clobetasol and Disease-Models--Animal

Motoneuronal loss is the main feature of amyotrophic lateral sclerosis, although pathogenesis is extremely complex involving both neural and muscle cells. In order to translationally engage the sonic hedgehog pathway, which is a promising target for neural regeneration, recent studies have reported on the neuroprotective effects of clobetasol, an FDA-approved glucocorticoid, able to activate this pathway via smoothened. Herein we sought to examine functional, cellular, and metabolic effects of clobetasol in a neurotoxic mouse model of spinal motoneuronal loss. We found that clobetasol reduces muscle denervation and motor impairments in part by restoring sonic hedgehog signaling and supporting spinal plasticity. These effects were coupled with reduced pro-inflammatory microglia and reactive astrogliosis, reduced muscle atrophy, and support of mitochondrial integrity and metabolism. Our results suggest that clobetasol stimulates a series of compensatory processes and therefore represents a translational approach for intractable denervating and neurodegenerative disorders.

Topics: Amyotrophic Lateral Sclerosis; Animals; Case-Control Studies; Cholera Toxin; Clobetasol; Databases, Genetic; Disease Models, Animal; Energy Metabolism; Glucocorticoids; Hedgehog Proteins; Humans; Inflammation Mediators; Male; Mice, 129 Strain; Mitochondria, Muscle; Motor Activity; Motor Neurons; Muscle, Skeletal; Neuronal Plasticity; Neuroprotective Agents; Open Field Test; Saporins; Signal Transduction; Smoothened Receptor; Spine

Targeting the nuclear receptor RORγt is thought to be effective in autoimmune disorders. Tertiary sulfonamide 1 was found to be a potent RORγt inverse agonist previously. However, the high hepatic clearance value limits its druggability. In this study, we designed and synthesized a series of N-sulfonamide-tetrahydroquinolines by molecular modeling and scaffold hopping strategy, aiming at improving the metabolic stabilities. Detailed SAR exploration led to identification of potent RORγt inverse agonists such as 13 with moderate binding affinity and inhibitory activity of Th17 cell differentiation. Binding mode of 13 with RORγt-LBD was revealed by molecular docking. Moreover, 13 showed lower intrinsic clearance in mouse liver microsomes compared with 1 and potent in vivo efficacy and safety in psoriasis models, which can be used as a good starting point for the further optimization.

Topics: Animals; Cell Differentiation; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Discovery; Female; Fluorescence Resonance Energy Transfer; Imiquimod; Mice; Mice, Inbred BALB C; Microsomes, Liver; Molecular Docking Simulation; Molecular Structure; Nuclear Receptor Subfamily 1, Group F, Member 3; Psoriasis; Quinolines; Structure-Activity Relationship; Sulfonamides; Th17 Cells

Topics: Administration, Cutaneous; Animals; Clobetasol; Disease Models, Animal; Drug Carriers; Drug Liberation; Female; Gels; Lipids; Male; Microscopy, Electron, Scanning; Nanoparticles; Particle Size; Psoriasis; Rats; Rats, Wistar; Rheology; Sebaceous Glands; Skin; Skin Absorption; Temperature

The goal of this study was to elucidate the anti-pruritic and anti-inflammatory efficacy of ruxolitinib cream in experimentally-induced dermatitis. Atopic dermatitis (AD), the most common chronic relapsing inflammatory skin disease, significantly impairs patients' quality of life, with pruritus being a common complaint. The sensation of itch results from the interplay between epidermal barrier dysfunction, upregulated immune signaling and the activation of the central nervous system. The Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway plays a central role in pro-inflammatory cytokine signaling in AD. Ruxolitinib cream is a potent and selective JAK1/2 inhibitor currently undergoing clinical evaluation in adults with mild-to-moderate AD (NCT03745638, NCT03920852 and NCT03745651). The efficacy of ruxolitinib cream was tested in murine models of acute and chronic dermatitis and was also characterized in an

Topics: Administration, Cutaneous; Animals; Betamethasone; Clobetasol; Cytokines; Dermatitis; Disease Models, Animal; Drug Eruptions; Drug Evaluation, Preclinical; Female; Fluorescein-5-isothiocyanate; Grooming; Humans; In Vitro Techniques; Interleukin-33; Janus Kinase Inhibitors; Lymphocyte Subsets; Mice; Mice, Inbred BALB C; Mice, Transgenic; Nitriles; Ointments; Organ Culture Techniques; Pruritus; Pyrazoles; Pyrimidines; Random Allocation; Signal Transduction; Skin; Specific Pathogen-Free Organisms; T-Lymphocytes, Helper-Inducer; Thymic Stromal Lymphopoietin; Transcriptome

Dillenia indica Linn. (Dilleniaceae) is traditionally used to treat skin inflammation.. This study evaluated the healing effect of Dillenia indica fruit extracts on induced psoriasis-like wounds in Wistar rats.. Extracts were standardized to betulinic acid, including an aqueous ethanolic extract (AEE), ethyl acetate extract (EAE) and petroleum ether extract. Effects against lipid peroxidation were assessed in vitro. Wounds were created at rat tails (n = 12). Topical treatments were applied once daily for 7 days (1 mL of AEE or EAE at 5 or 50 mg/mL). Maximal dose was defined by the extract solubility. A 10-fold lower dose was also tested. Positive and negative controls were treated with clobetasol (0.5 mg/mL) or excipient. Half of each group was euthanized for histology. The remaining animals were observed for 20 days for wound measurements.. Yields of AEE and EAE were 4.3 and 0.7%, respectively. Betulinic acid concentrations in AEE and EAE were 4.6 and 107.6 mg/g. Extracts neutralized lipid peroxidation in vitro at 0.02 μg/mL, accelerating healing at 50 mg/mL. Complete healing in mice treated with AEE occurred 16 days after wound induction. This time was 14 and 12 days in mice treated with EAE and clobetasol. Compared to orthokeratosis, parakeratosis was reduced by AEE (25%), EAE (45%) and clobetasol (55%). EAE caused superior protection against biomolecules oxidation of skin compared to AEE.. EAE exhibited activity closer to that of clobetasol. Betulinic acid may be an active constituent, which should be assessed in future studies.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Betulinic Acid; Biomarkers; Clobetasol; Dermatologic Agents; Dilleniaceae; Disease Models, Animal; Fruit; Lipid Peroxidation; Male; Malondialdehyde; Oxidative Stress; Pentacyclic Triterpenes; Phytotherapy; Plant Extracts; Plants, Medicinal; Protein Carbonylation; Psoriasis; Rats, Wistar; Skin; Solvents; Time Factors; Triterpenes; Ultraviolet Rays; Wound Healing

Topical corticosteroid and calcineurin inhibitor have similar therapeutic benefits in atopic dermatitis (AD), but the differences in therapeutic mechanisms of action of these agents against AD symptoms are not fully understood.. This study was performed to examine the different effects of topical betamethasone valerate (BMV), clobetasol propionate (CBP), and tacrolimus (TAC) on itch-related behavior and dermatitis in NC/Nga mice with AD-like symptoms.. AD-like dermatitis was induced in the dorsal skin of NC/Nga mice by repeated topical application of Dermatophagoides farinae body (Dfb) ointment twice weekly for three weeks. Mice with dermatitis scores over 5 were divided into five groups with equal dermatitis scores and treated with BMV, CBP, TAC, or Vaseline (Vas) once daily for two consecutive days, or were not treated (NT). Scratching behavior was analyzed using a SCLABA. After the second treatment, dermatitis showed significantly greater improvement in the CBP and TAC-treated groups than in the Vas-treated and NT groups. The numbers of scratching bouts were significantly lower in CBP and TAC-treated mice than in Vas-treated mice. TEWL was significantly lower in TAC-, but not in CBP-, treated mice than in Vas-treated mice. Immunohistochemical examination showed that BMV, CBP and TAC did not reduce the increased densities of epidermal protein gene product 9.5- and substance P-immunoreactive fibers. The numbers of dermal CD4-immunoreactive T cells were significantly lower in BMV and CBP-treated mice than in Vas-treated and NT mice. The numbers of dermal eosinophils were significantly lower in BMV, CBP and TAC-treated mice than in Vas-treated and NT mice, with CBP showing the strongest effect. CBP significantly reduced epidermal thickness compared with Vas and NT. There were no significant differences in the numbers of interleukin-31-immunoreactive cells and mast cells, or in expression of epidermal thymic stromal lymphopoietin among all five groups.. The therapeutic potency of TAC against AD-like symptoms, including pruritus, is equal to that of the corticosteroid CBP. Epidermal innervation of sensory nerves itself might not be related to the therapeutic effects of topical tacrolimus and corticosteroids in its early phase.

Topics: Administration, Topical; Adrenal Cortex Hormones; Animals; Betamethasone Valerate; Clobetasol; Cytokines; Dermatitis, Atopic; Dermatophagoides farinae; Disease Models, Animal; Emollients; Epidermis; Humans; Immunosuppressive Agents; Male; Mast Cells; Mice; Ointments; Petrolatum; Pruritus; Tacrolimus; Thymic Stromal Lymphopoietin; Treatment Outcome; Ubiquitin Thiolesterase

Neuromyelitis optica (NMO) is an inflammatory demyelinating disease of the central nervous system that can produce marked neurological deficit. Current NMO therapies include immunosuppressants, plasma exchange and B-cell depletion. Here, we evaluated 14 potential remyelinating drugs emerging from prior small molecule screens done to identify drugs for repurposing in multiple sclerosis and other demyelinating neurological diseases. Compounds were initially evaluated in oligodendrocyte precursor cell (OPC) and cerebellar slice cultures, and then in a mouse model of NMO produced by intracerebral injection of anti-AQP4 autoantibody (AQP4-IgG) and human complement characterized by demyelination with minimal axonal damage. The FDA-approved drug clobetasol promoted differentiation in OPC cultures and remyelination in cerebellar slice cultures and in mice. Intraperitoneal administration of 2 mg/kg/day clobetasol reduced myelin loss by ~60 %, even when clobetasol was administered after demyelination occurred. Clobetasol increased the number of mature oligodendrocytes within lesions without significantly altering initial astrocyte damage or inflammation. These results provide proof-of-concept for the potential utility of a remyelinating approach in the treatment of NMO.

Topics: Animals; Astrocytes; Cells, Cultured; Cerebellum; Clobetasol; Disease Models, Animal; Drug Evaluation, Preclinical; Male; Mice; Myelin Sheath; Neural Stem Cells; Neurogenesis; Neuromyelitis Optica; Neuroprotective Agents; Tissue Culture Techniques

Multiple sclerosis involves an aberrant autoimmune response and progressive failure of remyelination in the central nervous system. Prevention of neural degeneration and subsequent disability requires remyelination through the generation of new oligodendrocytes, but current treatments exclusively target the immune system. Oligodendrocyte progenitor cells are stem cells in the central nervous system and the principal source of myelinating oligodendrocytes. These cells are abundant in demyelinated regions of patients with multiple sclerosis, yet fail to differentiate, thereby representing a cellular target for pharmacological intervention. To discover therapeutic compounds for enhancing myelination from endogenous oligodendrocyte progenitor cells, we screened a library of bioactive small molecules on mouse pluripotent epiblast stem-cell-derived oligodendrocyte progenitor cells. Here we show seven drugs function at nanomolar doses selectively to enhance the generation of mature oligodendrocytes from progenitor cells in vitro. Two drugs, miconazole and clobetasol, are effective in promoting precocious myelination in organotypic cerebellar slice cultures, and in vivo in early postnatal mouse pups. Systemic delivery of each of the two drugs significantly increases the number of new oligodendrocytes and enhances remyelination in a lysolecithin-induced mouse model of focal demyelination. Administering each of the two drugs at the peak of disease in an experimental autoimmune encephalomyelitis mouse model of chronic progressive multiple sclerosis results in striking reversal of disease severity. Immune response assays show that miconazole functions directly as a remyelinating drug with no effect on the immune system, whereas clobetasol is a potent immunosuppressant as well as a remyelinating agent. Mechanistic studies show that miconazole and clobetasol function in oligodendrocyte progenitor cells through mitogen-activated protein kinase and glucocorticoid receptor signalling, respectively. Furthermore, both drugs enhance the generation of human oligodendrocytes from human oligodendrocyte progenitor cells in vitro. Collectively, our results provide a rationale for testing miconazole and clobetasol, or structurally modified derivatives, to enhance remyelination in patients.

Topics: Animals; Cell Differentiation; Cerebellum; Clobetasol; Demyelinating Diseases; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Germ Layers; Humans; Lysophosphatidylcholines; Male; MAP Kinase Signaling System; Mice; Miconazole; Mitogen-Activated Protein Kinases; Multiple Sclerosis; Myelin Sheath; Oligodendroglia; Phenotype; Pluripotent Stem Cells; Receptors, Glucocorticoid; Regeneration; Tissue Culture Techniques

The interleukin-23/interleukin 17A (IL-23/IL-17A) cytokine axis plays a critical role in the pathogenesis of psoriasis. In this study, we report the effects of topical calcipotriol, camptothecin, clobetasol and tazarotene on the treatment of imiquimod (IMQ)-induced psoriasis-like inflammation, the development of which is dependent on the IL-23/IL-17A axis. IMQ-induced epidermal hyperplasia and inflammation in the BALB/c mouse ear were significantly inhibited following clobetasol treatment but not calcipotriol, camptothecin or tazarotene treatments. Real-time polymerase chain reaction showed that the mRNA levels of IL-17A, IL-17F, IL-22, IL-1β, IL-6 and TNF-α in ear skin were significantly decreased by clobetasol. In addition, we observed that calcipotriol, camptothecin and tazarotene failed to show any inhibitory effects on the IL-23/IL-17A/IL-22 axis. We also found that clobetasol treatment inhibited the proliferation of γδ T cells and C-C chemokine receptor type 6 (CCR6) expression induced by IMQ. Calcipotriol, camptothecin and tazarotene not only failed to inhibit this proliferation but also enhanced retinoic acid-related orphan receptor γ (RORγ) expression in IMQ-induced psoriasis-like inflammation. In conclusion, we suggest that clobetasol induces the relief of IMQ-induced psoriasis-like inflammation in a mouse model but that calcipotriol, camptothecin and tazarotene cannot. Therefore, we suggest that more in-depth studies on pharmacological effects of tazarotene, camptothecin and calcipotriol should be carried out.

Topics: Adjuvants, Immunologic; Administration, Topical; Aminoquinolines; Animals; Anti-Inflammatory Agents; Calcitriol; Camptothecin; Clobetasol; Dermatologic Agents; Disease Models, Animal; Humans; Imiquimod; Mice; Mice, Inbred BALB C; Nicotinic Acids; Psoriasis; Topoisomerase I Inhibitors

Topical glucocorticoid (GC) therapy has been successfully used in the treatment of several common cutaneous diseases in clinical practice for a long time, and skin atrophy is one of the most typical cutaneous side effects of this therapy. The aim of this study was to evaluate the potential of noninvasive fluorescence spectroscopy (FS) technique in the detection and classification of GC-induced skin atrophy. A total of 20 male Wistar rats were used in the experimental protocol under controlled environmental conditions and with free access to food. One group received topical application of clobetasol propionate 0.05% for 14 days to induce cutaneous atrophy (atrophic group) and the other (control) group received only vehicle application following the same protocol and schedule. Histological analyses and FS measurements with laser excitation at both 532 nm and 408 nm were obtained on days 1 and 15. The FS results were classified as "normal" or "atrophic" according by histological analysis. Fluorescence spectra obtained with excitation at 408 nm allowed a clear distinction between the control and atrophic groups, and were more informative than the those obtained at 532 nm. Our results reveal that, if correctly applied, FS allows noninvasive evaluation of corticosteroid-induced skin atrophy, and thus represents an important step towards better monitoring of undesirable side effects of cutaneous therapy.

Topics: Animals; Atrophy; Clobetasol; Disease Models, Animal; Glucocorticoids; Male; Rats; Rats, Wistar; Skin; Spectrometry, Fluorescence

Although topical glucocorticoids (GCs) show potent anti-inflammatory activity in inflamed skin, they can also exert numerous harmful effects on epidermal structure and function. In contrast, topical applications of ligands of peroxisome proliferator-activated receptor-α (PPARα) not only reduce inflammation but also improve cutaneous barrier homeostasis. Therefore, we examined whether sequential topical GCs followed by topical Wy14643 (a ligand of PPARα) might be more effective than either alone for atopic dermatitis (AD) in a hapten (oxazolone (Ox))-induced murine model with multiple features of AD (Ox-AD). Despite expected anti-inflammatory benefits, topical GC alone induced (i) epidermal thinning; (ii) reduced expression of involucrin, loricrin, and filaggrin; and (iii) allowed outside-to-inside penetration of an epicutaneous tracer. Although Wy14643 alone yielded significant therapeutic benefits in mice with mild or moderate Ox-AD, it was less effective in severe Ox-AD. Yet, topical application of Wy14643 after GC was not only significantly effective comparable with GC alone, but it also prevented GC-induced structural and functional abnormalities in permeability barrier homeostasis. Moreover, rebound flares were largely absent after sequential treatment with GC and Wy14643. Together, these results show that GC and PPARα ligand therapy together is not only effective but also prevents development of GC-induced side effects, including rebound flares, in murine AD.

Topics: Adjuvants, Immunologic; Animals; Clobetasol; Dermatitis, Atopic; Disease Models, Animal; Drug Synergism; Drug Therapy, Combination; Epidermis; Female; Glucocorticoids; Haptens; Mice; Mice, Hairless; Naphthols; Oxazolone; Peroxisome Proliferators; PPAR alpha; Pyrimidines; Secondary Prevention; Triazines

Glucocorticoids (GCs) belong to the most widely used anti-inflammatory drugs at all. However, their topical use is limited by their side effect potential, with skin atrophy being the most prominent one. Thus, determining the atrophogenic potential of novel compounds is of importance for drug development. Currently, the most frequently performed model in the base and pharmaceutical research is the hr/hr rat model of GC-induced skin atrophy that lasts for 19 days. In this study, we analysed statistically skin atrophy experiments retrospectively to ascertain (i) the earliest time-point, at which skin atrophy is significantly induced; and (ii) whether the differences between the GC treatment groups change until the end of the experiment. We show here that the treatment duration of rat skin atrophy models might be reduced to 5 days for economical and ethical reasons.

Topics: Animals; Anti-Inflammatory Agents; Atrophy; Clobetasol; Disease Models, Animal; Glucocorticoids; Linear Models; Methylprednisolone; Mometasone Furoate; Pregnadienediols; Rats; Rats, Hairless; Skin; Time Factors

The active form of vitamin D(3) , calcitriol, is widely used for the treatment of psoriasis, with or without topical corticosteroids. Topical corticosteroids are known to disrupt permeability and antimicrobial barriers, even with short-term use. Yet, the effect of topical calcitriol on epidermal permeability and antimicrobial barriers disrupted by topical corticosteroids has not been determined.. To examine the effect of calcitriol on epidermal permeability and antimicrobial barrier function that has been impaired by corticosteroids, as well as to elucidate the mechanism of improvement.. Topical calcitriol or the control vehicle was applied to each flank of hairless mice 20 min after treatment with topical clobetasol propionate and repeated every 12 h for 3·5 days. Barrier function assessment, Nile red staining, electron microscopy, immunohistochemistry, Western blotting, and real-time reverse transcriptase-polymerase chain reaction studies were performed 24 h after the last application.. Epidermis co-treated with topical calcitriol showed an improvement of stratum corneum integrity and barrier recovery, more intense fluorescence staining with Nile red, and an increase in lamellar body (LB) maturation and density, as well as upregulation of major epidermal lipid synthesis-related enzymes (3-hydroxy-3-methylglutaryl-CoA, serine-palmitoyl transferase and fatty acid synthase), mouse beta-defensin 3, cathelin-related antimicrobial peptide and vitamin D receptor.. We found that topical calcitriol restored both the epidermal permeability and antimicrobial barrier that had been impaired by corticosteroids. This restoration was mediated by both an activation of the cutaneous vitamin D pathway and an increase of epidermal lipids and antimicrobial peptides, promoted by the formation of the LB and the activity of epidermal lipid synthesis-related enzymes.

Topics: Administration, Topical; Animals; Blotting, Western; Calcitriol; Calcium Channel Agonists; Clobetasol; Disease Models, Animal; Enzymes; Epidermis; Female; Glucocorticoids; Immunohistochemistry; Mice; Mice, Hairless; Microscopy, Electron; Oxazines; Permeability; Polymerase Chain Reaction; Skin Absorption; Up-Regulation

This study investigated the effect of potent intracanal corticosteroids on periodontal healing of replanted avulsed teeth and evaluated the systemic absorption of these corticosteroids. Sixty-seven extracted dog premolar roots were randomly assigned to one of the following groups: groups 1-3 filled with gutta-percha and replanted immediately and after 40 and 60 minutes, respectively; groups 4 and 5 filled with 0.05% clobetasol; and groups 6 and 7 filled with 0.05 % fluocinonide. Groups 4 and 6 were replanted after 40 minutes and groups 5 and 7 after 60 minutes. After 4 months, roots were evaluated histologically for signs of periodontal healing. Roots treated with clobetasol and fluocinonide healed more favorably than roots filled with gutta-percha and were different from each other at 60 minutes. No change in the systemic corticosteroid blood concentration was observed in any group. Corticosteroids were efficacious in the beagle model as intracanal medicaments for promoting favorable postavulsion periodontal healing.

Topics: Animals; Bicuspid; Clobetasol; Desiccation; Disease Models, Animal; Dogs; Fluocinonide; Glass Ionomer Cements; Glucocorticoids; Gutta-Percha; Periodontal Ligament; Random Allocation; Root Canal Filling Materials; Root Resorption; Time Factors; Tooth Avulsion; Tooth Replantation; Tooth Root; Wound Healing; Zinc Oxide-Eugenol Cement

Skin atrophy is one of the most frequent side-effects of the topical glucocorticoid. Skin barrier impairment has also been reported as a steroid-induced side effect. Although there have been various studies on preventing or minimizing this atrophogenic effect, little has been reported about preventing barrier impairment. This study was performed to determine the effects of a multilamellar emulsion (MLE) that had a well-ordered lamellar structure on the steroid-induced barrier impairment and epidermal atrophy. To confirm these effects of MLE, 0.05% clobetasol-17-propionate (CP) and 0.05% clobetasol-17-propionate in MLE (MLE/CP) were topically applied to both flanks of hairless mice for 9 days. The topically applied CP induced a significant impairment of the epidermal permeability barrier, and MLE/CP also did not have a preventive effect on this change. However, skinfold thickness studies and histological studies showed that MLE/CP significantly reduced the steroid-induced atrophy. The topical application of MLE/CP was also shown to have a preventive effect on the steroid-induced increase of the stratum corneum (SC) surface pH. In addition, the electron microscopic findings showed relatively well-conserved lamellar bilayers in the skin treated with MLE, as compared to CP only. The results showed that the topical application of MLE immediately after CP treatment prevented the glucocorticoid-induced transepidermal water loss values increase. Light microscopy measurements showed that the skin treated with MLE immediately after CP treatment for 1 week had a slightly lower decline of skin thickness than did the CP-treated skin. These results suggest that MLE should be effective for preventing glucocorticoid-induced epidermal atrophy and for repairing the barrier impairment.

Topics: Administration, Topical; Animals; Atrophy; Biopsy, Needle; Clobetasol; Disease Models, Animal; Emulsions; Epidermis; Female; Glucocorticoids; Immunochemistry; Male; Mice; Mice, Hairless; Permeability; Probability; Reference Values; Risk Factors; Sensitivity and Specificity; Skin Absorption; Skin Diseases

The permeabilities of normal human and normal, inflamed, or corticosteroid (CS) pretreated skin of young domestic pigs for pimecrolimus and tacrolimus were compared in vitro, using Franz-type diffusion cells. The test articles were either used as 1.0% solutions or as the marketed formulations (Elidel 1% cream, Protopic 0.1%, and 0.03% ointment). In normal human skin, the permeation rate of pimecrolimus from the 1% cream was about sixfold lower than that of tacrolimus from 0.1% ointment and by a factor of 4.3 lower compared with tacrolimus from Protopic 0.03%. In pigs, sodium laurylsulfate-induced irritant contact dermatitis resulted in significantly faster skin permeation of both drugs from applied solutions. The permeation rate for pimecrolimus was lower than that for tacrolimus. Thus, at 24 h, pimecrolimus concentrations in the receptor fluid were 2.8-fold lower than the tacrolimus levels. Compared with normal porcine skin, permeation of drugs through hydrocortisone (1.0%)-, mometasone (0.1%)-, or clobetasol-17-butyrate (0.05%)-pretreated skin was increased by factors of 3.6 (pimecrolimus, applied as 1% cream) and 1.7 (tacrolimus, applied as 0.1% ointment). In normal pig skin, the permeation rate of tacrolimus was found to be 11.2 times higher than that of pimecrolimus and 3.5- to 7.1-fold higher in CS-pretreated skin, independent of the potency of the CSs. The present in vitro data suggest that in patients with acute skin inflammation or after therapy with topical CSs, percutaneous absorption and, as a consequence, systemic drug exposure will be lower with Elidel 1% cream as compared with Protopic 0.1% and 0.03% ointment.

Topics: Adrenal Cortex Hormones; Animals; Anti-Inflammatory Agents, Non-Steroidal; Clobetasol; Dermatologic Agents; Disease Models, Animal; Humans; Hydrocortisone; Inflammation; Models, Animal; Mometasone Furoate; Pregnadienediols; Skin; Skin Physiological Phenomena; Swine; Tacrolimus

Previous studies have shown the antidotal efficacy of topical iodine at 15 and 30 min post-exposure to sulfur mustard (SM). Here we demonstrate efficacy at longer intervals (20, 30, 45, and 60 min, respectively, for data) using an improved topical povidone-iodine preparation termed N66, which contains steroidal and non-steroidal anti-inflammatory agents. In the mouse, N66 reduced severity of ear edema by 43, 47, 44, and 36%; ear epidermal ulceration by 74, 58, 45, and 58%; and epidermal necrosis by 54, 34, 26, and 31% at the respective time points. A similar effect was observed with encrustation. The healing marker, grade of acanthotic area, showed dramatic increases of 39.6-, 25.3-, 20.9-, and 22-fold. Severity of the dermal parameters, acute inflammation and dermal necrosis, was reduced by 63, 34, 34, and 38% and 80, 54, 54, and 59%, respectively. In guinea pig skin, topical treatment with N66 45 min post-exposure reduced the SM-induced ulceration area by 75%. The histological parameters subepidermal microblister formation, epidermal ulceration, epidermal necrosis, and encrustation were reduced by 63, 61, 41, and 41%, respectively. The healing marker, grade of acanthotic area, was elevated by 73%. N66 induced a statistically significant reduction in two dermal markers for tissue damage: acute inflammation (33%) and dermal necrosis (48%). Reduced skin damage was also observed in areas adjacent the treated sites. The pharmacologically active components of N66 showed additive effect. These findings suggest that the povidone-iodine preparation combined with anti-inflammatory agents functions as a potent antidote against skin lesions induced by SM at relatively long intervals between exposure and treatment.

Topics: Administration, Topical; Animals; Anti-Inflammatory Agents; Chemical Warfare Agents; Clobetasol; Disease Models, Animal; Drug Combinations; Ear Diseases; Edema; Guinea Pigs; Male; Mice; Mice, Inbred ICR; Mustard Gas; Piroxicam; Povidone-Iodine; Protective Agents; Skin; Skin Diseases; Skin Irritancy Tests; Time Factors

To understand further the possible clinical effects of tinidazole ointment at relatively high concentration (2%) for atopic dermatitis (AD), we performed a comparative study with readily available topical corticosteroids, clobetasol propionate (0.005 or 0.05%) and hydrocorotisone butyrate (0.1%) (hereafter referred as clobetasol and hydrocortisone, respectively), on inflammatory dermatitis in mice. We also observed the effects of combined application of tinidazole with clobetasol (0.005%, one tenth of the clinical use) in comparison with tinidazole itself, clobetasol (0.05%) or hydrocortisone (0.1%) on the animal model. All ointments suppressed inflammatory dermatitis induced by trinitrochlorobenzen (TNCB) or oxazolone. The rank order of the potency to suppress the ear edema was clobetasol (0.05%), tinidazole (2%) with clobetasol (0.005%) > clobetasol (0.005%) > tinidazole (2%) in TNCB-induced dermatitis, and hydrocortisone (0.1%), clobetasol (0.05%) > tinidazole (2%), tinidazole with clobetasol (0.005%) > clobetasol (0.005%) in case of oxazolone-induced dermatitis. We confirmed that tinidazole (2%) suppresses immediate and late phase reactions in mice passively sensitized with anti-DNP IgE Mab. In addition, tinidazole (2%) was much more potent than hydrocortisone (0.1%) in suppressing the amount of scratching, presumably due to itching, in passively sensitized mice. These results indicate that the advantage of using ointments of tinidazole would be that it has stronger anti-itching effects than corticosteroids.

Topics: Adjuvants, Immunologic; Administration, Topical; Animals; Antibodies, Monoclonal; Clobetasol; Dermatitis, Atopic; Dinitrophenols; Disease Models, Animal; Drug Therapy, Combination; Hydrocortisone; Immunoglobulin E; Male; Mice; Mice, Inbred Strains; Ointments; Ovalbumin; Oxazolone; Picryl Chloride; Tinidazole

Using the isolated perfused bovine udder as an in-vitro model of skin inflammation, the effects of topically administered arachidonic acid on prostaglandin and leukotriene synthesis have been shown previously. In this study, the effects of indometacin (indomethacin) and clobetasol-17-propionate (administered topically) as well as flunixin meglumine and meloxicam (administered via the perfusion fluid) have been studied. Compared with controls, arachidonic acid caused a significant increase in the dermal prostaglandin E2 (PGE2) and peptidoleukotriene (LTC4/D4/E4) concentration. Topical treatment with indometacin (1.6 mg cm(-2)) and clobetasol-17-propionate (90 microg cm(-2)), which were administered 60 min before arachidonic acid administration, inhibited the inflammatory reaction. Flunixin meglumine (1 microg mL(-1) perfusion fluid) was administered 30 min after and meloxicam (3 microg mL(-1) perfusion fluid) was administered 60 min before arachidonic acid application. Three hours after arachidonic acid administration, a significant inhibition of PGE2 synthesis was induced by flunixin. In contrast, meloxicam showed only a slight effect. The effect of flunixin was comparable with in-vivo results. It is known from animal studies that anti-inflammatory effects of meloxicam are obvious within up to 6 h after treatment. Therefore, the incomplete effect of meloxicam may be explained pharmacokinetically. In conclusion, the described in-vitro model seems to be suitable for studies of pharmacological effects on eicosanoid synthesis in the skin.

Topics: Administration, Topical; Animals; Anti-Inflammatory Agents; Anti-Inflammatory Agents, Non-Steroidal; Arachidonic Acid; Cattle; Clobetasol; Clonixin; Culture Techniques; Dinoprostone; Disease Models, Animal; Eicosanoids; Female; Glucocorticoids; Indomethacin; Mammary Glands, Animal; Meloxicam; Skin Physiological Phenomena; Thiazines; Thiazoles

In an earlier study we showed that tretinoin could prevent corticosteroid-induced skin atrophy in hairless mice. In this study, we examined the histochemical, biochemical, and immunochemical changes that accompanied the atrophy and its prevention. Mice were treated dorsally for 3 weeks in the morning and afternoon (AM:PM) as follows: 1) vehicle:vehicle, 2) steroid:vehicle, 3) steroid:tretinoin. Tretinoin concentration was 0.05% in an ethanol:propylene glycol vehicle. The steroid was clobetasol propionate (0.05%). The normally sparse dermal glycosaminoglycans were further reduced by steroid:vehicle treatment and increased to greater than vehicle:vehicle amounts by steroid:retinoid. Mast cells were similarly affected. Biochemical quantification of glycosaminoglycans confirmed the histochemical findings. Collagen, non-collagenous protein, and total protein content were reduced by the steroid. The latter two were returned to more normal levels by tretinoin whereas with collagen there was only a trend toward normal levels. Fibronectin, which was increased by the steroid:vehicle treatment, was reduced to more normal levels by steroid:tretinoin. We conclude that tretinoin has the ability to prevent the major steroid-induced biomechanical changes in hairless mouse dermal connective tissue that contribute to atrophy.

Topics: Adrenal Cortex Hormones; Animals; Atrophy; Clobetasol; Collagen; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Fibronectins; Glycosaminoglycans; Histocytochemistry; Image Processing, Computer-Assisted; Immunohistochemistry; Iron; Mast Cells; Mice; Mice, Hairless; Retinoids; Skin; Tolonium Chloride; Tretinoin

The immunosuppressive macrolide antibiotics FK 506 and rapamycin were tested for topical activity in experimental allergic contact dermatitis of farm pigs. This species was used because pig skin, in comparison to rodent skin, resembles human skin more closely. For comparison, cyclosporine A (CyA), which is orally but not topically active in patients with skin disease, dexamethasone, and clobetasol propionate were used. Treatment was performed twice, 30 min and 6 h after elicitation of challenge reaction. Topical application of 0.4 to 0.04% FK 506 caused a pronounced inhibition of inflammatory skin reactions of hypersensitivity to dinitrofluorobenzene. The treatment response was similar to the activity of 0.13% clobetasole. Dexamethasone (1.2%) was less active than clobetasol. In contrast, rapamycin and CyA were inactive at concentrations of 1.2 and 10%, respectively. Because the pig data on corticosteroids and cyclosporine A are in agreement with clinical findings, these studies indicate that immunosuppressive macrolides of the type FK 506 may be useful drugs for the topical treatment of human skin diseases that respond to local corticosteroids and oral treatment with cyclosporine A.

Topics: Animals; Clobetasol; Cyclosporine; Dermatitis, Contact; Dexamethasone; Disease Models, Animal; Female; Molecular Structure; Polyenes; Sirolimus; Swine; Tacrolimus

A mouse model for assessment of the anti-inflammatory effect of topical glucocorticoids is described. Mice, sensitized to picryl chloride, had both ears painted with the sensitizer followed 2 hours later by the application of the topical steroid to one ear and the corresponding vehicle to the other. The contact sensitivity reaction, measured by swelling of the ear, was recorded 24 hours later. Dilutions of the steroid formulations inhibited the ear swelling in a manner related to dose-response. Suppression of the contact sensitivity reaction of the vehicle-treated ear as well was regarded as a systemic effect of the glucocorticoid. There seems to be a good correlation between the efficacy of the topical steroids assessed in this mouse model and the vasoconstrictor test on intact human skin.

Topics: Administration, Topical; Animals; Anti-Inflammatory Agents; Biological Assay; Clobetasol; Dermatitis, Contact; Disease Models, Animal; Dose-Response Relationship, Immunologic; Ear, External; Glucocorticoids; Hydrocortisone; Male; Mice; Mice, Inbred C57BL; Mice, Inbred CBA; Picryl Chloride

Since the great majority of patients possess immune response to herpes simplex virus (HSV), the influence of a topical anti-inflammatory corticosteroid (0.1% clobetasone butyrate) on ulcerative herpetic keratitis was studied in rabbits with a previous HSV skin infection (immunised) and compared with that in normal rabbits. Corticosteroid treatment had a much greater ulceration-exacerbating effect in immunised than in normal animals. On day 7 the mean area of ulceration in immunised rabbits were 3 times greater in treated eyes. 0.01% clobetasone butyrate treatment had less effect on immunised rabbits; 0.001% had no effect. It is concluded that the immunised rabbit provides a useful experimental model for studying the relationship between concentration of topical anti-inflammatory agents and enhancement of herpetic ulceration.

Topics: Animals; Anti-Inflammatory Agents; Betamethasone; Clobetasol; Corneal Ulcer; Disease Models, Animal; Dose-Response Relationship, Drug; Immunization; Keratitis, Dendritic; Rabbits