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Item Avaliação da expressão salivar e tecidual das citocinas TGF-β e IL-10 em pacientes com carcinoma espinocelular de cavidade oral(Universidade Federal de Goiás, 2015-03-02) Arantes, Diego Antonio Costa; Costa, Nádia do Lago; Batista, Aline Carvalho; http://lattes.cnpq.br/0199082642322002; Batista, Aline Carvalho; Guerra, Eliete Neves da Silva; Fonseca, Simone Gonçalves daThe transforming growth factor β (TGF-β) and interleukin 10 (IL-10) are immunosuppressive cytokines which promote failure of the local anti-tumor immune response and, therefore, influence the proliferation and prognosis of malignant neoplasms. The aim of this study was to investigate the tissue and salivary expression of TGF-β and IL-10 in patients with oral squamous cell carcinoma (OSCC) and compare it with that of healthy subjects (Control). The association of these cytokines with clinical parameters of prognosis (staging, metastasis and survival) and histological grade of malignancy (WHO grading) was also investigated. Cytokines in the tissue (OSCC, n = 65; Control, n = 30) were identified using the immunohistochemistry technique (IHC) and in the saliva (OSCC, n = 22; Control, n = 23) the Enzyme-linked Immunosorbent Assay (ELISA) was used. The tissue expression of TGF-β and IL-10 in metastatic lymph nodes (n = 23) of OSCC patients was investigated. The expression of TGF-β and IL-10 in the tissue was measured using a semi-quantitative method in conjunction with staining intensity. Our findings demonstrated a high tissue expression of IL-10 and TGF-β2, and a low or absent expression of TGF-β1, in the majority of OSCC samples when compared to the group with clinically healthy oral mucosa (Control) (p < 0.05 for IL-10 and TGF-β2). The salivary concentration of IL-10 was also high, and distinguished the OSCC patients from their healthy counterparts (p = 0.04), while the salivary concentration of TGF-β1 was similar for both the OSCC and control groups (p = 0.97). The relationship between the cytokine expression and clinical and microscopic prognostic factors showed that the expression of IL-10 and TGF-β2 in neoplastic cells of the primary tumor was maintained by the metastatic neoplastic cells in the cervical lymph nodes. The expression of TGF-β1 remained low or absent in the metastatic lymph nodes. It was shown that there was an association between the high expression of IL-10 by tumor cells and the advanced clinical stages (T3-T4) of patients (p = 0.02). Although not statistically significant, the expression of TGF-β2 was higher in tumors at more advanced stages (p > 0.05). These findings demonstrate that OSCC provides an immunosuppressive environment conducive to tumor proliferation, with high expression of IL-10 and TGF-β2, which contributes to a worse clinical prognosis. In addition, of the immunosuppressive cytokines investigated, IL-10 has greater potential for becoming salivary biomarker when associated with an unfavorable clinical prognosis of OSCC patients.Item Ensaio clínico randomizado controlado triplo cego para avaliação da ansiedade e estresse de crianças submetidas à sedação com midazolam oral durante tratamento odontológico(Universidade Federal de Goiás, 2013-02-26) Gomes, Heloisa de Sousa; Costa, Luciane Ribeiro de Rezende Sucasas da; Batista, Aline Carvalho; http://lattes.cnpq.br/0535612924109069; Batista, Aline Carvalho; Alves Neto, Onofre; Sacono, Nancy TomokoThe fear and anxiety represent a barrier for dental care and can cause behavioral and physiological changes. These changes can be evaluated through behavioral scales and the measurement of salivary cortisol levels. The aim of this study was to evaluate the level of salivary cortisol in children during restorative dental treatment under moderate sedation with midazolam and taken a placebo and verify the correlation between this physiological measure and the assessment of the behavior conducted through the scale Ohio State University Behavioral Rating Scale (OSUBRS). A randomized controlled crossover triple-blind clinical trial was conducted with 18 healthy children from 2 to 5-year olds with necessity of at least two sessions of restorative dental treatment. Each child was undergo treatment under sedation with 1mg/kg oral midazolam in one session and the other with a placebo and in both sessions protective stabilization was associated. The assessment of child behavior was conducted from videos of clinical sessions using the scale of OSUBRS and the salivary cortisol level was evaluated in 4 moments on the two sessions (waking up, on arrival at the Dental School (DS), 25 minutes after the anesthesia and 25 minutes after finishing the procedure). The saliva samples were analyzed by the Enzyme Immunoassay test to get the mean salivary cortisol level. The results showed that the salivary cortisol level was lower when the children had received midazolam than when they had received the placebo at the moment of anesthesia (p=0.004). It was! Observed greater variation in cortisol level when the children received placebo than when they received midazolam. However, there were no differences between salivary cortisol levels observed in the four moments during treatment with sedation (p = 0.319) or placebo (p = 0.080). Regarding the child behavior it was not observed improvement during the treatment with sedation compared with placebo. The salivary cortisol levels showed no statistically significant correlation with the child’s behavior assessed by the scale OSUBRS during dental treatment under sedation or placebo. Therefore it was concluded that the oral midazolam dose of 1.0 mg/kg is effective in reducing the levels of salivary cortisol of children from 2 to 5-year olds during restorative dental treatment. However, this reduction in the level of cortisol in saliva did not reflect in better clinical behavior of these children.Item Avaliação de moléculas envolvidas no escape imunológico em desordens potencialmente malignas de boca(Universidade Federal de Goiás, 2017-02-20) Gonçalves, Andréia de Souza; Batista, Aline Carvalho; http://lattes.cnpq.br/0199082642322002; Batista, Aline Carvalho; Yamamoto, Fernanda Paula; Alves, Pollianna Muniz; Silva, Tarcilia Aparecida; Loyola, Patrícia Resende Alo NagibStudy Rationale: Oral potentially malignant disorders (OPMDs) consist of morphologically altered tissues, which present a greater risk of malignant transformation than normal tissue. The most frequently found OPMDs are leukoplakia (OL) and actinic cheilitis (AC). It is now known that mutated or genetically altered cells have developed immunomodulatory strategies which allow them to escape antitumor immune response. The immune escape mechanisms used by mutated cells include the expressions of HLA-G, HLA-E and PD-L1 molecules and the IL-10 and TGF-β cytokines. Objective: To evaluate tissue and salivary expressions of HLA-G, HLA-E and PD-L1 molecules and IL-10 and TGF-β1, -β2 e -β3 cytokines in OPMDs, and relate such immunomodulatory mediators with antitumor immune response and potential for malignant transformation of lesions. Methods: Samples of patients with OL (n= 80) were submitted to immunohistochemistry and ELISA techniques to evaluate tissue and salivary expressions of the HLA-G, HLA-E, PD-L1, IL-10, TGF-β1, TGF-β2 and TGF-β3. In addition, samples of patients with QA (n= 30) were submitted to immunohistochemistry technique to evaluation tissue expression of HLA-G, HLA-E, PD-L1 and IL-10. Control group (n= 20) consisted of saliva and tissue of healthy individuals. The immunostained tissue samples were measured using a semi-quantitative method in association with staining intensity. The expression of these molecules and cytokines were related with the malignant potential of the lesions (epithelial dysplasia grading, proliferation- Ki-67 and apoptosis index- mutated p53). Moreover, the association with the density of granzyme B (GB+) cells and regulatory T cells FOXP3+ was investigated. The statistical tests used were: Fisher’s exact or Pearson Chi-Squared, Mann-Whitney and Kruskal- Wallis tests. The significance level was set at P < 0.05. Results: Fifteen samples showed severe dysplasia, twenty moderate, thirty-two mild and thirteen non-dysplasia. Forty samples (50%) of OL presented combined high Ki-67/p53. Irrespective of the severity of epithelial dysplasia and proliferation/apoptosis index in OL, an overexpression of HLA-G, -E, PD-L1, IL-10, TGF-β2 and -β3 was found in OL when compared with control (P < 0.05). The number of GB+ and FOXP3+ cells in OL was similar to control. Salivary concentration of sHLA-G, IL-10 and TGF-β did not allow distinction between OL patients and healthy individuals (P > 0.05). As regards to AC, we showed that this lesion had an increase in expression of HLA-G, HLA-E, IL-10 and PD-L1 when compared to control; however this increase was statistically significant only for PD-L1 (P= 0.04). Conclusion: The OL showed a reduced cytotoxic immune response (low number of GB+ cells) associated with a high expression of immunomodulatory mediators; however, this expression was independent of epithelial dysplasia grading, proliferation and apoptosis index. Regarding AC we also showed an increase in expression of HLA-G, -E, IL-10 and PD-L1 when compared to the control. Thus, our findings suggest that this lesion present an immunosuppressive microenvironment which favors the escape of mutated keratinocytes in any stage dysplastic, proliferation or apoptosis which this disease finds itself.Item Avaliação da expressão da molécula HLA-G no microambiente tumoral e na saliva de pacientes com carcinoma espinocelular de cavidade oral(Universidade Federal de Goiás, 2014-01-31) Gonçalves, Andréia de Souza; Wastowski, Isabela Jubé; Batista, Aline Carvalho; http://lattes.cnpq.br/0199082642322002; Batista, Aline Carvalho; Bernardes, Vanessa de Fátima; Dias, Fátima Ribeiro; Wastowski, Isabela JubéHLA-G is a molecule of the major histocompatibility complex (MHC) class I nonclassical that plays an inhibitory effect on immunocompetent cells that are fundamental in the development of an antitumoral response. In this context, the aim of this study was to evaluate the expression of HLA-G in the microenvironment of oral cavity squamous cell carcinoma (OCSCC) as well as their relationship with clinical and microscopic parameters. In addition, we aimed also to compare the salivary concentration of soluble HLA-G (sHLA-G) in patients with OCSCC with healthy individuals (control group). The techniques of immunohistochemistry and Enzyme- Linked Immunosorbent Assay (ELISA) were used. The results revealed, with the immunohistochemistry technique, that expression of HLA-G by tumor cells was significantly higher in metastatic OCSCC (n=30) (72% of samples) when compared with non-metastatic OCSCC (n=30) (28% of samples) (P=0.01 ). Moreover, patients with a lower expression of HLA-G presented a tendency to survival longer (22 months) when compared with those with a higher expression of the molecule (16 months). With reference to other clinicopathological parameters analyzed, only the presence of lymph node metastasis (P=0.01) and depth of tumor invasion (P=0.02) were significantly associated with the expression of HLA-G. By ELISA technique, we showed that the salivary concentration of soluble HLA-G provided no differentiation between patients with OCSCC from healthy individuals, since there was no statistically significant difference between the analyzed groups (P=0.17). The findings of this study revealed that high expression of HLA-G in the microenvironment of metastatic OCSCC may represent a tumor escape mechanism which contributes to lower survival and an unfavorable clinical prognosis. However, concentration of soluble HLA-G in saliva of patients with OCSCC is similar to healthy individuals; thereby sHLA-G does not represent a salivary biomarker prognostic.Item Avaliação da expressão tecidual dos ligantes de morte programada-1 e -2, do receptor PD-1 e da resposta imunológica citotóxica no líquen plano oral(Universidade Federal de Goiás, 2020-05-29) Gonçalves, Julie Ane Maria; Costa, Nádia do Lago; http://lattes.cnpq.br/9120865567187887; Batista, Aline Carvalho; http://lattes.cnpq.br/0199082642322002; Costa, Nádia Do Lago; Mendonça, Elismauro Francisco de; Silveira, Ericka Janine Dantas daOral lichen planus (OLP) is a chronic autoimmune disease, mediated by T lymphocytes (TL) and characterized by apoptosis of basal/suprabasal keratinocytes. Although its etiopathogenesis is not completely elucidated, recent data reveal that blocking immunoregulatory molecules such as programmed death ligands (PD-Ls) and/or PD-1 receptor may promote the appearance of oral lichen planus-like lesions. It is classically established that this PD-Ls/PD-1 pathway contributes to evasion of neoplastic cells; however, it may be important in the regulation of helper and cytotoxic (CD8+) TL in autoimmune diseases. Objective: To investigate the tissue expression of PD-L1 and PD-L2 molecules, as well as PD-1+, CD8+ and granzyme B+ (GrB) cell populations in OLPs and whether there is a relationship between these immunoinhibitory proteins/cell populations and the severity of OLP. Material and methods: Samples of OLP patients (n = 23) were classified according to the histopathological criteria of the American Association of Oral and Maxillofacial Pathology (AAOMP/2016) and submitted to immunohistochemistry. Semi-quantitative (PD-L1+ and PD-L2+) and quantitative analysis (PD-1+, CD8+ and GrB+ cells) were performed. The severity of OLP was assessed by clinical subtype, symptomatology and response to corticosteroid therapy. Results: Most OLP samples were considered negative for PD-L1 (n = 14/22; 63.7%), however high PD-L2 expression (n = 19/22; 86.3%) by both keratinocytes and immunoinflammatory cells has been demonstrated. Low cytotoxic immune response (CD8/GrB ratio per mm2) was evidenced in OLP samples (subepithelial: 1047.4/140.6 and intraepithelial: 197.7/41.6). In addition, PD-1+/mm2 (subepithelial: 70.2 and intraepithelial: 7.4) cell density was reduced compared to CD8+/mm2 LT (subepithelial: 1047.4 and intraepithelial: 197.7) (p <0.01). There was a significantly lower number of GrB+ cells in the intraepithelial region in reticular OLP compared to erosive / bullous OLP (p = 0.03). Conclusions: The findings show that the PD-L1 / PD-1 pathway seems to be compromised in OLP due to low PD-L1 expression and PD-1 + cell scarcity in most samples. On the other hand, PD-L2 overexpression added to a possible regulation of cytotoxic immune response suggests an immune tolerance that may contribute to the chronic profile of OLP.Item Análise do perfil epidemiológico, clínico e patológico de pacientes com líquen plano oral(Universidade Federal de Goiás, 2020-01-27) Lima, Sara Lia Gonçalves de; Arantes, Diego Antônio Costa; http://lattes.cnpq.br/8341286790648954; Batista, Aline Carvalho; http://lattes.cnpq.br/0199082642322002; Batista, Aline Carvalho; Mendonça, Elismauro Francisco de; Leite, Angélica Ferreira Oton; Costa, Nádia do Lago; Ferreira, Alexandre BellottiINTRODUCTION: Oral lichen planus (OLP) is an immunomediated disease whose diagnosis and treatment can be challenging for clinicians and pathologists. OBJECTIVE: This case series aimed to evaluate the epidemiological, clinical and pathological profile of patients with OLP according to the criteria established by the American Academy of Oral and Maxillofacial Pathology (AAOMP/2016). METHODOLOGY: Sampling was initially established from the analysis of the annotations of the medical records and histopathological reports of patients diagnosed with OLP at the Centro Goiano de Doenças da Boca (CGDB) from 1998 to 2018. Subsequently, the clinical and demographic data of the patients lesions (location and clinical characteristics of the lesion) and the microscopic characterization of the specimens followed the criteria proposed by AAOMP / 2016. In addition, patients were invited to follow up and apply the Oral Health Impact Profile (OHIP-14) instrument to assess quality of life. Statistical analysis was descriptive with absolute and relative values and the Shapiro-Wilk test was used to assess the normality of the data collected in the OHIP-14 and then the groups were compared using the Mann-Whitney test. RESULTS: This case series (n=41) consisted mainly of women (n=29; 70.7%) with a mean age of 45 (±13.6) years. The buccal mucosa (68.2%) was the most affected site. Reticular (n=23; 56.1%) and erosive (n=14; 34.3%) OLP were the most frequent. According to OHIP-14, individuals with OLP at multiple sites of the oral cavity had worse values in the handicap domain (p=0.03). In addition, those patients who did not respond to corticosteroids had higher scores in the psychological discomfort domain (p=0.04). CONCLUSION: The findings of the present study using the AAOMP/2016 criteria are similar to other case series and retrospective studies reported in the literature, where OLP is more common in middle-aged women in reticular form. In addition, OLP, in its most severe clinical forms, seems to influence patients' quality of life.Item Avaliação da expressão de proteínas envolvidas na evasão tumoral no carcinoma mucoepidermóide de glândula salivar menor(Universidade Federal de Goiás, 2017-02-21) Mosconi, Carla; Batista, Aline Carvalho; http://lattes.cnpq.br/0199082642322002; Aline Carvalho Batista, Aline Carvalho Batista; Vêncio, Eneida Franco; Alves, Pollianna MunizBackground: The Human Leukocyte Antigen G and E (HLA-G and HLA-E) and Programmed Death Ligand-1 (PD-L1) are molecules which can provide tumor immunosuppression as well as the capacity of neoplastic evasion or escape to the immune system host. Objective: To identify the expression of HLA-G, HLA-E and PD-L1 in oral mucoepidermoid carcinomas (MEC) and assess whether such expressions are related to metastasis, survival, staging, localization and tumor grade. Design: This cross-sectional study selected samples of MEC minor salivary glands (n=30) and classified them according to the World Health Organization (WHO) grading as low grade (LG), intermediate grade (IG) and high grade (HG). HLA-G, HLA-E and PD-L1 were identified by immunohistochemistry and measured by the proportion of positive neoplastic (mucous, epidermoid, intermediate and clear) and stromal cells. The density of positive Cytotoxic T Lymphocytes (CTLs), Natural Killer (NK), and Granzyme B (GB) cells was also evaluated. The Kruskal-Wallis test was used with a 5% significance level. Results: Expressions of HLA-G, HLA-E and PD-L1 were identified in the majority of epidermoid, intermediate and clear cells, but not in the mucous cells of the MECs. Thus, the quantitative analysis of the total percentage of positive neoplastic cells showed an association between the expression of these proteins and histologic grading with the following median values: HLA-G (LG= 79%, IG= 96%, HG= 99%; p=0.0004), HLA-E (LG= 70%, IG= 96%, HG= 99%; p<0.0001) and PD-L1 (LG= 34%, IG= 79%, HG= 80%; p=0.01). In consonance with a microenvironment conducive to inhibiting CTLs and NK cells, low GB density was found in all MEC samples. There was no relationship between the percentage of HLA-G+, HLA-E+ and PD-L1+ cancer cells and the other clinical parameters evaluated. Conclusions: MEC expresses a set of proteins involved in neoplastic cell escape from the anti-tumor immune system. In addition, increased expression of HLA-G, HLA-E and of PD-L1 in the tumoral microenvironment is closely related to the predominant cell type, and consequently to the higher histologic grades of MEC.Item Avaliação do estresse, ansiedade e comportamento associados ao tratamento odontológico infantil sob sedação(Universidade Federal de Goiás, 2016-12-09) Rodrigues, Heloisa de Sousa Gomes; Costa, Luciane Ribeiro de Rezende Sucasas da; http://lattes.cnpq.br/9906371509661305; Batista, Aline Carvalho; http://lattes.cnpq.br/0199082642322002; Batista, Aline Carvalho; http://lattes.cnpq.br/0199082642322002; Costa, Paulo Sérgio Sucasas da; Rotta , Rejane Faria Ribeiro; Paiva, Saul Martins; Silva , Tarcília Aparecida daPaediatric dental treatment may stress children and their parents influencing the child behaviour and the maternal dental anxiety. It is important to understand those behavioural and physiological alterations to aid using of adequate sedatives techniques during paediatric dental procedure. The aim of this study was to evaluate the stress of children and their mothers during the paediatric dental treatment using different sedation protocols. Also, the maternal dental anxiety, the child’s behaviour and age and its associations were evaluated. This observational study is a secondary analysis of two randomised controlled clinical trials. Children aged 2-6 years old received one tooth restoration under moderate sedation according to the groups: [A] 18 children received oral midazolam (1.0 mg/kg) and [B] 18 children received placebo in a crossover design [Clinical Trials database (NCT01795222)]; [C] 14 children received oral midazolam (0.5 mg/kg) and oral ketamine (3.0 mg/kg) plus sevoflurane inhalation (0.3% - 0.4%) and [D] 13 children had oral midazolam (0.5 mg/kg) and oral ketamine (3.0 mg/kg) plus oxygen inhalation in a parallel design (NCT02284204). The sessions were video recorded for evaluation of child behaviour using OSUBRS scale (Ohio State University Behavioural Rating Scale) and mothers answered the Brazilian version of Dental Anxiety Scale (DAS). The saliva samples were collected on children and on their mothers at 4 moments: waking up (T0), upon arrival at Dental School (T1), 25 minutes after the local anaesthesia injection on child (T2) and 25 minutes after the end of procedure (T3). Salivary cortisol was measured using an immunoassay kit (ELISA). As the data presented non normal distribution (Shapiro Wilk, p>0,05), Kruskal-Wallis and Mann-Whitney tests were used for non paired comparisons and Mann-Whitney for associations among the variables. For paired comparisons, Friedman and Wilcoxon tests were used (p<0,05). The increase of cortisol levels from T1 to T2 (reactivity to stressful stimulus – local anaesthesia) was higher in children of group [B] [median (interquartile)] – [0.53 (0.60)] following by groups [D] – [0.21 (0.35)], [C] – [0.11 (0.49)] and [A] – [0.02 (0.59)] (p=0.02). The decrease of cortisol levels from T2 to T3 (regulation to stress) was higher in children of group [B] – [0.08 (0.29)] following by groups [A] [-0.02 (0.40)], [C] – [-0.18 (0.41)] and [D] – [-0.19 (0.8)] (p=0.02). Majority of mothers were not stressed during their child’s local anaesthesia injection (67.9%) and presented low/moderate anxiety (69.6%), while 25.0% of them presented high/severe anxiety (DAS scale). Mothers who reacted to stress (increasing of cortisol at least 10% from T1 to T2) had higher cortisol levels at the moments T2 [0.15 (0.48)] and T3 [0.16 (0.50)] compared to T1 [0.09 (0.17)] (p<0.01 and p<0.006, respectively). On the other hand, mothers who did not react to stress had higher cortisol levels at the moments: T1 [0.36 (0.18)] compared to T2 [0.16 (0.18)] (p<0.01) and T3 [0.10 (0.12)] (p<0.01) and T2 compared to T3 (p=0.01). There was no statistically significant association between maternal stress (salivary cortisol levels) with child behaviour (p=0.56), child’s age (p=0.48) and maternal dental anxiety (DAS) (p=0.69). The findings of this study allow to conclude that sedation protocol using oral ketamine caused higher liberation of salivary cortisol at the moments of local anaesthesia and at the end of procedure (higher reactivity and lower regulation) indicating a prolonged response to physiological stress in children, which was not observed during the use of oral midazolam. Although, there was any maternal dental anxiety most of mothers were not stressed during the dental treatment under sedation of their children. Also, maternal dental anxiety, child’s age and child behaviour did not influence the maternal stress.