BACKGROUND: The immortalized human keratinocyte line, HaCaT cells have been widely used as substitutes for normal epidermal keratinocytes. Recently, reconstruction of a skin equivalent using HaCaT cells showed a multilayered epithelium,but somewhat different tissue architecture as compared with normal epidermis. OBJECTIVE: In this study, using HaCaT cells we tried to reconstruct an epidermis resembling more closely to normal epidermis than the previous results. MATERIALS AND METHODS: HaCaT cells were cultured in air-liquid interface on a recently developed dermal substrated in our laboratory, de-epidermized dermis (DED) raised on fibroblast-populated collagen matrix and the result was compared with those on DED or fibroblast-populated collagen matrix alone. RESULTS: HaCaT cells on the new dermal substrate formed a multilayered epithelium with rete ridges, showing rather orderly cellular organization compared with those on fibroblast-populated collagen matrix. However, horny and granular layers were not observed contrary to normal epidermis. Immunohistochemical studies revealed that differentiation markers such as keratin 1, keratin 6 and involucrin showed the similar pattern to those in HaCaT cells cultured on fibroblast-populated collagen matrix. Markers of terminal differentiation, loricrin and filaggrin were not expressed contrary to normal epidermis. CONCLUSION: These results suggest that organotypic culture HaCaT cells on the dermal substrate combines DED with fivroblast-populated collagen matrix results in incomplete differentiation of HaCaT cells contrary to normal keratinocytes.
Antigens, Differentiation ; Collagen* ; Dermis* ; Epidermis ; Epithelium ; Humans ; Keratin-1 ; Keratin-6 ; Keratinocytes ; Skin

BACKGROUND: Melanocytes grown in pure monolayer culure lack many of the cellular interactions that exist in vivo. This can be partially overcome by growing melanocytes together with other epidermal cells in skin equivalent models. OBJECTIVE: The objective of the present study was to grow human melanocytes in human epidermis reconstructed on dermal substrates in vitro and to examine their response to UV radiation. METHODS: The skin equivalents were prepared by seeding cultured human keratinocytes together with cultured human melanocytes(in a ratio of 5%) onto de-epidermized dermis. After 7 days of culture, they were exposed to UVB irradiation(total 150m J/cm over 5days). On day 12 of air exposure the sections of the skin equivalents were prepared for histology. The structure of the skin equivalents was studied following staining with hematoxylin and eosin. Melanocytes were characterized by DOPA staining and by immunohistochemistry. RESULTS: Melanocytes were localized singly within the basal layer of the reconstructs. Melanin was also visible both in the melanocytes and in neighboring keratinocytes. There was an increase in melanocyte size and dendricity following UV irradiation. Melanocytes became positive to staining with HMB-45 antibody following UV irradiation. CONCLUSION: Our results indicate that melanocytes grown in reconstructed human epidermis are functional and capable of responding to UV irradiation.
Dermis ; Dihydroxyphenylalanine ; Eosine Yellowish-(YS) ; Epidermis* ; Hematoxylin ; Humans* ; Immunohistochemistry ; Keratinocytes ; Melanins ; Melanocytes* ; Skin

The hyperimmunoglobulinemia E (Jobs) syndrome (HIES) is characterized by marked elevated levels of IgE, recurrent cutaneous and systemic staphylococcal infections, atopic-like dermatitis, and defective neutrophil chemotaxis. Three cases of HIES have been reported in Korea, but not in the dermatology literature. We report a case of HIES with cutaneous infections and MRSA (methicillin-resistant Staphylococcus aureus). A 15-month-old girl presented with intractable pruritic excoriated papular pustular skin lesions and multiple subcutaneous abscesses. Surgical drainage of the abscesses and a course of antibiotic treatment in addition to topical steroids for about 7 weeks resulted in a remarkable improvement.
Abscess ; Chemotaxis ; Dermatitis ; Dermatology ; Drainage ; Female ; Humans ; Hypergammaglobulinemia* ; Immunoglobulin E ; Infant ; Job Syndrome* ; Korea ; Methicillin-Resistant Staphylococcus aureus ; Neutrophils ; Skin ; Staphylococcal Infections ; Staphylococcus ; Steroids

BACKGROUND: Neurofibiomat,osis type 1(NF-1) is a multisystemic disorder of genetic ori gin, affecting one in every 3000 to 4000 people. It is clinically important in the aspect of dermatology, pediatrics, orthopedic surgery, neurology, neurosurgery and ophthalmology. OBJECTIVE: The purpore of this study was to elucidate the clinical characteristics of NF-1 in Korean people. METHODS: We carried out a retrospective study on 112 patients which were compatible to the diagnostic criteria of Riccardi and Neurofibromatosis Conference Statement. The results were compared with other western studies. RESULTS: The age of onset, sex ratio, family history of neurofibromatosis, and clinica features of cafe-au-lait spot, neurofibroma, and axillary freckinings did not differed from western countries. However, some characterist,ics of NF 1(e.g. Lisch nodule) were not as sessed in the most of the cases and incomplete evaluations of the systemic diseases wen found. CONCLUSION: In this study t.he clinial features of NF-1 did not differ from western coun tries in many aspects. A more intensive evaluation of patient,s status is needed to manag; NF-1 patients appropritely.
Age of Onset ; Cafe-au-Lait Spots ; Dermatology ; Humans ; Neurofibroma ; Neurofibromatoses* ; Neurofibromatosis 1* ; Neurology ; Neurosurgery ; Ophthalmology ; Orthopedics ; Pediatrics ; Retrospective Studies ; Sex Ratio

BACKGROUND: A number of in vitro skin models have been developed for the purpose of the screening of cosmetics, pharmaceuticals, and environmental chemicals. To mimic the skin in vivo, a model should resemble morphologically and biochemically the parent, tissue. OBJECTIVE: The purpos of this study is to study the differentiation and organization of the artificial epidermis in comparsion with epidermis in vivo based on the expression of epidermal protein antigens and basement membrane components. METHODS: Human keratinocytes were cultured on deepidermidized dermis (RE-DED) or on fibroblast-populated collag-,n matrix (LSE). After 10 days culture, the sections of RE-DED and LSE were stained with hematoxylin and eosin. An immunohistochemical study was also performed with the sections of RE-DED and LSE using antibodies recognizing proliferating cell nuclear antigens (PCNA), epidermal growth factor receptor (EGFR), keratin 1, involucrin, filaggrin, loricrin, keratin 13, type IV collagen, and laminin. RESULTS: In both culture systems(RE-DED and LSE) a multilayered epidermis with a horny layer was observed. In the human epidermis reconstructed by both culture systems, differentiation markers appeared but with a topography slightly different from that of epidermis in vivo, and components of the basement membrane was also expressed. CONCLUSION: Our findings suggest the epidermis obtained in both culture systems(RE-DED and LSE) resembled in vivo epidermis morphologically and biochemically, although it was not the same.
Antibodies ; Antigens, Differentiation ; Basement Membrane* ; Collagen Type IV ; Dermis ; Eosine Yellowish-(YS) ; Epidermis* ; Hematoxylin ; Humans* ; Keratin-1 ; Keratin-13 ; Keratinocytes ; Laminin ; Mass Screening ; Parents ; Proliferating Cell Nuclear Antigen ; Receptor, Epidermal Growth Factor ; Skin

BACKGROUND: A number of in vitro skin models have been developed for the purpose of the screening of cosmetics, pharmaceuticals, and environmental chemicals. To mimic the skin in vivo, a model should resemble morphologically and biochemically the parent, tissue. OBJECTIVE: The purpos of this study is to study the differentiation and organization of the artificial epidermis in comparsion with epidermis in vivo based on the expression of epidermal protein antigens and basement membrane components. METHODS: Human keratinocytes were cultured on deepidermidized dermis (RE-DED) or on fibroblast-populated collag-,n matrix (LSE). After 10 days culture, the sections of RE-DED and LSE were stained with hematoxylin and eosin. An immunohistochemical study was also performed with the sections of RE-DED and LSE using antibodies recognizing proliferating cell nuclear antigens (PCNA), epidermal growth factor receptor (EGFR), keratin 1, involucrin, filaggrin, loricrin, keratin 13, type IV collagen, and laminin. RESULTS: In both culture systems(RE-DED and LSE) a multilayered epidermis with a horny layer was observed. In the human epidermis reconstructed by both culture systems, differentiation markers appeared but with a topography slightly different from that of epidermis in vivo, and components of the basement membrane was also expressed. CONCLUSION: Our findings suggest the epidermis obtained in both culture systems(RE-DED and LSE) resembled in vivo epidermis morphologically and biochemically, although it was not the same.
Antibodies ; Antigens, Differentiation ; Basement Membrane* ; Collagen Type IV ; Dermis ; Eosine Yellowish-(YS) ; Epidermis* ; Hematoxylin ; Humans* ; Keratin-1 ; Keratin-13 ; Keratinocytes ; Laminin ; Mass Screening ; Parents ; Proliferating Cell Nuclear Antigen ; Receptor, Epidermal Growth Factor ; Skin

Dermatofibrosarcoma protuberans (DFSP) is a dermal spindle cell neoplasm of intermediate malignancy. It typically forms a brown indurated plaque on which firm nodules subsequently arise, sometimes with ulceration. Atypical DFSP presentations are not unusual, including atrophic, pedunculated, morphea-like and angioma-like forms. However, subcutaneous variant of DFSP that may either arise without dermal involvement or with minimal dermal involvement is very rare. A 36-year-old man presented with a palpable nodule without surface change around the right nipple. Microscopically, the neoplasm was composed of spindle cells with monomorphic storiform arrangement. The superficial part of the neoplasm was located in the subcutaneous tissue. Immunohistochemical staining showed strong cytoplasmic positivity for CD34, but not for factor XIIIa. Dermatologists should pay careful attention to these unusual variant of DFSP, which can be confused with other soft tissue tumors.
Adult ; Breast ; Cytoplasm ; Dermatofibrosarcoma ; Factor XIIIa ; Humans ; Nipples ; Subcutaneous Tissue ; Ulcer

The goal of this study was to evaluate the function of hyaluronic acid (HA) on the active oxygen release from polymorphonuclear leukocytes (PMNs) and the protective effect of bovine corneal endothelial cells (BCEC) from activated PMNs. We used HA with three different molecular weights (MW 700, 000, 2, 000, 000, and 4, 000, 000) and five different concentrations (0, 0.1, 1, 2, and 3 mg/ml). We evaluated the amount of released superoxide from activated PMNs by using dismutase-inhibitable ferricytochrome C reduction. To compare the property and protective effect of HA with those of other viscoelastic substances, we used the same concentration of methylcellulose. HA suppressed superoxide release from PMNs and protected BCEC from activated PMNs in a dose-dependent, rather than a molecular weight-dependent, manner. The effect of HA reached almost a plateau at concentration above 2 mg/ml. However, methylcellulose, another viscoelastic substance, showed a similar effect. Therefore, it seems that the suppression of superoxide released from PMNs is not a property that is unique to HA, but is a general property of viscoelastic substances. Our results indicate that the action mechanism of HA proceeds not only through cell surface HA-receptor. We think that HA also acts as a physical barrier and/or a scavenger of superoxide.
Animals ; Cattle ; Cell Survival ; Cells, Cultured ; Comparative Study ; Cytochromes c/metabolism ; Cytoprotection ; Dose-Response Relationship, Drug ; Endothelium, Corneal/cytology/*drug effects ; Humans ; Hyaluronic Acid/*pharmacology ; Methylcellulose ; Molecular Weight ; *Neutrophil Activation ; Neutrophils/*drug effects/metabolism ; Superoxides/*metabolism

No abstract available.
Immunoglobulin A* ; Immunoglobulins*

BACKGROUND: Ultraviolet B (UVB) irradiation can induce apoptosis of melanocytes and melanoma cells. However, mechanism of UVB-induced apoptosis of melanoma cells is not clarified yet. OBJECTIVE: Our purpose was to study the molecular mechanism of UVB-induced apoptosis of melanoma cells. METHODS: G361 lightly pigmented melanoma cells were analyzed for apoptotic mechanism by flow cytometry and western blotting. RESULTS: G361 melanoma cells showed apoptotic features with gradual increment of UVB doses by MTT and flow cytometry. Western blotting disclosed activation of caspase-3 and poly (ADP-ribose) polymerase (PARP) after UVB irradiation. CONCLUSION: In this study, we showed that UVB-induced apoptosis of melanoma cells is mediated by PARP activation which is induced by caspase cascade.
Apoptosis* ; Blotting, Western ; Caspase 3 ; Caspases* ; Cell Line* ; Flow Cytometry ; Humans* ; Melanocytes ; Melanoma*