Central giant cell granulomas of the jaws stromal cells harbour mutations and have osteogenic differentiation capacity, in vivo and in vitro.

BACKGROUND: Central giant cell granulomas (CGCG) of the jaws are osteolytic lesions that may behave aggressively and respond poorly to surgery. Microscopically, in addition to giant cells, there is a mononuclear cell population composed of macrophage/monocytic cells and spindle-shaped cells of mesenchymal origin. Seventy two percent of these tumours harbour mutually exclusive TRPV4, KRAS and FGFR1 mutations. We aimed to assess the mutational status of mononuclear and giant cells and the osteogenic potential of stromal cells in vitro and in vivo. METHODS AND RESULTS: We screened CGCG for signature mutations and used laser-capture microdissection to demonstrate that the mutations are restricted to the mononuclear cells. Additionally, we established CGCG primary cell culture and observed that the cells retained the mutations throughout passages. By flow cytometry, we observed predominance of CD14- CD51- CD61- cells, consistent with the expected profile for stromal cells. Considering the mesenchymal origin of stromal cells, we assessed the osteogenic differentiation potential of CGCG cells in culture by cytochemistry (von Kossa and alizarin red staining), alkaline phosphatase (ALP) activity assay and gene expression of osteogenic markers. CGCG cells presented self-capacity to increase ALP levels in a time-dependent manner and under osteogenic induction presented increasing number of calcium deposits, and overall higher expression of osteocalcin, RUNX2, ALPL and osteopontin than cells without osteogenic induction. A patient-derived xenograft model for CGCG was established, and osteoid material deposition was observed. CONCLUSION: Collectively, the results confirm that the signature mutations are restricted to stromal cells in CGCG, and the in vitro and in vivo results support that these cells have the capacity to differentiate into osteoblasts, in line with the bone formation often observed in the stroma of these lesions.

Unveiling metabolic changes in marsupialized odontogenic keratocyst: A pilot study.

OBJECTIVE: We aimed to assess which metabolic pathways would be implicated in the phenotypic changes of the epithelial lining of odontogenic keratocyst after marsupialization, comparing pre- and post-marsupialized lesions with adjacent oral mucosa. MATERIALS AND METHODS: Eighteen formalin-fixed and paraffin-embedded tissues from six subjects were divided into three paired groups: odontogenic keratocyst pre- (n = 6) and post-marsupialization (n = 6), and adjacent oral mucosa (n = 6). The metabolic pathways found in these groups were obtained by high-performance liquid chromatography-mass spectrometry-based untargeted metabolomics performed. RESULTS: Through putative metabolite annotation followed by pathway enrichment and predictive analysis with automated algorithms (Mummichog and Gene Set Enrichment Analysis), we found differences in many cellular processes that may be involved in inflammation, oxidative stress response, keratinocyte-basal membrane attachment, differentiation, and proliferation functions, all relevant to odontogenic keratocyst pathobiology and the phenotype acquired after marsupialization. CONCLUSION: Our study was able to identify several metabolic pathways potentially involved in the metaplastic changes induced by marsupialization of odontogenic keratocysts. An improved comprehension of this process could pave the way for the development of targeted therapies.

KRAS mutations in brown tumor of the jaws in hyperparathyroidism.

BACKGROUND: Brown tumors are giant cell-rich lesions that result from abnormal bone metabolism in hyperparathyroidism, one of the most common endocrine disorders worldwide. Brown tumors occasionally affect the jaws and, despite well-known clinical and microscopic features, their molecular pathogenesis remains unclear. We investigated the presence of pathogenic activating mutations in TRPV4, FGFR1, and KRAS in a cohort of brown tumors since these have recently been reported in giant-cell lesions of the jaws and non-ossifying fibromas of the bones (FGFR1 and KRAS), which are histologic mimics of brown tumors. METHODS: We target sequenced 13 brown tumors of the jaws associated with primary or secondary hyperparathyroidism. As mutations in these genes are known to activate the MAPK/ERK signaling pathway, we also assessed the immunostaining of the phosphorylated form of ERK1/2 (pERK1/2) in these lesions. RESULTS: KRAS pathogenic mutations were detected in seven cases (p.G12V n = 4, p.G12D n = 1, p.G13D n = 1, p.A146T n = 1). KRAS variants of unknown significance (VUS), p.A134T and p.E37K, were also detected. All samples showed wild-type sequences for FGFR1 and TRPV4 genes. The activation of the MAPK/ERK signaling pathway was demonstrated by pERK1/2 immunohistochemical positivity of the brown tumors´ mononuclear cells. CONCLUSION: Mutations in KRAS and activation of the MAPK/ERK signaling pathway were detected in brown tumors of hyperparathyroidism of the jaws, expanding the spectrum of giant cell lesions whose molecular pathogenesis involve RAS signaling.

Adenoid ameloblastoma in the posterior maxilla: a case report and review of the literature.

INTRODUCTION: Adenoid ameloblastoma is a rare benign odontogenic tumor that favors a slight predilection for male patients, fourth and fifth decades of life, and posterior regions of the jaws. To date, less than 40 cases have been reported in the English language literature. The radiographic aspects of adenoid ameloblastoma vary from unilocular and well-defined lesions to diffuse and multilocular lesions. Most of the lesions exhibit a radiolucent image and are usually large, with a mean size of 3.5 cm. Microscopically, pseudoductal structures composed of columnar cells in a palisaded arrangement formed from the parenchyma of the tumor were observed. CASE PRESENTATION: We describe a case of adenoid ameloblastoma in a 54-year-old woman, who presented with no symptoms. Panoramic radiography showed a well-circumscribed, unilocular radiolucency in the left posterior maxilla. CONCLUSION: As odontogenic tumors are rare, some entities are infrequently encountered, making the diagnosis more difficult. Clinicians, oral and maxillofacial surgeons and oral pathologists should be familiar with the adenoid ameloblastoma and its differential diagnosis for accurate diagnosis and management.

MicroRNA profiling reveals dysregulated microRNAs and their target gene regulatory networks in cemento-ossifying fibroma.

BACKGROUND: Cemento-ossifying fibroma (COF) is a benign fibro-osseous neoplasm of uncertain pathogenesis, and its treatment results in morbidity. MicroRNAs (miRNA) are small non-coding RNAs that regulate gene expression and may represent therapeutic targets. The purpose of the study was to generate a comprehensive miRNA profile of COF compared to normal bone. Additionally, the most relevant pathways and target genes of differentially expressed miRNA were investigated by in silico analysis. METHODS: Nine COF and ten normal bone samples were included in the study. miRNA profiling was carried out by using TaqMan® OpenArray® Human microRNA panel containing 754 validated human miRNAs. We identified the most relevant miRNAs target genes through the leader gene approach, using STRING and Cytoscape software. Pathways enrichment analysis was performed using DIANA-miRPath. RESULTS: Eleven miRNAs were downregulated (hsa-miR-95-3p, hsa-miR-141-3p, hsa-miR-205-5p, hsa-miR-223-3p, hsa-miR-31-5p, hsa-miR-944, hsa-miR-200b-3p, hsa-miR-135b-5p, hsa-miR-31-3p, hsa-miR-223-5p and hsa-miR-200c-3p), and five were upregulated (hsa-miR-181a-5p, hsa-miR-181c-5p, hsa-miR-149-5p, hsa-miR-138-5p and hsa-miR-199a-3p) in COF compared to normal bone. Eighteen common target genes were predicted, and the leader genes approach identified the following genes involved in human COF: EZH2, XIAP, MET and TGFBR1. According to the biology of bone and COF, the most relevant KEGG pathways revealed by enrichment analysis were proteoglycans in cancer, miRNAs in cancer, pathways in cancer, p53-, PI3K-Akt-, FoxO- and TGF-beta signalling pathways, which were previously found to be differentially regulated in bone neoplasms, odontogenic tumours and osteogenesis. CONCLUSION: miRNA dysregulation occurs in COF, and EZH2, XIAP, MET and TGFBR1 are potential targets for functional analysis validation.

DNA methylation patterns of genes related to immune response in the different clinical forms of oral lichen planus.

BACKGROUND: The oral lichen planus is a chronic inflammatory disease. Although its aetiology is not well understood, the role of T lymphocytes in its inflammatory events is recognised. Identifying the epigenetic mechanisms involved in the pathogenesis of this immune-mediated condition is fundamental for understanding the inflammatory reaction that occurs in the disease. The purpose of this work was to evaluate the methylation pattern of 21 immune response-related genes in the different clinical forms of oral lichen planus. METHODS: A cross-sectional study was performed to analyse the DNA methylation patterns in three distinct groups of oral lichen planus: (i) reticular/plaque lesions; (ii) erosive lesions; (iii) normal oral mucosa (control group). After DNA extraction from biopsies, the samples were submitted to digestions by methylation-sensitive and methylation-dependent enzymes and double digestion. The relative percentage of methylated DNA for each gene was provided using real-time polymerase chain reaction arrays. RESULTS: Hypermethylation of the STAT5A gene was observed only in the control group (59.0%). A higher hypermethylation of the ELANE gene was found in reticular/plaque lesions (72.1%) compared to the erosive lesions (50.0%). CONCLUSION: Our results show variations in the methylation profile of immune response-related genes, according to the clinical type of oral lichen planus after comparing with the normal oral mucosa. Further studies are necessary to validate these findings using gene expression analysis.

Oral paracoccidioidomycosis: Retrospective analysis of 55 Brazilian patients.

Paracoccidioidomycosis (PCM) is a rare fungal infection in the world, but endemic and acquired exclusively in Latin America, with the highest prevalence in South America and Brazil, particularly. Changes in oral cavity are common and constitute the first clinical manifestation in many patients. The aim of this study was to describe the prevalence of oral PCM and analyse the profile of the disease and patients. Retrospective research, consisting of information present in the medical records in the period 1998-2015, whose histopathological diagnosis was oral PCM. Fifty-five oral PCM cases were confirmed. Of these patients, 90.9% were males and 9.1% were females. The average age was 49.66 years and the most reported occupation was rural workers. The painful symptomatology was present in 61.82% of patients. Erythematous lesions were predominant in 73% of them. In single lesions (22 cases), the most common locations were jugal mucosa and tongue. In multiple involvement (30 cases), the most affected regions were lips, jugal mucosa and alveolar ridge. Epidemiology of PCM, was similar to several other studies, especially in Brazil. This is the most important fungal infection in Latin America and the recognition of oral lesions is extremely important, as is often the first and in many cases the only manifestation of the disease.

Mucosal leishmaniasis in Brazilian patients: two case reports with similar clinical presentation and different approaches.

Mucosal leishmaniasis is a chronic infection that affects the upper respiratory tract and/or the oral mucosa caused by Leishmania protozoan parasites. We present two cases of oral leishmaniasis and discuss the different diagnostic strategies and treatment. In both cases, the patients were male, 60 and 94 years of age, and presented with lesions on the soft palate. In the first patient, the final diagnosis was made on the basis of histopathologic examination. In the second case, polymerase chain reaction and Montenegro skin test were necessary to confirm the diagnosis. The first patient was treated with meglumine antimoniate (Glucantime), and the lesions healed after 2 months. In the second case, the patient received treatment with liposomal amphotericin B but later died as a result of generalized infection. Mucosal leishmaniasis is a highly disfiguring disease. Early diagnosis is important to prevent a lethal outcome.

Desmoplastic fibroblastoma (collagenous fibroma) of the oral cavity.

UNLABELLED: Desmoplastic fibroblastoma is benign soft tissue tumor, with fibroblastic or myofibroblastic origin, that rarely occurs in oral cavity. We reported the case of a 56-year-old man who presented a tumor in the left mandibular alveolar ridge, with slow and asymptomatic growth, with no osseous involvement. The tumor was sessile with lobulated surface, covered by healthy mucosa with erythematous areas. The lesion was excised and specimens sent to histopathology and immunohistochemistry. Histopathological exam showed a non-encapsulated fibroblastic proliferation, characterized by myofibroblasts, spindle and stellate fibroblasts with large or oval nuclei and bi or tri nucleation, immersed in an abundant hypocellular dense collagen stroma. Tumor cells were positive for vimentin, HHF35, α-smooth muscle actin and factor XIIIa. The diagnosis of desmoplastic fibroblastoma was based in the clinical history of absence of trauma related to the growth in the alveolar ridge, associated with macroscopic, microscopic and immunohistochemical features. The patient is free-diseases by eight months. KEY WORDS: Collagenous fibroma, desmoplastic fibroblastoma, neoplasm of connective and soft tissue.

Synovial Chondromatosis of the Temporomandibular Joint Successfully Treated by Surgery.

Synovial chondromatosis (SC) is a chronic process, defined as a reactive cartilaginous proliferation, characterized by formation of cartilaginous nodules, usually loose in the joint space of the synovial membrane. It mainly affects large joints such as knee, hip, shoulder, and elbow, commonly in male patients. However, its manifestation in the Temporomandibular joint (TMJ), is a rare finding, occurring predominantly in females. This paper reports a case of a woman who presented to the service of Stomatology complaining of mouth opening limitations and pain in her left pre-auricular region. After clinical and radiographic analyses, the condition was diagnosed as SC of the TMJ. The loose bodies within the TMJ were removed under general anesthesia. Histological and follow-up features of this lesion are also discussed. To our knowledge, this is the second report of SC of the TMJ in Brazil.

Clonal nature of odontogenic tumours.

BACKGROUND: Although clonal origin is an essential step in the comprehension of neoplasias, there have been no studies to examine whether odontogenic tumours are derived from a single somatic progenitor cell. The purpose of this study was to investigate the clonal origin of odontogenic tumours. METHODS: Fresh samples of seven ameloblastomas, two odontogenic mixomas, two adenomatoid odontogenic tumour, one calcifying odontogenic cyst, one calcifying epithelial odontogenic tumour (CEOT) and six odontogenic keratocyst (OKC) of female patients were included in this study. After DNA extraction, the HUMARA gene polymorphism assay was performed. RESULTS: Most of the informative odontogenic lesions studied (12 out of 16) showed a monoclonal pattern. Among the polyclonal cases, two were OKC, one CEOT and one odontogenic mixoma. CONCLUSIONS: Our results suggest that most odontogenic tumours are monoclonal.

Calcifying odontogenic cyst associated with an orthokeratinized odontogenic cyst.

Odontogenic tumors composed of two or more distinct types of lesions are unusual. In this paper, a case of an odontogenic lesion characterized by simultaneous occurrence of areas of calcifying odontogenic cyst (COC) and orthokeratinized odontogenic cyst (OOC) is described. The lesion was asymptomatic and presented at the radiographic examination as a unilocular well-delimited radiolucency extending from left incisor to right premolar area in the mandible. To date, this is the first report of COC associated with an OOC.