been repeatedly irradiated to the lesional facial skin of a patient for 1 month, many mini-punch grafts were removed from the lesional skin and the same numbers of punch biopsies taken from normal scalp skin were placed into the holes of the recipient skin. Repigmentation started soon, and excellent color-matched repigmentation was observed after 12 months. Our easy and speedy mMG seems to be useful for treatment of vitiligo.     

碱性成纤维细胞生长因子和内皮素-1体外协同培养正常人黑素细胞

目的探讨在不含霍乱毒素的MCDB153培养基中添加碱性成纤维细胞生长因子(bFGF)和内皮素-1(ET-1)能否体外培养正常人黑素细胞。方法取健康青少年包皮环切术后的新鲜包皮标本,经修剪、分离、消化和离心获得表皮细胞,用含bFGF和ET-1的MCDB153培养基重悬细胞,于常规条件下培养;倒置显微镜下观察细胞形态;L-Dopa染色及S-100免疫细胞化学法鉴定黑素细胞;生长曲线法记录黑素细胞生长状况;MTT法测定黑素细胞活性。结果黑素细胞生长良好,1周后即进入指数生长期;细胞形态正常,具有2～6个树突以及2～3级树突,细胞纯度较高,无角质形成细胞和/或成纤维细胞污染。多巴染色使黑素细胞胞浆和树突呈棕褐色,胞核呈棕红色,S-100免疫组化DAB显色使黑素细胞胞浆呈棕褐色,证实为正常人黑素细胞;细胞活性良好,随时间延长活性逐渐升高。结论采用bFGF和ET-1协同培养人黑素细胞是一种较好的方法,可用于实验研究。     

Comprehensive analysis of melanogenesis and proliferation potential of melanocyte lineage in solar lentigines

BackgroundSolar lentigines (SLs) are characterized by hyperpigmented macules, commonly seen on sun-exposed areas of the skin. Although it has been reported that an increase in the number of melanocytes and epidermal melanin content was observed in the lesions, the following questions remain to be answered: (1) Is acceleration of melanogenesis in the epidermis caused by an increased number of melanocytes or the high melanogenic potential of each melanocyte? (2) Why does the number of melanocytes increase?ObjectiveTo elucidate the pathogenic mechanism of SLs by investigating the number, melanogenic potential and proliferation status of the melanocyte lineage in healthy skin and SL lesions.MethodsImmunostaining for melanocyte lineage markers (tyrosinase, MART-1, MITF, and Frizzled-4) and a proliferation marker, Ki67, was performed on skin sections, and the obtained images were analyzed by image analysis software.ResultsThe expression level of tyrosinase to MART-1 of each melanocyte was significantly higher in SL lesions than healthy skin. The numbers of melanocytes in the epidermis, melanoblasts in the hair follicular infundibulum and melanocyte stem cells in the bulge region were increased in SL; however, no significant difference was observed in the Ki67-positive rate of these cells.ConclusionThe melanogenic potential of each melanocyte was elevated in SL lesions. It was suggested that the increased number of melanocytes in the SL epidermis might be attributed to the abnormal increase of melanocyte stem cells in the bulge.     

中药对人黑素细胞酪氨酸酶活性及黑素合成的影响效果

目的：探讨刺蒺藜、黄芩苷对体外培养人黑素细胞（Melanocyte,MC）及其与角质形成细胞（Keratinoeyte,KC）共培养体系中酪氨酸酶（Tyrosinase,TYR）活性及黑素合成的影响效果。方法培养人表皮黑素细胞及黑素细胞与角质形成细胞共培养体系,利用MTT法、多巴氧化法及NaOH法测定刺蒺藜、黄芩苷对黑素细胞的增殖、酪氨酸酶活性及黑素合成的影响,以补骨脂素作为对照。结果刺蒺藜、黄芩苷均可促进酪氨酸酶活性及增加黑素合成,但对共培养体系中黑素细胞的作用强于体外纯黑素细胞的作用（P<0.05）;在对酪氨酸酶活性的影响中,黄芩苷的作用强于刺蒺藜,其差异具有统计学意义（P<0.05）;在对黑素合成的作用上,刺蒺藜强于黄芩苷（P<0.05）。结论刺蒺藜、黄芩苷对人黑素细胞酪氨酸酶活性及黑素合成均有较强的促进作用,且对共培养体系的作用强于对纯黑素细胞的作用。     

Ultrastructural evidence of cell communication between epithelial dark cells and melanocytes in vestibular organs of the human inner ear

Background: The possibility of interaction between epithelial dark cells and melanocytes in the mammalian inner ear has been pointed out because of their morphological and biochemical characteristics, although very few studies have dealt directly with communication between these two types of cells. We investigated the dark cell area of human vestibular organs in order to clarify the ultrastructual evidence for cell interaction between epithelial dark cells and melanocytes. Methods: All of the material was obtained from vestibular schwannoma operations. Paraffin sections were stained with hematoxylin and eosin (H&E) and by the Fontana-Masson technique. Other paraffin sections were also stained immunohistochemically for S-100 protein. Glutaraldehyde fixed specimens were investigated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Results: Light microscopy revealed melanin pigment granules in the cytoplasm of epithelial dark cells. Melanocytes in the subepithelial layer stained positively for S-100 protein. The presence of intraepithelial melanocytes was confirmed by the presence of cell profiles with a large number of melanin pigment granules and S-100 protein in the cytoplasm. SEM showed that the dark cells had a pentagonal surface with microvilli on the apical surface edge. They had complicated structures at the basal portion of their cytoplasm. Melanocytes extending cytoplasmic processes to adjacent areas were observed under the dark cells. TEM showed that the dark cells were tightly linked by junctional complexes in the upper lateral portion of their cytoplasmic membrane and interdigitated by lateral infoldings. Compound melanosomes (phagosomes or secondary lysosomes) found in the cytoplasm of the dark cells contained poorly pigmented melanosomes with a periodic internal structure. Gap junctions were clearly showed between adjacent melanocytes in the subepithelial layer. Conclusions: The characteristic substructures of dark cells and melanocytes suggested the presence of intimate cell interaction between these two types of cells in the vestibular organs of the human inner ear, although it is not clear at this stage whether such cell interaction is specific only for patients with vestibular schwannoma. Dark cells and melanocytes form a cell community that serves to maintain homeostasis in vestibular organs through communication in which cell information obtained by both dark cells and melanocytes serves to facilitate the system. © 1995 Wiley-Liss, Inc.     

体外培养人黑素细胞HLA—DR抗原的表达

为了解正常人表皮黑素细胞是否表达ＨＬＡ－ＤＲ抗原,黑素细胞经体外培养后,是否因细胞增减的添加或培养时间的长短而影响ＬＨＡ－ＤＲ抗原表达而应用免疫酶染色法进行检测。结果发现表皮基底层的黑素细胞不表达ＨＬＡ－ＤＲ抗原;接种于含ＴＰＡ增减液的黑素细胞,在传代后２周,黑素细胞表达ＨＬＡ－ＤＲ抗原,７周后ＨＬＡ－ＤＲ抗原失表达;接种于不含ＴＰＡ培养液的黑素细胞,在传代后２周和７周时均不表达ＨＬＡ－ＤＲ抗原是     

Lack of Association between ctla-4 A49G Polymorphism and Vitiligo

Background: Vitiligo is an acquired skin disorder that selectively destroys melanocytes in epidermis with an unknown etiology.   Objective: To investigate the exon 1 A49G polymorphism of cytotoxic T lymphocyte antigen-4 (ctla-4) gene in vitiligo patients.   Methods: The A49G polymorphism was detected by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method in 101 patients and 208 normal healthy age/ethnicity matched individuals.   Results: The frequencies of heterozygote genotypes in patients and controls were found to be 42 (41.6%) of 101 and 85 (40.9%) of 208, respectively. The frequencies of homozygote A and G genotypes were 49 (48.5%) and 10 (9.9%) in 101 patients, whereas, these frequencies in 208 control individuals were 103 (49.5%) and 20 (9.6%), respectively. There was no significant difference between the genotype (P = 0.98) and allele (P = 0.86) frequencies of A49G polymorphism in patients and normal healthy individuals.   Conclusion: Our results indicate that in contrast to several immune mediated disorders, there is no association between ctla-4 A49G gene polymorphism and vitiligo.