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Introduction: We recently developed a non-invasive sampling procedure for oral squamous cell carcinoma (OSCC) detection based on DNA methylation analysis of a panel of 13 genes. Oral cancer, as well as acute and chronic inflammatory diseases, may influence the methylation level of several genes in the oral cavity. In the present study, we evaluated the presence of periodontal disease (PD) and the methylation status using our 13-gene panel. Methods: Oral brushing specimens were collected from three different patient groups: 23 gingival OSCC patients, 15 patients affected by PD, and 15 healthy volunteers lacking evidence of PD. DNA methylation analysis was performed and each sample was determined to be positive or negative based on a predefined cut-off value. Results: Positive results were found for 23/23 OSCC patients, 3/15 PD patients, and 0/15 samples from healthy volunteers. The GP1BB and MIR193 genes in the PD group exhibited mean methylation levels similar to OSCC patients. ZAP70 showed different methylation levels among three groups. Conclusion: Preliminary data identified shared epigenetic alterations between PD and OSCC patients in two inflammatory genes (GP1BB and MIR193). This study may help to identify potential links between the two diseases and serve as a starting point for the future research focused on pathogenesis.

In a recent study our research group described a non-invasive sampling procedure based on [...]

Background The aim of this Italian multicenter study was to evaluate the diagnostic performance of a minimally invasive method for the detection of oral squamous cell carcinoma (OSCC) based on 13-gene DNA methylation analysis in oral brushing samples. Methods Oral brushing specimens were collected in 11 oral medicine centers across Italy. Twenty brushing specimens were collected by each center, 10 from patients with OSCC, and 10 from healthy volunteers. DNA methylation analysis was performed in blindness, and each sample was determined as positive or negative based on a predefined cutoff value. Results DNA amplification failed in 4 of 220 (1.8%) samples. Of the specimens derived from patients with OSCC, 93.6% (103/110) were detected as positive, and 84.9% (90/106) of the samples from healthy volunteers were negative. Conclusion These data confirmed the diagnostic performance of our novel procedure in a large cohort of brushing specimens collected from 11 different centers and analyzed in blindness.

Introduction The outbreak and diffusion of the novel SARS‐CoV2 coronavirus have caused an emergency status in the dental education system. Materials and methods An anonymous survey composed of 34 questions was delivered to students of the Master Degree Programme in Dentistry and Dental Prosthodontics of the Universities of Emilia‐Romagna, the fifth Italian region most affected by the pandemic. The psychological impact of COVID‐19 was assessed by means of the Generalised Anxiety Disorder‐7 scale (GAD‐7). Numerically recoded data were analysed using the Analysis of Variance (ANOVA), whilst to investigate the association between quantitative variables, the Pearson correlation coefficient (R) was computed. Results The questionnaire was completed by 399 students (75%) out of 532. Most students experienced difficulties in working at the thesis during the COVID‐19 emergency. For over half of them, online teaching could only partially replace traditional face‐to‐face lessons. The negative impact on the study career was judged as particularly high by sixth‐year students. Clinical training activities were considered as exposing to the risk of contracting COVID‐19 infection by the majority of the students. The level of concern of contracting COVID‐19 infections during future university activities was positively correlated to risk perception related to clinical training. Conclusion The results of this survey could be used to train students to a correct risk assessment. Students reported experiencing concern whilst thinking of COVID‐19 and 6.5% of them showed symptoms related to high levels of anxiety. These data may guide Universities in trying to reduce students' anxiety by means of correct communication strategies.

Objectives: To analyze psychological profiles, pain and oral symptoms in patients with oral lichen planus (OLP). Materials and methods: 300 patients with keratotic OLP (K-OLP; reticular, papular, plaque-like subtypes), 300 patients with predominant non-keratotic OLP (nK-OLP; erythematosus atrophic, erosive, ulcerative, bullous subtypes) and 300 controls were recruited in 15 universities. The number of oral sites involved and oral symptoms were recorded. The Numeric Rating Scale (NRS), Total Pain Rating Index (T-PRI), Hamilton Rating Scales for Depression and for Anxiety (HAM-D and HAM-A), Pittsburgh Sleep Quality Index (PSQI) and Epworth Sleepiness Scale (ESS) were administered. Results: The OLP patients, especially the nK-OLP, showed higher scores in the NRS, T-PRI, HAM-D, HAM-A and PSQI compared with the controls (p-value

Background: The wellbeing of oral lichen planus patients (OLPs) may be strongly influenced by a poor quality of sleep (QoS) and psychological impairment. The aims were to analyze the prevalence of sleep disturbance, anxiety, and depression in OLPs and to validate the Pittsburgh Sleep Quality Index (PSQI) in OLPs. Methods: Three hundred keratotic OLPs (K-OLPs), 300 with predominant non-keratotic OLP (nK-OLPs), and 300 controls were recruited in 15 Italian universities. The PSQI, Epworth Sleepiness Scale (ESS), Hamilton Rating Scales for Depression and Anxiety (HAM-D and HAM-A), Numeric Rating Scale (NRS), and Total Pain Rating Index (T-PRI) were administered. Results: Oral lichen planus patients had statistically higher scores than the controls in the majority of the PSQI sub-items (p-values5) in K-OLPs and nK-OLPs, respectively. The PSQI’s validation demonstrated good internal consistency and reliability of both the total and subscale of the PSQI. Conclusions: The OLPs reported an overall impaired QoS, which seemed to be an independent parameter according to the regression analysis. Hence, clinicians should assess QoS in OLPs and treat sleep disturbances in order to improve OLPs management.

Purpose: To evaluate the factors affecting peri-implant marginal bone level of single platform-switched implants with a smooth neck placed at gingival level (tissue level) using a flapless technique. Materials and Methods: Consecutive healthy patients requiring dental implant rehabilitations were enrolled in this study. Titanium implants with a zirconium-oxide-blasted surface and a platform-switch neck tulip configuration were used. Loading was performed 3 months after insertion with a provisional resin crown and after approximately 15 days with a definitive ceramic crown. Peri-implant marginal bone level (MBL) was measured on periapical radiographs at 1, 3, 6, 12, 24, 36, and 48 months by a blinded assessor. The following parameters were evaluated: location (maxillary/mandibular), position (anterior/posterior), sex (male/female), smoke (yes/no), implant placement timing (immediate, early, delayed), gingival thickness (thin/thick), endodontically treated adjacent teeth (yes/no), and diameter (3.8/4.25/5.0 mm). Multilevel analyses exploring factors associated with MBL at 36 and 48 months were performed. Results: A total of 76 patients (42 women, 34 men; mean age: 55.6 ± 10.7 years) received 128 implant rehabilitations. The survival rate was 98.4%. MBL displayed an initial increase during the first months from insertion (preload period). Cumulative mean MBL at T48 was 0.99 ± 0.68, which was not statistically significant from the values at T24 to T36 (P >.05). Mandibular location, delayed implants, and presence of adjacent endodontically treated teeth showed higher bone loss at 36 months (P .05). Conclusion: Platform-switched implants placed nonsubmerged with a flapless approach showed a reduced bone loss progression in the first 4 years, as MBL remained stable at longer times (36 and 48 months). Implants placed with early and immediate timing showed reduced bone loss compared with delayed implants.

BACKGROUND: A non-invasive sampling procedure for the early detection of Oral Squamous Cell Carcinoma (OSCC) based on DNA methylation analysis of a panel of 13 genes was applied in 4 different OSCC risk-group of patients. Aim of the study is to evaluate the between-group differences and the variables related to the methylation profile of each group. METHODS: Oral brushing samples were collected from 54 healthy subjects, 31 Oral Leukoplakia (OL) patients, 18 Oral Lichen Planus (OLP) patients and 26 patients previously treated for OSCC. Each sample was considered positive or negative in relation to a predefined cut-off value. RESULTS: None of the samples from 54 healthy subjects were positive, whereas 22/31 OL, 3/18 OLP and 8/26 surgically treated OSCC samples showed positive values with respect to the cut-off. In OL patients, dysplasia was the only variable significantly related to positive values: 10/10 OLs with high-grade dysplasia were positive with respect to 12/21 OLs without dysplasia (Chi 6.039, p< 0.05). CONCLUSION: DNA methylation analysis in epithelial cells collected by oral brushing seems to be a promising genetic method to distinguish lesions at high risk of developing OSCC. Larger population studies and an adequate follow-up period are necessary to confirm these preliminary data.

Genetic heterogeneity displayed by tumour cells (intratumoural heterogeneity, ITH) represents a diagnostic challenge when assessing tumour mutational profile. In oral squamous cell carcinoma (OSCC), ITH may be found both in tumour cells and in adjacent mucosa. Genetic heterogeneity of the adjacent mucosa can be interpreted as evidence of the field cancerization (field heterogeneity, FH). The aim of the study was to investigate the impact of intratumoural and intrafield heterogeneity on locoregional control. Ten OSCC patients (5 recurrent and 5 nonrecurrent) were studied. Multiple areas were sampled from the bulk of the tumour and the adjacent nonneoplastic mucosa. A panel of 10 tumour-specific OSCC driver genes was analysed for each sample and was used to calculate heterogeneity. Values were compared among recurrent and nonrecurrent OSCC. Mutational analysis highlighted that a single tumour sample has limited accuracy in assessing the genetic profiles of tumours. High values of ITH considering shared mutations between specimens were found in both recurrent and non-recurrent OSCC (p = 0.095). On the contrary, the intrafield genetic heterogeneity was significantly less frequently in the non-recurrent OSCC group (p = 0.032). Heterogeneity within each specimen calculated with variant allele frequency confirmed that there was better discrimination between recurrent and nonrecurrent groups using nonneoplastic adjacent mucosa than tumour tissue (p value 0.0006 and 0.0048 respectively). In agreement with the theory of field cancerization, intrafield genetic heterogeneity correlates with a higher risk of developing loco-regional recurrences and second primaries. In order to reduce the ITH effects, analysis of multiple tumour areas should be encouraged.

Background: Prognosis of oral squamous cell carcinoma (OSCC) is difficult to exactly assess on pre-operative biopsies. Since OSCC DNA methylation profile has proved to be a useful pre-operative diagnostic tool, the aim of the present study was to evaluate the prognostic impact of DNA methylation profile to discriminate OSCC with high and low aggressive potential. Methods: 36 OSCC cases underwent neoplastic cells collection by gentle brushing of the lesion, before performing a pre-operative biopsy. The CpG islands methylation status of 13 gene (ZAP70, ITGA4, KIF1A, PARP15, EPHX3, NTM, LRRTM1, FLI1, MiR193, LINC00599, MiR296, TERT, GP1BB) was studied by bisulfite Next Generation Sequencing (NGS). A Cox proportional hazards model via likelihood-based component-wise boosting was used to evaluate the prognostic power of the CpG sites. Results: The boosting estimation identified five CpGs with prognostic significance: EPHX3-24, EPHX3-26, ITGA4-3, ITGA4-4, and MiR193-3. The combination of significant CpGs provided promising results for adverse events prediction (Brier score = 0.080, C-index = 0.802 and AUC = 0.850). ITGA4 had a strong prognostic power in patients with early OSCC. Conclusions: These data confirm that the study of methylation profile provides new insights into the molecular mechanisms of OSCC and can allow a better OSCC prognostic stratification even before surgery.

Aim: This prospective clinical study analyzed the 24-month outcome of conventional apical surgery retro-filled with calcium-silicate cement versus apical surgery with simultaneous orthograde retreatment by means of clinical and radiographic criteria. Materials and methods: This study included 83 teeth affected by persistent periapical lesions in 68 patients. Mean age was 52 years (median = 51 years; range 19–81 years). Twenty-eight cases were treated with apical surgery, 16 cases with apical surgery with simultaneous orthograde retreatment and 39 cases with orthograde retreatment in previously treated teeth established as control group. Periapical index score (PAI) was used as radiographic criteria. Teeth were examined at 6 months, 1 and 2 years and classified as healed (without any symptoms and PAI ≤ 2), healing (without any symptoms and PAI = 3) or diseased (with symptoms or PAI ≥ 4 and not functional) on the basis of radiographic and clinical criteria. At 24 months evaluation, healed and healing were considered as success and diseased and fracture as failure. Multilevel GLM model and an ordered logistic regression as statistical analysis was made with level of significance set at p

Recent studies have emphasized the role of podoplanin in oral lesions at risk of malignant transformation. We investigated a group of oral leukoplakias (OLs) to determine a possible relation between altered podoplanin expression and dysplasia, and to compare the results with those obtained by other, widely used biomarkers.The population consisted of 40 consecutive patients with a clinical and histological diagnosis of OL. Thirty-two OLs did not show dysplasia, whereas eight lesions presented with dysplasia. Immunohistochemical expression of podoplanin, p53 and Ki67 was analyzed in all samples.All three biomarkers were positive in seven of eight dysplastic OLs. Among the 32 OLs without dysplasia, Ki67 and p53 showed positive values in 21 and 10 samples respectively, whereas podoplanin was positive in only one case. Multiple logistic regression showed that podoplanin was the most powerful variable (Chi square 9.77; p .01) statistically related to the presence of dysplasia. In addition, podoplanin showed a higher specificity value (96.87%) than Ki67 (34.37%) and p53 (68.75%).Podoplanin seems to be a reliable means of discriminating lesions with epithelial dysplasia and could be introduced in routine practice as a marker to discriminate OLs at risk of developing cancer.

Objectives A novel classification based on molecular methods to assess clonality defines three types of secondary oral squamous cell carcinoma (OSCC): second primary tumour (SPT) independent from the index tumour, local recurrence (LR), clonally related to the primary tumour, and second field tumour (SFT), derived from the same genetically altered mucosal field as the primary tumour. The present study applied mtDNA analysis in a group of patients experiencing a second loco-regional neoplastic manifestation. The purpose was to differentiate secondary tumours into LRs, SPTs and SFTs and evaluate the prognostic impact in terms of survival rate. Material and methods The study population comprised 23 patients who experienced a second neoplastic lesion after a surgical resection of primary OSCC. mtDNA D-loop analysis was applied in paired neoplastic lesions and in clinically and histologically normal mucosa. On the basis of mtDNA results, the second OSCC was classified as LR or SPT or SFT. Disease-free survival was defined as the duration between the appearance of the second neoplastic lesion and death of disease, or last follow-up visit. Results Seven secondary tumours were classified as LR, 12 as SFT, 4 as SPT. An altered mucosal field proved a variable significantly related to a better survival rate (p

O b j e c t i v e The objective of this study was to evaluate radiographic, clinical and esthetic parameters of a new type of nonsubmerged 2-piece implant placed in patients in need of single-tooth replacement. M a t e r i a l s a n d m e t h o d s Fifty-four consecutive patients requiring single-tooth replacement received 62 2-piece nonsubmerged flapless implants characterized by an innovative hyperbolic neck. The implant placement timing was as follows: 15 immediately post-extraction (immediate), 18 after 8–12 weeks (early) and 29 after 10–12 months (delayed). Customized abutments with an abutment–implant connection approximately 1–2 mm above the soft-tissue level were positioned after 3 months, loaded with provisional crowns and 20 days later with definitive crowns. Gingival biotype (thin or thick) was investigated in all patients. Periimplant marginal bone level (MBL; mm) was measured single-blinded on periapical radiographs at 1, 3, 6 and 12 months (T1, T3, T6, T12). Papilla index (PI), plaque score and bleeding on probing (BoP) were evaluated as clinical parameters of soft tissue. Pink Esthetic Score (PES) was calculated as the esthetic parameter. R e s u l t s The survival rate was 100%. The dropout rate was 1.85%. The mean MBL was - 0.01 ± 0.26 at T1, - 0.17 ± 0.38 at T3, - 0.28 ± 0.32 at T6 and - 0.37 ± 0.41 at T12. The PES (0–14) was 7.30 ± 2.80 at T0 (preoperatively), 11.06 ± 0.97 at T6 and 11.95 ± 1.04 at T12. At (T12), delayed implants showed a greater (P < 0.05) bone loss compared with early and immediate implants. Implants placed in thin biotype tissue showed the greatest bone loss at 12 months with a significant (P < 0.01) difference with respect to that at (T6). PES and PI increased from T0 to T12. C o n c l u s i o n These implants allow preservation of a good MBL and offer a new approach to soft- and hard-tissue management, allowing a reduced healing time with minimally invasive surgery, no additional re-entry and fewer complications.

Purpose: To evaluate the survival rate and marginal bone loss (MBL) of a calcium phosphate-blasted titanium implant inserted by a flap or flapless technique and to study the morphochemical characteristics of the implant surface. Materials and Methods: Sixty out of 85 patients who received one or more implants were selected as eligible for this prospective longitudinal cohort clinical study. Titanium implants (n = 132) were placed in human healthy subjects using either a flap or flapless technique, selected on the basis of the best practice. The survival rate and MBL were evaluated after 3 months (preloading stress-free healing period) and after 6, 12, 24, and 36 months in relation to implant diameter, location, sex, and smoking habits. Surface morphochemical analyses were performed by environmental scanning electron microscopy connected with energy-dispersive x-ray (ESEM/EDX). Results: The overall survival rate was 97.72%. After 3 and 36 months, MBL was 0.36 ± 0.66 mm and 1.09 ± 1.10 mm for the flapless group and 0.29 ± 0.51 mm and 1.03 ± 0.92 mm for the flap group. MBL showed a statistically significant increase with time (P = .0001), and differences were found at all times since 6 months. No statistical differences between the flapless and flap groups and sex were observed at any time, while MBL was significantly higher in the maxillary versus mandibular location (6, 12, and 24 months) and in smokers versus nonsmokers (6 months). Higher MBL in both groups was found for a smaller diameter (3.5 mm) than larger diameter (4.1, 5 mm). The surface showed a Ti-Al-V alloy displaying a uniform nanotexture. Conclusion: The implant system showed a high survival rate and markedly lower MBL considering the limits for acceptable progression. Flapless and flap techniques demonstrated similar results of MBL at the preloading healing period and at the 6 months to 3 years postloading periods. Both surgical procedures induced an early MBL during the preloading stress-free period. Implant diameter, mandibular/ maxillary location, preloading stress-free period, and smoking habits affect MBL more than the type of surgery after both short- and long-term follow-up.

MicroRNAs have recently been proposed as non-invasive biomarkers in Oral Squamous Cell Carcinoma (OSCC). The aim of this study was to analyze the expression of a panel of miRNAs in epithelial cells collected by oral brushing from OSCCs from regenerative areas after OSCC surgical resection and from their respective normal distant mucosa. Oral brushing specimens were collected from 24 healthy donors, 14 OSCC patients with specimens from tumour and normal distant mucosa, and from 13 patients who had OSCC resection, with samples from regenerative areas after OSCC resection and normal distant mucosa. Expression levels of eight targets (miR-21, miR-375, miR-345, miR-181b, miR-146a, miR-649, miR-518b, and miR-191) were evaluated by real-time Polymerase Chain Reaction (PCR). A highly significant between-group difference was found for miR-21 (F = 6.58, p &lt, 0.001), miR-146a (F = 6.974, p &lt, 0.001), and miR-191 (F = 17.07, p &lt, 0.001). The major difference was observed between samples from healthy donors and from OSCC brushing, whereas no significant differences were observed between areas infiltrated by OSCC and their respective normal distant mucosa. Furthermore, altered expression of miR-146a and miR-191 was also observed in regenerative areas after OSCC resection. Conclusions: Oral brushing could be proposed as a noninvasive method to study microRNA expression in oral mucosa in OSCC patients.

Purpose Improvements in sequencing technologies have shown that genetic differences among neoplastic cells can reflect clonal expansion. Intratumor heterogeneity (ITH) has been suggested to explain differences in prognosis and treatment response, indicating that personalized medicine is the goal of the future. This study evaluated ITH in 5 patients with recurrent metastatic oral squamous cell carcinoma (OSCC) and tracked the evolution from non-neoplastic tissue to neoplastic events developing after primary tumor formation. Patients and Methods Representative regions were macrodissected from specimens obtained from patients with OSCC of the tongue (n = 4) and floor of the mouth (n = 1). ITH and tumor evolution were explored by analyzing DNA mutations disclosed by next-generation sequencing of specific driver genes combined with changes in the mtDNA D-loop hypervariable region. Phylogenetic trees were generated employing MAFFT tool with UPGMA/Jukes-Cantor serving as the substitute model. Results High levels of heterogeneity were observed within and among tumors. ITH emerged as metastatic and recurrent events progressed, but the evolutionary patterns differed. In some patients, specific subclones persisted during tumor relapse. Neighboring tissue also was heterogeneous at the premalignant level. Conclusions A multiregion approach yielded more representative data than did single samples when tumors were subjected to molecular investigation. Persistent mutations that might be targeted by individualized medicine were thus exposed. Mitochondrial DNA is a useful adjunct tool when studying the phylogenetic evolution of subclones. The clinical implications of “field” heterogeneity should be studied in depth.

Objectives: To investigate the quality of sleep and the psychological profiles of a large cohort of Italian patients with burning mouth syndrome (BMS) and to clarify the relationships between these variables and pain. Methods: In this case–control study, 200 patients with BMS vs an equal number of age- and sex-matched healthy controls, recruited in 10 universities, were enrolled. The Pittsburgh Sleep Quality Index (PSQI), Epworth Sleepiness Scale (ESS), Hamilton Rating Scale for Depression (HAM-D), Hamilton Rating Scale for Anxiety (HAM-A), Numeric Pain Intensity Scale (NRS) and Total Pain Rating Index (T-PRI) were administered. Descriptive statistics, including the Mann–Whitney U test and hierarchical multiple linear regression analysis, were used. Results: Poor sleep quality (PSQI≥5) was present in 78.8% (160) patients with BMS. BMS patients had statistically higher scores in all items of the PSQI and ESS than the healthy controls (p&nbsp

Oral leukoplakia (OL) is the most common potentially malignant lesion of the oral cavity. Immunohistochemical analysis of p53 and Ki67 proteins is a simple and inexpensive method widely used in non-dysplastic OLs to reveal lesions predicted to develop oral cancer. The present longitudinal study evaluated the predictive role of p53 and Ki67 proteins alone or in combination in a group of OLs without dysplasia followed for many years. Seventy-seven OL patients referred to our Department between January 2006 and October 2013 underwent histochemical analysis of p53 and Ki67 expression. OLs were considered at high risk in the presence of either high p53 expression (>20%), or low/normal p53 expression associated with high Ki67 expression (Ki67/p53 ratio >3). Seven OLs evolved to OSCC during the follow-up period. Three cases had p53 overexpression, while four had a high Ki67/p53 ratio. Statistical significance was reached when samples with p53 overexpression were combined with samples with high Ki67/p53 ratio (Chi square 5.3; p

Background Oral squamous cell carcinoma (OSCC) is usually diagnosed at an advanced stage and is commonly preceded by oral premalignant lesions. The mortality rates have remained unchanged (50% within 5 years after diagnosis), and it is related to tobacco smoking and alcohol intake. Novel molecular markers for early diagnosis are urgently needed. The purpose of this study was to evaluate the diagnostic value of methylation level in a set of 18 genes by bisulfite next-generation sequencing. Methods With minimally invasive oral brushing, 28 consecutive OSCC, one squamous cell carcinoma with sarcomatoid features, six high-grade squamous intraepithelial lesions (HGSIL), 30 normal contralateral mucosa from the same patients, and 65 healthy donors were evaluated for DNA methylation analyzing 18 target genes by quantitative bisulfite next-generation sequencing. We further evaluated an independent cohort (validation dataset) made of 20 normal donors, one oral fibroma, 14 oral lichen planus (OLP), three proliferative verrucous leukoplakia (PVL), and two OSCC. Results Comparing OSCC with normal healthy donors and contralateral mucosa in 355 CpGs, we identified the following epigenetically altered genes: ZAP70, ITGA4, KIF1A, PARP15, EPHX3, NTM, LRRTM1, FLI1, MIR193, LINC00599, PAX1, and MIR137HG showing hypermethylation and MIR296, TERT, and GP1BB showing hypomethylation. The behavior of ZAP70, GP1BB, H19, EPHX3, and MIR193 fluctuated among different interrogated CpGs. The gap between normal and OSCC samples remained mostly the same (Kruskal-Wallis P values

AIM: The aim of this study was to compare the clinical and aesthetic outcomes of two different surgical approaches for the treatment of deep gingival recession affecting the mandibular incisors. METHODS: Fifty patients with Miller class I and II gingival recessions (≥ 3 mm) in the lower incisors were enrolled. Twenty-five patients were randomly assigned to the control group and the other 25 patients to the test group. All defects were treated with the coronally advanced flap + connective tissue graft (CAF + CTG) and in the test group the labial submucosal tissue (LST) was removed. Post-operative morbidity was evaluated at 1 week. Clinical and aesthetic evaluations were made at 1 year. RESULTS: Statistically greater recession reduction, probability of CRC (adjusted OR 7.94 95% CI = 1.88-33.50, p = 0.0024) and greater increase in GT were observed in the test group. Greater graft exposure and increase in KTH were demonstrated in the control group. Better aesthetics outcomes were observed in the test group. No statistically significant between groups differences were demonstrated in patient analgesic assumption and post-operative discomfort and bleeding. CONCLUSIONS: LST removal during CAF + CTG surgery is indicated to provide better root coverage and aesthetic outcomes in the treatment of gingival recessions affecting the lower incisors.

Patients with head and neck squamous cell carcinoma (SCC) are at risk of developing additional tumors in the head and neck. The detection of a late lymph node metastasis poses a problem to the clinician: it could be a delayed regional metastasis or a new metastasis from a yet unknown second primary tumor. Differentiation between metastasis and recurrence of primary tumors versus second primary tumor may be difficult because all lesions have the histologic appearance of SCC. Differentiation between these possibilities, however, carries important differences in therapeutic and prognostic consequences. In the following case report we present an unusually late regional lymph node metastasis in a patient who was treated 4 years earlier for an SCC in the inferior alveolar ridge. The purpose of the present study was to apply mitochondrial DNA D-loop analysis to assess the clonal relationship between oral tumor and node metastasis.

Background Histological and clinical criteria are generally used to differentiate second primary tumors (SPTs) from local recurrences. The purpose of the present study was to apply mitochondrial DNA (mtDNA) D-loop analysis to differentiate SPTs from local recurrences and to validate the clinical classification. Methods The study population consisted of 20 consecutive patients presenting multiple oral neoplastic lesions for a total of 25 paired lesions. The mtDNA D-loop analysis was performed by direct sequencing and phylogenetic clusterization. Results Agreement between mtDNA analysis and clinical classification was found in 19 cases. Discrepancies arose in 6 cases in which the clinical criteria based only on the spatial or temporal distance of the second lesion from the index tumor had led to a diagnosis of SPT (2 cases) or local recurrence (4 cases). Conclusion The present data highlight the value of mtDNA analysis in establishing the clonal relationship between the index tumor and the second neoplastic lesion. © 2013 Wiley Periodicals, Inc. Head Neck 36: 94–100, 2014

Purpose The aggressive behavior of oral squamous cell carcinoma (OSCC) has recently been related to the mucosa surrounding the primary mass, consisting of genetically altered cells that might be responsible for cancer progression. The aim of the study was to determine whether an abnormal cell turnover in clinically and histologically “normal” mucosa distant from the primary tumor is associated with a poor prognosis in terms of locoregional control (LRC) of disease and disease-specific survival (DSS). Material and methods This prospective study monitored 55 OSCC patients. Cell turnover in areas clinically and histologically distant from the tumor mass was evaluated by immunohistochemical expression of Ki67. A Ki67 value >20% was considered “high.” Results The mean follow-up period of the population studied was 53.7 ± 32.4 months (range 12–110 months). Multivariate analysis showed that the Ki67 value in distant mucosa was a powerful independent prognostic factor for LRC. In addition, high Ki67 expression in distant mucosa was the only variable statistically related to worse LRC (χ 2 = 9.5; p = 0.002) and DSS (χ 2 = 5.51; p = 0.02) in T1-2N0 OSCCs. Conclusions The present study confirmed the role of Ki67 in tumor-distant areas as a prognostic marker for OSCC patients.

BACKGROUND: A method to predetermine the maximum root coverage level (MRC) was recently demonstrated to be reliable in predicting the position of the soft tissue margin after root coverage surgery. The aim of the present study is to suggest a decision-making process for treating non-carious cervical lesions (NCCLs) associated with gingival recessions based upon the topographic relationship between the MRC and NCCL and to assess patient and independent-periodontist esthetic evaluations. METHODS: Five treatments were performed in 94 patients with NCCLs associated with a single gingival recession: 1) coronally advanced flap (CAF); 2) bilaminar procedure; 3) coronal odontoplasty plus restoration plus root odontoplasty plus CAF; 4) restoration plus CAF; and 5) restorative therapy. Clinical and esthetic evaluations made by the patient and an independent periodontist were done 1 year after treatments. RESULTS: The satisfaction of the patient and periodontist with esthetics was very high in all NCCL treatments and Miller Class gingival recessions. The patient satisfaction and evaluation of root coverage and the periodontist evaluation of root coverage were statistically correlated with color-match evaluations and not with the amount of root coverage clinically achieved in each patient. CONCLUSION: The proposed approaches provided good esthetic appearance and correct emergence profile for the great majority of NCCLs associated with gingival recessions.

Foschini M P, Morandi L, Marchetti C, Cocchi R, Eusebi L H, Farnedi A, Badiali G, Gissi D B, Pennesi M G & Montebugnoli L (2011) Histopathology58, 361–367 Cancerization of cutaneous flap reconstruction for oral squamous cell carcinoma: report of three cases studied with the mtDNA D-loop sequence analysis Aims: Tissue defects, resulting from surgical resection of oral squamous cell carcinoma (OSCC), are reconstructed routinely with skin grafts. OSCC arising from the grafted skin has been described; however, it is still unclear whether primary and second tumours have a common clonal origin. The aim of this study was to evaluate the clonal relationship between the primary OSCC and secondary neoplastic changes appearing in the skin graft in three patients, by screening the mitochondrial DNA D-loop region (mtDNA). Methods and results: In all three cases, the neoplastic lesions arising in the skin graft showed a clonal relationship with the previous OSCC and, on the basis of the results obtained by mtDNA analysis, could be considered to be a recurrence of the primary OSCC rather than a second primary OSCC. Conclusions: Starting from a field of genetically altered cells in the oral mucosa, the spread of the clonal cell population to the cutaneous flap might be stimulated by cytokines produced by the grafted skin. More studies are needed to evaluate the molecular relationship between primary and second OSCC to identify patients at higher risk of developing a second tumour in the skin graft.

Montebugnoli L, Cervellati F, Cocchi R, Farnedi A, Pennesi M G, Flamminio F & Foschini M P (2010) Histopathology57, 528–534 Immunohistochemical expression of p16INK4A protein as a helpful marker of a subset of potentially malignant oral epithelial lesions: study on a series with long-term follow-up Aim: To examine a group of lesions that progressed to oral squamous cell carcinoma (OSCC) to determine whether p16INK4A expression is an early finding during malignant transformation, and whether immunohistochemical evaluation of p16INK4A is an appropriate prognostic marker. Methods and results: Twenty cases of OSCC were investigated. All cases had had a biopsy on the same site as OSCC performed at least 1 year before OSCC (range 1–11 years; mean 3.15 ± 3.1 years). Twenty specimens from normal oral mucosa served as controls. p16INK4A expression was evaluated by immunohistochemical analysis and cases showing >5% of stained cells were defined as ‘positive’. All 20 control cases were negative for p16INK4A. Oral lesions were p16INK4A-positive in nine cases and negative in 11. No significant relationship was found between p16INK4A positivity and the presence/absence of dysplasia. Among OSCC, nine tumours showed p16INK4A positivity and 11 showed negativity. A significant relationship (χ2 = 7.1; P

This in vivo study evaluated the effects of topical fluoride application on enamel by repeated scanning electron microscopy analysis of replicas. Baseline fluid droplets were employed as qualitative indication of enamel permeability. CaF(2)-like globules were detected in vivo after fluoride application and were not found after professional brushing, ultrasound action, or chemical extraction. Absence of water permeability of enamel was demonstrated even after removal of CaF(2)-like globules. Droplets reappeared within 1 h in sodium fluoride-treated teeth, but they did not reappear even after 1 week following topical enamel treatment with acidulated phosphate fluoride. Teeth treated with an acidulate fluoride-free solution showed lack of CaF(2)-like globules and no droplets for at least 1 week as detected in acidulate phosphate fluoride-treated teeth. The caries-preventing action of fluoride may be due to its ability to decrease permeability and diffusion pathways. CaF(2)-like globules seem to be indirectly involved in enamel protection over time maintaining an impermeable barrier, and phosphoric acid seemed to play an unexpected fluoride-independent preventive role.

Laugier-Hunziker syndrome is a rare benign condition characterized by diffuse oral hyperpigmentation associated with pigmentation of the nails. The syndrome must be included in the differential diagnosis of diffuse oral pigmentation to exclude other conditions with systemic implications. We describe a 43-year-old white woman with the clinical and histological features of Laugier-Hunziker syndrome associated with toenail pigmentation. The correct clinical identification avoids the need for detailed investigations and treatment. We also review the potential causes of oral pigmentation.

p53 over-expression has been proposed as a reliable marker associated to oral carcinogenesis, although only about 50% of oral carcinomas (OSCC) are associated with p53 over-expression and even p53-negative lesions can progress to OSCC. The aim of the study was to determine whether the combination of p53 over-expression and p53 low-expression associated with Ki67 over-expression (high Ki67/p53 ratio) could lead to a more sensitive parameter. Immunohistochemical expression of Ki67 and p53 was measured in 54 specimens from OSCC; 27 specimens from moderate/severe epithelial dysplasia; 32 specimens from oral leukoplakias without epithelial dysplasia, and 13 specimens with normal epithelium. p53 over-expression was found in 31 (53%) samples from OSCC, in 10 (37%) samples from severe dysplasias, and in 5 (15%) samples from non-dysplastic lesions, while the combination of high p53 values with high Ki67/p53 ratio was observed in 93% of OSCC, in 81% of dysplastic lesions, and in 50% of non-dysplastic lesions. This parameter may have a clinical implication to detect early lesions with an impairment of p53 pathway, and probably at risk of progress to OSCC.

Purpose Oral squamous cell carcinoma (OSCC) is commonly preceded by oral potentially malignant lesions (OPML). The aim of the present study was to assess, by bisulfite next-generation sequencing (NGS), the methylation status of a list of candidate genes obtained from oral brushings to early detect OPML and OSCC. Material and methods Oral brushing specimens from 11 OSCC, 11 high-grade squamous intraepithelial lesions (HG-SIL), 9 low-grade SIL (LG-SIL), 9 oral lichen planus (OLP), and 8 healthy donors were included in this study. We investigated, by means of bisulfite NGS, the promoter of GP1BB, ZAP70, KIF1A, p16[CDKN2A], CDH1, miR137, and miR375. Statistical significance between lesions and a pool of healthy donors were evaluated with the Mann–Whitney U test. Results ZAP70 was found to be hypermethylated in 100% of OSCC and HG-SIL and in 28.6% of LG-SIL. GP1BB hypomethylation was detected in 90.9% OSCC and HG-SIL and in 37.5% of LG-SIL. MiR137 was hypermethylated in 100% of OLP, 44.4% of OSCC, 40% HG-SIL, and 25% LG-SIL. KIF1A hypermethylation was found to be associated with TP53 mutations ( p Conclusion In the present preliminary cohort of patients, DNA methylation analysis of GP1BB and ZAP70 seems to be a promising noninvasive tool for early detection of OSCC and HG SIL from oral brushing specimens.

Objectives The study makes a comparison between two surgical approaches for the treatment of oral leukoplakia (OL) in terms of recurrence in a well-defined cohort of patients with a long-term follow-up period. Methods The cohort consisted of 77 OL patients divided into 2 groups. Group 1: 47 patients treated with laser evaporation using a Nd:YAG laser. Group 2: 30 patients treated with a CO 2 laser for excision. Clinical and histological examinations were performed for the diagnosis of OL before treatment. We included OLs with or without dysplasia. The mean follow-up period was 60 ± 32.49 months. Results Of the 77 patients, 22 (28.5%) showed recurrence during the follow-up period. No significant difference was found between the two treatments (χ 2 = 2.6; p = 0.2). However, CO 2 laser excision resulted in better results than the Nd:YAG laser evaporation, considering the non-homogeneous OLs (χ 2 = 3.9; p = 0.04) and OLs with mild dysplasia (χ 2 = 4.6; p = 0.03). Discussion The study makes a comparison between our results and articles from the literature, and suggests when each of the two surgical approaches is most appropriate.

This study investigated the long-term clinical outcome of root canal treatment. 240 root-treated teeth (n = 61 patients) were initially classified on the basis of radiographic presence/absence of initial apical periodontitis (IAP) and clinical data. The final outcome measure was the periapical healing (healed/disease). The outcome at 6–9 months was correlated with the outcome at 10 years following treatment. Prognostic factors for the periapical healing were assessed. Extraction data were recorded. Univariate and multivariate logistic regression analysis was used to identify risk indicators for apical periodontitis (AP) development. Chi-square analysis was performed to evaluate a possible relationship between the 6–9 months outcome and the final outcome related to IAP. Mean observation time was 14 ± 3.7 years. Survival rate was 84.6 % and healing rate was 79 % (10–19 years). Predictors of outcome (p

This study was aimed to evaluate the possible alert signals of paraesthesia by local anaesthetics, focusing on those used in dentistry. A case/non-case study of spontaneous adverse events recorded in FAERS (FDA Adverse Event Reporting System) between 2004 and 2011 was performed. Cases were represented by the reports of reactions grouped under the term ‘Paraesthesias and dysaesthesias’ involving local anaesthetics (ATC: N01B*); non-cases were all other reports of the same drugs. Reporting odds ratios (ROR) with the relevant 95% confidence intervals (95CI) were calculated. Alert signal was considered when number of cases >3 and lower limit of ROR 95CI > 1. To estimate the specificity of signals for dentistry, the analysis was restricted to the specific term “Oral Paraesthesia” and to reports concerning dental practice. Overall, 528 reports of ‘Paraesthesias and dysaesthesias’ were retrieved, corresponding to 573 drug–reaction pairs (247 lidocaine, 99 bupivacaine, 85 articaine, 30 prilocaine, 112 others). The signal was significant only for articaine (ROR=18.38; 95CI = 13.95–24.21) and prilocaine (2.66; 1.82–3.90). The analysis of the specific term “Oral Paraesthesia” retrieved 82 reports corresponding to 90 drug–reaction pairs (37 articaine, 19 lidocaine, 14 prilocaine, 7 bupivacaine, 13 others) and confirmed the signal for articaine (58.77; 37.82–91.31) and prilocaine (8.73; 4.89–15.57). The analysis of reports concerning dental procedures retrieved a signal for articaine, both for any procedures (8.84; 2.79–27.97) and for non-surgical ones (15.79; 1.87–133.46). In conclusion, among local anaesthetics, only articaine and prilocaine generated a signal of paraesthesia, especially when used in dentistry.

A recent study conducted in humans demonstrated for the first time that bone loss in the immediate period after implant insertion before loading did not significantly differ in organ transplant recipients with respect to normal subjects.The purpose of this study is to evaluate bone and periodontal response and peri-implant microflora in a group of organ-transplanted patients 1 year after prosthetic loading.The study population included 13 consecutive organ-transplanted (11 hearts, two livers) patients and 13 normal (healthy) control subjects who received 29 and 28 submerged dental implants, respectively. Crestal bone level, peri-implant probing depth (PIPD), and bleeding on probing were evaluated at prosthetic loading and after 1 year. Samples for microbiological testing were taken from the subgingival microbiota of each implant 1 year after loading and analyzed with polymerase chain reaction. All samples were examined for the presence of five microorganisms: Treponema denticola, Porphyromonas gingivalis, Prevotella intermedia, Tannarella forsythensis, and Actinobacillus actinomycetemcomitans.A mean bone loss of 0.17 ± 0.10 and 0.20 ± 0.10 mm at 1 year was observed in the group of transplant recipients and in controls, respectively (N.S.). Similar results were obtained considering PIPD changes at 1 year (respectively 0.06 ± 0.71 mm in transplants vs 0.11 ± 0.74 mm in controls). Detection frequencies of pathogens were not statistically different between normal and transplanted patients.The present pilot study seems to indicate that bone and periodontal response and microbiological status around submerged dental implants in immunocompromised organ-transplanted patients do not differ 1 year after loading from those observed in control patients and that this particular population of patients may be successfully rehabilitated with dental implants.

Ghost cell odontogenic carcinomas are rare neoplasms that arise in the maxillary bones either from a calcifying odontogenic cyst or de novo. They are aggressive locally and can metastasize. We report herein a case of a ghost cell odontogenic carcinoma arising in the mandible of a Caucasian male 86 years of age. We have described the clinical and radiographic features, histological characteristics, immunohistochemistry findings, and surgical treatment. We especially focused on how Ki-67 expression guides the treatment choice. Finally, we reviewed 32 cases described in the literature and compared them with the cases described up until 2014 to help clinicians identify the diagnostic characteristics of and select appropriate treatment modalities for ghost cell odontogenic carcinomas.

The study aimed to assess the proliferative activity and karyotype in Oral Lichen Planus (OLP) lesions. G-banding chromosomal analysis of short-term primary cultures, and immunohistochemical expression of Ki67 and p53 were applied in 30 consecutive OLP patients divided into two groups according to clinical presentation of the lesions, and in nine subjects as negative controls. Mean values of Ki67 and p53 expression differed significantly (P < .01) between controls and patients groups with reticular or atrophic-erosive forms of OLP, whereas there was no significant difference between the two groups of patients with reticular or atrophic-erosive lesions. Six OLP patients showed clonal chromosome alterations, four of them associated with p53 overexpression. In conclusion, OLP is characterized by a high cellular turnover in most patients irrespective of clinical disease presentation. The genetic instability found in some patients should be interpreted as a consequence of the enhanced epithelial turnover, although we cannot rule out the possibility that some of the cytogenetic non-random anomalies observed represent early steps in cancer development.