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Accurate diagnosis of oral soft tissue lesions is critical for effective treatment, yet conventional histopathological examination, the gold standard, faces limitations. These include two-dimensional (2D) visualization and malorientation, which can obscure critical diagnostic features, like epithelial-connective tissue interfaces. Micro-computed tomography (µCT) offers a non-destructive, high-resolution three-dimensional (3D) imaging alternative to address these challenges. Still, its use for soft tissue visualization is limited. We tested a method with specific radio-opaque staining and µCT scanning settings to visualize oral soft tissue biopsies as a proof of concept. Biopsies from 12 patients with different oral mucosa lesions were stained with Lugol's iodine, scanned at 3µm resolution with 70kV energy, and the resulting volumes were compared to histopathological sections by specialists in oral radiology and oral pathology. µCT produced 2D images with tissue architecture comparable to hematoxylin and eosin (H&E)-stained sections, distinguishing epithelium, connective tissue, and keratin, while 3D reconstructions revealed topographic details, such as ulceration depth and vascular patterns, unattainable in histopathology. These findings highlight µCT potential as a complementary diagnostic tool, enhancing topographic rendering while preserving tissue integrity. Standardized protocols and broader validation, particularly for precancerous and malignant lesions, are essential for clinical adoption, promising improved diagnostic accuracy in oral pathology.

Purpose Oral cancer (OC) patients are at high risk to develop recurrent disease or secondary primary cancers with no available biomarkers to detect these events until a visible lesion is readily present and diagnosed by biopsy. Exosomes secreted by cancer cells are involved in tumor growth, invasion and metastasis. We aimed to determine morphological and molecular differences between oral fluid (OF)-derived exosomes of OC patients and those isolated from healthy individuals (HI). Methods OF from OC patients ( n  = 36) and HI ( n  = 25) was initially assessed by nanoparticle tracking analysis (NTA). Following ultracentrifugation, exosomal pellets of OC patients and HI were morphologically examined by transmission electron microscopy and atomic force microscopy (AFM). Enzyme-linked immunosorbent assay (ELISA) and western blotting (WB) were used to analyze the expression of exosomal markers—CD9, CD81 and CD63. Results NTA showed that OC samples of OF had a significantly higher concentration of nanoparticles/ml ( p  = 0.01) and modal nanoparticle size ( p  = 0.002) compared to HI. The difference in size was structurally highlighted by AFM three-dimensional images applied on exosomal pellets. ELISA and WB showed differential expression of exosomal markers in OC exosomes compared to HI: lower expression of CD81 and CD9 in contrast to a higher expression of CD63 (~53 kDa). Conclusions OF-derived exosomes from OC patients differ both morphologically and molecularly from exosomes present in HI. This study is a baseline that provides a starting point for finding exosomal biomarkers for early detection of malignant changes in high-risk patients without overt clinical signs/lesions.

Expression of angiotensin-converting enzyme (ACE)-2 and co-factors like furin, play key-roles in entry of SARS-CoV-2 into host cells. Furin is also involved in oral carcinogenesis. We investigated their expression in oral pre-malignant/malignant epithelial pathologies to evaluate whether ACE2 and furin expression might increase susceptibility of patients with these lesions for SARS-CoV-2 infection. Study included normal oral mucosa (N = 14), epithelial hyperplasia-mild dysplasia (N = 27), moderate-to-severe dysplasia (N = 24), squamous cell carcinoma (SCC, N = 34) and oral lichen planus (N = 51). Evaluation of ACE2/furin membranous/membranous-cytoplasmic immunohistochemical expression was divided by epithelial thirds (basal/middle/upper), on a 5-tier scale (0, 1-weak, 1.5 -weak-to-moderate, 2-moderate, 3-strong). Total score per case was the sum of all epithelial thirds, and the mean staining score per group was calculated. Real time-polymerase chain reaction was performed for ACE2-RNA. Statistical differences were analyzed by One-way ANOVA, significance at p<0.05. All oral mucosa samples were negative for ACE2 immuno-expression and its transcripts. Overall, furin expression was weakly present with total mean expression being higher in moderate-to-severe dysplasia and hyperplasia-mild dysplasia than in normal epithelium (p = 0.01, each) and SCC (p = 0.008, p = 0.009, respectively). Oral mucosa, normal or with epithelial pathologies lacked ACE2 expression. Furin was weak and mainly expressed in dysplastic lesions. Thus, patients with epithelial pathologies do not seem to be at higher risk for SARS-CoV-2 infection. Overall, results show that oral mucosae do not seem to be a major site of SARS-CoV-2 entry and these were discussed vis-à-vis a comprehensive analysis of the literature.

ObjectivesThe aim of the present study was to assess macroscopically the time-related dimensional pattern of excisional palatal mucoperiosteal wound closure in rats, concomitantly with microscopic analysis of the density of inflammatory infiltrate and myofibroblasts.Materials and methodsExcisional palatal wounds, 4.2 mm in diameter, were made in twenty-one 2-month-old male Wistar rats. The total area and anteroposterior/laterolateral dimensions of the wounds were measured macroscopically at 1, 2, and 3 weeks post-operatively. In addition, histomorphometry was used for assessment of the intensity of inflammation and density of myofibroblasts in the wound area.ResultsA significant decrease was found in the total area and anteroposterior but not the laterolateral dimension of the wounds during the 2nd week. These changes were less prominent in the 3rd week. Three weeks post-operatively, the density of inflammatory infiltrate remained high in the central part of the wound concomitant with a significant increase in the number of myofibroblasts.ConclusionsWe concluded that the second week was the most significant in wound closure, with wound contraction first occurring in an anteroposterior plane followed by the laterolateral plane. The increased inflammatory reaction and changes in the density of myofibroblasts may explain the macroscopic decrease in wound dimensions in a time-related manner.Clinical relevanceThese findings emphasize the importance of the amount of soft tissue left at surgery, and suggest that the most appropriate time for the use of healing promoters would be the second post-operative week.

Background Pericytic tumors are a group of soft tissue neoplasms characterized by a hemangiopericytoma (HPC)-like pattern. The underlying genetic lesion of pericytic neoplasms is still obscure; however, recent advances in molecular pathology have enabled a mechanistic understanding and accurate diagnosis of these tumors. Nevertheless, a subset of unclassified pericytic tumors continues to present substantial diagnostic challenges, potentially impacting clinical decision-making and therapeutic strategies. Methods Using Ion AmpliSeq Comprehensive Cancer Panel (Thermo Fisher Scientific, Inc.) assay, we searched for HPC-like-associated mutations in a newborn female with a congenital large sublingual unclassified pericytic tumor that recurred after resection and was refractory to chemotherapy treatment. Further analysis of a series of other tumors with distinctive pericytic features was performed via Sanger sequencing and the results were validated via the auto genomics INFINITI ® and Biocartis Assays. Results A BRAF V600D mutation was identified in the tumor tissue of the newborn patient. Treatment with the BRAF inhibitor (BRAFi) dabrafenib resulted in a dramatic response and regression of the tumor. Sequencing of 15 additional HPCs revealed the presence of a BRAF V600E mutation in 6 samples. Conclusions Our findings suggest that BRAFi may be a useful therapy for unclassified pericytic tumors. Genetic testing for BRAFV600 mutations and other actionable oncogenic variants should be part of the evaluation for pericytic neoplasms, especially with the currently available targeted drug therapies.

Objective: To apply the technique of deep learning on a small dataset of panoramic images for the detection and segmentation of the mental foramen (MF). Study design: In this study we used in-house dataset created within the School of Dental Medicine, Tel Aviv University. The dataset contained randomly chosen and anonymized 112 digital panoramic X-ray images and corresponding segmentations of MF. In order to solve the task of segmentation of the MF we used a single fully convolution neural network, that was based on U-net as well as a cascade architecture. 70% of the data were randomly chosen for training, 15% for validation and accuracy was tested on 15%. The model was trained using NVIDIA GeForce GTX 1080 GPU. The SPSS software, version 17.0 (Chicago, IL, USA) was used for the statistical analysis. The study was approved by the ethical committee of Tel Aviv University. Results: The best results of the dice similarity coefficient ( DSC), precision, recall, MF-wise true positive rate (MFTPR) and MF-wise false positive rate (MFFPR) in single networks were 49.51%, 71.13%, 68.24%, 87.81% and 14.08%, respectively. The cascade of networks has shown better results than simple networks in recall and MFTPR, which were 88.83%, 93.75%, respectively, while DSC and precision achieved the lowest values, 31.77% and 23.92%, respectively. Conclusions: Currently, the U-net, one of the most used neural network architectures for biomedical application, was effectively used in this study. Methods based on deep learning are extremely important for automatic detection and segmentation in radiology and require further development.

Background Oral erythroplakia (OE) is a rare oral potentially malignant disorder, that has a high rate of malignant transformation. The definition of OE still lacks uniformity. In particular, lesions that look clinically like erythroplakias, but are histopathologically diagnosed as squamous cell carcinomas are still sometimes called erythroplakias. The purpose of this study is to present demographic and clinicopathologic features of a series of OEs and clinically oral erythroplakia -like squamous cell carcinomas (OELSCC), to study their differences and to discuss the definition of OE. Methods A multicenter retrospective case series of OEs and OELSCCs. Descriptive statistics were used to analyze the data. Results 11 cases of OEs and 9 cases of OELSCCs were identified. The mean age of the OE patients was 71 years and 72.7% were female, while the mean age of the OELSCC patients was 69 years, and all were female. 9% of the OE and 22% of the OELSCC patients had smoked or were current smokers. 72.7% of the OEs and 55.5% of OELSCCs were uniformly red lesions. 63.6% of the OE and 22% of the OELSCC patients had a previous diagnosis of oral lichenoid disease (OLD). The malignant transformation rate of OE was 9% in a mean of 73 months. Conclusions OE and OELSCC may arise de novo or in association with OLD. Tobacco and alcohol use were not prevalent in the present cases. The clinical features of OEs and OELSCC are similar, but symptoms, uneven surface and ulceration may be more common in OELSCCs than in OEs. Clinical recognition of OE is important since it may mimic other, more innocuous red lesions of the oral mucosa. The diagnosis of OE requires biopsy and preferably an excision. Clarification of the definition of OE would aid in clinical diagnostics.

AimThis study aims to examine the composition of lining and masticatory mucosa at the pre- and post-soft tissue augmentation procedures with a volume-stable cross-linking collagen matrix (VCMX) in humans.Materials and methodsIn 12 patients, single implant sites were augmented with a VCMX. Biopsies were obtained including masticatory (MM) and lining (LM) mucosa before augmentation and at 12 weeks post-augmentation procedures. Rete pegs density (RPD), length (RPL), and blood vessel density (BVD) were histomorphometrically analyzed at both time points. Picrosirius red staining under polarized light microscopy was used to evaluate collagen fiber organization. The effects of time and tissue type were evaluated by ANOVA with repeated measures.ResultsBoth MM and LM areas demonstrated an increase in mean RPL following augmentation, 382.6 µm ± 95.1 vs. 290.5 µm ± 79.3 and 335.6 µm ± 94.2 vs. 292.9 µm ± 77.0, respectively (p < .05). There was a significant difference in the numbers of RP per 1 mm length (RPD) between the MM (9.2 ± 1.7) and LM (6.1 ± 2.8) mucosa but not between the pre- and post-VCMX augmentation time points. The mean BVD in the LM was greater than in the MM (5.5 ± 2.4 and 6.3 ± 2.4 vs. 3.4 ± 3.3 and 3.7 ± 1.8, respectively, p < .05) but not between time points. The collagen fiber arrangements pre- and post-augmentation were not significantly different.ConclusionAugmentation with VCMX did not alter the composition of lining and masticatory mucosa at implant sites.Clinical relevanceA thick soft tissue phenotype around the implant neck is an important factor to maintain peri-implant health. A non-autogenous cross-linking collagen matrix is proposed as an alternate graft substitute in soft tissue augmentation procedures in order to improve implant soft tissue phenotype.