Expression of spindle assembly checkpoint proteins BubR1 and Mad2 expression as potential biomarkers of malignant transformation of oral leukoplakia: an observational cohort study.

BACKGROUND: The Spindle Assembly Checkpoint (SAC) is a surveillance mechanism essential to ensure the accuracy of chromosome segregation during mitosis. Our aim was to evaluate the expression of SAC proteins in oral carcinogenesis, and to assess their potential in predicting malignant transformation of oral leukoplakia. MATERIAL AND METHODS: We analysed the immunoexpression of BubR1, Mad2, Bub3, and Spindly proteins in 64 oral biopsies from 52 oral leukoplakias and 12 normal tissues. Univariate and multivariate analysis were performed to evaluate predictive factors for malignant transformation (MT). RESULTS: We observed that BubR1 and Mad2 were more highly expressed in high dysplasia grade lesions than in low grade or normal tissues (P<0.05). High expression of Spindly was significantly correlated with a high Ki-67 score (P=0.004). Six (11.5%) oral leukoplakias underwent malignant transformation. In univariate analysis, the binary dysplasia grade (high grade) (P<0.001) was associated with a higher risk of malignant transformation as well as high BubR1 (P<0.001) and high Mad2 (P=0.013) expression. In multivariate analysis, high expression of BubR1 and Mad2 when combined showed an increased risk for malignant transformation (P=0.013; HR of 4.6, 95% CI of 1.4-15.1). CONCLUSIONS: Our findings reveal that BubR1 and Mad2 were associated with an increased risk for malignant transformation independently of histological grade and could be potential and useful predictive risk markers of malignant transformation in oral leukoplakias.

A histological evaluation of the surgical margins from human oral fibrous-epithelial lesions excised with CO2 laser, Diode laser, Er:YAG laser, Nd:YAG laser, electrosurgical scalpel and cold scalpel.

BACKGROUND: We aim to evaluate the presence of histological artefacts in the surgical margins of human oral fibro-epithelial hyperplasias excised with lasers of different wavelengths, and also electrosurgical scalpel and cold scalpel. Moreover, we aim to determine if some of these instruments could impair the normal histological diagnosis of these lesions. MATERIAL AND METHODS: We included 130 consecutive surgical samples of 80 females and 50 males (mean age of 53.82±16.55) with a histological diagnosis of an oral benign fibrous-epithelial hyperplasias. The samples were categorized into 6 groups according to the type of instrument used: CO2 laser group, diode laser group, Er:YAG laser group, Nd:YAG laser group, electrosurgical scalpel group and cold scalpel group. Histological instrument-induced changes were microscopic evaluated and related with clinical and pathological variables. RESULTS: The instrument with highest tissue damage extension (TDE) was the electrosurgical scalpel (1002.2µm±434.92), followed by diode laser (913.73 µm±322.45), Nd:YAG (899.83µm±327.75), CO2 laser (538.37µm±170.50), Er:YAG laser (166.47µm±123.85), and at last with fewer alterations the cold scalpel group (2.36µm±7.27) (P < 0.001). The most regular incision was observed in CO2 laser group, followed by Er:YAG laser, Nd:YAG laser, electrosurgical scalpel and diode laser group with the less regular incision using cold scalpel as comparison (P < 0.001). A correlation was found between the incision score and TDE (P < 0.001). Regarding histological diagnosis, no case showed any limitation of diagnosis related with the use of any instrument evaluated. CONCLUSIONS: Our results suggest that lasers can be used for the excision of oral benign fibrous-epithelial hyperplasias, without hispathological diagnosis limitations, as long as the physical properties of each laser are known and respected. Er:YAG laser have shown to be a laser with few tissue damage extension and with good incision regularity, been a possible instrument of choice for the surgical removal of these lesions.

Type of surgical treatment and recurrence of oral leukoplakia: A retrospective clinical study.

BACKGROUND: Oral leukoplakia (OL) is the most typical potentially malignant disorder of the oral mucosa. We aimed to evaluate the clinical outcome of oral leukoplakia treated with several types of lasers and with the use of quantic molecular resonance (QMR) lancet, in terms of recurrence rate. MATERIAL AND METHODS: Eighty-seven previously untreated OL (52 occurring in females and 35 in males, mean age of 59.4 ± 13.9 years) were consecutively submitted to surgical treatment at University Hospital of Parma, Italy, and Hospital de Valongo, Portugal, (1999 to 2012). Interventions were subclassified into 5 groups according to the instrument used for the surgical removal of OL (cold blade - 17; Nd:YAG 1064nm laser - 14; Er:YAG 2940nm laser - 33; CO2 10600nm laser - 15; and QMR scalpel - 8). The mean follow-up period after treatment was 21.6 months (range 1-151 months). The outcome of treatment was scored through the same clinical protocol in the two participating units. Statistical analysis were carried by univariate analysis using chi-square test (or Pearson's test when appropriate). RESULTS: Recurrences were observed in 24 cases of OL (27.6%). Malignant transformation occurred in one patient (1.1%) after a period of 35 months. Statistical comparison of the 5 surgical treatment modalities showed no differences in clinical outcomes nor in the recurrence rate of OL. However, when Er:YAG laser group was compared with traditional scalpel, a significantly better outcome in cases treated with Er:YAG laser (P = 0.015) was highlighted. CONCLUSIONS: Our results suggests that Er:YAG laser could be a promising option for the treatment of OL.

Coffee is protective against oral and pharyngeal cancer: A systematic review and meta-analysis.

OBJECTIVES: Coffee is one of the most popular and consumable drinks worldwide. However, there are conflicting results on the influence of this drink in oral and pharyngeal cancer risk. To clarify this, we aimed to systemically review and carry out a meta-analysis of the relevant literature on the association between coffee and oral and pharyngeal cancer. STUDY DESIGN: We carried out an electronic search of publications up to August 2016 from PubMed, National Library of Medicines Medline, Embase, Science Direct and the Cochrane Central Register. The Newcastle-Ottawa scale was used to address the quality of the studies a meta-analysis was carried out using random-effects models. RESULTS: From the 22,515 entries identified in the search, 13 case-control and 4 cohort studies were selected. With regards to quality on the Newcastle-Ottawa scale, an overall value of 6.06 was obtained. The analysis for oral and pharyngeal cancer grouped together indicated a pooled OR of .69 (95% CI of .57-.84; p<.001) for high versus low coffee consumption with a moderate heterogeneity (I2: 50.3%; p=.009). Regarding studies on oral cavity cancers we observed a pooled OR of 0.82; 95% CI =.58-1.16; p=.257) and for pharyngeal cancers a pooled OR of .72 (95% CI of 0.54-.95; p=.019). There was no significant publication bias. CONCLUSION: The results show an inverse association between high coffee consumption and the risk of oral and pharyngeal cancers, which indicates that coffee may have a protective role against these cancers. Further larger prospective observational cohort studies are needed to address any effect of other possible co-factors.

Prognostic significance of CD44v6, p63, podoplanin and MMP-9 in oral squamous cell carcinomas.

OBJECTIVES: To analyse the expression of the CD44v6, p63, podoplanin and MMP-9, and their prognostic significance in patients with oral squamous cell carcinomas (OSCC). MATERIAL AND METHODS: Immunohistochemistry technique was performed on 60 OSCC for detection of CD44v6, p63, podoplanin and MMP-9 proteins. Extent and intensity of staining were evaluated in tumour cells and were compared with patients' clinical-pathological characteristics and survival. RESULTS: CD44v6 expression was detected at the membrane of tumour cells of 94% cases. Nuclear expression of p63 protein was present in 96.5%. Podoplanin was observed at the membrane of tumour cells of 94% cases. MMP-9 was found in the cytoplasm of tumour cells in 83.7% cases. A high level of expression (67%-89%) in all four proteins was noted. Podoplanin was associated with the expression of MMP-9 (P = 0.010) and both were associated with lymph node metastasis (P = 0.011 and P = 0.018, respectively). Co-expression of podoplanin/MMP-9 was an adverse independent prognostic factor for cancer-specific survival (P = 0.008) and recurrence-free survival (P = 0.042). CONCLUSION: Podoplanin and MMP-9 together could contribute to tumour progression and dissemination of OSCC. Their combined overexpression showed an adverse effect on survival, suggesting that they could be regarded as important prognostic biomarkers in OSCC.

Helicobacter pylori identification: a diagnostic/confirmatory method for evaluation.

The Helicobacter pylori extra gastric reservoir is probably the oral cavity. In order to evaluate the presence of this bacterium in patients with periodontitis and suspicious microbial cultures, saliva was collected from these and non-periodontitis subjects. PCRs targeting 16S rRNA gene and a 860 bp specific region were performed, and digested with the restriction enzyme DdeI. We observed that the PCR-RFLP approach augments the accuracy from 26.2 % (16/61), found in the PCR-based results, to 42.6 % (26/61), which is an excellent indicator for the establishment of this low-cost procedure as a diagnostic/confirmatory method for H. pylori evaluation.

Analysis of peri-implant bone defects by using cone beam computed tomography (CBCT): an integrative review.

The objective of this work was to perform an integrative review of the inspection of peri-implant bone defects using cone beam computed tomography (CBCT). An electronic search was performed in the PubMed database using the following scientific terms: CBCT or Cone Beam computed tomography; dental implant; peri-implant; bone loss; defects. The survey identified 267 studies, of which 18 were considered relevant to this study. These studies provided important data taking into account the accuracy of cone beam computed tomography in the detection and measurement of peri-implant bone defects such as fenestrations, dehiscence and intraosseous circumferential defects. The effectiveness of CBCT in aiding in geometric bone calculations and in the diagnosis of peri-implant defects was influenced by factors such as artefacts, defect size, bone wall thickness, implant material, adjustment of acquisition parameters and observer experience. A not insignificant number of studies compared intraoral radiography to CBCT in the detection of peri-implant bone loss. CBCT was clearly superior to intraoral radiography in the detection of all peri-implant bone defects, except for defects located in the interproximal zone. In general, studies have shown that peri-implant bone measurements adjacent to the implant surface can be correctly determined, as well as the diagnosis of peri-implant bone defects with an average discrepancy of less than 1 mm from the actual measurement of the defect.

Relationship between dental and periodontal health status and the salivary microbiome: bacterial diversity, co-occurrence networks and predictive models.

The present study used 16S rRNA gene amplicon sequencing to assess the impact on salivary microbiome of different grades of dental and periodontal disease and the combination of both (hereinafter referred to as oral disease), in terms of bacterial diversity, co-occurrence network patterns and predictive models. Our scale of overall oral health was used to produce a convenience sample of 81 patients from 270 who were initially recruited. Saliva samples were collected from each participant. Sequencing was performed in Illumina MiSeq with 2 × 300 bp reads, while the raw reads were processed according to the Mothur pipeline. The statistical analysis of the 16S rDNA sequencing data at the species level was conducted using the phyloseq, DESeq2, Microbiome, SpiecEasi, igraph, MixOmics packages. The simultaneous presence of dental and periodontal pathology has a potentiating effect on the richness and diversity of the salivary microbiota. The structure of the bacterial community in oral health differs from that present in dental, periodontal or oral disease, especially in high grades. Supragingival dental parameters influence the microbiota's abundance more than subgingival periodontal parameters, with the former making a greater contribution to the impact that oral health has on the salivary microbiome. The possible keystone OTUs are different in the oral health and disease, and even these vary between dental and periodontal disease: half of them belongs to the core microbiome and are independent of the abundance parameters. The salivary microbiome, involving a considerable number of OTUs, shows an excellent discriminatory potential for distinguishing different grades of dental, periodontal or oral disease; considering the number of predictive OTUs, the best model is that which predicts the combined dental and periodontal status.

Treatment of Papillon-Lefèvre syndrome periodontitis.

BACKGROUND, AIMS: Conventional mechanical treatment of Papillon-Lefèvre syndrome periodontitis has a poor prognosis. This report describes an effective antimicrobial treatment of rapidly progressing periodontitis in an 11-year old girl having Papillon-Lefèvre syndrome. METHOD: Clinical examination included conventional periodontal measurements and radiographic analysis. Occurrence of major suspected periodontopathic bacteria was determined by selective and non-selective culture and by polymerase chain reaction (PCR) identification. Presence of cytomegalovirus and Epstein-Barr type 1 virus was determined by a nested-PCR detection method. Therapy included scaling and root planing, oral hygiene instruction, and systemic amoxicillin-metronidazole therapy (250 mg of each/3 times daily/10 days) which, based on follow-up microbiological testing, was repeated after 4 months. Supportive periodontal therapy took place at 2 visits during a 16-month period. RESULTS: At baseline, 10 of 22 available teeth demonstrated severe periodontal breakdown. At 16 months, probing and radiographic measurements revealed no teeth with additional attachment loss, and several teeth exhibited significant reduction in gingivitis and pocket depth, increase in radiographic alveolar bone height and clinical attachment level, and radiographic evidence of crestal lamina dura. Baseline subgingival microbiota included Actinobacillus actinomycetemcomitans (3.4% of total isolates), Prevotella nigrescens (16.4%), Fusobacteriumnucleatum (14.3%) and Peptostreptococcus micros (10.6%), as well as cytomegalovirus and Epstein-Barr type 1 virus. At termination of the study, culture and PCR examinations showed absence of A. actinomycetemcomitans, P. micros and herpesviruses, and P. nigrescens and F.nucleatum each comprised less than 0.1 % of subgingival isolates. CONCLUSION: This study suggests that controlling the periodontopathic microbiota by appropriate antibiotic and conventional periodontal therapy can arrest Papillon-Lefèvre syndrome periodontitis.

Microbiological features of Papillon-Lefèvre syndrome periodontitis.

Papillon-Lefevre syndrome patients exhibit hyperkeratosis palmo-plantaris and severe periodontitis. The syndrome is an autosomal recessive trait, but the mechanism of periodontal destruction is not known. This report presents the clinical and microbiological features of an 11-year old girl with Papillon-Lefèvre syndrome. Clinical examination included conventional periodontal measurements and radiographic analysis. In samples from 3 deep periodontal lesions, the occurrence of major suspected periodontopathic bacteria was determined by selective and non-selective culture and polymerase chain reaction (PCR) identification, and the presence of cytomegalovirus and Epstein-Barr type 1 virus by a nested-PCR detection method. 10 of 22 available teeth demonstrated severe periodontal breakdown. Major cultivable bacteria included Actinobacillus actinomycetemcomitans (3.4% of total isolates), Prevotella nigrescens (16.4%), Fusobacterium nucleatum (14.3%) and Peptostreptococcus micros (10.6%). A. actinomycetemcomitans, P. nigrescens, Porphyromonas gingivalis and Eikenella corrodens were identified by PCR analysis. The patient's non-affected parents and older brother revealed several periodontal pathogens but not A. actinomycetemcomitans. The viral examination demonstrated cytomegalovirus and Epstein-Barr type 1 virus in the subgingival sample of the Papillon-Lefèvre syndrome patient. The father and brother yielded subgingival cytomegalovirus but not Epstein-Barr type 1 virus. We hypothesize that human herpesviruses in concert with A. actinomycetemcomitans play important rôles in the development of Papillon-Lefèvre syndrome periodontitis.