Human Papillomavirus-Associated Oral Epithelial Dysplasia: A Practical Approach to Make the Diagnosis.

Background: High-risk Human Papillomavirus (HPV) genotypes are found in malignant oral epithelial lesions, and HPV infection is proposed as a risk factor for initiating Squamous cell carcinoma (SCC) in the head and neck region. This study suggests a practical approach to detect HPV in HPV-associated oral epithelial dysplasia (HAOED). Methods: Fifty-four oral epithelial dysplasia specimens were examined, comprising twenty-seven cases diagnosed with high-grade dysplasia and twenty-seven cases diagnosed with low-grade dysplasia using a binary grading system. To assess the cases for HPV, the specimens were examined for p16 protein using an immunohistochemical (IHC) study, and then, the Chromatin In Situ Hybridization (CISH) test was performed for all positive cases. Chromatin Immunoprecipitation-Polymerase Chain Reaction (ChIP-PCR) was performed on CISH-positive specimens to assess the outcome. This cross-sectional study was conducted in 2020 at Tehran University of Medical Science. SPSS software version 22.0 was used to perform the Chi square or Fisher's exact test to examine the relationship between variables (statistically significant level P<0.05). Results: The expression of p16 protein was not associated with the severity of epithelial dysplasia (81.5% in low-grade and 59.2% in high-grade cases) (P=0.16). Moreover, according to the CISH test result, 9.25% of all specimens were positive (P>0.99), and in the nine cases, undergone the ChIP-PCR study, two cases (22.2%) showed positivity for HPV-16, while one case (11.1%) demonstrated positivity for HPV-51. Conclusion: Regarding HAOED, here, we proposed a step-by-step combination approach using different diagnostic methods, including IHC for p16 protein, CISH, and ChIP-PCR based on a complementary algorithm.

Oral palisaded encapsulated neuroma; a diagnosis seldom suspected clinically.

Key Clinical Message: Palisaded encapsulated neuroma (PEN) is generally seen in the head and neck area as an asymptomatic nodule with the same color as the surrounding skin and rarely occurs in the oral cavity. The exact etiology of PEN is not known, but there is evidence supporting the role of trauma as its etiological factor. Abstract: Palisaded encapsulated neuroma (PEN) is one of the benign nerve sheath tumors of Schwann cell origin, which is commonly found in the skin of the head and neck area, and rarely occurs in the oral cavity. Its exact etiology is unknown, but there is evidence that supports the role of trauma as an etiological factor. Here we present a case of PEN in the hard palate of a 30-year-old patient and review the differential diagnoses of these nerve sheath tumors of the oral cavity.

Keratoameloblastoma of the lower jaw: A rare challenging entity.

Ameloblastoma is the second most common benign odontogenic tumor with various histopathologic features. Except for the unicystic type of ameloblastoma, the different microscopic patterns of this tumor show no significant correlation with long-term clinical behavior. During recent decades, additional challenging subtypes of ameloblastoma, including "Keratoameloblastoma" (KA), have been introduced in the literature. Here, we present a case of KA and discuss the important diagnostic microscopic features.

Cysts of the jaws: A multicentre study.

OBJECTIVES: To determine the relative frequency, demographic and pathologic profiles of patients diagnosed with cysts of the jaws. MATERIALS AND METHODS: Biopsy records of the participating institutions from 2000 to 2020 were reviewed for lesions diagnosed in the cyst category. Demographic data, the location of the cysts and pathologic diagnoses were collected. Data were analyzed by appropriate statistics using IBM SPSS software version 28.0. RESULTS: From 148,353 accessioned cases, 25,628 cases (17.28%) were diagnosed in the cyst category. Mean age of the patients ± SD = 42.62 ± 19.36 years. Paediatric patients (aged ≤ 16 years) accounted for 9.63%, while geriatric patients (aged ≥ 65) comprised 14.22% of all the patients. The male-to-female ratio was 1.27:1. The majority of the lesions were encountered in the mandible. The most prevalent cyst was radicular cyst followed by dentigerous cyst and odontogenic keratocyst. In the paediatric group, dentigerous cyst was the most prevalent, whereas in the geriatric group, radicular cyst was the most common. CONCLUSIONS: In general, the results of this study are in accordance with previous studies. This study provides an invaluable database for clinicians when formulating clinical differential diagnoses as well as for pathologists in rendering the final diagnosis.

Adenoid 'ameloblastoma': Clinicopathological description of 4 additional BRAF-negative cases.

OBJECTIVE: Adenoid ameloblastoma (AA) is an epithelial odontogenic tumor that was recognized as a separate entity in the last odontogenic classification of WHO in 2022. The etiology is unknown, and the pathogenesis remains controversial. The objective of this study is to contribute the clinicopathological features of 4 additional BRAF-negative cases to the existing literature, aiming to enhance the molecular understanding of this unique tumor in the forthcoming classification. MATERIALS AND METHODS: This study consists of a case series of four patients diagnosed with AA. The patients' demographic and clinical information were collected from the universities' medical achieves. Histopathologically, all cases were reexamined according to the latest update of the WHO odontogenic tumor classification. In addition to H&E and immunohistochemical stains, cytogenetics was also evaluated. RESULTS: Well-defined unilocular radiolucent lesions were observed in all cases. Ameloblastoma-like components exhibited reserved nuclear polarity, suprabasal stellate reticulum-like epithelium, duct-like structure, whorls/morules, and cribriform architecture were common features. Variable immunoreactivity to CK7, CK19, CK14, p63, and p40 were determined, and proliferative activity was greater than 15%. The BRAF molecular study revealed no mutations. CONCLUSIONS: When diagnosing AA, the essential histopathological characteristics must be rigorously applied, and a significant portion of the lesion should contain these features. Additionally, despite limited molecular data, since the BRAF mutation commonly observed in ameloblastomas is not present in the majority of AA cases, we propose changing the term "ameloblastoma" to "ameloblastic" and referring to it as "adenoid ameloblastic tumor" in the forthcoming classification.

Solid variant of odontogenic keratocyst in a 22-year-old man: Report of a case.

Key Clinical Message: In conclusion, we can focus on histologic features such as stellate reticulum, reverse polarization of basal cell layer nuclei, and luminal lamellated keratinization as distinguishing factors of ameloblastoma and SKCO. If there is any clinically doubt, molecular testing could be helpful. Abstract: "Solid odontogenic keratocyst" is a rare variant of odontogenic keratocyst, which usually involves mandible. This case was presented as a unique variant of odontogenic keratocyst in an unusual site of left maxilla with extension to the maxillary sinus.

Disulfiram-Loaded Niosomes Reduces Cancerous Phenotypes in Oral Squamous Cell Carcinoma Cells.

OBJECTIVE: Surgery and chemotherapy are the most common therapeutic strategies proposed for oral squamous cell carcinoma (OSCC). However, some of the disadvantages associated with the current methods like unwanted side effects and poor drug response lead the scientist to seek for novel modalities and delivery approaches to enhance the efficacy of treatments. The study aimed to assess the effectiveness of disulfiram (DSF)-loaded Niosomes on cancerous phenotypes of the OSCC cells. MATERIALS AND METHODS: In this experimental study, an optimum formulation of DSF-loaded Niosomes was developed for the treatment of OSCC cells to reduce drug doses and improve the poor stability of DSF in the OSCC environment. The design expert software was utilized to optimize the particles in terms of size, polydispersity index (PDI), and entrapment efficacy (EE). RESULTS: Acidic pH increased the release rate of DSF from these formulations. The size, PDI, and EE of Niosomes were more stable at 4°C compared to 25°C. The results indicated that DSF-loaded Niosomes could induce apoptosis (P=0.019) in the OSCC cells compared to the control group. Moreover, it could reduce colony formation ability (P=0.0046) and also migration capacity of OSCC cells (P=0.0015). CONCLUSION: Our findings indicated that the application of proper dose of DSF-loaded Niosomes (12.5 µg/ml) increases apoptosis, decreases colony formation capacity and declines the migration ability of OSCC cells.

High levels of histone H3 K27 acetylation and tri-methylation are associated with shorter survival in oral squamous cell carcinoma patients.

Background: Acetylation and trimethylation of histone H3 lysine 27 (H3K27ac and H3K27me3) generally activate and repress transcription, respectively. Concurrent activation of H3K27ac and H3K27me3 has been reported to correlate with poor prognosis in hepatocellular carcinoma. A high level of H3K27me3 has been shown to be associated with advanced oral squamous cell carcinoma (OSCC) tumour stage, but prognostic impact of H3K27ac level alone/or in combination with H3K27me3 in OSCC patients had not yet been reported. Material and methods: In this study, immunohistochemistry using specific antibodies against H3K27ac and H3K27me3 was performed on a series of 72 OSCC samples to investigate the association between H3K27ac and H3K27me3 levels and OSCC patient's survival. For each mark, the proportion of labelled cells (percentage) and the intensity of the labelling were measured, and a score of percentage x intensity was calculated. Results: A high percentage of H3K27me3 positive cells was significantly associated with survival in univariate and multivariate analyses (logrank p-value < 0.05). Patients with high total scores of H3K27ac and H3K27me3 labelling also showed significantly shorter survival probabilities (logrank p-value < 0.05) compared to patients with low total scores of labelling for these histone posttranslational modifications (PTMs). Conclusion: Our findings suggest that the detection of both H3K27ac and H3k27me3 could help evaluating prognosis in OSCC patients.

Central oral pulse granuloma: a case report of huge mandibular jaw lesion.

Oral pulse granuloma is a rare lesion of the oral cavity with unclear etiology. Some authors believe this lesion is a foreign body reaction to implanted food particles. In the oral cavity, most cases are found in the posterior regions of the mandible. The edentulous mandible was involved in 20 cases with oral pulse granuloma. In these cases, the premolar-molar site was the most common region. Here we present a case of a 70-year-old male with huge unilateral swelling of the mandible on the left side. This paper aims to present a case of oral pulse granuloma with wide extension, detailed clinicohistopathologic features with 2-year follow-up and a short review of reported cases.

Combination of Binary System with Ki67 Immunohistochemical Staining: a Reliable Grading Method for Oral Epithelial Dysplasia.

Statement of the Problem: Oral epithelial dysplasia (OED) is a common potentially malignant lesion of oral cavity that should be managed to prevent likely malignant transformation. Purpose: Here, we present a combination of binary grading system with complementary immunohistochemical (IHC) staining for Ki67 biomarker to provide a reproducible OED grading system. Materials and Method: In this cross-sectional study, seventy out of one hundred OED specimens, which were accompanied by IHC stained microscopic slides for Ki67 antigen were evaluated by four independent oral pathologists. Both three-tier and binary grading systems based on WHO microscopic criteria were employed , blindly in a four-step method with at least two-week interval between each observation. Intra- and inter-observational reliability was assessed using Kappa statistical analysis. Results: OED diagnosis based on binary system showed significant intra-observer reliability comparing to three-tier system without biomarker. Moreover, OED diagnosis based on binary system using Ki67 biomarker showed significant inter-observer reliability comparing to diagnosis in three tier system and based on binary system without Ki67 biomarker showed significant inter-observer reliability comparing to diagnosis based on three-tier system without Ki67. Conclusion: Here, we found that application of IHC staining for Ki67 biomarker in binary system might provide a more reliable grading method for oral pathologist form different educational background.

Effect of Light-Emitting Diode Phototherapy on Allograft Bone After Open Sinus Lift Surgery: A Randomized Clinical Trial (Concurrent Parallel).

Introduction: Phototherapy with a light-emitting diode (LED) is used in medicine due to its potential bio-stimulatory effects on the human body. However, controversy still exists regarding the efficacy of low-level laser therapy (LLLT) and phototherapy with LED. This in vivo study aimed to quantitatively and qualitatively assess the newly formed bone following LED phototherapy of the human maxillary sinuses. Methods: This randomized clinical trial (concurrent parallel) was conducted on 44 patients in two groups (n=22) at the Implant Department of Tehran University of Medical Sciences. Randomization was done by a random sequence generator program. The inclusion criteria were absence of chronic sinusitis and chronic bone marrow conditions, no history of surgery at the site, absence of diabetes mellitus, no history of chemotherapy or radiotherapy, maxillary premolar edentulism, and signing informed consent forms. Group A underwent LED phototherapy with 620 ± 2 nm wavelength for 20 minutes daily for a total of 21 days after sinus lift surgery. Group B served as the control group and did not receive phototherapy. After 6 months, the grafted sites were re-opened for implant placement, and bone biopsy samples were obtained using a trephine bur. The samples were stained with hematoxylin and eosin and inspected under a light microscope. The results were statistically analyzed using the Mann-Whitney U test. Both the surgeon and pathologist were blinded to the group allocation of patients. Results: Forty tissue specimens were analyzed. Insignificant differences existed between the two groups in terms of the degree of inflammation, bone quality, and maturity of collagen. Histological analyses revealed no significant difference in the mineralized areas of bone between the two groups (P >0.05). Conclusion: The results indicated that LED phototherapy cannot significantly enhance osteogenesis after sinus lift surgery. No side effects were observed in the experimental group.

Conventional osteosarcoma of the mandible: Report of a rare case.

Osteosarcoma of the jaw can be successfully treated by a single aggressive surgical operation with good prognosis. Timely diagnosis, with a complete surgical removal of the tumor with clear margins, can fulfill favorable clinical results.

The combined detection of Amphiregulin, Cyclin A1 and DDX20/Gemin3 expression predicts aggressive forms of oral squamous cell carcinoma.

BACKGROUND: Large-scale genetic and epigenetic deregulations enable cancer cells to ectopically activate tissue-specific expression programmes. A specifically designed strategy was applied to oral squamous cell carcinomas (OSCC) in order to detect ectopic gene activations and develop a prognostic stratification test. METHODS: A dedicated original prognosis biomarker discovery approach was implemented using genome-wide transcriptomic data of OSCC, including training and validation cohorts. Abnormal expressions of silent genes were systematically detected, correlated with survival probabilities and evaluated as predictive biomarkers. The resulting stratification test was confirmed in an independent cohort using immunohistochemistry. RESULTS: A specific gene expression signature, including a combination of three genes, AREG, CCNA1 and DDX20, was found associated with high-risk OSCC in univariate and multivariate analyses. It was translated into an immunohistochemistry-based test, which successfully stratified patients of our own independent cohort. DISCUSSION: The exploration of the whole gene expression profile characterising aggressive OSCC tumours highlights their enhanced proliferative and poorly differentiated intrinsic nature. Experimental targeting of CCNA1 in OSCC cells is associated with a shift of transcriptomic signature towards the less aggressive form of OSCC, suggesting that CCNA1 could be a good target for therapeutic approaches.

A fast-growing schwannoma of the tongue in a 15-year-old Iranian male: Review of literature and case report.

Schwannoma can be included in the list of differential diagnoses of tongue masses but seems to be a rare finding in the Iranian population. The current case was presented as an exophytic nodular sessile mass which was growing fast.

The effect of aflibercept and arsenic trioxide on the proliferation, migration and apoptosis of oral squamous cell carcinoma in vitro.

Aflibercept and arsenic trioxide drugs apply a cytotoxic effect on some human cancer cell lines. However, no more study has followed the effects of both drugs, especially arsenic trioxide, on oral squamous cell carcinoma (OCC). We used three OCC lines as a model to show the effect of these drugs on the genetically complex disease and investigate its targeted therapy. In this study, three human OCC cell lines were used from different patients. We treated cell lines with both medications to detect the effect and relevant molecular basis. First, methyl thiazolyl tetrazolium (MTT) assay was performed to detect the cytotoxicity effect and cell growth. Second, flow cytometry, gene and protein expression were performed to evaluate the anti-angiogenic effect on OCC lines. Next apoptosis was analyzed by flow cytometry. Finally, clonogenesis capacity and cell migration were assessed by colony formation assay and wound healing, respectively. Aflibercept had no cytotoxic effect on the three OCC cell lines but decreased cell growth rate. Arsenic trioxide had a significant cytotoxic effect on three cell lines. Our results demonstrated that both drugs significantly decreased endoglin, VEGFA, and VEGFB expression. In addition, Migration and colony formation assays confirmed that these drugs have significant anti-proliferative and anti-migration effect on oral carcinoma cells. These results revealed that both medications might be a potential drug for the management of oral cancer patients.

Evaluation of the presence of myofibroblasts and matrix metalloproteinase 1 expression in the stroma of oral verrucous hyperplasia and verrucous carcinoma.

BACKGROUND: Oral verrucous carcinoma is a low-grade subtype of oral squamous cell carcinoma that should be differentiated from oral verrucous hyperplasia, a premalignant lesion. Stromal activated myofibroblasts known as cancer-associated fibroblasts have an active role in the initiation and progression of the cancers via secretion of different molecules including matrix metalloproteinases. AIMS: This study is designed to understand the differences in the presence of myofibroblasts and expression of matrix metalloproteinase-1 in the adjacent stroma of verrucous carcinoma and oral verrucous hyperplasia (OVH). SETTINGS AND DESIGN: Cross-sectional study. MATERIAL AND METHODS: Twenty-seven OVH, 19 oral verrucous carcinoma (OVC), and 8 cutaneous verrucous carcinoma (CVC) specimens were analyzed for immunohistochemical (IHC) expression of α-smooth muscle actin (αSMA) and MMP-1. RESULTS: IHC studies for αSMA expression in nonvascular stromal cells of the adjacent stroma revealed mild or no expression in 81.4%, 73.7%, and 62.5% of the cases of OVH, OVC, and CVC groups, respectively. No significant difference was seen in αSMA expression index between OVH and OVC groups (Adj. Sig. = 0.220) and between OVC and CVC groups (Adj. Sig. = 1.00). Pairwise analysis revealed a significant difference in MMP-1 expression index between the groups. No significant correlation was observed between MMP-1 expression index and αSMA expression index in OVH (pv = 0.358) and OVC (pv = 0.388) groups. CONCLUSION: The differences in MMP-1 expression between OVH and OVC can be used as an adjunctive aid in challenging cases including disoriented or inadequate samples.

Starvation promotes histone lysine butyrylation in the liver of male but not female mice.

AIMS: Post-translational modifications (PTMs) of histones are regulated by the availability of their respective acyl-CoAs. Among these histone PTMs, the metabolic origin of histone butyrylation (Kbu) is still poorly understood. MATERIAL AND METHODS: The impact of starvation on the levels of Kbu was determined by western blotting on histones extracted from the liver of fed and fasted C57BL/6 mice and immunohistochemistry on liver paraffin sections. KEY FINDINGS: Using animal model we provide evidence that the stimulation of ketogenesis following starvation, in addition to histone beta-hydroxybutyrylation (Kbhb), also leads to an increase in histone butyrylation (Kbu). Using an immunohistochemistry (IHC) approach we report first that hepatocytes contained butyrylated histones with important cell-to-cell heterogeneity. More importantly, our investigations based on western blotting and IHC also proposed that the basal levels of Kbu differ between male and female mice, with female mouse hepatocytes containing higher levels of butyrylated histones. Starvation enhanced solely histone Kbu levels in the liver of males but not females. SIGNIFICANCE: This is the first demonstration of a sex-dependent large-scale stimulation of histone acylation. Our data also point to different basal metabolic conditions of the male and female liver cells with a sex-dependent impact on the hepatocytes' epigenome.

Relative Expression of OCT4, SOX2 and NANOG in Oral Squamous Cell Carcinoma Versus Adjacent Non- Tumor Tissue.

Objective: Oral squamous cell carcinoma (OSCC) accounts for over 90% of oral neoplasms. Finding molecular markers for predicting prognosis is a high priority. The core transcription factors, OCT4, SOX2, and NANOG that regulate embryonic stem cell pluripotency have been implicated in progression of various malignancies. The predictive value of these markers and their role in the development of OSCC is still controversial. In this study, we therefore evaluated their expression in OSCCs and adjacent non-tumor tissue. Methods: A total of 60 frozen tumor and adjacent non-tumor tissue samples from 30 patients with OSCC were examined using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Clinical and pathological data of patients including tumor stage, lymph node metastasis and tumor grade were also recorded. Results: Expression of SOX2 was significantly higher in adjacent non-tumor as compared to tumor tissue (P=0.04). No statistically significant differences were found for expression of OCT4 (P=0.50) and NANOG (P=0.68). Also, there was no significant association between expression of OCT4, SOX2, and NANOG and clinical or pathological data (P>0.05), although slightly higher values were noted in patients without lymph node metastasis. Conclusion: Based on the present data, decreased expression of SOX2 is correlated with carcinogenesis in the oral cavity and development of OSCC.

PTEN and miR-26b: Promising prognostic biomarkers in initiation and progression of Oral Squamous Cell Carcinoma.

BACKGROUNDS: Oral Squamous Cell Carcinoma (OSCC) is the most common malignancy of the oral cavity. Phosphatase and TENsin homolog (PTEN) is a well-known tumor suppressive gene regulated by several biomarkers including a small single-stranded molecule, microRNA26b (miR-26b). Here, we studied the expression of PTEN and miR-26b in OSCC specimens in comparison with adjacent normal mucosa. METHODS: The expressions of PTEN and miR-26b genes were evaluated at mRNA level in OSCC and adjacent normal fresh frozen tissues in 49 patients using Quantitative Real-Time PCR and analyzed their associations with clinicopathological factors. RESULTS: The expression level of PTEN was significantly lower in OSCC specimens comparing with adjacent normal tissues (P-value = 0.000). The expression of PTEN was associated with T stage (P-value = 0.006) and N stage (P-value = 0.043). A nonsignificant decrease in miR-26b expression level was also observed in OSCC tissues. Additionally, in patients with more aggressive tumoral behavior, including vascular invasion (P-value = 0.012) and positive N stage (P-value = 0.02), significant decreases were found. CONCLUSIONS: These findings suggest that inactivation of PTEN may have an impact on initiation and progression of OSCC. Additionally, miR-26b might have a tumor suppressive role in OSCC.

Benign Cementoblastoma Involving Deciduous and Permanent Mandibular Molars: A Case Report.

Cementoblastomas are rare benign odontogenic tumors. Diagnosis of these lesions must be made by an association of clinical, radiographic, and histopathological findings. Cementoblastomas rarely occur in both primary and permanent dentitions. We describe the sixth case of cementoblastoma in the literature with the involvement of both deciduous and permanent teeth. The aim of this case report is to present the clinicoradiopathologic features of a cementoblastoma in a 4.5-year-old boy with an unusual recurrence. The first clinical and radiographic features appeared on the deciduous mandibular second molar. The second lesion occurred 1 year after treatment at 5.5 years old, involving the permanent mandibular first molar, and a subsequent lesion was seen at age 8 years in the edentulous region of the extracted mandibular first molar. After the last surgery, there was no recurrence of the lesion at 6 months' follow-up. Follow-ups of patients with cementoblastomas are highly recommended for an early detection of recurrences.