Oral lichen planus: case series and experience in a tertiary dermatology service in Brazil

Abstract Background: Lichen planus is an inflammatory disease that can affect both the skin and mucous membranes, including the oral mucosa. There is very little original Brazilian dermatology literature about oral lichen planus. Objective: To describe the clinical, pathological, and treatment data of 201 patients diagnosed with oral lichen planus followed at the Stomatology Outpatient Clinic of Hospital das Clínicas, Universidade de São Paulo, from 2003 to 2021. Method: The patients demographic profile, the morpho-topographic features of the lesions, the treatment employed, and the possible presence of squamous cell carcinoma were analyzed. Results: The disease was more common in women over 50 years of age, tending to be chronic, with a large number of cases showing cicatricial sequelae in the mucosa. Topical treatment with potent corticosteroids was shown to be effective in the vast majority of cases. Squamous cell carcinoma in oral lichen planus cicatricial sequelae was observed in eight cases. Study limitations: Retrospective study of medical records, with gaps regarding the filling out of data; unequal observation time among the studied cases. Conclusions: This is the largest Brazilian dermatology series on oral lichen planus. The response to topical corticoid therapy was excellent in the vast majority of cases. The high prevalence of atrophic lesions, demonstrating the chronicity and tissue destruction potential of this disease, may explain the large number of cases of squamous cell carcinoma.

Relevant proteins for the monitoring of engraftment phases after allogeneic hematopoietic stem cell transplantation

Abstract Introduction Hematopoietic Stem Cell Transplant (HSCT) has been successfully used as standard therapy for hematological disorders. After conditioning therapy, patients undergoing allogeneic HSCT, present three different phases of engraftment: early pre-engraftment, early post-engraftment, and late engraftment. Severe complications are associated with morbidity, mortality, and malignancies in these phases, which include effects on the oral cavity. Objectives The changes in the salivary composition after HSCT may contribute to identifying relevant proteins that could map differences among the phases of diseases, driven for personalized diagnostics and therapy. Methods Unstimulated whole saliva was collected from patients submitted to HSCT. The samples were submitted to trypsin digestion for a Mass spectrometry analysis. MaxQuant processed the Data analysis, and the relevant expressed proteins were subjected to pathway and network analyses. Results Differences were observed in the most identified proteins, specifically in proteins involved with the regulation of body fluid levels and the mucosal immune response. The heatmap showed a list of proteins exclusively expressed during the different phases of HSCT: HBB, KNG1, HSPA, FGB, APOA1, PFN1, PRTN3, TMSB4X, YWHAZ, CAP1, ACTN1, CLU and ALDOA. Bioinformatics analysis implicated pathways involved in protein processing in the endoplasmic reticulum, complement and coagulation cascades, apoptosis signaling, and cholesterol metabolism. Conclusion The compositional changes in saliva reflected the three phases of HSCT and demonstrated the usefulness of proteomics and computational approaches as a revolutionary field in diagnostic methods.

Immunostaining study of cytokeratins in human hair follicle development

Abstract Background: The hair follicle is a unique structure, one of the most dynamic structures in mammalians, which can reproduce in every new cycle all the mechanism involved in its fetal development. Although a lot of research has been made about the human hair follicle much less has been discovered about the importance of the cytokeratins (CKs) in its development. Objective: Study the immunohistochemical pattern of epithelial CKs during human hair follicle development. Methods: We performed an immunohistochemical study using fresh post-mortem skin biopsies of human fetuses between 4 and 25 weeks of gestational age to study the expression of cytokeratins (CKs): CK1, CK10, CK13, CK14, CK16 and CK20 during human hair follicle fetal development. Study limitations: Restrospective study with a good number of makers but with a small population. Results/conclusion: We found that, the CKs were expressed in an intermediate time during follicular development. The epithelial CKs (CK1, CK14, CK10, CK13) and the epithelial CKs with a proliferative character such as CK16 were expressed first, as markers of cellular maturation and follicular keratinization. At a later phase, CK20 was expressed in more developed primitive hair follicles as previously discussed in literature.

Invasión ósea del carcinoma de células escamosas de la cavidad oral, análisis clínico - patológico de 62 casos / Bone invasion of squamous cell carcinoma of the oral cavity, clinical-pathological analysis of 62 cases

RESUMEN Introducción: El pronóstico de pacientes con carcinoma de células escamosas (CEC) de la cavidad oral con presencia de invasión ósea continua sombrío. El patrón de invasión ósea puede ser usado como indicador de agresividad y correlacionado con el comportamiento clínico del tumor. Objetivo: Evaluar los patrones histopatológicos de invasión ósea en pacientes con CEC de la cavidad oral y correlacionarlos con la tasa de sobrevida. Materiales y métodos: La muestra fue constituida por 62 pacientes con presencia de CEC en la cavidad oral e invasión ósea. Las características epidemiológicas e histopatológicas fueron tabuladas y analizadas. Los Test Chi- cuadrado y exacto de Fischer fueron empleados para verificar las asociaciones estadísticas entre los datos. Resultados: Fueron afectados más hombres en la quinta década de vida y observada una fuerte asociación con tabaquismo y etilismo crónico. Fueron relatadas muertes en 58% de los pacientes, en un periodo de 5 años. El patrón histológico más prevalente fue el infiltrativo, asociado a menores tasas de sobrevida. Conclusión: El análisis del patrón histopatológico de los CEC de la cavidad oral, puede ser usado como factor pronóstico auxiliando en la decisión del abordaje del tratamiento oncológico.

SUMMARY Introduction: Prognosis for patients with squamous cell carcinoma (SCC) in oral cavity with bone invasion presence is concerning. Bone invasion patterns can be used to indicate aggressiveness and can be correlated with tumor clinical behavior. Objective: To evaluate bone invasion histopathological patterns in patients with oral cavity SCC and correlate them with the survival rate. Materials and methods: 62 patients with presence of SCC in the oral cavity as well as bone invasion were recruited for this study. Epidemiological and histopathological characteristics were tabulated and analyzed. Fischer's Square and Exact Tests were used to verify any statistical associations between the data. Results: More men were affected, especially in the fifth decade of life, and a strong association with smoking and chronic alcoholism was observed. Deaths were reported in 58% of patients, over a period of 5 years. The most prevalent histological pattern was the infiltrative, associated with lower survival rates. Conclusion: An analysis of the histopathological patterns of oral cavity SCC can be used as a prognostic factor thereby assisting with the decision of which oncological treatment approach to use.

Ameloblastoma with distinctive granular cell pattern: an 8 case study

Ameloblastoma with granular cell pattern (AGC) is classified as an unusual histological subtype of solid/multicystic ameloblastoma, characterized by granular changes in stellate-like cells located within the inner portion of the epithelial follicles. Studies have revealed that lysosomal overload causes cytoplasmic granularity; however, the mechanism involved remains poorly understood. Here we report on eight cases of granular cell ameloblastoma, in the posterior region of the mandible. The age of the patients included in this case series ranged from 35 -64 years old and 87.5% of cases occurred in non-Caucasians, with a slight gender predilection for men (62.5%). There was no evidence of recurrence, and the majority of the cases were treated with surgical resection (87.5%). All tumors displayed histopathological features consistent with the diagnosis of ameloblastoma with granular cell pattern.

Profile of apoptotic proteins in oral squamous cell carcinoma: A cluster analysis of 171 cases

Oral squamous cell carcinoma (OSCC) is the eighth most prevalent cancer worldwide. In recent large-scale studies, by immunohistochemistry and cluster analysis, several markers were associated with patient survival in various tumors. The aim of this study was to analyze the expression profiles of 23 proteins that have been linked to the inhibition (Bcl-2, Bcl-x, Bcl-xL, Bcl-2-related protein A1, BAG-1, and survivin) and promotion (Bak, Bax, Bim/Bod, Bim-Long, Bad, Bid, PUMA, Apaf-1, caspase-2, caspase-3, caspase-6, caspase-7, caspase-8, caspase-9, caspase-10, Smac/DIABLO, and cytochrome c) of apoptosis in OSCC. METHODS: Two-hundred and twenty nine cases of OSCC, arranged in a tissue microarray, were immunohistochemically analyzed, and the results were quantified on an automated imaging system. The data were analyzed using a random forest clustering method. RESULTS: Overall protein expression patterns defined two chief clusters: an anti-apoptotic cluster (142 cases) and a pro-apoptotic cluster (29 cases). These groups could not be explained by any clinical or pathological characteristic, and overall and disease-free survival did not differ between them. CONCLUSIONS: Although there was no association with survival, the cluster analysis demonstrated specific protein profiles that could be of interest for using targeted therapies: in one of the clusters, the expression of pro-apoptotic proteins was more prominent, demonstrating a pro-apoptotic profile and highlighting the importance of apoptosis during OSCC development.

Effects of periodontal treatment on primary sjogren's syndrome symptoms

Abstract The aim of this longitudinal prospective study was to evaluate the effects of periodontal treatment on the clinical, microbiological and immunological periodontal parameters, and on the systemic activity (ESSDAI) and subjective (ESSPRI) indexes in patients with primary Sjögren’s Syndrome (pSS). Twenty-eight female patients were divided into four groups: pSS patients with or without chronic periodontitis (SCP, SC, respectively), and systemically healthy patients with or without chronic periodontitis (CP, C, respectively). Periodontal clinical examination and immunological and microbiological sample collection were performed at baseline, 30 and 90 days after nonsurgical periodontal treatment (NSPT). Levels of interleukin IL-1β, IL-8 and IL-10 in saliva and gingival crevicular fluid (GCF) were evaluated by ELISA, as well as the expression of Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans, (Aa) Tannerella forsythia (Tf), and Treponema denticola (Td), by qPCR. Systemic activity and pSS symptoms were evaluated by ESSDAI and ESSPRI. NSPT resulted in improved periodontal clinical parameters in both SCP and CP groups (p>0.05). Pg, Aa, and Tf levels decreased after NSPT only in CP patients (p<0.05). Significantly greater levels of IL-10 in GCF were verified in both SCP and CP groups (p<0.05). SCP patients showed increased salivary flow rates and decreased ESSPRI scores after NSPT. In conclusion, NSPT in pSS patients resulted in improved clinical and immunological parameters, with no significant effects on microbiological status. pSS patients also showed increased salivary flow and lower ESSPRI scores after therapy. Therefore, it can be suggested that NSPT may improve the quality of life of pSS patients.

BPI-fold (BPIF) containing/plunc protein expression in human fetal major and minor salivary glands

Abstract The aim of this study was to determine expression, not previously described, of PLUNC (palate, lung, and nasal epithelium clone) (BPI-fold containing) proteins in major and minor salivary glands from very early fetal tissue to the end of the second trimester and thus gain further insight into the function of these proteins. Early fetal heads, and major and minor salivary glands were collected retrospectively and glands were classified according to morphodifferentiation stage. Expression of BPI-fold containing proteins was localized through immunohistochemistry. BPIFA2, the major BPI-fold containing protein in adult salivary glands, was detected only in the laryngeal pharynx; the lack of staining in salivary glands suggested salivary expression is either very late in development or is only in adult tissues. Early expression of BPIFA1 was seen in the trachea and nasal cavity with salivary gland expression only seen in late morphodifferentiation stages. BPIFB1 was seen in early neural tissue and at later stages in submandibular and sublingual glands. BPIFA1 is significantly expressed in early fetal oral tissue but BPIFB1 has extremely limited expression and the major salivary BPIF protein (BPIFA2) is not produced in fetal development. Further studies, with more sensitive techniques, will confirm the expression pattern and enable a better understanding of embryonic BPIF protein function.

Cytokeratin immunoprofile of primary and metastatic adenoid cystic carcinoma of salivary glands: a report of two cases

Distant metastases from salivary gland tumors are considered infrequent: the incidence of distant metastases ranges from 24% to 61% according to different histotypes and to the site of the primary mass. The most common site of distant metastases due to salivary gland malignancies is the lung. From the pathology point of view, cytokeratins (CK) are important differentiation markers in salivary gland tumors, which are often used for the diagnostic process. Their employment also may be useful to identify and confirm the diagnosis of their distant metastases. We report the expression of CK in two cases of primary and metastatic adenoid cystic carcinoma (ACC) and their CK profiles of the primary and metastatic masses. Both patients-one male and one female-were diagnosed with an ACC cribriform and tubular, respectively, with lung metastases. In case 1, the metastatic mass presented the same histotype and CK profile of the primary tumor. For case 2, the metastatic lung mass was distinct from the primary mass (a solid ACC) and presented a different CK profile. Although salivary gland metastatic disease presents a poor prognosis, both patients reported herein are alive despite the presence of the disease in long-term follow-up. Therefore, the modifications seen in the CK profiles do not appear to be predictive of tumor behavior and outcome. The use of a CK profile seems to be useful to identify the nature of a distant mass and its possible correlations with a primary salivary gland tumor.

Oral mucosal diseases in children - casuistics from the Department of Dermatology - University of São Paulo - Brazil

Abstract There are no studies about pediatric oral mucosal diseases performed by dermatologists in Brazil. This study presents the casuistics of oral mucosal diseases in children examined at the Oral Diseases Clinic at the Department of Dermatology - University of São Paulo - Brazil. Cases were retrospectively studied from the hospital records from 2003 to 2015. A hundredsix children have been examined. Commoner lesions examined included mucoceles and aphthae. Rare and difficult cases were also seen and have been published; this clinic is based in a tertiary hospital center that deals mostly with complex cases.

Aspectos relevantes da síndrome de Sjõgren para o cirurgião-dentista / Sjogren’s syndrome’s relevant aspects for the dental surgeon

A síndrome de Sjõgren (SS), conhecida como síndrome sicca, é uma doença autoimune caracterizada pela hipofunção das glândulas salivares e lacrimais, cuja prevalência na população mundial é de aproximadamente 0,5% a 1%. Por ser uma doença autoimune complexa e de difícil diagnóstico, é sub-diagnosticada e sub-tratada segundo o consenso realizado em 2012 pelo Colégio Americano de Reumatologia (ACR). O Cirurgião-Dentista pode desempenhar papel importante na detecção de possíveis alterações compatíveis com a síndrome, além de auxiliar no tratamento de diversas patologias orais decorrentes da síndrome. Este trabalho tem como objetivo explanar aspectos importantes referentes ao diagnóstico e tratamento da síndrome aqui discutida. A SS apesar de ser considerada uma doença de evolução lenta, em estágios avançados pode ser fatal, principalmente por aumentar as chances dos pacientes virem a desenvolver linfoma não Hodking. O tratamento odontológico dos pacientes com SS deve principalmente ser profilático, com a recomendação do uso de repositores de saliva e controle rígido da higiene bucal.

Sjogren’s syndrome (SS), known as the sicca syndrome, is an autoimmune disease characterized by salivary and lacrimal glands hypofunction which prevalence in the world population is approximatel y around 0,5% to 1%. For being a complex autoimmune disease and with difficult diagnosis, it is sub diagnosed and miss treated according to the consensus occurred in 2012 by the American College of Rheumatology (ACR). The surgeon-dentist (SD) may play a important role on the detection of possible changes compatible to the syndrome, besides can help in the treatment of many oral pathologies caused by the syndrome. This work has the main purpose to explain the important aspects regards to the correct diagnosis and treatment of this syndrome.The SS besides been considered a slow evolution disease, in advanced stages it can be fatal,mainly for increasing the patient’s chances of developing non-Hodking lymphoma. The dental treatment of patients with SS must be prophylactic, with the recomedations of the use of salivary replenishing and careful control of the oral hyigiene.

Recomendações para o tratamento da síndrome de Sjögren / Recommendations for the treatment of Sjögren's syndrome

RESUMOAs recomendações propostas pela Comissão de Síndrome de Sjögren da Sociedade Brasileira de Reumatologia para tratamento da síndrome de Sjögren foram baseadas em uma revisão sistemática da literatura nas bases de dados Medline (PubMed) e Cochrane até outubro de 2014 e opinião de especialistas na ausência de artigos sobre o assunto. Foram incluídos 131 artigos classificados de acordo com Oxford & Grade. Essas recomendações foram elaboradas com o objetivo de orientar o manejo adequado e facilitar o acesso aos tratamentos para aqueles pacientes com adequada indicação de recebê-los, considerando o contexto socioeconômico brasileiro e os medicamentos disponíveis no país.

ABSTRACTThe recommendations proposed by the Sjögren's Syndrome Committee of the Brazilian Society of Rheumatology for the treatment of Sjögren's syndrome were based on a systematic review of literature in Medline (PubMed) and the Cochrane databases until October 2014 and on expert opinion in the absence of studies on the subject. 131 articles classified according to Oxford & Grade were included. These recommendations were developed in order to guide the management and facilitate the access to treatment for those patients with an appropriate indication, considering the Brazilian socioeconomic context and pharmacological agents available in this country.

Facial lesions caused by renal osteodystrophy in a patient with chronic renal insufficiency: a case report / Lesões faciais por osteodistrofia renal em paciente com insuficiência renal crônica: relato de caso

PURPOSE: Chronic renal insufficiency (CRI) is the last stage of a chronic renal condition in which the kidney loses its filtration and endocrine functions. Chronic endocrine hypofunction causes generalized damage to the body known as Uremic Syndrome, which affects the central nervous system as well as the cardiovascular, hematologic, dermatologic, ophthalmic, endocrine, respiratory, gastrointestinal and skeletal systems. The present study reports the case of a female patient with CRI who presented facial osteodystrophy of the osteitis fibrosa type, and highlights the main features of this condition. CASE DESCRIPTION: A 24-year old, female, Caucasian patient presented chronic glomerulonephritis recurrence and lost the transplanted kidney five years before, undergoing arteriovenous fistula hemodialysis three times a week. She presented swelling of the left masseter area with a hard consistency on palpation, covered by intact skin, swelling at the bottom of the left atrium, with a hard consistency on palpation, a mucosa-like color and absence of inflammation signs, suggesting expansive bone lesions on the face. These features were compatible with hyperparathyroidism brown tumor and/or osteodystrophy. The CT scan showed expansive bone lesions of heterogeneous appearance on the left jaw, maxilla/nasal floor, and right frontotemporal suture areas. The clinical and histopathological characteristics of the lesion, in association with PHT hormone high serum levels led to renal osteodystrophy diagnosis. The patient was referred to the nephrology services. CONCLUSION: Osteodystrophic bone alterations have a high prevalence in renal disease patients, and the dentist must take these alterations into consideration in bone lesion diagnosis for this specific group of patients.

OBJETIVO: A Insuficiência Renal Crônica (IRC) é o último estágio da doença renal crônica, na qual o rim perde sua função de filtração e endócrina. A hipofunção endócrina renal ocasiona danos generalizados ao organismo, que no conjunto são conhecidos como Síndrome Urêmica, sendo composta por danos que comprometem o sistema nervoso central, assim como os sistemas cardiovasculares, hematológicos, dermatológicos, oftálmicos, endócrinos, respiratórios, gastrointestinais e ósseos. O presente estudo irá relatar o caso de uma paciente portadora de IRC apresentando osteodistrofia em face do tipo osteíte fibrosa, evidenciando as principais características dessa doença. DESCRIÇÃO DO CASO: Uma paciente de 24 anos, sexo feminino, caucasiana, apresentou glomerulonefrite recorrente após perda do rim transplantado, estando sob hemodiálise três vezes por semana. Apresentou inchaço da área do m. masseter esquerdo com consistência dura à palpação, coberto por pelo de aspecto normal, inchaço do átrio esquerdo, com consistência dura e ausência de sinais inflamatórios, sugerindo lesões ósseas na face. Estas características eram compatíveis com tumor marrom de hiperparatireoidismo e/ou osteodistrofia. A imagem de tomografia computadorizada mostrou lesões ósseas expansivas de aparência heterogênea na mandíbula esquerda, assoalho da maxila e nasal, e nas áreas de sutura frontotemporal direita. As características clínicas e histopatológicas da lesão, em associação com níveis séricos elevados de hormônio PHT conduziram ao diagnóstico de osteodistrofia renal. A paciente foi encaminhada para o serviço de nefrologia. CONCLUSÃO: As alterações ósseas osteodistróficas são de grande prevalência em pessoas com doença renal, tendo o cirurgião-dentista o dever de levá-las em consideração no diagnóstico de lesões ósseas neste grupo específico de pessoas.

Perfil epidemiológico e histopatológico de 642 casos de carcinoma basocelular de cabeça e pescoço operados em uma instituição terciária / Epidemiological and histopathological profile of 642 cases of basal cell carcinoma of head and neck surgery in a tertiary institution

Introdução: Carcinoma basocelular (CBC) é o câncer de pelemais frequente. A maioria dos CBCs tem comportamentoindolente, com cura em 95% dos casos após tratamentolocal. Uma pequena parcela corresponde a lesões de altamorbidade tratadas pela Cirurgia de Cabeça e Pescoço (CCP).A caracterização desses casos é pouco encontrada. Objetivo:Avaliar o perfil epidemiológico dos pacientes operados ecaracterísticas anatomopatológicas de CBCs em uma instituiçãoterciária. Método: Prontuários de 642 pacientes operados porCBC foram analisados, sendo comparados os dados de 322casos tratados pela CCP (grupo CCP) com 320 pela Dermatologia(grupo Derma). Sexo, idade, localização do tumor primário,zona de acometimento, tamanho, subtipo histológico e invasõesperineural e angiolinfática foram avaliados. Resultados: 51,8%dos pacientes do grupo CCP e 45,9% do grupo Derma eramhomens (p=0,13). A idade média dos grupos CCP e Dermaforam de 58,8±11,4 e 76,1±10,1 anos (p<0,01). A zona H foisede de 78,9% dos tumores do grupo CCP e 71,9% do grupoDerma (p=0,04). A mediana do tamanho tumoral foi de 2,5cm(1,45-4,0) no grupo CCP e 0,6cm (0,4-1,0) no grupo Derma(p<0,01). Os subtipos esclerodermiforme e adenoide forammais frequentes no grupo CCP (p<0,001), enquanto nodular eesclerosante foram no grupo Derma(p<0,001). Invasão perineuralfoi observada em 22,5% no grupo CCP e 13,2% no grupo Derma(p<0,01). Conclusão: A caracterização dos casos tratados pelaCCP permite maior conhecimento sobre lesões agressivas epossivelmente a identificação precoce dos casos de maior risco.

Líquen plano oral / Oral lichen planus

O líquen plano da mucosa oral (LPO) é afecção relativamente comum, que pode aparecer isolado ou associado ao líquen plano cutâneo, havendo, no entanto, significantes diferenças clínicoevolutivas: o LPO tende a ser crônico, recidivante e de difícil tratamento, levando a importante morbidade, principalmente em sua forma erosiva. Novas formas clínicas agressivas têm sido salientadas na literatura, como a forma gingivo-vulvar. Este artigo revisa a etiopatogenia, as formas clínicas, a diagnose diferencial e laboratorial, a prognose e o tratamento do LPO, além de mencionar, brevemente, a experiência dos autores com esta enfermidade, vivida no Ambulatório de Estomatologia da Divisão de Dermatologia do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo.

Oral lichen planus (OLP) is a relatively common mucosal disease that can present isolated or associated with cutaneous lichen planus. Contrarily to its cutaneous counterpart, though, OLP tends to be chronic, relapsing, and difficult to treat. Severe morbidity is related to erosive forms, and more aggressive presentations have been described, such as the "gingivo-vulvar syndrome". This article reviews the current knowledge about the pathogenesis, clinical picture, differential and laboratorial diagnosis, prognosis, and treatment of OLP.

Ultrastructural evaluation of human keratinocyte growth and differentiation on a fibrin substrate / Avaliação ultraestrutural do crescimento e da diferenciação de queratinócitos sobre um substrato de fibrina

PURPOSE: In order to circumvent several difficulties that have been met in the routine use of the in vitro keratinocyte cultures using the standard procedure described by Rheinwald and Green, and obtain a more resilient and the least possible immunogeneic skin substitute for a future clinical application, this work studied a new keratinocyte culture system, which envisages the utilization of a fibrin substrate in association with high densities of human keratinocytes. METHODS: Through light and transmission electron microscopy and immunohistochemical assays, long-term proliferative and differentiative characteristics of keratinocytes cultured onto a fibrin gel under immerse and air-liquid interface culture conditions were evaluated. RESULTS: Despite the absence of a dermal substitute, the results demonstrated that the proposed composite was constituted of a transparent and elastic fibrin film covered by a well-attached, multistratified epithelium with morphological characteristics that resemble human epidermis, including the neoformation, albeit incomplete, of the basement membrane. CONCLUSIONS: Increased mechanical resistance due to the presence of an easy handling substrate, the delivery of nonclonfluent keratinocytes as well as the removal of animal-derived cells from the culture system suggest its potential use for future transplantation purposes.

OBJETIVO: Com o intuito de contornar diversas dificuldades encontradas no uso rotineiro de queratinócitos cultivados in vitro pela técnica descrita por Rheinwald e Green, e obter um substituto cutâneo mais resistente e o menos imunogênico possível para futuras aplicações clínicas, este trabalho avaliou um novo sistema de cultura de queratinócitos que prevê a utilização de um substrato de fibrina em associação com queratinócitos humanos em alta densidade. MÉTODOS: Através de microscopia óptica e eletrônica e análise imunohistoquímica, foram avaliadas as características proliferativas e de diferenciação em longo prazo de queratinócitos cultivados em condição imersa e na interface ar-líquido. RESULTADOS: Apesar da ausência de um substituto dérmico, foi demonstrado que o composto proposto constituiu-se de um substrato de fibrina transparente e elástico coberto por epitélio multi-estratificado, bem aderido, com características morfológicas semelhantes à epiderme humana, incluindo a neo-formação, embora incompleta, da membrana basal. CONCLUSÕES: A maior resistência mecânica com a presença de um substrato de fácil manuseio, a possível liberação de queratinócitos não-confluentes, e a remoção de células com origem animal dos sistemas de cultura sugerem que o composto proposto neste estudo apresenta grande potencial para uso clínico futuro.

Melanoma primário da mucosa oral como modelo de investigação em patologia geral e aplicada: análise clínico-morfológica, pesquisa de marcadores biológicos e estabelecimento de células cultivadas in vitro / Primary melanoma of oral mucosa as a model for scientific investigation on general and applied pathology: clinical-pathological analysis, immunohistochemistry of biological markers and establishment of a cell culture

Introdução: O melanoma primário da cavidade oral é raro. Pode ocorrer em qualquer faixa etária e os locais de maior ocorrência são o palato e a gengiva superior. Histologicamente os meia no mas orais são compostos de melanócitos neoplásicos fusiformes, plasmocitóides ou epitelióides. Melanina é encontrada em 90% das lesões. Os mecanismos precisos envolvidos no desenvolvimento dos melanomas ainda não estão bem estabelecidos mas alguns fatores são considerados importantes: 1) diminuição de receptores importantes para a comunicação e adesão aos queratinócitos; 2) aumento de receptores e moléculas de adesão ausentes em melanócitos normais; 3) perda da ancoragem à membrana basal devido a alterações na expressão de moléculas de adesão envolvidas nesse processo. Esses fatores estão diretamente relacionados aos níveis de expressão de moléculas de adesão e têm impacto em vias sinalizadoras como por exemplo na via do p16. Objetivos: O presente estudo contemplou a análise clínico-morfológica de 35 melanomas primários da mucosa oral e a pesquisa de moléculas de adesão e de fatores da via do p16. Ainda estabeleceu uma cultura primária de células derivadas de um melanoma oral para futuros ensaios funcionais. Material e métodos: Para a realização do estudo investigação de melanócitos em espécimes de fetos humanos em diferentes fases gestacionais foi realizada. A análise dos melanomas primários da mucosa oral incluiu estudo dos dados dos prontuários de 35 casos diagnosticados como melanoma primário da mucosa oral foram analisados. Análise histológica dos casos também foi realizada. Os blocos de parafina contendo fragmentos dos tumores foram selecionados e áreas representativas de cada caso foram organizados em tissue microarray para a investigação imunoistoquímica das moléculas de adesão (caderinas, integrinas, imunoglobulinas e claudinas). Fragmentos provenientes de biopsia de um melanoma primário...

Introduction: Primary oral mucosal melanoma is arare neoplasm. It occurs in a wide age range and palate and superior gingiva are the most common anatomic sites affected. Histologically these tumours comprise spindle, plasmacytoid and epithelioid neoplastic melanocytes, melanin is detected in 90% of the lesions. The precise mechanisms involved in meia no ma development are not well-established but some aspects of basic biology are considered relevant: 1) downregulation of keratinocytes receptors; 2) upregulation of some types of adhesion molecules; 3) 1055 of cell/extracellular matrix receptors. These mechanisms are directly related to the expression of adhesion molecules with impact in the signalling casca de of p16. Aims: 1) To analyse the clinical and morphological aspects of 35 primary oral mucosal melanomas and to study the expression of adhesion molecules and p16 cascade. 2) To establish an in vitro model for further functional studies of primary oral mucosal melanomas. Material and Methods: Immunohistochemistry analysis of normal melanocytes in both developing and fully developed oral mucosa was performed. Analysis of the clinical and histopathological aspects of 35 primary oral mucosal melanomas was then carried out. Immunohistochemistry evaluation of adhesion molecules (cadherins, integrins, immunoglobulins and claudins), as well as study of p16 cascade components was performed in a tissue microarray containing fragments of the 35 cases of primary oral melanomas included in the study. A fresh oral mucosal melanoma was sampled and the tissue was prepared to establish a primary culture derived from the tumour. Results: Melanocytes were detected in oral mucosa from the 20th weeks of gestation. Primary oral melanomas were detected between the 5th and 7th decades of life and were more frequent in Caucasian...