Introduction: The hair follicle bulge has recently been added to a growing list of human tissue compartments that exhibit a complex combination of immunosuppressive mechanisms, termed immune privilege (IP), which appear to restrict immune mediated injury in specific locations. As epithelial hair follicle stem cells (eHFSC) reside in the hair follicle bulge region it is conceivable that these IP mechanisms protect this vital compartment from immune-mediated damage, thereby ensuring the ongoing growth and cyclic regeneration of the hair follicle. Lichen planopilaris (and variants) are inflammatory hair disorders that result in hair follicle destruction and permanent alopecia. Growing evidence suggests that eHFSC destruction is a key factor in the permanent follicle loss seen in these conditions, and that IP collapse may predispose these cells to immune mediated injury. Aims: The overall aim of this project was to generate immunohistomorphometric, gene profiling, and limited functional evidence to probe the 'bulge immune privilege collapse' hypothesis in a carefully selected model disease for inflammation-induced epithelial stem cell death, lichen planopilaris (LPP). Methods: Adult patients with LPP (or variant frontal fibrosing alopecia) were recruited. Biopsies from lesional and non-lesional scalp skin were performed and either snap frozen in liquid nitrogen, fixed in formalin for paraffin embedding or transferred immediately for hair follicle organ culture. Both frozen and paraffin embedded tissue was processed for immunohistochemistry (IHC) analysis using various immune privilege, hair bulge (eHFSC) and immune cell markers. Cultured samples were supplemented with various chemicals know to influence hair follicle immune privilege with analysis performed using IHC. Further, additional paired lesional and non-lesional samples were sectioned horizontally for laser capture microdissection of bulge cells. Following extraction of RNA, reverse transcription and amplification of cDNA from these selected bulge cells; gene expression profiling was performed comparing lesional with non-lesional samples. Selected, differentially regulated genes were validated using IHC and quantitative real-time PCR. Results: Bulge IP collapse is present in active LPP, as suggested by increased expression of MHC class I, β2microglobulin and MHC class II, along with reduced expression of the locally produced immunosuppressant TGFβ2, at both the gene and protein level. Microarray pathway analysis supports these data with the antigen processing canonical pathway being prominently enriched. Cell mediated immune responses are prominent in active disease, suggested by significantly increased numbers of activated and cytotoxic T-cells infiltrating the bulge epithelium, along with greater numbers of mast cells and macrophages in the peri-follicular connective tissue sheath. Bulge cell eHFSC loss is suggested by loss of bulge cell markers (e.g. keratin 15) on IHC, microarray and qPCR, and supported by microarray analysis showing virtually global loss of recognised bulge eHFSC signatures. Hair follicle organ culture experiments confirm bulge IP collapse in a separate cohort of patients, and demonstrate that the pro-inflammatory cytokine, interferon-γ can further collapse IP in the bulge epithelium of cultured hair follicles. Conclusion: These data identify collapse of immune privilege in bulge cells in active LPP, and identify prominent cell-mediated immune responses and loss of eHFSC signatures in active disease. The pro-inflammatory cytokine, interferon-γ also appears to play a prominent role in IP collapse and contributes to immune cell trafficking into affected tissue. Future study is required to ascertain triggering factors of IP collapse and pursue other identified candidates from gene expression analysis.

Primary scalp alopecia is classically divided into cicatricial (scarring) and non-cicatricial (non-scarring). Challenging cases are assessed with a scalp biopsy. The use of both horizontal and vertical sections (HoVert sections) has dramatically improved the accuracy of histopathological diagnosis. In this work, we have used immunostaining to address diagnostic difficulties, which persist despite all currently available tools. We performed an immunostain panel (CD3, CD4, CD8 and CD20) in order to distinguish pattern hair loss from alopecia aerate in cases which do not have the usual peribulbar lymphocytic infiltrate and showed that CD3+ T-lymphocytes within the empty fibrous follicular tracts favor a diagnosis of alopecia areata. We performed CD123 in order to distinguish lichen planopilaris from alopecia lupus erythematosus in cases with only a superficial lymphocytic infiltrate and an uninvolved interfollicular epidermis and showed that clusters of CD123+ plasmacytoid dendritic cells favor a diagnosis of lupus erythematosus. We performed cytokeratin 15 in order to assess whether the loss of the follicular bulge stem cells has diagnostic value in cicatricial alopecia and demonstrated that the loss of cytokeratin 15+ bulge stem cells is identified in lichen planopilaris, frontal fibrosing alopecia, and lupus erythematous, so cytokeratin 15 has no diagnostic value. We have attempted to integrate the new concepts and our findings into the traditional classifications of alopecia and proposed a new diagnostic algorithm. In conclusion, immunostaining combined with HoVert grossing advances the accuracy of histopathological diagnosis of primary scalp alopecia. / L’alopécie primitive du cuir chevelu est habituellement classée en cicatricielle et non-cicatricielle. Dans les cas difficiles, la biopsie du cuir chevelu peut aider au diagnostic. L’utilisation de coupes, à la fois verticales et horizontales sur le même spécimen (technique HoVert), a radicalement amélioré le diagnostic histopathologique. Dans ce travail, nous avons utilisé l’immunohistochimie pour évaluer les difficultés diagnostiques qui persistent malgré tous les outils actuels. Nous avons utilisé les CD3, CD4, CD8 et CD20 pour différencier l’alopécie androgénique de la pelade dépourvue de l’infiltrat lymphocytaire péribulbaire habituel et nous avons démontré que la présence de lymphocytes CD3+ dans les travées folliculaires fibreuses est en faveur de la pelade. Nous avons utilisé le CD123 pour différencier le lichen plan pilaire du lupus érythémateux alopécie avec infiltrat lymphocytaire superficiel et sans atteinte de l’épiderme interfolliculaire et nous avons démontré que la présence d’amas de cellules dendritiques plasmacytoïdes CD123+ est en faveur du lupus érythémateux. Nous avons utilisé la cytokératine 15 pour évaluer si la perte des cellules souches du bulge a une valeur diagnostique dans l’alopécie cicatricielle et nous avons démontré que cette perte s’observait de manière identique dans le lichen plan pilaire, l’alopécie frontale fibrosante comme dans le lupus érythémateux et n’avait donc aucune valeur diagnostique. Nous avons tenté d’intégrer les nouveaux concepts et nos données dans les classifications traditionnelles des alopécies et nous avons élaboré un nouvel algorithme diagnostique. L’association des immunomarquages avec la technique HoVert ouvre de nouvelles perspectives dans le diagnostic histopathologique des alopécies primaires du cuir chevelu. / Doctorat en Sciences médicales (Médecine) / info:eu-repo/semantics/nonPublished

Orientador: Silvana Artioli Schellini / Resumo: OBJETIVO: Avaliar a eficiência da injeção de ácido hialurônico (AH) ou de soro fisiológico (SF), aplicados no subcutâneo da pálpebra inferior de portadores de ectrópio cicatricial, visando à correção não cirúrgica do mal posicionamento palpebral. MÉTODOS: este foi um estudo prospectivo, intervencionista, envolvendo 23 pálpebras de 15 portadores de ectrópio cicatricial, divididos aleatoriamente em dois grupos: G1 (13 pálpebras) no qual foram feitas quatro aplicações de 4ml de SF (Cloreto de Sódio 0,9%, Equiplex, Goiás, Brasil), associado a 1ml de cloridrato de lidocaína a 2,0% sem vasoconstritor (Xylestesin®, Cristália, São Paulo, Brasil), em intervalos de uma semana entre as aplicações; e G2 (10 pálpebras) que receberam aplicação de 1 ml de AH (Restylane® Lidocaine, Q med, Uppsala, Suécia) em aplicação única. Foram avaliadas as variáveis demográficas dos participantes, as queixas, o grau do ectrópio, o grau de flacidez palpebral, a localização do ectrópio, além de avaliações quantitativas realizadas utilizando-se a fotodocumentação sistematizada dos olhos dos pacientes, 30 dias após a primeira aplicação no G1 e 7 e 30 dias após as aplicações do G2. As imagens obtidas foram transferidas para um computador e avaliadas utilizando-se o programa Image J, avaliando-se a distância da pálpebra inferior até o reflexo corneano (DMR2), distância limbo-margem (LM), ângulo da comissura interna (AI) e externa (AE), área total (AT), lateral (AL) e medial (AM). As áreas a serem avaliadas for... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: PURPOSE: Evaluate the efficiency of injection of hyaluronic acid (HA) or saline solution (SF) applied to the lower eyelid subcutaneous, aiming at the non-surgical correction of cicatricial ectropion. METHODS: This was a prospective study involving 23 eyelids of 15 patients with cicatricial ectropion, randomly divided into two groups: G1 (13 eyelids) with four applications of 4ml of SF (Sodium Chloride 0.9%, Equiplex, Goiás , Brazil), associated to 1ml of lidocaine hydrochloride 2% without vasoconstrictor (Xylestesin®, Cristália, São Paulo, Brazil), with a one-week interval between applications; and G2 (10 eyelids) which received single application of 1 ml of AH (Restylane® Lidocaine, Q med, Uppsala, Sweden). Demographic variables, complaints, ectropion degree, eyelid flaccidity degree, ectropion location, and quantitative assessments using the systematized photodocumentation of patients eyelids 30 days after the first application in G1 and 7 and 30 days after applications in G2 were studied. The images were transferred to a computer and evaluated using the Image J program and the distance between the lower eyelid margin to corneal reflex (DMR2), limbus margin distance (LM), internal angle (IA) and external angle (EA), total (TA), lateral (LA) and medial areas (MA) were analyzed. The area measurements were delimited by a line between the two commissures and along the lid margin of the lower eyelid. The evaluations were done without traction and with traction downward of the lo... (Complete abstract click electronic access below) / Mestre

Submitted by Laryssa Kataki De Oliveira Veloso (laryssakataki@hotmail.com) on 2018-06-15T13:25:57Z No. of bitstreams: 1 DISSERTAÇÃO LARYSSA FINAL.pdf: 1251310 bytes, checksum: 43a08fca6a961ece3b6833de3671f737 (MD5) / Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2018-06-18T20:28:55Z (GMT) No. of bitstreams: 1 veloso_lko_me_bot.pdf: 1251310 bytes, checksum: 43a08fca6a961ece3b6833de3671f737 (MD5) / Made available in DSpace on 2018-06-18T20:28:55Z (GMT). No. of bitstreams: 1 veloso_lko_me_bot.pdf: 1251310 bytes, checksum: 43a08fca6a961ece3b6833de3671f737 (MD5) Previous issue date: 2018-05-30 / OBJETIVO: Avaliar a eficiência da injeção de ácido hialurônico (AH) ou de soro fisiológico (SF), aplicados no subcutâneo da pálpebra inferior de portadores de ectrópio cicatricial, visando à correção não cirúrgica do mal posicionamento palpebral. MÉTODOS: este foi um estudo prospectivo, intervencionista, envolvendo 23 pálpebras de 15 portadores de ectrópio cicatricial, divididos aleatoriamente em dois grupos: G1 (13 pálpebras) no qual foram feitas quatro aplicações de 4ml de SF (Cloreto de Sódio 0,9%, Equiplex, Goiás, Brasil), associado a 1ml de cloridrato de lidocaína a 2,0% sem vasoconstritor (Xylestesin®, Cristália, São Paulo, Brasil), em intervalos de uma semana entre as aplicações; e G2 (10 pálpebras) que receberam aplicação de 1 ml de AH (Restylane® Lidocaine, Q med, Uppsala, Suécia) em aplicação única. Foram avaliadas as variáveis demográficas dos participantes, as queixas, o grau do ectrópio, o grau de flacidez palpebral, a localização do ectrópio, além de avaliações quantitativas realizadas utilizando-se a fotodocumentação sistematizada dos olhos dos pacientes, 30 dias após a primeira aplicação no G1 e 7 e 30 dias após as aplicações do G2. As imagens obtidas foram transferidas para um computador e avaliadas utilizando-se o programa Image J, avaliando-se a distância da pálpebra inferior até o reflexo corneano (DMR2), distância limbo-margem (LM), ângulo da comissura interna (AI) e externa (AE), área total (AT), lateral (AL) e medial (AM). As áreas a serem avaliadas foram delimitadas por uma linha que une as duas comissuras e outra linha que coincide com a margem da pálpebra inferior. As avaliações foram feitas sem tração e com tração da pálpebra inferior para baixo. Todos os dados foram transferidos para a planilha Excel, sendo realizada a análise comparativa antes e após as aplicações nos dois grupos. RESULTADOS: Os grupos foram semelhantes quanto a idade, sexo, cor da pele e grau do ectrópio. Houve melhora signficativa dos sintomas nos indivíduos de G2 após a aplicação de AH. Apos a injeção, o G1 apresentou redução significativa de AI com e sem tração, AE com tração, LM sem tração e AM com tração. No G2, após as injeções as medidas AI sem tração, AE, DMR2, AT e AM com e sem tração, LM e AL com tração apresentaram alterações significativas (p<0,05). A comparação das medidas realizadas no G2 após sete e 30 dias mostrou estabilidade dos resultados obtidos com a injeção de AH. CONCLUSÃO: A aplicação de AH na pálpebra inferior de portadores de ectrópio cicatricial mostrou melhora parcial do mal posicionamento palpebral. A aplicação de SF não foi efetiva no tratamento do ectrópio cicatricial. / PURPOSE: Evaluate the efficiency of injection of hyaluronic acid (HA) or saline solution (SF) applied to the lower eyelid subcutaneous, aiming at the non-surgical correction of cicatricial ectropion. METHODS: This was a prospective study involving 23 eyelids of 15 patients with cicatricial ectropion, randomly divided into two groups: G1 (13 eyelids) with four applications of 4ml of SF (Sodium Chloride 0.9%, Equiplex, Goiás , Brazil), associated to 1ml of lidocaine hydrochloride 2% without vasoconstrictor (Xylestesin®, Cristália, São Paulo, Brazil), with a one-week interval between applications; and G2 (10 eyelids) which received single application of 1 ml of AH (Restylane® Lidocaine, Q med, Uppsala, Sweden). Demographic variables, complaints, ectropion degree, eyelid flaccidity degree, ectropion location, and quantitative assessments using the systematized photodocumentation of patients eyelids 30 days after the first application in G1 and 7 and 30 days after applications in G2 were studied. The images were transferred to a computer and evaluated using the Image J program and the distance between the lower eyelid margin to corneal reflex (DMR2), limbus margin distance (LM), internal angle (IA) and external angle (EA), total (TA), lateral (LA) and medial areas (MA) were analyzed. The area measurements were delimited by a line between the two commissures and along the lid margin of the lower eyelid. The evaluations were done without traction and with traction downward of the lower eyelid. All data were transferred to the Excel spreadsheet and comparative analysis was performed before and after the applications in the two groups. RESULTS The groups were similar according to age, sex, skin color and degree of ectropion. There was a significant improvement in G2 individuals after the application of AH. After injections G1 presented a significant reduction of AI with and without traction, EA with traction, LM without traction and AM with traction. The G2 after injections showed AI measurements without traction, EA, DMR2, TA and MA with and without traction, LM and LA with traction with significant alterations (p <0.05). The comparison of the G2 measurements obtained after 7 and 30 days showed stability of the results obtained with the AH injection. CONCLUSION: The application of AH in the lower eyelid of patients with cicatricial ectropion resulted in partial improvement of palpebral malposition. The application of SF was not effective in the treatment of cicatricial ectropion.

Submitted by ISABELA CRISTINA DE SOUZA MARQUES null (isabelamarques@terra.com.br) on 2016-03-09T16:07:22Z No. of bitstreams: 2 tese isabela cristina de souza marques final.pdf: 6465517 bytes, checksum: 55023e7b2380abb38ca39b874f034c05 (MD5) tese isabela cristina de souza marques final.pdf: 6465517 bytes, checksum: 55023e7b2380abb38ca39b874f034c05 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-03-09T18:03:43Z (GMT) No. of bitstreams: 1 marques_ics_dr_jabo.pdf: 6465517 bytes, checksum: 55023e7b2380abb38ca39b874f034c05 (MD5) / Made available in DSpace on 2016-03-09T18:03:43Z (GMT). No. of bitstreams: 1 marques_ics_dr_jabo.pdf: 6465517 bytes, checksum: 55023e7b2380abb38ca39b874f034c05 (MD5) Previous issue date: 2016-02-05 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O presente trabalho, teve como objetivo avaliar o potencial cicatricial do óleo de semente de maracujá. O óleo de semente de maracujá é um fitoterápico, inovador com grandes perspectivas para para acelerar o processo cicatricial de lesões cutâneas. Nesse estudo foram avaliados feridas de equinos e ratos Wistar. Foram feitas avaliações macroscópicas e microscópicas nos equinos e avaliação imunuistoquímica nos ratos Wistar. Nas primeiras avaliações macroscópicas e microscópicas nos equinos, pudemos notar a diferença entre o grupo controle e o grupo tratado com óleo de semente de maracujá, os resultados fora estatisticamente significativos, em relação ao grupo tratado com óleo de semente de maracujá. Quando feita a imunoistoquímica no material biológico coletado dos ratos, obtivemos um resultado significativo, estatisticamente comprovado, avaliando COX-2, TGFβ, VEGF, Alfa Actina, Fator VIII, CD68. São marcadores de extrema importância na evolução cicatricial. Com os resultados obtidos, pudemos comprovar que o fitoterápico, óleo de semente de maracujá é eficiente e a acelera o processo cicatricial. / This study aimed to evaluate the healing potential of passion fruit seed oil. The passion fruit seed oil is a herbal, innovative with great prospects for to accelerate the healing process of skin lesions. In this study were evaluated wounds of horses and Wistar rats. Macroscopic and microscopic evaluations were done in horses and imunuistoquímica assessment in Wistar rats. In the first macroscopic and microscopic evaluations in horses, could tell the difference between the control group and the group treated with passion fruit seed oil, the results had been statistically significant, compared to the group treated with passion fruit seed oil. When done immunohistochemistry biological material collected from the mice, we obtained a significant result, demonstrated statistically evaluating COX-2, TGFb, VEGF, Actin alpha, Factor VIII, CD68. Markers are very important in scar evolution. With the results, we could confirm that the herbal medicine, passion fruit seed oil is efficient and speeds up the healing process. / CNPq: 141919/2013

Made available in DSpace on 2014-08-20T14:38:04Z (GMT). No. of bitstreams: 1 dissertacao_mariana_tillmann.pdf: 1164818 bytes, checksum: 36d068fbc96d4569dd3f8db7e6b7f838 (MD5) Previous issue date: 2011-02-21 / The healing process comprises a dynamic cellular process that ensures the restoration tissue and the healing treatments assist in this process. The objectives of this work were evaluated the healing evolution and quality of surgical wounds treated with antiseptic and open skin wounds treated with creams containing different concentrations of Triticum vulgare. The evaluated of antiseptics was realized in dogs submitted to ovariohysterectomy and treated with 10% polyvinyl-pyrrolidon iodine (PVPI), 0,5% chlorhexidine or 0,9% physiological solution, during 10 days. The evaluated of phitotherapy was realized in rabbits being studied 135 wounds at days seven, 14 and 21 and divided in groups with ten wounds of each treatment for clinical and histopathological analysis and 15 for tensiometer analysis, the treatments were realized with aqueous extract of T. vulgare in the concentration 2mg/ml and 10mg/ml and nonionic cream, during 21 days. The surgical and open wounds were analyzed as the healing quality and climical aspect in: exudate type and quantity, epithelialization and granulation tissue formation, being that in open skin wounds was added the concentration analysis. In the open skin wounds were still realized analysis histopathological, tensile breaking strength and strength tensiometer. In the surgical wounds no observed significative difference between the treatments in clinical evaluations over the studied period. Purulent exudates occurred in all the treatments during the experimental period, although in the day 10 the wound treated with 0,9% physiological solution differed from the other treatments. (p=0,0429). The wounds treated with physiological solution allowed the stitches removal within 7 days, differed from others treatments (p=0,0357). In the evaluated of healing quality the wound treated with 0,5% chlorhexidine presented more percentage of normotrophy wounds (70%) and in to the skin anatomic limits(40%). The treatment with PVPI differed of other groups because has not wound into the skin anatomic limits (p=0,0085). In the work with open skin wounds treated with different concentration of Triticum vulgare and cream nonionic was demonstrated that the clinical analysis evolution, histopathological (phases of healing process and collagen) and tensile breaking strength have not significant difference between the groups. In the strength tensiometer analysis the treatment with Triticum vulgare 2mg/ml statistically different from the control group. (p=0,0295). In the study condition concluded that there was no difference in healing of surgical wound treated with 10% polyvinyl-pyrrolidon-iodine, 0,5% chlorhexidine or 0,9% physiological solution, although the chlorhexidine has determined highest percentage of normotrophy wound and into the normal limits. Have not difference in the healing process of open wounds treated with the two concentrations of creams containing aqueous extract of Triticum vulgare and nonionic cream, but the wound treated with cream containing aqueous extract of Triticum vulgare 2mg/ml presented better results in the strength tensiometer evaluation. / O processo cicatricial compreende um dinâmico processo celular que garante a restauração tissular, os tratamentos para a cicatrização auxiliam nesse processo. Na medicina veterinária os anti-sépticos são utilizados rotineiramente para a cicatrização de feridas e atualmente os fitoterápicos têm sido bastante estudados. Os objetivos desse trabalho foram avaliar a evolução e a qualidade da cicatrização de feridas cirúrgicas tratadas com anti-sépticos e feridas cutâneas abertas tratadas com cremes contendo concentrações diferentes de Triticum vulgare. A avaliação dos anti-sépticos foi realizada em 30 fêmeas caninas submetidas à ovariosalpingohisterectomia e tratados com: polivinil-pirrolidona iodo (PVPI) 10%, clorexidine 0,5% ou solução fisiológica 0,9%, em dias alternados durante dez dias. A avaliação do fitoterapico foi realizada em coelhos, sendo estudadas 135 feridas aos dias sete, 14 e 21 e divididas em grupos de dez feridas de cada tratamento para análise clínica e histopatológica e 15 para a tensiométrica, os tratamentos utilizados foram com extrato aquoso de T. vulgare na concentração de 2mg/ml e de 10mg/ml e creme não iônico, durante 21 dias. As feridas cirúrgicas e abertas foram analisadas quanto a qualidade cicatricial e ao aspecto clínico em: tipo e quantidade de exsudato, formação do tecido de granulação e epitelização, sendo que nas feridas cutâneas abertas foi acrescentando a análise de contração. Nas feridas cutâneas abertas ainda foram realizadas análise histopatológica, força de ruptura e tensiométrica. Nas feridas cirúrgicas não foi observada diferença significativa entre os tratamentos ao longo do período estudado nas avaliação clínicas. Exsudato purulento ocorreu em todos os tratamentos durante o período experimental, embora no dia 10 as feridas tratadas com solução fisiológica 0,9% diferiram dos demais tratamentos (p=0,0429). As feridas tratadas com solução fisiológica permitiram a retirada de pontos em até sete dias, diferindo dos demais tratamentos (p=0,0357). Na avaliação da qualidade cicatricial, as feridas tratadas com clorexidine 0,5% apresentaram maior percentual de feridas normotróficas (70%) e dentro dos limites anatômicos da pele (40%). O tratamento com PVPI diferiu dos demais grupos devido não possuir feridas dentro dos limites anatômicos da pele (p=0,0085). Nas feridas cutâneas abertas tratadas com concentrações diferenciadas de T. vulgare e creme não iônico foi demonstrado que na evolução da análise clínica, histopatológica (fases do processo cicatricial e colágeno) e da força de ruptura não houve diferença significativa entre os grupos. Na análise tensiométrica o tratamento com T. vulgare 2mg/mI diferenciou-se estatisticamente do grupo controle (p=0,0295). Nas condições do estudo concluí-se que não houve diferença na cicatrização de feridas cirúrgicas tratadas com polivinil pirrolidona iodo 10%, clorexidine 0,5% e solução fisiológica 0,9%, embora o clorexidine tenha determinado maior percentual de feridas normotróficas e dentro dos limites normais. Não houve diferença no processo cicatricial de feridas abertas tratadas com os as duas concentrações de cremes contendo extrato aquoso de T. vulgare e creme não iônico, mas as feridas tratadas com o creme contendo extrato aquoso de T. vulgare 2 mg/ml apresentaram melhor resultado na avaliação tensiométrica