Background: Skin diseases characterized by epithelial barrierdysfunction show altered sphingolipid metabolism,which results in changes in the stratum corneum intercellularlipid components and structure. Under pathological conditions,1-deoxysphingolipids form as atypical sphingolipidsfrom de novo sphingolipid biosynthesis. Objective: Thisstudy investigated the potential role of 1-deoxysphingolipidsin skin barrier dysfunction secondary to X-ray and ultravioletB (UVB) irradiation in vitro and in vivo. It was also evaluatedchanges in the expression of 1-deoxysphingolipids in lesionalhuman skin of atopic dermatitis. Methods: In thisstudy, the changes in these 1-deoxysphingolipids levels ofskin and serum samples were investigated in skin barrier dysfunctionassociated with X-ray and UVB irradiation in vitroand in vivo. Results: Increased 1-deoxysphingolipids were observed in cultured normal human epidermal keratinocytesafter X-ray irradiation. X-ray or UVB irradiation increased theproduction of 1-deoxysphingosine in a reconstituted 3-dimensional(3D) skin model. Interestingly, treatment with aphysiological lipid mixture (multi-lamellar emulsion containedpseudoceramide), which can strengthen the epidermalpermeability barrier function, resulted in decreased1-deoxysphingosine formation in a reconstituted 3D skinmodel. Further investigation using a hairless mouse modelshowed similar preventive effects of physiological lipid mixtureagainst 1-deoxysphingosine formation after X-ray irradiation.An increased level of 1-dexoysphingosine in the stratumcorneum was also observed in lesional skin of atopic dermatitis. Conclusion 1-deoxysphingosine might be a novelbiomarker of skin barrier dysfunction and a physiological lipidmixture treatment could prevent 1-deoxysphingosine productionand consequent skin barrier dysfunction.

BACKGROUND AND OBJECTIVES: This study compared two types of skin graft reconstruction for the defect of the radial forearm free flap. SUBJECTS AND METHOD: Ten cases of split-thickness skin graft (STSG) harvested from the thigh were analyzed. Also, ten cases of full-thickness skin graft (FTSG) harvest from the inguinal area applied with vacuum-assisted closure (VAC) system were analyzed. RESULTS: The defect size of the radial forearm was increased more in the STSG group than in the FTSG group (p<0.05). Skin grafts were recovered completely sooner in the FTSG group than in the STSG group although it was not statistically significant (p=0.082). Five complications (pruritus, hypertrophic scar) were found in the donor site in the STSG group (p<0.05). FTSG gave better scores according to the Vancouver Scar Scale in terms of pigmentation, pliability, and height (p<0.05). CONCLUSION FTSG harvested from the inguinal area with the application of VAC system has many advantages for the defect of the radial forearm free flap although it is usually used for smaller size defects than for STSGs.

BACKGROUND AND OBJECTIVES: This study compared two types of skin graft reconstruction for the defect of the radial forearm free flap. SUBJECTS AND METHOD: Ten cases of split-thickness skin graft (STSG) harvested from the thigh were analyzed. Also, ten cases of full-thickness skin graft (FTSG) harvest from the inguinal area applied with vacuum-assisted closure (VAC) system were analyzed. RESULTS: The defect size of the radial forearm was increased more in the STSG group than in the FTSG group (p<0.05). Skin grafts were recovered completely sooner in the FTSG group than in the STSG group although it was not statistically significant (p=0.082). Five complications (pruritus, hypertrophic scar) were found in the donor site in the STSG group (p<0.05). FTSG gave better scores according to the Vancouver Scar Scale in terms of pigmentation, pliability, and height (p<0.05). CONCLUSION: FTSG harvested from the inguinal area with the application of VAC system has many advantages for the defect of the radial forearm free flap although it is usually used for smaller size defects than for STSGs.
Cicatrix ; Forearm ; Free Tissue Flaps ; Humans ; Methods ; Negative-Pressure Wound Therapy ; Pigmentation ; Pliability ; Reconstructive Surgical Procedures ; Skin Transplantation ; Skin ; Thigh ; Tissue Donors ; Transplants

BACKGROUND: Many inflammatory mediators, including various cytokines (e.g. interleukins and tumor necrosis factor [TNF]), inflammatory proteases, and histamine are released following mast cell activation. However, the endogenous modulators for mast cell activation and the underlying mechanism have yet to be elucidated. Endogenous cannabinoids such as palmitoylethanolamide (PEA) and N-arachidonoylethanolamine (anandamide or AEA), were found in peripheral tissues and have been proposed to possess autacoid activity, implying that cannabinoids may downregulate mast cell activation and local inflammation. OBJECTIVE: In order to investigate the effect of cannabinoid receptor-1 (CB1R) agonists on mast cell activation, AEA-derived compounds were newly synthesized and evaluated for their effect on mast cell activation. METHODS: The effects of selected compounds on FcepsilonRI-induced histamine and beta-hexosaminidase release were evaluated in a rat basophilic leukemia cell line (RBL-2H3). To further investigate the inhibitory effects of CB1R agonist in vivo, an oxazolone-induced atopic dermatitis mouse model was exploited. RESULTS: We found that CB1R inhibited the release of inflammatory mediators without causing cytotoxicity in RBL-2H3 cells and that CB1R agonists markedly and dose-dependently suppressed mast cell proliferation indicating that CB1R plays an important role in modulating antigen-dependent immunoglobulin E (IgE)-mediated mast cell activation. We also found that topical application of CB1R agonists suppressed the recruitment of mast cells into the skin and reduced the level of blood histamine. CONCLUSION: Our results indicate that CB1R agonists down-regulate mast cell activation and may be used for relieving inflammatory symptoms mediated by mast cell activation, such as atopic dermatitis, psoriasis, and contact dermatitis.
Animals ; Basophils ; beta-N-Acetylhexosaminidases ; Cannabinoid Receptor Agonists ; Cannabinoids ; Cell Line ; Cytokines ; Dermatitis, Atopic* ; Dermatitis, Contact ; Histamine ; Immunoglobulin E ; Immunoglobulins ; Inflammation ; Interleukins ; Leukemia ; Mast Cells* ; Mice ; Peptide Hydrolases ; Psoriasis ; Rats ; Skin ; Tumor Necrosis Factor-alpha

PURPOSE: High-affinity receptor I (FcεRI) on mast cells and basophils plays a key role in the immunoglobulin E (IgE)-mediated type I hypersensitivity mediated by allergen cross-linking of the specific IgE-FcεRI complex. Thus, prevention of IgE binding to FcεRI on these cells is an effective therapy for allergic disease. We have developed a strategy to disrupt IgE binding to FcεRI using an antibody targeting FcεRIα. MATERIALS AND METHODS: Fab fragment antibodies, which lack the Fc domain, with high affinity and specificity for FcεRIα and effective inhibitory activity against IgE-FcεRI binding were screened. IgE-induced histamine, β-hexosaminidase and Ca2+ release in basophils were determined by ELISA. A B6.Cg-Fcer1a(tm1Knt) Tg(FCER1A)1Bhk/J mouse model of passive cutaneous anaphylaxis (PCA) was used to examine the inhibitory effect of NPB311 on allergic skin inflammation. RESULTS: NPB311 exhibited high affinity to human FcεRIα (KD=4 nM) and inhibited histamine, β-hexosaminidase and Ca2+ release in a concentration-dependent manner in hFcεRI-expressing cells. In hFcεRIα-expressing mice, dye leakage was higher in the PCA group than in controls, but decreased after NPB311 treatment. NPB311 could form a complex with FcεRIα and inhibit the release of inflammation mediators. CONCLUSION: Our approach for producing anti-FcεRIα Fab fragment antibody NPB311 may enable clinical application to a therapeutic pathway in IgE/FcεRI-mediated diseases.
Animals ; Antibodies ; Antibody Affinity ; Basophils ; Enzyme-Linked Immunosorbent Assay ; Histamine ; Humans ; Hypersensitivity, Immediate ; Immunoglobulin E* ; Immunoglobulins* ; Inflammation Mediators ; Inflammation* ; Mast Cells ; Mice ; Passive Cutaneous Anaphylaxis ; Sensitivity and Specificity ; Skin

PURPOSE: High-affinity receptor I (FcεRI) on mast cells and basophils plays a key role in the immunoglobulin E (IgE)-mediated type I hypersensitivity mediated by allergen cross-linking of the specific IgE-FcεRI complex. Thus, prevention of IgE binding to FcεRI on these cells is an effective therapy for allergic disease. We have developed a strategy to disrupt IgE binding to FcεRI using an antibody targeting FcεRIα. MATERIALS AND METHODS: Fab fragment antibodies, which lack the Fc domain, with high affinity and specificity for FcεRIα and effective inhibitory activity against IgE-FcεRI binding were screened. IgE-induced histamine, β-hexosaminidase and Ca2+ release in basophils were determined by ELISA. A B6.Cg-Fcer1a(tm1Knt) Tg(FCER1A)1Bhk/J mouse model of passive cutaneous anaphylaxis (PCA) was used to examine the inhibitory effect of NPB311 on allergic skin inflammation. RESULTS: NPB311 exhibited high affinity to human FcεRIα (KD=4 nM) and inhibited histamine, β-hexosaminidase and Ca2+ release in a concentration-dependent manner in hFcεRI-expressing cells. In hFcεRIα-expressing mice, dye leakage was higher in the PCA group than in controls, but decreased after NPB311 treatment. NPB311 could form a complex with FcεRIα and inhibit the release of inflammation mediators. CONCLUSION: Our approach for producing anti-FcεRIα Fab fragment antibody NPB311 may enable clinical application to a therapeutic pathway in IgE/FcεRI-mediated diseases.
Animals ; Antibodies ; Antibody Affinity ; Basophils ; Enzyme-Linked Immunosorbent Assay ; Histamine ; Humans ; Hypersensitivity, Immediate ; Immunoglobulin E* ; Immunoglobulins* ; Inflammation Mediators ; Inflammation* ; Mast Cells ; Mice ; Passive Cutaneous Anaphylaxis ; Sensitivity and Specificity ; Skin

PURPOSE: Chronic use of topical hypotensive agents induces several side effects caused by preservatives. The purpose of this study was to evaluate the effects of prostaglandin analogs with varying concentrations of benzalkonium chloride (BAC), preservative-free (PF), and alternative preservatives on mouse corneal tissue. METHODS: Thirty-five, 8- to 10-week-old female C57BL/6 mice (five mice for each group) were used for this study. To the control group, we applied normal saline, and to each drug-treated group we applied 0.02% BAC, bimatoprost 0.01% (with BAC 0.02%), latanoprost 0.005% (with BAC 0.02%), travoprost 0.004% (with 0.001% polyquad) or tafluprost 0.0015% with/without 0.001% BAC, once a day (9 p.m.) for 4 weeks. Corneal fluorescein staining was evaluated in all groups. After harvest, the corneal tissues were embedded in paraffin and then Hematoxylin-Eosin stain was performed for histopathological examination. Immunofluorescence staining was done against TNF-alpha, IL-6, HLA DR, pJNK, and pAkt. RESULTS: In corneal fluorescein staining, severe punctate epithelial keratitis was seen in the groups of 0.02% BAC, 0.02% BAC containing bimatoprost 0.01% and latanoprost 0.005%. The surface desquamation, irregular surface, loss of cell borders, anisocytosis and stromal shrinkage were observed in the groups of BAC-containing eye drops. Moreover, the groups treated with BAC-containing eye drops have high inflammatory markers, significantly decreased cell viability-related signal, pAkt, and higher apoptosis-inducing signal, pJNK, than the control group. On the other hand, travoprost 0.004% and PF tafluprost 0.0015% have less cellular morphologic changes, lower inflammation, and higher cellular viability than BAC-containing formulations. CONCLUSIONS: Corneal damage, increased inflammation and apoptosis and low cell viability were observed in BAC-containing groups. PF or alternatively preserved glaucoma medications seem to be a reasonable and viable alternative to those preserved with BAC.
Animals ; Cell Survival ; Conjunctiva/drug effects/*pathology ; Disease Models, Animal ; Epithelium, Corneal/drug effects/*pathology ; Female ; Glaucoma/*drug therapy/pathology ; Mice ; Mice, Inbred C57BL ; Microscopy, Fluorescence ; Ophthalmic Solutions ; Preservatives, Pharmaceutical ; Prostaglandins, Synthetic/*administration & dosage

PURPOSE: Recognition of microbes is important to trigger the innate immune system. Mycolic acid (MA) is a component of the cell walls of mycobacteria such as Mycobacterium bovis Bacillus Calmette-Guerin. MA has immunogenic properties, which may modulate the innate and adaptive immune response. This study aimed to investigate whether a novel synthetic MA (sMA) inhibits allergic inflammatory responses in a mouse model of asthma. METHODS: BALB/c mice were injected intraperitoneally with sMA followed by sensitization and challenge with ovalbumin (OVA). Mice were examined for bronchial hyperresponsiveness (BHR), the influx of inflammatory cells into the lung tissues, histopathological changes in the lungs and CD4+CD25+Foxp3+ T cells in the spleen, and examined the response after the depleting regulatory T cells (Tregs) with an anti-CD25mAb. RESULTS: Treatment of mice with sMA suppressed the asthmatic response, including BHR, bronchoalveolar inflammation, and pulmonary eosinophilic inflammation. Anti-CD25mAb treatment abrogated the suppressive effects of sMA in this mouse model of asthma and totally depleted CD4+CD25+Foxp3+ T cells in the spleen. CONCLUSIONS: sMA attenuated allergic inflammation in a mouse model of asthma, which might be related with CD4+CD25+Foxp3+ T cell.
Adaptive Immunity ; Animals ; Asthma* ; Bacillus ; Cell Wall ; Eosinophils ; Immune System ; Inflammation* ; Lung ; Mice* ; Mycobacterium bovis ; Mycolic Acids* ; Ovalbumin ; Spleen ; T-Lymphocytes ; T-Lymphocytes, Regulatory

BACKGROUND: Topical steroid treatment induces diverse local Wand systemic adverse effects. Several approaches have been tried to reduce the steroid-induced adverse effects. Simultaneous application of physiological lipid mixture is also suggested. OBJECTIVE: Novel vehicles for topical glucocorticoids formulation were evaluated for the efficacy of reducing side-effects and the drug delivery properties of desonide, a low potency topical steroid. METHODS: Transcutaneous permeation and skin residual amount of desonide were measured using Franz diffusion cells. The in vivo anti-inflammatory activity was evaluated using murine model. RESULTS: Topical steroids formulation containing desonide, in either cream or lotion form, were prepared using multi-lamellar emulsion (MLE), and conventional desonide formulations were employed for comparison. MLE formulations did not affect the anti-inflammatory activity of the desonide in phobol ester-induced skin inflammation model, compared with conventional formulations. While the penetrated amounts of desonide were similar for all the tested formulations at 24 hours after application, the increased lag time was observed for the MLE formulations. Interestingly, residual amount of desonide in epidermis was significantly higher in lotion type MLE formulation. Steroid-induced adverse effects, including permeability barrier function impairment, were partially prevented by MLE formulation. CONCLUSION: Topical desonide formulation using MLE as a vehicle showed a better drug delivery with increased epidermal retention. MLE also partially prevented the steroid-induced side effects, such as skin barrier impairment.
Desonide ; Diffusion ; Epidermis ; Glucocorticoids ; Inflammation ; Permeability ; Retention (Psychology) ; Skin ; Steroids

PURPOSE: Various therapeutic approaches have been suggested for preventing or reducing the adverse effects of topical glucocorticoids, including skin barrier impairment. Previously, we have shown that impairment of skin barrier function by the highest potency topical glucocorticoid, clobetasol 17-propinate (CP), can be partially prevented by co-application of a physiological lipid mixture containing pseudoceramide, free fatty acids, and cholesterol (multi-lamellar emulsion [MLE]). Skin atrophic effects of CP were also partially reduced by MLE. In this study, the preventive effects of MLE on the lowest potency topical glucocorticoid, hydrocortisone (HC), were investigated using animal models. METHODS: Anti-inflammatory activity of topical HC was evaluated using a 12-O-tetradecanoylphobol-13-acetate-induced skin edema model. Topical steroid induced adverse effects were evaluated using hairless mouse. RESULTS: The results showed that the anti-inflammatory activity was not altered by co-application of either MLE or hydrobase. However, co-application of MLE and 1.0% HC showed less impairment in the epidermal permeability barrier function, skin hydration, and skin surface pH compared with hydrobase. Stratum corneum integrity, evaluated by measuring trans-epidermal water loss after repeated tape stripping, showed less damage with MLE co-application. Long-term application of topical HC induced skin atrophy, measured by a reduction in skinfold and epidermal thickness and in the number of epidermal proliferating cell nucleus antigen (PCNA)-positive keratinocytes. Co-application of MLE did not affect the skinfold or epidermal thickness, but the number of PCNA-positive keratinocytes was less decreased with MLE use. CONCLUSIONS: These results suggest that co-application of MLE is effective in reducing the local adverse effects of low-potency topical glucocorticoids and supports the therapeutic efficacy of physiological lipid mixtures on skin barrier function.
Animals ; Atrophy ; Cell Nucleus ; Cholesterol ; Clobetasol ; Edema ; Fatty Acids, Nonesterified ; Glucocorticoids ; Hydrocortisone ; Hydrogen-Ion Concentration ; Keratinocytes ; Permeability ; Skin ; Steroids ; Water Loss, Insensible