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1.
An. bras. dermatol ; 96(4): 454-457, July-Aug. 2021. graf
Article in English | LILACS | ID: biblio-1285099

ABSTRACT

Abstract Currently, pagetoid dyskeratosis is believed to involve an accelerated keratinization process, possibly induced by mechanical trauma. It represents, in almost its totality, incidental histological findings of specific cells, except when it occurs in the hands, where it usually occurs simultaneously with skin lesions and local dyschromia. These are large, rounded keratinocytes, with pale cytoplasm and a pyknotic nucleus surrounded by a clear halo, which can be easily mistaken by other skin diseases. Its etiology is not completely elucidated, and the correct identification of this entity can be of great importance in the differential diagnosis of skin disorders and the understanding of the keratinization process of the epidermis.


Subject(s)
Skin Neoplasms , Carcinoma in Situ , Paget Disease, Extramammary , Keratinocytes , Epidermis
3.
Braz. j. med. biol. res ; 54(9): e10700, 2021. tab, graf
Article in English | LILACS | ID: biblio-1249336

ABSTRACT

It was previously demonstrated that the methanol fraction of Sideroxylon obtusifolium (MFSOL) promoted anti-inflammatory and healing activity in excisional wounds. Thus, the present work investigated the healing effects of MFSOL on human keratinocyte cells (HaCaT) and experimental burn model injuries. HaCaT cells were used to study MFSOL's effect on cell migration and proliferation rates. Female Swiss mice were subjected to a second-degree superficial burn protocol and divided into four treatment groups: Vehicle, 1.0% silver sulfadiazine, and 0.5 or 1.0% MFSOL Cream (CrMFSOL). Samples were collected to quantify the inflammatory mediators, and histological analyses were performed after 3, 7, and 14 days. The results showed that MFSOL (50 μg/mL) stimulated HaCaT cells by increasing proliferation and migration rates. Moreover, 0.5% CrMFSOL attenuated myeloperoxidase (MPO) activity and also stimulated the release of interleukin (IL)-1β and IL-10 after 3 days of treatment. CrMFSOL (0.5%) also enhanced wound contraction, promoted improvement of tissue remodeling, and increased collagen production after 7 days and VEGF release after 14 days. Therefore, MFSOL stimulated human keratinocyte (HaCaT) cells and improved wound healing via modulation of inflammatory mediators of burn injuries.


Subject(s)
Humans , Female , Burns/drug therapy , Sapotaceae , Proline , Keratinocytes , Plant Leaves , Methanol
4.
Int. j. morphol ; 38(6): 1700-1706, Dec. 2020. graf
Article in English | LILACS | ID: biblio-1134501

ABSTRACT

SUMMARY: The treatment of chronic wounds has become a public health issue in recent years mainly due to comorbidities associated with an older population and bacterial resistance. Honey has emerged as an alternative treatment for chronic wounds but lack of knowledge of its mechanism of actionin the treated tissue and low quality of evidence in clinical triads has distanced the medical community from honey as a possible treatment. One of the main processes that is altered in chronic wounds is re-epithelialization mediated by keratinocytes, where proliferation and migration processes are altered. Markers of proliferation, migration and activation of keratinocytes, such as adhesion molecules, growth factors, membrane receptors, signal translating proteins, transcription factors, microRNAs, among others are deregulated in this process. In general, honeys from different floral origins have a positive effect on markers of proliferation and migration in keratinocytes. In conclusion there are still few studies that focus on the molecular action of honey in keratinocytes and fail to report details on the honey used not allowing to achieve the same results.


RESUMEN: El tratamiento de heridas crónicas (HC) se ha vuelto un tema de salud pública en los últimos años, principalmente debido a comorbilidades asociadas a una población de mayor edad y a la resistencia bacteriana. La miel ha surgido como un tratamiento alternativo para HC pero la falta de conocimiento de su mecanismo de acción en el tejido tratado y de la baja calidad de la evidencia en triadas clínicas, ha distanciado a la comunidad médica de la miel como posible tratamiento. Uno de los principales procesos que se ve alterado en las HC es la re-epitelización mediada por queratinocitos, donde se ven alterados los procesos de proliferación y migración. Marcadores de proliferación, migración y activación de queratinocitos, como moléculas de adhesión, factores de crecimiento, receptores de membrana, proteínas traductores de señales, factores de transcripción, microARNs, entre otras, se ven desreguladas en éste proceso. De manera general las mieles de diferentes orígenes florales tienen un efecto positivo en marcadores de proliferación y migración en queratinocitos. En conclusión aún existen pocos estudios que se enfoquen en la acción molecular de la miel en queratinocitos y los pocos que existen fallan en la entrega de información en relación a la miel utilizada que pueda hacer reproducibles los resultados.


Subject(s)
Wound Healing/physiology , Keratinocytes/physiology , Re-Epithelialization/physiology , Honey , Wound Healing/genetics , MicroRNAs/physiology , MicroRNAs/genetics , Re-Epithelialization/genetics
5.
Gac. méd. Méx ; 156(5): 426-431, sep.-oct. 2020. tab, graf
Article in Spanish | LILACS | ID: biblio-1249941

ABSTRACT

Resumen Introducción: El jabón para el aseo cutáneo es de empleo común entre la población, sin embargo, es posible que cause daño a las células de la piel y modifique la barrera cutánea. Objetivo: Determinar el efecto citotóxico de los jabones en queratinocitos cultivados in vitro y correlacionarlo con la irritación clínica. Método: Se realizó una encuesta para conocer los jabones comerciales más utilizados y su cantidad; posteriormente, se evaluó su citotoxicidad en cultivos de queratinocitos humanos mediante el método de resazurina. Los jabones con mayor y menor citotoxicidad se aplicaron en piel de voluntarios sanos para evaluar su efecto en la barrera cutánea mediante ensayos de colorimetría y pérdida transepidérmica de agua. Resultados: De los jabones analizados, 37 % demostró ser tóxico para los queratinocitos in vitro. El jabón con mayor toxicidad indujo el mayor índice de eritema y pérdida transepidérmica de agua, en comparación con el jabón menos tóxico y el vehículo empleado como solución control. Conclusión: Los jabones comercializados para el aseo cutáneo pueden incluir ingredientes químicos que dañan los queratinocitos humanos y causan irritación subclínica de la barrera cutánea. Su utilización puede agravar dermatosis preexistentes, generar dermatitis xerósica o de contacto irritativa y causar atrofia y dermatoporosis.


Abstract Introduction: The use of soap for skin cleansing is common among the population. However, it is possible that it causes damage to skin cells and disrupts the skin barrier. Objective: To determine the cytotoxic effect of soaps on in vitro-cultured keratinocytes and to correlate it with clinical irritation. Method: A survey was conducted to find out the most widely used commercial soaps and their number. Subsequently, their cytotoxicity was evaluated in human keratinocyte cultures using the resazurin assay. The soaps with the highest and lowest cytotoxicity were applied to the skin of healthy volunteers to assess their effect on the skin barrier using colorimetry and transepidermal water loss (TEWL) assays. Results: Of the analyzed soaps, 37 % were shown to be toxic to keratinocytes in vitro. The soap with the highest toxicity induced the highest rate of erythema and TEWL, in comparison with the least toxic soap and the vehicle used as the control solution. Conclusion: Soaps marketed for skin cleansing can contain chemical ingredients that damage human keratinocytes and cause skin barrier subclinical irritation. Their use can worsen preexisting dermatoses, generate xerotic or irritant contact dermatitis, and cause atrophy and dermatoporosis.


Subject(s)
Humans , Soaps/adverse effects , Keratinocytes/drug effects , Skin Irritancy Tests , Irritants/adverse effects , Skin/drug effects , Soaps/chemistry , Body Water , Cells, Cultured , Dermatitis, Irritant/etiology , Colorimetry , Erythema/chemically induced , Healthy Volunteers , Hydrogen-Ion Concentration
6.
Rev. argent. dermatol ; 101(1): 11-20, mar. 2020. graf
Article in Spanish | LILACS | ID: biblio-1092405

ABSTRACT

RESUMEN Presentamos un caso típico de Dermatosis Terra Firma-Forme en un adolescente sano de 13 años de edad, visto recientemente en el Servicio de Dermatología de nuestro hospital. Con este caso queremos mostrar las características clínicas de esta dermatosis que con frecuencia no es correctamente diagnosticada o bien pasa desapercibida durante años lo que origina preocupación y ansiedad en el paciente además de pruebas diagnósticas innecesarias. Su diagnóstico es clínico apoyado en la dermatoscopía y el tratamiento sencillo, presentando escasas recidivas.


SUMMARY We report a typical case of a Terra Firma-Forme Dermatosis in a 13-year-old healthy male recently seen in the Dermatology Department of our hospital. The aim of the authors is to show clinical features of this frequently misdiagnosed and underreported dermatosis causing concern and anxiety in the patient as well as unnecessary diagnostic tests. Its diagnosis is clinical supported by dermoscopy and its simple treatment presents few recurrences.


Subject(s)
Humans , Male , Adolescent , Skin Diseases/diagnosis , Hyperpigmentation/diagnosis , Keratinocytes/pathology , Hyperpigmentation/therapy , Diagnosis, Differential
7.
An. bras. dermatol ; 95(2): 244-246, Mar.-Apr. 2020. graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS | ID: biblio-1130856

ABSTRACT

Abstract The clinical diagnosis of Kyrle's disease may sometimes be challenging, due to the clinical similarity of lesions to other pruritic dermatosis. Although the dermoscopy is being increasingly used in daily practice, there is insufficient data in literature describing the dermoscopic patterns of Kyrle's disease, since only one report has been published to date. Herein we report our dermoscopic observation with additional diagnostic tips in a case who was diagnosed with Kyrle's disease histopathologically.


Subject(s)
Humans , Female , Dermoscopy/methods , Darier Disease/pathology , Darier Disease/diagnostic imaging , Biopsy , Keratinocytes/pathology , Reproducibility of Results , Middle Aged
8.
An. bras. dermatol ; 95(2): 238-240, Mar.-Apr. 2020. graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS | ID: biblio-1130853

ABSTRACT

Abstract Some epidermal alterations in measles has been described, such as keratinocytes apoptotic, parakeratosis, giant-cell formation, intranuclear and cytoplasmic inclusions, dyskeratosis, spongiosis, and intracellular edema. The authors report for the first time in human a case of measles with the presence of multinucleated giant cells in the hair follicle and dyskeratosis in acrosyringium.


Subject(s)
Humans , Male , Child , Hair Follicle/pathology , Epidermis/pathology , Measles/pathology , Parakeratosis/pathology , Biopsy , Giant Cells/pathology , Keratinocytes/pathology
9.
São José dos Campos; s.n; 2020. 119 p. il., graf., tab..
Thesis in Portuguese | LILACS, BBO | ID: biblio-1224659

ABSTRACT

O objetivo neste estudo foi produzir hidrogel de quitosana (CH) com PCL e fitoterápicos para uso preventivo de úlcera de pressão. Os hidrogéis de CH foram produzidos com glicerofosfato (GP) e com xantana (X), associados ao PCL e foram caracterizados por estereomicroscopio, intumescimento, molhabilidade e MEV. Posteriormente foram submetidos ao teste de viabilidade (MTT) com fibroblastos HFF-1 e queratinócitos HaCat. O hidrogel que apresentou melhor resultado foi escolhido para continuar na pesquisa. Posteriormente, extratos de Pfaffia panculata K, Juglans regia L, Rosmarinus officinalis L, Zingiber officinale, Própolis e Hamamelis foram colocados em contato com cepas de Staphylococcus aureus (S.a) (ATCC 6538), Streptococcus pyogenes (S.p) (ATCC 19615), Staphylococcus epidermidis (S.e) (ATCC 12228), Pseudomonas aeruginosa (P.a) (ATCC 15442), Escherichia coli (E.c) (ATCC 25922) e Klebsiella Pneumoniae (K.p) (ATCC 4352) na forma planctônica nos testes de CIM e CMM. Os dois melhores extratos fitoterápicos foram avaliados quanto ao sinergismo no teste checkerboard e posteriormente associados ao hidrogel anteriormente eleito. A seguir, o comportamento da HaCat e HFF-1 com os hidrogéis foi analisado por MTT, proteína total, ELISA, genotoxicidade e formação de biofilme monotípico com suspensões padronizadas (107 cel/mL) de S.a, S.e, S.p, P.a, E.c e K.p. Na caracterização e viabilidade o hidrogel CHX PCL apresentou os melhores resultados. Os extratos selecionados após CIM, CMM e checkerboard foram gengibre (G) e própolis (P). O extrato G se destacou na CIM com inibição de K. p e P. a. Os extratos de G e P demonstraram ação microbicida para K. p e P. a e somente o extrato P obteve ação microbicida para S. a na CMM. Houve ação aditiva dos extratos associados no checkerboard para S.p e ação aditiva e sinérgica para S. e. Os grupos de hidrogéis foram compostos por: quitosana xantana (CHX), CHX própolis (CHXP), CHX gengibre (CHXG) e CHX própolis e gengibre associados (CHXPG), todos associados ao PCL. Todos os hidrogéis demonstraram viabilidade celular acima de 70% do grupo controle, permitindo metabolismo celular observado na proteína total. Houve quantificação de IL-6 maior no grupo CHX nas duas linhagens de células enquanto a quantificação de IL-10 não exibiu diferença estatística entre os grupos. Todos os hidrogéis promoveram redução acentuada de biofilme de K.p e E.c. Os grupos CHX, CHXP e CHXG reduziram biofilme de S.e. O grupo CHXG reduziu biofilme de S.p. Para S.a e P.a o grupo CHXPG foi mais eficaz reduzindo biofilme. Concluímos que os hidrogéis apresentaram resultados satisfatórios e promissores, trazendo inovação por associação de biopolímeros e associação de extratos fitoterápicos pouco estudados. Os resultados positivos justificam a continuidade dos estudos com esse biomaterial(AU)


The aim of this study was to produce chitosan hydrogel (CH) with PCL and herbal medicines for preventive use of pressure ulcers. The CH hydrogels were produced with glycerophosphate (GP) and xanthan (X), associated with PCL and were characterized by stereomicroscope, swelling, wettability and SEM. Subsequently, they were submitted to a viability test (MTT) with HFF-1 fibroblasts and HaCat keratinocytes. The hydrogel that presented the best result was chosen to continue the research. Subsequently, extracts of Pfaffia panculata K, Juglans regia L, Rosmarinus officinalis L, Zingiber officinale, Propolis and Hamamelis were placed in contact with strains of Staphylococcus aureus (Sa) (ATCC 6538), Streptococcus pyogenes (Sp) (ATCC 19615), epidermidis (Se) (ATCC 12228), Pseudomonas aeruginosa (Pa) (ATCC 15442), Escherichia coli (Ec) (ATCC 25922) and Klebsiella Pneumoniae (Kp) (ATCC 4352) in planktonic form in CIM and CMM tests. The two best herbal extracts were evaluated for synergism in the checkerboard test and subsequently associated with the previously elected hydrogel. Next, the behavior of HaCat and HFF-1 with hydrogels was analyzed by MTT, total protein, ELISA, genotoxicity and monotypic biofilm formation with standardized suspensions (107 cel / mL) of Sa, Se, Sp, Pa, Ec and Kp In the characterization and viability the CHX PCL hydrogel presented the best results. The extracts selected after MIC, CMM and checkerboard were ginger (G) and propolis (P). The G extract stood out in the MIC with inhibition of K. p and P. a. The extracts of G and P showed microbicidal action for K. p and P. a and only the extract P obtained microbicidal action for S. a in CMM. There was an additive action of the associated extracts on the checkerboard for S.p and an additive and synergistic action for S. e. The hydrogel groups were composed of: xanthan chitosan (CHX), CHX propolis (CHXP), CHX ginger (CHXG) and CHX propolis and ginger associated (CHXPG), all associated with PCL. All hydrogels demonstrated cell viability above 70% of the control group, allowing cellular metabolism observed in the total protein. There was a greater quantification of IL-6 in the CHX group in the two cell lines while the quantification of IL-10 did not show statistical difference between the groups. All hydrogels promoted a marked reduction in the biofilm of K.p and E.c. The CHX, CHXP and CHXG groups reduced S.e biofilm. The CHXG group reduced S.p. For S.a and P.a, the CHXPG group was more effective in reducing biofilm. We conclude that the hydrogels presented satisfactory and promising results, bringing innovation through association of biopolymers and association of phytotherapic extracts little studied. The positive results justify the continuity of studies with this biomaterial(AU)


Subject(s)
Chitosan/therapeutic use , Keratinocytes/immunology , Biofilms , Hydrogels/administration & dosage , Phytotherapeutic Drugs , Nanofibers/adverse effects , Fibroblasts/microbiology
10.
Braz. oral res. (Online) ; 34: e033, 2020. graf
Article in English | LILACS | ID: biblio-1089391

ABSTRACT

Abstract The aim of our study was to isolate populations of keratinocyte stem cells based on the expression of cell surface markers and to investigate whether the culture could affect their phenotype. keratinocytes from human oral mucosa were sorted based on the expression of the epithelial stem cell markers p75NTR and CD71. We also examined the co-expression of other epithelial stem markers such as integrins β1 and α6 and their stem cell-like proprieties in in vitro assays. Three passages after being sorted by MACS, more than 93% of the p75NTR+ve cells lost the expression of p75NTR, while 5.46% of the p75NTR-ve gained it. Within the small population of the p75NTR+ve cells, 88% co-expressed other epithelial stem cell markers such as integrins β1 and α6, while only 28% of p75NTR-ve cells co-expressed these markers. These results were confirmed by sorting cells by FACS. Additionally, when double staining was used for sorting cells, 99% of the p75NTR+veCD71-ve and 33% of the p75NTR-veCD71+ve cells expressed both integrins, but just one week after culture, only 1.74% of the p75NTR+veCD71-ve cells still expressed p75NTR and only 0.32% still expressed CD71. Similar results were obtained when co-culturing p75NTR+ve and p75NTR-ve populations before analysis. Our results suggest that phenotype changes may be part of an intrinsic cellular mechanism to conserve levels of protein expression as they may found in the human body. In addition, in vitro culture may not offer ideal conditions for epithelial stem cell maintenance due to phenotype changes under standard culture conditions.


Subject(s)
Humans , Phenotype , Stem Cells/cytology , Keratinocytes/cytology , Cell Culture Techniques/methods , Epithelial Cells/cytology , Mouth Mucosa/cytology , Receptors, Transferrin/analysis , Biomarkers/analysis , Antigens, CD/analysis , Cell Separation/methods , Reproducibility of Results , Receptors, Nerve Growth Factor/analysis , Flow Cytometry/methods , Nerve Tissue Proteins/analysis
11.
Biol. Res ; 53: 48, 2020. graf
Article in English | LILACS | ID: biblio-1142415

ABSTRACT

BACKGROUND: Psoriasis is a common chronic inflammatory skin disease. Keratinocytes hyperproliferation and excessive inflammatory response contribute to psoriasis pathogenesis. The agents able to attenuate keratinocytes hyper-proliferation and excessive inflammatory response are considered to be potentially useful for psoriasis treatment. Daphnetin exhibits broad bioactivities including anti-proliferation and anti-inflammatory. This study aims to evaluate the anti-psoriatic potential of daphnetin in vitro and in vivo, and explore underlying mechanisms. METHODS: HaCaT keratinocytes was stimulated with the mixture of IL-17A, IL-22, oncostatin M, IL-1α, and TNF-α (M5) to establish psoriatic keratinocyte model in vitro. Cell viability was measured using Cell Counting Kit-8 (CCK-8). Quantitative Real-Time PCR (qRT-PCR) was performed to measure the mRNA levels of hyperproliferative marker gene keratin 6 (KRT6), differentiation marker gene keratin 1 (KRT1) and inflammatory factors IL-1ß, IL-6, IL-8, TNF-α, IL-23A and MCP-1. Western blotting was used to detect the protein levels of p65 and p-p65. Indirect immunofluorescence assay (IFA) was carried out to detect p65 nuclear translocation. Imiquimod (IMQ) was used to construct psoriasis-like mouse model. Psoriasis severity (erythema, scaling) was scored based on Psoriasis Area Severity Index (PASI). Hematoxylin and eosin (H&E) staining was performed to examine histological change in skin lesion. The expression of inflammatory factors including IL-6, TNF-α, IL-23A and IL-17A in skin lesion was measured by qRT-PCR. RESULTS: Daphnetin attenuated M5-induced hyperproliferation in HaCaT keratinocytes. M5 stimulation significantly upregulated mRNA levels of IL-1ß, IL-6, IL-8, TNF-α, IL-23A and MCP-1. However, daphnetin treatment partially attenuated the upregulation of those inflammatory cytokines. Daphnetin was found to be able to inhibit p65 phosphorylation and nuclear translocation in HaCaT keratinocytes. In addition, daphnetin significantly ameliorate the severity of skin lesion (erythema, scaling and epidermal thickness, inflammatory cell infiltration) in IMQ-induced psoriasis-like mouse model. Daphnetin treatment attenuated IMQ-induced upregulation of inflammatory cytokines including IL-6, IL-23A and IL-17A in skin lesion of mice. CONCLUSIONS: Daphnetin was able to attenuate proliferation and inflammatory response induced by M5 in HaCaT keratinocytes through suppression of NF-κB signaling pathway. Daphnetin could ameliorate the severity of skin lesion and improve inflammation status in IMQ-induced psoriasis-like mouse model. Daphnetin could be an attractive candidate for future development as an anti-psoriatic agent.


Subject(s)
Humans , Animals , Mice , Rabbits , Psoriasis/chemically induced , Psoriasis/drug therapy , Umbelliferones/pharmacology , Adjuvants, Immunologic/adverse effects , Imiquimod/adverse effects , Inflammation/drug therapy , Anti-Inflammatory Agents/pharmacology , Keratinocytes , Cell Proliferation , Mice, Inbred BALB C
12.
Annals of Dermatology ; : 122-129, 2020.
Article in English | WPRIM | ID: wpr-811086

ABSTRACT

BACKGROUND: Loss-of-function mutations in the filaggrin gene (FLG), which encodes an epidermal protein crucial for the formation of a functional skin barrier, have been identified as a major predisposing factor in the etiopathogenesis of atopic dermatitis (AD). Recent reports of relatively low frequencies of FLG-null mutations among specific ethnic groups with AD necessitated analysis of the epigenetic regulation which may control FLG expression without altering its DNA sequence.OBJECTIVE: The study aimed to identify DNA methylation-dependent regulation of FLG expression.METHODS: Quantitative polymerase chain reaction was performed to determine the restoration of FLG mRNA expression in normal human epidermal keratinocyte (NHEK) cells after treatment with epigenetic modulating agents. Bisulfite genomic sequencing and pyrosequencing analyses of the FLG promoter region were conducted to identify the citical CpG sites relevant to FLG expression. We performed small-scale pilot study for epidermal tissues obtained from Korean patients with severe AD.RESULTS: We here show that DNA methylation in the FLG with non-CpG island promoter is responsible for the transcriptional regulation of FLG in undifferentiated NHEK cells. The methylation frequencies in a single CpG site of the FLG promoter were significantly higher in lesional epidermis than those in matched nonlesional epidermis of subjects with severe AD.CONCLUSION: Our in vitro and clinical studies point to this unique CpG site as a potential DNA methylation marker of FLG, which can be a promising therapeutic target in the complications of filaggrin-related skin barrier dysfunction as well as in AD.


Subject(s)
Base Sequence , Causality , Dermatitis, Atopic , DNA , DNA Methylation , Epidermis , Epigenomics , Ethnic Groups , Gene Expression , Humans , In Vitro Techniques , Keratinocytes , Methylation , Pilot Projects , Polymerase Chain Reaction , Promoter Regions, Genetic , RNA, Messenger , Skin
13.
Article in Chinese | WPRIM | ID: wpr-826359

ABSTRACT

To observe the cell origin of N-methyl-D-aspartic acid(NMDA)receptor expression in skin after chronic ischemic pain modeling in rats and explore the role of NMDA receptor in type Ⅰ complex regional pain syndrome. Forty-two adult male Sprague-Dawley rats were randomly divided into five groups:sham operation group(=12),chronic post ischemia pain(CPIP)group(=12),CPIP+normal saline(NS)group(=6),CPIP+NMDA group(=6),and CPIP+MK801 group(=6).Six rats in the sham operation group and CPIP group were sacrificed under deep anesthesia one day after modeling.The plantar skin and L3-L5 spinal cord tissue were used for NR1(NMDA receptor)subunit immunofluorescence detection and for Western blotting of NR1,interleukin(IL)-1β,and tumor necrosis factor(TNF)-α.For the remaining rats,the mechanical withdrawal threshold(MWT)values on the 2nd,6th,10th and 14th day after ischemia were recorded,and the corresponding drugs were injected subcutaneously from the 6th day after ischemia.The skin and L3-L5 spinal cords were collected on the 14th day,and the same detection methods were applied. Compared with the sham operation group,the CPIP group had significantly higher expressions of NR1(1.708±0.064;=12.120, <0.001),IL-1β(2.575±0.305;=5.158, =0.003),and TNF-α(2.691±0.217;=7.786, <0.001)in the skin on the first day after modeling.After intervention with NMDA and MK801,the MWT value was [(20.37±0.95)g] in the CPIP+NS group,which was significantly higher than that in CPIP+NMDA group [(15.85±1.09)g;=10.920, <0.001] but significantly lower than that in CPIP+MK801 group[(22.95±0.96)g;=6.421, <0.001] 10 days after modeling.On the 14th day,compared with the MWT of the CPIP+NS group [(21.57±0.96)g],the CPIP+NMDA group had significantly decreased MWT value [(16.53±1.63)g;=12.190, <0.001],and the CPIP+MK801 group had significantly increased MWT value [(23.27±1.28)g;=4.094, =0.025].Compared with the sham operation group,the CPIP group had significantly increased NR1 expression(1.708±0.064;=10.910, <0.001)and the CPIP+NS group had significantly increased expressions of IL-1β(2.518±0.147;=11.010, <0.001)and TNF-α(1.949±0.184;=10.870, <0.001).Compared with the CPIP+NS group,the CPIP+NMDA group had significantly increased expressions of IL-1β(4.816±0.607;=16.670, =0.003)and TNF-α(2.629±0.349;=7.790, <0.001)and the CPIP+MK801 group had significantly decreased expressions of IL-1β(1.048±0.257;=10.660, =0.003)and TNF-α(0.790±0.165;=13.280, <0.001). NMDA receptor activation in skin keratinocytes after chronic ischemia in rats hinders the expression of inflammatory cytokines such as IL-1β and TNF-α,which may be involved in central sensitization and pain conduction of type Ⅰ complex regional pain syndrome.


Subject(s)
Animals , Interleukin-1beta , Keratinocytes , Male , Pain , Rats , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate , Spinal Cord , Tumor Necrosis Factor-alpha
14.
An. bras. dermatol ; 94(6): 677-683, Nov.-Dec. 2019. tab, graf
Article in English | LILACS | ID: biblio-1054900

ABSTRACT

Abstract Background: Psoriasis is a skin-articular disease with unclear etiopathogenesis. It has been suggested that the disease is immune-mediated by T-lymphocytes, predominantly Th17 cells. Similar to psoriasis, geographic tongue is an inflammatory disease with participation of Th17 cells and direct correlation with psoriasis. Objective: To investigate and compare the inflammatory responses and the Th17 pathway in psoriasis and geographic tongue. Methods: This was a cross-sectional study with 46 participants that were categorized into three groups: (A) patients with psoriasis vulgaris; (B) patients with geographic tongue and psoriasis; (C) patients with geographic tongue without psoriasis. All patients underwent physical examination, and a skin and oral biopsy for histopathological examination and immunohistochemical analysis with anti-IL6, anti-IL17, and anti-IL23 antibodies. Results: Histological analysis of all lesions showed mononuclear inflammatory infiltrate. However, moderate intensity was prevalent for the patients with geographic tongue and psoriasis and geographic tongue groups. Immunopositivity for the antibodies anti-IL6, anti-IL17, and anti-IL23 revealed cytoplasmic staining, mainly basal and parabasal, in both psoriasis and geographic tongue. Regarding IL-6, in patients with geographic tongue and psoriasis cases the staining was stronger than in patients with geographic tongue without psoriasis cases. IL-17 evidenced more pronounced and extensive staining when compared to the other analyzed interleukins. IL-23 presented similar immunopositivity for both geographic tongue and psoriasis, demonstrating that the neutrophils recruited into the epithelium were stained. Study limitation: This study was limited by the number of cases. Conclusion: The inflammatory process and immunostaining of IL-6, IL-17, and IL-23 were similar in geographic tongue and psoriasis, suggesting the existence of a type of geographic tongue that represents an oral manifestation of psoriasis.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Psoriasis/pathology , Th17 Cells/pathology , Glossitis, Benign Migratory/pathology , Psoriasis/immunology , Biopsy , Severity of Illness Index , Immunohistochemistry , Keratinocytes/pathology , Cross-Sectional Studies , Interleukin-6/immunology , Interleukin-17/immunology , Interleukin-23/immunology , Th17 Cells/immunology , Glossitis, Benign Migratory/immunology , Antibodies/analysis
15.
An. bras. dermatol ; 94(4): 434-441, July-Aug. 2019. tab, graf
Article in English | LILACS | ID: biblio-1038295

ABSTRACT

Abstract: Background: In-vitro studies showed that Leucine-rich glioma inactivated 3 (LGI3) is a keratinocyte-derived cytokine that stimulates melanin synthesis and is increased after ultra violet B (UVB) irradiation. So, we postulated that LGI3 may be involved in vitiligo aetiopathogenesis and may participate in narrow band ultra violet B (NB-UVB) induced pigmentation in vitiligo. Objectives: To assess this hypothesis, lesional LGI3 immunohistochemical expression of vitiligo patients before and after NB-UVB phototherapy was studied, and its correlation with repigmentation was evaluated. Methods: Forty vitiligo patients and 20 age, sex, and skin phenotype-matched controls were enrolled. Patients were treated with NB-UVB thrice weekly for 12 weeks. VASI score was evaluated before and after NB-UVB sessions. For vitiligo patients, baseline LGI3 immunohistochemical staining was estimated, and compared to that of controls and to its post-treatment data in those patients. Results: Baseline LGI3 immunohistochemical studied parameters (expression, intensity, percentage and H score) were significantly lower in vitiligo cases than controls (p=0.003, 0.013, 0.001 and 0.001 respectively). After 12 weeks of NB-UVB phototherapy, these LGI3 immunohistochemical parameters were up-regulated and became comparable to that of controls (p >0.05 for all). There was a significant positive correlation between the improvement of both VASI score and LGI3 H score mean values (r=-0.349 , p=0.027). Study limitations: Small number of investigated subjects. Conclusions: Decreased LGI3 protein may play an active role in vitiligo pathogenesis and its up-regulation after NB-UVB phototherapy, may actively participate in NB-UVB photo-induced melanogenesis.


Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Young Adult , Ultraviolet Therapy/methods , Vitiligo/pathology , Vitiligo/radiotherapy , Proteins/analysis , Cytokines/analysis , Reference Values , Time Factors , Severity of Illness Index , Immunohistochemistry , Case-Control Studies , Keratinocytes/radiation effects , Treatment Outcome , Statistics, Nonparametric , Melanocytes/radiation effects
16.
Pesqui. vet. bras ; 39(4): 292-298, Apr. 2019. ilus
Article in English | LILACS, VETINDEX | ID: biblio-1002809

ABSTRACT

The importance of the hoof to the horse health is clear, and the current knowledge regarding the cellular aspects of hoof keratinocytes is poor. Studies on equine keratinocyte culture are scarce. Developing keratinocyte cultures in vitro is a condition for studies on molecular biology, cell growth and differentiation. Some methods have already been established, such as those for skin keratinocyte culture. However, few methodologies are found for lamellar keratinocytes. The objective of this study was to standardize the equine hoof keratinocyte isolation and cultivation, and then characterize the cell immunophenotype. For this, the primary culture method used was through explants obtained from three regions of the equine hoof (medial dorsal, dorsal, and lateral dorsal). After the cell isolation and cultivation, the cell culture and its explants were stained with anti-pan cytokeratin (pan-CK) (AE1/AE3), vimentin (V9), p63 (4A4), and Ki-67 (MIB-1) antibodies. Cells were grown to third passage, were positive for pan-CK, p63 and Ki-67, and few cells had vimentin positive expression. As for the explants, the epidermal laminae were not stained for vimentin or Ki-67. However, some cells presented positive pan-CK and p63 expression. This study demonstrated the viability of lamellar explants of equine hooves as a form of isolating keratinocytes in primary cultures, as well as characterized the proliferation ability of such keratinocytes in monolayers.(AU)


É notória a importância do casco na saúde dos equinos, mas o conhecimento em nível celular é pouco entendido. Estudos envolvendo o cultivo de queratinócitos equinos são escassos. Sabe-se que o desenvolvimento de cultivos de queratinócitos in vitro é uma condição para estudos sobre a biologia molecular, crescimento e diferenciação celular. Alguns métodos já estão estabelecidos, como para cultivo de queratinócitos de pele, mas poucas metodologias são encontradas para queratinócitos lamelares. O objetivo desse estudo foi padronizar o cultivo de queratinócitos provenientes de casco equino visando futuramente associar ao estudo da medicina regenerativa para assim estabelecer um modelo experimental in vitro e indicar o uso criterioso de terapias regenerativas para a laminite equina. Desta forma, o cultivo em monocamada e a caracterização de queratinócitos lamelares foram realizados. Para isso, o método de cultura primária utilizado foi através de explantes obtidos de três regiões do casco (dorso-medial, dorsal e dorso-lateral). As células foram caracterizadas para os marcadores anti pan-cytokeratin (AE1/AE3), vimentin (V9), p63 (4A4) e Ki-67 (MIB-1) nos cultivos e nos explantes. As células foram cultivadas até terceira passagem, tendo marcação positiva para pan-CK, p63 e Ki-67 e fraca marcação para vimentina. Já as lâminas epidermais não tiveram marcação de vimentin e Ki-67, porém marcaram acentuadamente para pan-CK e p63. Este estudo demonstrou a exiquibilidade do uso de explantes lamelares do casco de equinos, como forma de isolamento de queratinócitos em cultivos primários, bem como caracterizou a habilidade de proliferação desses queratinócitos em monocamada.(AU)


Subject(s)
Animals , Primary Cell Culture/veterinary , Foot Diseases/veterinary , Hoof and Claw/pathology , Horse Diseases/pathology , Horse Diseases/therapy , Keratinocytes/cytology
18.
Zagazig univ. med. j ; 25(3): 326-334, 2019. ilus
Article in English | AIM, AIM | ID: biblio-1273859

ABSTRACT

Background: Vitiligo is a skin disease with complex, multifactorial pathogenesis. Abnormalities in surrounding keratinocytes may cause melanocyte death due to deprivation of growth factors. Narrow band ultraviolet B (NB-UVB) is an effective therapeutic option especially in patients with generalized disease.Objective: The aim of this study was to identify histopathological changes in lesional and perilesional skin of vitiligo patients and the effect of NB-UVB therapy on them.Methods: Twenty patients were enrolled in this study. They received NB-UVB twice weekly on non-consecutive days for a total of 40 sessions. Skin biopsies from lesional and perilesional skin were obtained from each patient before and after therapy.Results: After therapy, 10% of patients showed excellent clinical response, 10% showed good response, 40% showed moderate response, 35% showed poor response and 5% showed progressive disease. Before therapy, 50% of patients showed a basal lymphocytic infiltrate with a perivascular lymphocytic infiltrate in both lesional and perilesional skin. 40% of them showed additional hydropic degeneration of lower epidermis with apoptotic keratinocytes in 20% of them. After therapy, these inflammatory changes were significantly reduced (p=0.04).Conclusion: NB-UVB is an effective method of treatment of vitiligo. This may be due to its immunosuppressive effects. Also, keratinocyte apoptosis may have a role in pathogenesis of vitiligo


Subject(s)
Apoptosis , Egypt , Keratinocytes , Ultraviolet Therapy/methods , Vitiligo/therapy
19.
Chonnam Medical Journal ; : 75-85, 2019.
Article in English | WPRIM | ID: wpr-763279

ABSTRACT

Psoriasis is a chronic, recurrent, heterogeneous, cutaneous inflammatory skin disease for which there is no cure. It affects approximately 7.5 million people in the United States. Currently, several biologic agents that target different molecules implicated in the pathogenic processes of psoriasis are being assessed in diverse clinical studies. However, relapse usually occurs within weeks or months, meaning there is currently no cure for psoriasis. Therefore, recent studies have discovered diverse new potential treatments for psoriasis: inhibitors of bacteria such as Staphylococcus aureus, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and neuropilin 1 (NRP1). A promising approach that has recently been described involves modifying antimicrobial peptides to develop new cutaneous anti-bacterial agents that target inflammatory skin disease induced by Staphylococcus. Increased expression of TRAIL and its death receptors DR4 and DR5 has been implicated in the pathogenesis of plaque psoriasis. In addition, TRAIL has the ability to inhibit angiogenesis by inducing endothelial cell death and by negative regulation of VEGF-induced angiogenesis via caspase-8-mediated enzymatic and non-enzymatic functions. Since NRP1 regulates angiogenesis induced by multiple signals, including VEGF, ECM and semaphorins, and also initiates proliferation of keratinocytes through NF-κB signaling pathway in involved psoriatic skin, targeting NRP1 pathways may offer numerous windows for intervention in psoriasis. In this review, we will focus on the current knowledge about the emerging role of synthetic antimicrobial peptides, TRAIL and NRP1 blocking peptides in the pathogenesis and treatment of psoriasis.


Subject(s)
Anti-Bacterial Agents , Bacteria , Biological Factors , Endothelial Cells , Keratinocytes , Necrosis , Neuropilin-1 , Peptides , Psoriasis , Receptors, Death Domain , Recurrence , Semaphorins , Skin , Skin Diseases , Staphylococcus , Staphylococcus aureus , Therapeutic Uses , TNF-Related Apoptosis-Inducing Ligand , United States , Vascular Endothelial Growth Factor A
20.
Article in English | WPRIM | ID: wpr-763046

ABSTRACT

Rab25, a member of the Rab11 small GTPase family, is central to achieving cellular polarity in epithelial tissues. Rab25 is highly expressed in epithelial cells of various tissues including breast, vagina, cervix, the gastrointestinal tract, and skin. Rab25 plays key roles in tumorigenesis, mainly by regulating epithelial differentiation and proliferation. However, its role in skin physiology is relatively unknown. In this study, we demonstrated that Rab25 knock-out (KO) mice show a skin barrier dysfunction with high trans-epidermal water loss and low cutaneous hydration. To examine this observation, we investigated the histology and epidermal differentiation markers of the skin in Rab25 KO mice. Rab25 KO increased cell proliferation at the basal layer of epidermis, whereas the supra-basal layer remained unaffected. Ceramide, which is a critical lipid component for skin barrier function, was not altered by Rab25 KO in its distribution or amount, as determined by immunohistochemistry. Notably, levels of epidermal differentiation markers, including loricrin, involucrin, and keratins (5, 14, 1, and 10) increased prominently in Rab25 KO mice. In line with this, depletion of Rab25 with single hairpin RNA increased the expression of differentiation markers in a human keratinocyte cell line, HaCaT. Transcriptomic analysis of the skin revealed increased expression of genes associated with skin development, epidermal development, and keratinocyte differentiation in Rab25 KO mice. Collectively, these results suggested that Rab25 is involved in the regulation of epidermal differentiation and proliferation.


Subject(s)
Animals , Antigens, Differentiation , Breast , Carcinogenesis , Cell Line , Cell Proliferation , Cervix Uteri , Epidermis , Epithelial Cells , Female , Gastrointestinal Tract , GTP Phosphohydrolases , Humans , Immunohistochemistry , Keratinocytes , Mice , RNA , Skin Physiological Phenomena , Skin , Vagina , Water
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