Análise das diferentes técnicas cirúrgicas de recobrimento radicular utilizando tecido conjuntivo

TCC (graduação) - Universidade Federal de Santa Catarina. Centro de Ciências da Saúde. Odontologia.O crescente interesse pela estética, cada vez mais requisitada pelos pacientes, juntamente com a exigência de resultados funcionais favoreceram o desenvolvimento das técnicas de recobrimento radicular. A recessão gengival é uma patologia de etiologia multifatorial, e pode acarretar em problemas funcionais (hipersensibilidade, cárie de raiz, acúmulo de placa e evolução de recessões já existentes) e estéticos. As técnicas de enxerto de tecido conjuntivo subepitelial , as mais utlizadas para recobrimento radicular segundo a liretatura, possibilitam a recuperação do conforto funcional e a devolução de estética ao sorriso. O objetivo deste trabalho foi, através de uma revisão de literatura, avaliar as diferentes técnicas cirúrgicas de recobrimento radicular utilizando tecido conjuntivo. Concluiu-se que as diferentes técnicas de enxerto de tecido conjuntivo subepitelial avaliadas apresentam alta previsibilidade de sucesso devido às duas fontes de nutrição ao tecido, mas não há técnica ideal. As indicações de cada técnica, assim como as limitações anatômicas e biológicas de cada caso clínico devem ser consideradas durante a escolha

Cell proliferation markers at the invasive tumor front of oral squamous cell carcinoma: comparative analysis in relation to clinicopathological parameters of patients

Objectives To evaluate the number of AgNORs per nucleus and the expression of Ki-67 at the tumor invasion front (TIF) in relation to clinical parameters (TNM), TIF classification and the prognosis of oral squamous cell carcinomas in an Uruguayan population. Material and Methods This study was conducted through a retrospective survey from 2000 to 2010 at the National Institute of Cancer Montevideo, Uruguay and included 40 patients. The samples were obtained from the resection of the tumor and the TIF was defined according with Bryne, et al.5 (1992). Expression of Ki-67 was assessed by the percentage of positive tumor cells and the AgNOR was recorded as the mean AgNOR (mAgNOR) and the percentage of AgNOR per nucleus (pAgNOR). All analyzes were performed by a blinded and calibrated observer. Results No statistically significant association was observed between immunostaining of Ki-67 and AgNOR with the different types of TIF, regional metastasis and patients prognosis, however it was observed an increase in Ki-67 expression associated with worse patient’s clinical staging, although not statistically significant. Conclusions Our results suggest that proliferation markers as AgNOR and Ki-67 are not prognostic markers at the tumor invasive front of carcinoma of oral squamous cell

Chemoprevention in oral leukoplakia: challenges and current landscape

Oral potentially malignant disorders have the potential to transform into oral cancer. Oral leukoplakia is a prevalent OPMD with a 9.8% malignant transformation rate. The standard management for OL involves surgical excision, but its efficacy in preventing clinical recurrence and malignant transformation is limited. Therefore, alternative strategies such as chemoprevention modalities have emerged as a promising approach to inhibit the carcinogenesis process. The aim of this review was to identify human studies that investigated the effectiveness of chemopreventive agents in preventing the progression of oral leukoplakia and to provide guidance for future research. Several systemic and topical agents have been evaluated for their potential chemopreventive effects in oral leukoplakia. Systemic agents that have been investigated include vitamin A, lycopene, celecoxib, green tea extract, ZengShengPing, Bowman Birk inhibitor, beta-carotene, curcumin, erlotinib, and metformin. In addition, topical agents tested include bleomycin, isotretinoin, ONYX-015 mouthwash, ketorolac, and dried black raspberry. Despite numerous agents that have already been tested, evidence supporting their effectiveness is limited. To improve the search for an ideal chemopreventive agent for oral leukoplakia, we propose several strategies that can be implemented. Oral leukoplakia chemoprevention presents a promising opportunity for decreasing the incidence of oral cancer. Identifying new chemopreventive agents and biomarkers for predicting treatment response should be a focus of future research

Cell proliferation markers at the invasive tumor front of oral squamous cell carcinoma: comparative analysis in relation to clinicopathological parameters of patients

Objectives: To evaluate the number of AgNORs per nucleus and the expression of Ki-67 at the tumor invasion front (TIF) in relation to clinical parameters (TNM), TIF classification and the prognosis of oral squamous cell carcinomas in an Uruguayan population.Material and Methods: This study was conducted through a retrospective survey from 2000 to 2010 at the National Institute of Cancer Montevideo, Uruguay and included 40 patients. The samples were obtained from the resection of the tumor and the TIF was defined according with Bryne, et al.5 (1992). Expression of Ki-67 was assessed by the percentage of positive tumor cells and the AgNOR was recorded as the mean AgNOR (mAgNOR) and the percentage of AgNOR per nucleus (pAgNOR). All analyzes were performed by a blinded and calibrated observer.Results: No statistically significant association was observed between immunostaining of Ki-67 and AgNOR with the different types of TIF, regional metastasis and patients prognosis, however it was observed an increase in Ki-67 expression associated with worse patient’s clinical staging, although not statistically significant.Conclusions:Our results suggest that proliferation markers as AgNOR and Ki-67 are not prognostic markers at the tumor invasive front of carcinoma of oral squamous cell.Objectives: To evaluate the number of AgNORs per nucleus and the expression of Ki-67 at the tumor invasion front (TIF) in relation to clinical parameters (TNM), TIF classification and the prognosis of oral squamous cell carcinomas in an Uruguayan population.Material and Methods: This study was conducted through a retrospective survey from 2000 to 2010 at the National Institute of Cancer Montevideo, Uruguay and included 40 patients. The samples were obtained from the resection of the tumor and the TIF was defined according with Bryne, et al.5 (1992). Expression of Ki-67 was assessed by the percentage of positive tumor cells and the AgNOR was recorded as the mean AgNOR (mAgNOR) and the percentage of AgNOR per nucleus (pAgNOR). All analyzes were performed by a blinded and calibrated observer.Results: No statistically significant association was observed between immunostaining of Ki-67 and AgNOR with the different types of TIF, regional metastasis and patients prognosis, however it was observed an increase in Ki-67 expression associated with worse patient’s clinical staging, although not statistically significant.Conclusions:Our results suggest that proliferation markers as AgNOR and Ki-67 are not prognostic markers at the tumor invasive front of carcinoma of oral squamous cell.Objectives: To evaluate the number of AgNORs per nucleus and the expression of Ki-67 at the tumor invasion front (TIF) in relation to clinical parameters (TNM), TIF classification and the prognosis of oral squamous cell carcinomas in an Uruguayan population.Material and Methods: This study was conducted through a retrospective survey from 2000 to 2010 at the National Institute of Cancer Montevideo, Uruguay and included 40 patients. The samples were obtained from the resection of the tumor and the TIF was defined according with Bryne, et al.5 (1992). Expression of Ki-67 was assessed by the percentage of positive tumor cells and the AgNOR was recorded as the mean AgNOR (mAgNOR) and the percentage of AgNOR per nucleus (pAgNOR). All analyzes were performed by a blinded and calibrated observer.Results: No statistically significant association was observed between immunostaining of Ki-67 and AgNOR with the different types of TIF, regional metastasis and patients prognosis, however it was observed an increase in Ki-67 expression associated with worse patient’s clinical staging, although not statistically significant.Conclusions:Our results suggest that proliferation markers as AgNOR and Ki-67 are not prognostic markers at the tumor invasive front of carcinoma of oral squamous cell

Organotypic Co-Cultures as a Novel 3D Model for Head and Neck Squamous Cell Carcinoma

Background: Head and neck squamous cell carcinomas (HNSCC) are phenotypically and molecularly heterogeneous and frequently develop therapy resistance. Reliable patient-derived 3D tumor models are urgently needed to further study the complex pathogenesis of these tumors and to overcome treatment failure. Methods: We developed a three-dimensional organotypic co-culture (3D-OTC) model for HNSCC that maintains the architecture and cell composition of the individual tumor. A dermal equivalent (DE), composed of healthy human-derived fibroblasts and viscose fibers, served as a scaffold for the patient sample. DEs were co-cultivated with 13 vital HNSCC explants (non-human papillomavirus (HPV) driven, n = 7; HPV-driven, n = 6). Fractionated irradiation was applied to 5 samples (non-HPV-driven, n = 2; HPV-driven n = 3). To evaluate expression of ki-67, cleaved caspase-3, pan-cytokeratin, p16INK4a, CD45, ∝smooth muscle actin and vimentin over time, immunohistochemistry and immunofluorescence staining were performed Patient checkup data were collected for up to 32 months after first diagnosis. Results: All non-HPV-driven 3D-OTCs encompassed proliferative cancer cells during cultivation for up to 21 days. Proliferation indices of primaries and 3D-OTCs were comparable and consistent over time. Overall, tumor explants displayed heterogeneous growth patterns (i.e., invasive, expansive, silent). Cancer-associated fibroblasts and leukocytes could be detected for up to 21 days. HPV DNA was detectable in both primary and 3D-OTCs (day 14) of HPV-driven tumors. However, p16INK4a expression levels were varying. Morphological alterations and radioresistant tumor cells were detected in 3D-OTC after fractionated irradiation in HPV-driven and non-driven samples. Conclusions: Our 3D-OTC model for HNSCC supports cancer cell survival and proliferation in their original microenvironment. The model enables investigation of invasive cancer growth and might, in the future, serve as a platform to perform sensitivity testing upon treatment to predict therapy response

Estudo Study of the effect of smoking and its abandonment on the speed of proliferation of cells of the oral mucosa: longitudinal evaluation

OBJETIVO: avaliar através da citopatologia, associada a técnica AgNOR, o reflexo da cessação do fumo sobre a velocidade de proliferação das células epiteliais descamadas de borda de língua (BL) e assoalho de boca (AB), ao longo do tempo. MÉTODOS: Foram incluídos na amostra inicial 138 indivíduos. As coletas foram realizadas em um momento inicial (T1), após 8-15 meses (T2) e após 16-30 meses (T3). Os participantes foram divididos em três grupos de acordo com os seguintes critérios: Grupo Controle (GC) (n=71): indivíduos atendidos na Faculdade de Odontologia da UFRGS, que nunca fumaram, e que ingeriam até 30g de álcool por semana. Grupo Abandono de Fumo (GAF) (n=26): indivíduos em acompanhamento no Grupo dos Fumantes do Hospital de Clínicas de Porto Alegra (HCPA), que fumavam no momento do inicial do trabalho, e que ao longo do tempo cessaram o hábito tabágico. Alcoolistas ou não. Grupo Fumo (GF) (n=41): indivíduos em acompanhamento no Grupo dos Fumantes do HCPA, que fumavam no momento do inicial do trabalho, e que não cessaram o hábito tabágico ao longo da pesquisa. Alcoolistas ou não. As amostras foram submetidas à técnica de AgNOR e quantificadas por três examinadores previamente calibrados. A média de AgNOR por núcleo (mAgNOR) e a porcentagem de células com mais de 1, 2 E 3 AgNORs (pAgNOR>1, pAgNOR>2 e pAgNOR>3) foram calculados nas 50 primeiras células, nucleadas, não sobrepostas, e bem distendidas. RESULTADOS: ao final dos tempos experimentais os grupos tinham 9 individuos em cada. Não foram observada diferença estatística do mAgNOR e pAgNOR>3 entre os grupos avaliados, nos diferentes tempos em ambos os sítios. Foi observada uma tendência à diminuição na velocidade de proliferação, no tempo intermediário, e uma tendência a aumentar no tempo final, em todos os grupos Em T1, os grupos expostos ao fumo (GAF e GF) apresentavam velocidade de proliferação mais elevada, se comparados ao GC no BL. No AB o GC apresenta valores maiores que GAF. Em T2, os indivíduos do GAF apresentaram uma redução da taxa de proliferação celular, em ambos os sítios analisados, maior do que GC e GF. Em T3 os valores retornam ao equilíbrio, exibindo valores maiores que os em T1. No BL o pAgNOR>1 e 2 mostrou uma diferença estatística entre os grupos em T1 e T2. O sítio AB apresentou diferença estatística no pAgNOR>1, em T2, mostrando que indivíduos do GC e GAF apresentam velocidade de proliferação menor quando comparados ao GF. CONCLUSÕES: O estudo mostrou que o GAF apresenta maior flutuação, de mAgNOR, que o GC e GF, ao longo do tempo. Estudo futuros ampliando o tempo de controle, controlando os agentes carcinógenos, e observando indivíduos com características populacionais seriam convenientes para buscar a padronização deste modelo de prevenção de câncer.OBJECTIVE: evaluate, through cytopathology associated with AgNOR technique, the reflection of smoking cessation on the speed of proliferation of desquamated epithelial cells of tongue edge (TE) and mouth floor (MF) over time. METHODS: We included in the initial sample 138 individuals. Samples were collected at an early time (T1), after 8-15 months (T2) and after 16-30 months (T3). Participants were divided into three groups, according to the following criteria: control group (CG) (n = 71): patients seen at the Faculty of Dentistry at UFRGS, who had never smoked and who drank till 30g of alcohol a week. Abandonment group Smoke (AS) (n = 26): individuals in monitoring the Group of Smokers of Hospital de clínicas dePorto Alegre (HCPA), who smoked at the time of the initial work, and that over time ceased the smoking habit. Alcoholic or not. Tobacco group (TG) (n = 41): follow-up individuals in the group of smokers HCPA, who smoked at the time of the initial work, and that did not stop the smoking habit during the research. Alcoholic or not. Samples were subjected to AgNOR technique and quantified for three calibrated examiners. The average AgNOR per core (Magnor) and the percentage of cells with 1, 2 and 3 AgNOR (pAgNOR> 1, pAgNOR> 2 and pAgNOR> 3) were calculated on the first 50 cells, nucleated, nonoverlapping, and well- distended. RESULTS: At the end of the experimental time groups had 9 individuals in each. There were no statistical difference of mAgNOR and pAgNOR> 3 between the groups in different times on both sites. A tendency to decrease in the proliferation rate was observed, at the intermediate time, and a trend of increase in the end time for all groups in T1, the smoke exposed groups (AS and TG) showed higher proliferation rate, compared the CG in the TE. MF in the CG presents values greater than AS. In T2, AS individuals showed a reduction in cell proliferation rate in both sites evaluated greater than TG and CG. In T3 values return to equilibrium, exhibiting values greater than the T1. TE in the pAgNOR> 1 and 2 showed a statistical difference between groups in T1 and T2. The site MF showed statistical difference in pAgNOR> 1, T2, showing that CG and the AS show less proliferation of speed when compared to the TG. CONCLUSIONS: The study showed that the AS has greater fluctuation mAgNOR that CG and TG, over time. Future study extending the time control, controlling carcinogens, and observing individuals with population characteristics would be convenient to seek standardization of this cancer prevention model

A metilação do DNA é um evento comum que regula a expressão gênica no câncer de cabeça e pescoço

Head and neck squamous cells carcinoma (HNSCC) represents one of the most prevalent and lethal human malignancies worldwide. Despite the advances in early detection and implementation of interdisciplinary therapy, the aggressive and invasive tumor growth pattern in addition to the resistance to well established treatment modalities, remains the main cause for poor outcome. Thus, a better understanding of molecular features involved in early and late HNSCC steps is an urgent need for the improvement of patients management and outcome. In this study, we utilize multiscale omics based on whole-exome sequencing, global DNA methylation and gene expression profile to investigate cancer-related genes in tumor initiation and progression processes. The combination of genetic and epigenetic alterations showed the effect of DNA methylation in regulating two different genes (RYR2 and SOX2) during tumor initiation and progression. The detection of DNA methylation and respective gene expression might help to predict patients risk for malignant transformation and worse outcome.O carcinoma de células escamosas de cabeça e pescoço (HNSCC) representa um dos mais prevalentes e malignidades humanas letais em todo o mundo. Apesar dos avanços na detecção precoce e implementação da terapia interdisciplinar, o padrão de crescimento tumoral agressivo e invasivo em além da resistência a modalidades de tratamento bem estabelecidas, continua sendo a principal causa de resultado. Assim, uma melhor compreensão das características moleculares envolvidas no HNSCC precoce e tardio passos é uma necessidade urgente para a melhoria da gestão dos pacientes e resultados. Neste estudo, utilizamos ômicas multiescala com base no sequenciamento do exoma inteiro, metilação global do DNA e genes perfil de expressão para investigar genes relacionados ao câncer na iniciação e progressão do tumor processos. A combinação de alterações genéticas e epigenéticas mostrou o efeito do DNA metilação na regulação de dois genes diferentes (RYR2 e SOX2) durante a iniciação do tumor e progressão. A detecção da metilação do DNA e a respectiva expressão gênica podem ajudar a prever o risco de pacientes para transformação maligna e pior resultado

Estudo Study of the effect of smoking and its abandonment on the speed of proliferation of cells of the oral mucosa: longitudinal evaluation

OBJETIVO: avaliar através da citopatologia, associada a técnica AgNOR, o reflexo da cessação do fumo sobre a velocidade de proliferação das células epiteliais descamadas de borda de língua (BL) e assoalho de boca (AB), ao longo do tempo. MÉTODOS: Foram incluídos na amostra inicial 138 indivíduos. As coletas foram realizadas em um momento inicial (T1), após 8-15 meses (T2) e após 16-30 meses (T3). Os participantes foram divididos em três grupos de acordo com os seguintes critérios: Grupo Controle (GC) (n=71): indivíduos atendidos na Faculdade de Odontologia da UFRGS, que nunca fumaram, e que ingeriam até 30g de álcool por semana. Grupo Abandono de Fumo (GAF) (n=26): indivíduos em acompanhamento no Grupo dos Fumantes do Hospital de Clínicas de Porto Alegra (HCPA), que fumavam no momento do inicial do trabalho, e que ao longo do tempo cessaram o hábito tabágico. Alcoolistas ou não. Grupo Fumo (GF) (n=41): indivíduos em acompanhamento no Grupo dos Fumantes do HCPA, que fumavam no momento do inicial do trabalho, e que não cessaram o hábito tabágico ao longo da pesquisa. Alcoolistas ou não. As amostras foram submetidas à técnica de AgNOR e quantificadas por três examinadores previamente calibrados. A média de AgNOR por núcleo (mAgNOR) e a porcentagem de células com mais de 1, 2 E 3 AgNORs (pAgNOR>1, pAgNOR>2 e pAgNOR>3) foram calculados nas 50 primeiras células, nucleadas, não sobrepostas, e bem distendidas. RESULTADOS: ao final dos tempos experimentais os grupos tinham 9 individuos em cada. Não foram observada diferença estatística do mAgNOR e pAgNOR>3 entre os grupos avaliados, nos diferentes tempos em ambos os sítios. Foi observada uma tendência à diminuição na velocidade de proliferação, no tempo intermediário, e uma tendência a aumentar no tempo final, em todos os grupos Em T1, os grupos expostos ao fumo (GAF e GF) apresentavam velocidade de proliferação mais elevada, se comparados ao GC no BL. No AB o GC apresenta valores maiores que GAF. Em T2, os indivíduos do GAF apresentaram uma redução da taxa de proliferação celular, em ambos os sítios analisados, maior do que GC e GF. Em T3 os valores retornam ao equilíbrio, exibindo valores maiores que os em T1. No BL o pAgNOR>1 e 2 mostrou uma diferença estatística entre os grupos em T1 e T2. O sítio AB apresentou diferença estatística no pAgNOR>1, em T2, mostrando que indivíduos do GC e GAF apresentam velocidade de proliferação menor quando comparados ao GF. CONCLUSÕES: O estudo mostrou que o GAF apresenta maior flutuação, de mAgNOR, que o GC e GF, ao longo do tempo. Estudo futuros ampliando o tempo de controle, controlando os agentes carcinógenos, e observando indivíduos com características populacionais seriam convenientes para buscar a padronização deste modelo de prevenção de câncer.OBJECTIVE: evaluate, through cytopathology associated with AgNOR technique, the reflection of smoking cessation on the speed of proliferation of desquamated epithelial cells of tongue edge (TE) and mouth floor (MF) over time. METHODS: We included in the initial sample 138 individuals. Samples were collected at an early time (T1), after 8-15 months (T2) and after 16-30 months (T3). Participants were divided into three groups, according to the following criteria: control group (CG) (n = 71): patients seen at the Faculty of Dentistry at UFRGS, who had never smoked and who drank till 30g of alcohol a week. Abandonment group Smoke (AS) (n = 26): individuals in monitoring the Group of Smokers of Hospital de clínicas dePorto Alegre (HCPA), who smoked at the time of the initial work, and that over time ceased the smoking habit. Alcoholic or not. Tobacco group (TG) (n = 41): follow-up individuals in the group of smokers HCPA, who smoked at the time of the initial work, and that did not stop the smoking habit during the research. Alcoholic or not. Samples were subjected to AgNOR technique and quantified for three calibrated examiners. The average AgNOR per core (Magnor) and the percentage of cells with 1, 2 and 3 AgNOR (pAgNOR> 1, pAgNOR> 2 and pAgNOR> 3) were calculated on the first 50 cells, nucleated, nonoverlapping, and well- distended. RESULTS: At the end of the experimental time groups had 9 individuals in each. There were no statistical difference of mAgNOR and pAgNOR> 3 between the groups in different times on both sites. A tendency to decrease in the proliferation rate was observed, at the intermediate time, and a trend of increase in the end time for all groups in T1, the smoke exposed groups (AS and TG) showed higher proliferation rate, compared the CG in the TE. MF in the CG presents values greater than AS. In T2, AS individuals showed a reduction in cell proliferation rate in both sites evaluated greater than TG and CG. In T3 values return to equilibrium, exhibiting values greater than the T1. TE in the pAgNOR> 1 and 2 showed a statistical difference between groups in T1 and T2. The site MF showed statistical difference in pAgNOR> 1, T2, showing that CG and the AS show less proliferation of speed when compared to the TG. CONCLUSIONS: The study showed that the AS has greater fluctuation mAgNOR that CG and TG, over time. Future study extending the time control, controlling carcinogens, and observing individuals with population characteristics would be convenient to seek standardization of this cancer prevention model