Transmission electron microscopy in oral cytopathology: a pilot study / Microscopia eletrônica de transmissão em citopatologia bucal: um estudo piloto

Introduction: Cytopathology is a collection method that allows cell analysis through the different techniques. The oral mucosa exfoliated cells observation demonstrates morphological, biochemical and/or molecular aspects depending on the type of processing of the sample. Aim: This study tested the use of oral cytopathology associated with transmission electron microscopy (TEM) to observe the morphology of cells, mainly in relation to the cell nucleus, the cytoplasmic membrane, and cell junctions. Materials and Methods: Exfoliated epithelial cells from the oral mucosa were analyzed by TEM from individuals exposed to tobacco and alcohol, with leukoplakia or with a histopathological diagnosis of squamous cell carci-noma. Results: The cytoplasmic cell-cell junctions in the malignant samples lost the characteristic irregular pattern formed by the numerous interdigitations and the junctional process of normal cells and started to present a straight cytoplasmic surface. The nuclei of cells from leukoplakia and squamous cell carcinoma samples showed heterogeneous staining, while non-lesional cells were homogeneous. Discussion: The analysis of oral cytopathological smears by TEM contributes to the un-derstanding of the changes that occur during the process of malignancy of the oral mucosa, especially with regard to the cytoplasmic membrane and intercellular junctions. Conclusion: TEM may be a good analytical method to investigate morphological changes in exfoliated cells of the oral epithelium.

Introdução: A citopatologia é um método de coleta que permite a análise celular por meio de diferentes técnicas. A observação das células esfoliadas da mucosa bucal demonstra aspectos morfológicos, bioquímicos e/ou moleculares dependendo do tipo do processamento empregado. Objetivo: Este estudo testou o emprego da técnica de citopatologia bucal associada à microscopia eletrônica de transmissão (MET) para observar a morfologia das células, principalmente com relação à membrana citoplasmática, as junções celulares e ao núcleo da célula. Materiais eMétodos: Células epiteliais esfoliadas da mucosa bucal foram analisadas por MET de indivíduos expostos a tabaco e álcool, apresentando leucoplasia ou com diagnóstico de carcinoma espinocelular. Resultados:As junções citoplasmáticas célula-célula nas amostras malignas perderam o padrão irregular característico formado pelas inúmeras interdigitações e o processo juncional das células normais e passaram a apresentar uma superfície citoplasmática reta. O núcleo das células das amostras de leucoplasia e do carcinoma espinocelular apresentou coloração heterogênea, enquanto as células não lesionais foram homogêneas. Discussão: A análise de esfregaços citopatológicos bucais por MET contribui para o entendimento das alterações que ocorrem durante o processo de malignidade da mucosa bucal, principal-mente no que diz respeito à membrana citoplasmática e as junções intercelulares. Conclusão: A MET pode ser um bom método analítico para investigar alterações morfológicas em células esfoliadas do epitélio bucal.

Perfil dos usuários e necessidades odontológicas dos pacientes atendidos nas clínicas odontológicas da Universidade Franciscana ­ Santa Maria, RS / Users profile and dental needs of patients attending the dentistry clinics of Universidade Franciscana ­ Santa Maria, RS

Objetivo: o modelo de uso de serviços de saúde de um determinado grupo populacional é, em grande parte, explicadopor seu perfil de necessidades em saúde. O conhecimento deste é importante para estabelecer o modocomo a atenção odontológica será ofertada. Métodos: realizou-se um estudo retrospectivo a partir de dados secundários(prontuários) (n=381) de pacientes atendidos nas clínicas odontológicas da Universidade Franciscana(UFN) de Santa Maria, RS. Os dados deste estudo foram obtidos através da análise dos prontuários de pacientesatendidos nas clínicas odontológicas da UFN entre os anos de 2005 e 2019. Apenas prontuários com a assinaturado paciente ou responsável foram inseridos neste trabalho. Para a obtenção dos dados, foi construído uminstrumento de coleta, sendo obtidas informações sociodemográficas (idade, sexo, cor, estado civil, profissão elocal de residência) e sobre o estado de saúde (doenças sistêmicas, uso de medicação, perfil de saúde bucal eCPO-D). Cada sujeito selecionado recebeu uma codificação que garantiu seu anonimato. A análise estatísticafoi do tipo descritiva e foi realizada em software estatístico. Resultados: a maioria dos pacientes atendidos erado sexo feminino (68,5%), sendo que a faixa etária mais prevalente foi entre 31 e 50 anos (44,5%). A maioriados pacientes eram estudantes (20,3%), seguidos de trabalhadores do lar (10,9%) e aposentados (7,8%).A queixa principal mais reportada foi a dor de dente (25,2%). Nos resultados deste trabalho, a presença dedoença preexistente foi reportada por 87 (22,8%) pacientes, sendo todas classificadas como doenças crônicas,de acordo com a classificação proposta por Murow e Oglesby13 (1996); a mais prevalente foi problemas alérgicos,69 (18,1%), seguidos por problemas respiratórios, 42 (11%), e problemas gástricos, 39 (10%). A utilizaçãode medicação de uso contínuo foi relatada por 150 (39,3%) pacientes. Conclusão: os pacientes das clínicasodontológicas da UFN de Santa Maria, RS, são, em sua maioria, estudantes, aposentados e trabalhadores do lar,sendo a dor e a fratura dentária as principais queixas dos usuários. Dessa maneira, acredita-se que os discentesda instituição devem estar aptos a realizarem atendimentos de caráter emergencial. O conhecimento do manejoodontológico de pacientes com doenças crônicas não transmissíveis deve ser uma preocupação para daquialguns anos, visto que o envelhecimento da população é esperado para os próximos 30 anos.(AU)

Objective: the health services use model of a given population group is largely explained by its profile of health needs. The knowledge of this is important to establish how dental care will be offered. Methods: in view of this, a retrospective study was carried out using secondary data (medical records) (n = 381) of patients seen at the dental clinics at the Franciscana University (UFN) in Santa Maria, RS. The data of this study were obtained through the analysis of the medical records of patients seen at dental clinics at Universidade Franciscana, between the years 2005 and 2019. Only medical records with the signature of the patient or guardian were included in this work. To obtain the data, an instrument for collecting and collecting sociodemographic information (age, sex, color, marital status, profession and place of residence) and health status (systemic diseases, use of medication, oral health profile and CPO-D) were obtained. Each selected subject received a code that guaranteed their anonymity. The statistical analysis was descriptive and was performed using statistical software. Results: most of the patients seen were female (68.5%), with the most prevalent age group between 31 and 50 years old (44.5%). Most patients were students (20.3%), followed by household workers (10.9%) and retired (7.8%). The main complaint most reported was toothache (25.2%). In the results of this study, the presence of pre-existing disease was reported by 87 (22.8%) patients, all of whom were classified as chronic diseases, according to the classification proposed by Murow and Oglesby13 (1996). The most prevalent were allergic problems, 69 (18.1%), followed by respiratory problems, 42 (11%), and gastric problems, 39 (10%). The use of medication for continuous use was reported by 150 (39.3%) patients. Conclusion: patients at dental clinics at UFN in Santa Maria / RS are mostly students, retirees and home workers, with dental pain and fracture being the main complaints of users. Thus, it is believed that the institution's students must be able to provide emergency care. Knowledge of the dental management of patients with chronic non-communicable diseases should be a concern for some years to come, since the aging of the population is expected for the next 30 years.(AU)

Pericoronal follicles revealing unsuspected odontogenic cysts and inflammatory lesions: A retrospective microscopy study.

AIM: This study aimed to analyze the prevalence of diseases related to pericoronal follicles, and assess the rate of concordance between clinical and histopathological diagnoses. METHODS: Histologically, we analyzed 1,298 tissue samples surrounding the crowns of teeth that were diagnosed clinically as pericoronal follicles. In addition, we determined associations among histopathological diagnosis, patients' age and sex, tissue site, presence of nests of odontogenic epithelium, presence of reduced enamel epithelium, and presence of diffuse inflammation. RESULTS: Odontogenic pathologies were present in 35% of the samples, and rate of concordance between clinical and histopathological diagnoses was 0.54. Probability of developing odontogenic pathologies was high in the mandibular molars (odds ratio: 2.13) and in the tissues with odontogenic epithelial remnants (odds ratio: 1.2), reduced enamel epithelium (odds ratio: 1.3), and diffuse inflammation. (odds ratio: 10.5). CONCLUSIONS: The findings of this study highlight the clinical relevance of histopathological examination of the pericoronal tissue in unerupted and partially erupted teeth for early diagnosis of pathologies because this study demonstrated the odontogenic cysts and inflammatory lesions in tissues clinically diagnosed as pericoronal follicles.

Optimizing Fixation Protocols to Improve Molecular Analysis from FFPE Tissues.

Most Departments of Pathology around the world have a considerable archive of formalin-fixed paraffin-embedded (FFPE) tissue suitable for molecular assessment. This article points out the potential DNA damage that may occur if basic steps are not followed during processing and storage of these samples. Furthermore, it hopes to establish parameters to optimize quality and quantity of DNA extracted from FFPE tissues.

Expression of E-cadherin and involucrin in leukoplakia and oral cancer: an immunocytochemical and immunohistochemical study.

To assess the immunocytochemical and immunohistochemical correlation of adhesion (E-cadherin) and cell differentiation (involucrin) molecules in oral leukoplakia and oral squamous cell carcinoma. Cytological samples and biopsies were obtained from male and female patients aged over 30 years with oral leukoplakia (n = 30) and oral squamous cell carcinoma (n = 22). Cell scrapings and the biopsy were performed at the site of the lesion and histological slides were prepared for the immunocytochemical analysis of exfoliated oral mucosal cells and for the immunohistochemical analysis of biopsy tissues using E-cadherin and involucrin. Spearman's correlation and kappa coefficients were used to assess the correlation and level of agreement between the techniques. Immunostaining for E-cadherin and involucrin by both techniques was similar in the superficial layers of the histological sections compared with cell scrapings. However, there was no statistical correlation and agreement regarding the immunocytochemical and immunohistochemical expression of E-cadherin and involucrin in oral leukoplakia (R = 0.01, p = 0.958) (Kappa = 0.017, p = 0.92) or in oral squamous cell carcinoma (R = 0.26, p = 0.206) (Kappa = 0.36, p = 0.07). The immunoexpression of E-cadherin and involucrin in tissues is consistent with the expression patterns observed in exfoliated oral mucosal cells, despite the lack of a statistically significant correlation. There is an association of the histopathological characteristics of leukoplakia with the expression E-cadherin and of the microscopic aspects of oral squamous cell carcinoma with immunohistochemical expression of involucrin.

Optimizing Fixation Protocols to Improve Molecular Analysis from FFPE Tissues

Abstract Most Departments of Pathology around the world have a considerable archive of formalin-fixed paraffin-embedded (FFPE) tissue suitable for molecular assessment. This article points out the potential DNA damage that may occur if basic steps are not followed during processing and storage of these samples. Furthermore, it hopes to establish parameters to optimize quality and quantity of DNA extracted from FFPE tissues.

Resumo A maioria dos Departamentos de Patologia em todo o mundo têm um considerável acervo de tecidos embebidos em parafina e fixados em formalina, que são passíveis para análises moleculares. Este artigo apresenta os danos ao DNA que podem ocorrer se passos básicos não forem seguidos durante o processamento e armazenamento destas amostras. Além disso, procura estabelecer parâmetros para otimizar a qualidade e quantidade do DNA extraído de tecidos FFPE.

Expression of E-cadherin and involucrin in leukoplakia and oral cancer: an immunocytochemical and immunohistochemical study

Abstract To assess the immunocytochemical and immunohistochemical correlation of adhesion (E-cadherin) and cell differentiation (involucrin) molecules in oral leukoplakia and oral squamous cell carcinoma. Cytological samples and biopsies were obtained from male and female patients aged over 30 years with oral leukoplakia (n = 30) and oral squamous cell carcinoma (n = 22). Cell scrapings and the biopsy were performed at the site of the lesion and histological slides were prepared for the immunocytochemical analysis of exfoliated oral mucosal cells and for the immunohistochemical analysis of biopsy tissues using E-cadherin and involucrin. Spearman’s correlation and kappa coefficients were used to assess the correlation and level of agreement between the techniques. Immunostaining for E-cadherin and involucrin by both techniques was similar in the superficial layers of the histological sections compared with cell scrapings. However, there was no statistical correlation and agreement regarding the immunocytochemical and immunohistochemical expression of E-cadherin and involucrin in oral leukoplakia (R = 0.01, p = 0.958) (Kappa = 0.017, p = 0.92) or in oral squamous cell carcinoma (R = 0.26, p = 0.206) (Kappa = 0.36, p = 0.07). The immunoexpression of E-cadherin and involucrin in tissues is consistent with the expression patterns observed in exfoliated oral mucosal cells, despite the lack of a statistically significant correlation. There is an association of the histopathological characteristics of leukoplakia with the expression E-cadherin and of the microscopic aspects of oral squamous cell carcinoma with immunohistochemical expression of involucrin.

Oral Lesions in Pycnodysostosis Syndrome: Eleven Years of Follow-Up.

Pycnodysostosis is a rare genetic disease that is characterized by osteosclerosis, short stature, and bone fragility. There are not cases of gnathic bones lesions reported on the international literature. This study aims to describe a clinical case of a 10-year-old girl with pycnodysostosis syndrome and an uncommon association with 4 distinct lesions (dentigerous cyst, central giant cell lesions, and 2 fibro-osseous lesions).

Influence of factors in the oral mucosa maturation pattern: a cross-sectional study applying multivariate analyses

Aim: To evaluate the association between oral health status, socio-demographic and behavioralfactors with the pattern of maturity of normal epithelial oral mucosa. Methods: Exfoliative cytologyspecimens were collected from 117 men from the border of the tongue and floor of the mouth onopposite sides. Cells were stained with the Papanicolaou method and classified into: anucleated,superficial cells with nuclei, intermediate and parabasal cells. Quantification was made by selectingthe first 100 cells in each glass slide. Sociodemographic and behavioral variables were collected froma structured questionnaire. Oral health was analyzed by clinical examination, recording decayed,missing and filled teeth index (DMFT) and use of prostheses. Multivariable linear regression modelswere applied. Results: No significant differences for all studied variables influenced the patternof maturation of the oral mucosa except for alcohol consumption. There was an increase of cellsurface layers of the epithelium with the chronic use of alcohol. Conclusions: It is appropriate to usePapanicolaou cytopathological technique to analyze the maturation pattern of exposed subjects,with a strong recommendation for those who use alcohol - a risk factor for oral cancer, in which achange in the proportion of cell types is easily detected.

Immunocytochemistry associated with oral exfoliative cytology: methodological analysis..

OBJECTIVE: To evaluate different immunocytochemical protocol variations to find the most effective protocol for the analysis of involucrin, epidermal growth factor receptor (EGFR), and E-cadherin antibodies. Exfoliative cytology is a noninvasive method used to monitor and screen for early changes in the oral mucosa of patients exposed to carcinogens such as tobacco and alcohol. It has been postulated that its association with immunocytochemistry may improve the effectiveness of the screening process. STUDY DESIGN: Four graduate students from Porto Alegre in southern Brazil had oral smears collected from the border of the tongue using a cytobrush. The following variables were analyzed: cell membrane permeability, antigen retrieval method (microwave oven or water bath), antibody incubation time (overnight or 1 hour), detection system used (Envision or LSAB), and chromogen incubation time (10 seconds or 5 minutes). RESULTS: Best results were obtained with the following combinations: (1) for involucrin: water bath, 1-hour incubation for primary antibody, Envision, and chromogen incubation for 10 seconds; (2)for EGFR: microwave, overnight incubation, LSAB, and chromogen incubation for 5 minutes; and (3) for E-cadherin: water bath, over-night incubation, Envision, and chromogen incubation for 5 minutes. CONCLUSION: Our findings suggest that each antibody requires a specific immunocytochemical protocol to guarantee optimal results with oral smears.