Adenomatoid odontogenic tumor: Features of ameloblastic-like epithelial cells differentiation, secretion, and the nature of tumor cells products.

BACKGROUND: This study aimed to investigate the differentiation of ameloblastic-like cells and the nature of the secreted eosinophilic materials in adenomatoid odontogenic tumors. METHODS: We studied histological and immunohistochemical characteristics of 20 cases using: cytokeratins 14 and 19, amelogenin, collagen I, laminin, vimentin, and CD34. RESULTS: Rosette cells differentiated into ameloblastic-like cells positioned face-to-face, displaying collagen I-positive material between them. Epithelial cells of the rosettes can differentiate into ameloblastic-like cells. This phenomenon probably occurs due to an induction phenomenon between these cells. The secretion of collagen I is probably a brief event. Amelogenin-positive areas were interspersed by epithelial cells in the lace-like areas, outside the rosettes and distant from the ameloblastic-like cells. CONCLUSIONS: There are at least two types of eosinophilic material in different areas within the tumor, one in the rosette and solid areas and another in lace-like areas. The secreted eosinophilic material in the rosettes and solid areas is probably a product of well-differentiated ameloblastic-like cells. It is positive for collagen I and negative for amelogenin, whereas some eosinophilic materials in the lace-like areas are positive for amelogenin. We hypothesize that the latter eosinophilic material could be a product of odontogenic cuboidal epithelial or intermediate stratum-like epithelial cells.

Oral epithelial dysplasia grading: Comparing the binary system to the traditional 3-tier system, an actuarial study with malignant transformation as outcome.

BACKGROUND: Establishing the risk of malignant transformation (MT) in oral leukoplakia is usually based on grading oral epithelial dysplasia (OED) on biopsy tissue, for which two systems are proposed: a 3-tier and a binary system. Only very few actuarial studies have tested the accuracy of such methods in predicting MT, especially for the binary system. This study aimed to assess the accuracy of the two grading systems in predicting MT in a cohort of oral leukoplakia (OL) from Brazil, with follow-up data. METHODS: The sample comprised 878 individuals diagnosed with OL from 2005 to 2018. Follow-up data were obtained both locally and from the regional cancer registry. All lesions were graded using both the 3-tier and the binary systems. Kaplan-Meier curves (Log-rank Mantel-Cox) were used to assess risk and kappa to assess interobserver agreement. RESULTS: Thirty-five individuals underwent MT (4%). Both systems demonstrated prognostic value, though the 3-tier system proved superior, with OR 9.23 (3.42-23.69), PPV 0.152, NPV 0.98, compared to binary OR 3.49 (1.79-6.79), PPV 0.079, NPV 0.976. Interobserver agreement was also superior in the 3-tier system (0.47, p < 0.05) compared to the binary system (0.139, p = 0.39). Combining the two systems enhanced prognostic values (OR 14.28, PPV 0.217, NPV 0.981). CONCLUSION: The 3-tier system presented superior prognostic value to the binary system. Combining both systems to double-grade intermediate lesions might enhance risk assessment.

Microvesicles derived from squamous cell carcinoma induce cell death, autophagy, and invasion of benign myoepithelial cells.

BACKGROUND: There has been great interest recently in the mechanisms of cell-to-cell communication through microvesicles (MV). These structures are produced by many different cell types and can modulate cellular activity by induction of epigenetic alterations. These vesicles may promote tumor mass increase either by stimulating cell proliferation via growth factors or by inhibiting apoptosis, which reinforces the role of such vesicles as important modulators of tumor progression. METHODS: The present in vitro study aimed to characterize MV derived from malignant neoplastic epithelial cell cultures (EP) and their effect on the expression of apoptosis/autophagy and invasion related genes of benign myoepithelial (Myo) cell cultures. RESULTS: The results revealed round structures with a mean size of 153.6 (±0.2) nm, with typical MV morphology. CD63 quantification indicated that EP cell culture at 70%-80% confluence secreted 3.088 × 108 MV/mL. Overall, Myo exposed to MVs derived from EP showed both up- and downregulation of tumorigenesis promoting genes. MVs from EP cells promoted cell death of Myo cells and positively modulate BAX, SURVIVIN, LC3B, MMP-2, and MMP-9 expression. Furthermore, an increasing of MMP-2 and MMP-9 secretion by Myo was observed after MV exposure. CONCLUSIONS: These findings suggest that MVs from EP modulate autophagy of Myo cells, which may, in part, explain the disappearance of these cells in in situ areas of invasive carcinoma ex-pleomorphic adenoma. Additionally, the overexpression of MMPs contributes to the development of an invasive phenotype of Myo cells, which could favor the dissolution of the basement membrane during tumorigenesis process.

Benign odontogenic ghost cell lesions revisited and new considerations on dysplastic dentin.

OBJECTIVES: This study aimed to revisit benign odontogenic ghost cell lesions (BOGCL) by hematoxylin and eosin staining and immunohistochemistry. MATERIALS AND METHODS: Thirty cases of calcifying odontogenic cyst (COC) and 6 cases of dentinogenic ghost cell tumor (DGCT) were selected for histopathological and immunohistochemical analysis. Sections stained for cytokeratin (K) 14, K-19, amelogenin, collagen type 1 (COL-1), and dentin matrix acidic phosphoprotein 1 (DMP-1) were evaluated using qualitative analysis. Sections stained for Ki-67 and minichromosome maintenance protein-2 (MCM-2) were evaluated using semi-quantitative analysis. RESULTS: A morphologic overlap was noticed in all BOGCL. Moreover, no differences were detected in the expression of K-14 and K-19. The expression of proliferative markers Ki-67 and MCM-2 was similar between cystic and tumor lesions (p > .05). The presence of COL-1 and absence of amelogenin in the so-called dysplastic dentin, associated with its histologic pattern, suggest that this is in fact an enameloid-like tissue. CONCLUSIONS: The dysplastic dentin should be considered an enameloid-like tissue in these lesions. CLINICAL RELEVANCE: The similarity in histology, protein expression, and proliferative marker indices between COC and DGCT suggest that they are a sole entity and likely represent types of the same neoplasia.

Microvessel density and cell proliferation in juvenile ossifying fibroma: A comparative study with central ossifying fibroma.

Considered as an aggressive counterpart of central ossifying fibroma (OF), juvenile ossifying fibroma (JOF) is a benign fibro-osseous neoplasm characterized by an unpredictable destructive behavior, elevated morbidity, mutilating treatment and high potential for local recurrences. The aim of this study is to compare the analysis for cell proliferation and vascular markers between JOF and OF. Cell proliferation index was measured by Ki-67 and Mcm-2 expression and microvessel density (MVD) was obtained by the immunoexpression of CD34/CD105. We observed a reduced expression of vascular markers, where MVD for CD34 was significantly higher in JOF than in OF (p = 0.009), but no statistical difference was found for CD105. JOF and OF showed low expression for Ki-67 and Mcm-2 and no difference was noted between both, suggesting that other mechanisms such as anti-apoptotic and/or pro-autophagic pathways or even increased expression of matrix metalloproteinases may be responsible for the aggressiveness of JOF.

Mammaglobin and DOG-1 expression in polymorphous low-grade adenocarcinoma: an appraisal of its origin and morphology.

BACKGROUND: Polymorphous low-grade adenocarcinoma (PLGA) remains a diagnostic challenge for most pathologists due to its large spectrum of histological patterns. In this study, the expression of two new markers recently described for salivary gland tumors was studied in PLGA. METHODS: The morphology of 33 cases of PLGA was carefully evaluated using hematoxylin-and-eosin-stained sections and confirmed by immunohistochemistry for cytokeratin 7, vimentin, and S-100. Periodic acid-Schiff with diastase digestion was also used. The expression of mammaglobin and DOG-1 was carried out using the EnVision System. Mammaglobin was assessed according to the percentage of positively stained tumor cells, while DOG-1 was evaluated according to its presence and site. For MCM-2 and Ki-67, markers of proliferation, the labeling index of cell nuclei positivity was evaluated using total cell number. The ETV6-NTRK3 fusion was examined by fluorescence in situ hybridization analysis. RESULTS: The histological patterns of the tumor were classified as lobular or non-lobular. For the non-lobular pattern, tubular, cribriform, glomeruliform, trabecular, and papillary patterns were observed. Mammaglobin was present in all PLGA cases, and its expression was stronger (P = 0.01) in the lobular than in the non-lobular pattern. The expression of DOG-1 was present in the apical portion and cytoplasm of the cells. Proliferation markers were low for all cases independent of histological pattern. CONCLUSIONS: Polymorphous low-grade adenocarcinoma has been confirmed to originate from the intercalated duct and to feature high expression of mammaglobin in its lobular pattern resembling that of mammary secretory analogue carcinoma, except for the ETV6 gene rearrangement.

Immunoexpression of growth factors and receptors in polymorphous low-grade adenocarcinoma.

BACKGROUND: Polymorphous low-grade adenocarcinoma (PLGA) is a rare malignant tumor that usually arises in the minor salivary glands. Growth factors are cell-secreted peptides that regulate biological processes such as growth, proliferation, and differentiation. In salivary gland tumors, immunoexpression of growth factors and their receptors is associated with cell proliferation, malignant transformation, and tumor invasion. This study analyzed the expression of growth factors and receptors in PLGA, in other to better understand the mechanisms involved in the process of neoplastic cell proliferation and tumor progression. METHODS: The expression of growth factors FGF-2, PDGF-A, PDGF-B and receptors FGFR-1, FGFR-2, PDGFR-α, and EGFR was analyzed in 24 PLGA samples in comparison with normal salivary glands, by immunohistochemistry. A semi-quantitative analysis determined cell positivity in all stained sections. Scores were assigned according to percentage of reactive cells: score 0 < 10%; score 1-10 to 25%; score 2-25% to 50%; score 3->50%. The level of significance was set at 5%. RESULTS: Most of the growth factors and receptors, apart from FGFR-2, were significantly reactive in PLGA. Comparing to salivary acini, all of the reactive growth factors and receptors were significantly stronger in PLGA. Comparing to salivary ducts, the expression of FGF-2, PDGF-B, FGFR-1, and EGFR was significantly stronger in the nuclei and/or cytoplasm of the neoplastic cells. CONCLUSIONS: The increased expression of the growth factors and receptors in the PLGA, compared to normal salivary glands, may be related to cell proliferation, somehow participating in the oncogenic process.

Tooth alterations in areas of bisphosphonate-induced osteonecrosis.

OBJECTIVE: Osteonecrosis of the jaw is a potential side effect when using bisphosphonates. Most studies on the effects of bisphosphonates on teeth have been conducted in vitro or in animal models of tooth development. Therefore, the aim of this study was to describe alterations found in human teeth extracted from areas of bisphosphonate-induced osteonecrosis. MATERIALS AND METHODS: Using a retrospective study design, 16 teeth from 13 patients were extracted from areas of bisphosphonate-induced osteonecrosis during surgical debridement. The specimens were decalcified and embedded in paraffin. A series of 5-µm sections were prepared, stained with hematoxylin and eosin (H&E) and observed under a light microscope. RESULTS: The majority of the patients were female (53.85 %), with a mean age of 60.23 ± 13.18 years. Zoledronate (IV) was the most common bisphosphonate used (92.3 %), over a mean period of 2 years. The commonest alteration observed was hypercementosis (87.5 %), followed by pulpar necrosis (81.25 %), pulp stones attached to the dentine and loose pulp stones in the pulp chamber and root canals in addition to linear calcifications (68.75 %), dentinoid/osteoid material formation (18.75 %), and dental ankylosis (6.25 %). CONCLUSIONS: Patients undergoing bisphosphonate therapy present diverse tooth alterations, which should be closely monitored by clinicians to prevent complications. CLINICAL RELEVANCE: It is paramount that the teeth involved in oral lesions are always examined. Attention should be drawn to the need to establish preventive measures, in terms of dental treatment, for patients prior to starting bisphosphonate therapy.

Peripheral odontogenic fibroma: an uncommonly overviewed lesion.

Peripheral odontogenic fibroma is considered a gingival tumor characterized by a proliferation of relatively cellular fibrous or fibromyxomatous connective tissue which exhibits variable amounts of odontogenic epithelium and sometimes foci of calcification in the form of dentinoid, cementicles, or bone. It is considered the extraosseous counterpart of central odontogenic fibroma. This lesion usually is presented as a focal swelling in the gingiva, occurring in a wide age range, and the anterior region of the gingiva is the most frequent anatomic site. Conservative local excision is the treatment frequently adopted and its recurrence rate varies widely, and its biologic behavior is still unknown. In this study, the authors discuss 3 cases of peripheral odontogenic fibroma, and present their clinical and histopathological features and management.

Oral foregut cyst in a neonate.

Oral foregut cysts are congenital choristomas that arise in the oral cavity during embryonic development from remnants of foregut-derived epithelium. This is an unusual report of a neonate with a large congenital sublingual cystic lesion, extending superficially from the left ventral tongue to the anterior floor of the mouth, impeding breast-feeding. The differential diagnosis included dermoid cyst, epidermoid cyst, mucous retention cyst, and oral lymphangioma. The treatment of choice was enucleation under general anesthesia. Histology showed a cystic lesion with a ciliated pseudostratified columnar epithelium with numerous goblet cells. Immunohistochemistry was positive for cytokeratin 7 and thyroid transcription factor 1 and negative for cytokeratin 20, resulting in a final diagnosis of an oral foregut cyst. Three weeks after surgery, the tongue had healed with good mobility, and breast-feeding could be established. No recurrence was present at 6 months of follow-up.

Central Mucoepidermoid Carcinoma Radiographically Mimicking an Odontogenic Lesion.

Central mucoepidermoid carcinoma (CMEC) is a rare pathological entity with only a few case reports in the literature. The present case reported an uncommon occurrence of CMEC mimicking an odontogenic lesion in a young patient. A 17-year-old female patient sought dental care due to a slight swelling located in the posterior region of the mandible on the left side. Radiographic exams revealed an osteolytic lesion with defined limits in relation to proximity to the pericoronal follicle of tooth #38. The clinical and radiographic diagnostic hypothesis was an odontogenic lesion. Histological sections showed the presence of a neoplasm of glandular origin, not encapsulated, with a predominantly cystic growth pattern. The neoplasm consisted of mucous, intermediate, and squamous cells. In the immunohistochemical staining, the neoplastic cells were positive for cytokeratin 7. Mucous cells were positive for PAS with diastase digestion. The final diagnosis consisted of mucoepidermoid carcinoma. The tumor was removed surgically, and the patient has shown no signs of relapse nor recurrence. In conclusion, CMEC may mimic radiographic features of various pathologies, but despite its rarity, clinicians and oral radiologists should consider CMEC as a diagnostic hypothesis for jaw lesions.

In vitro evaluation of the suppressor potential of conditioned medium from benign myoepithelial cells from pleomorphic adenoma in malignant cell invasion.

Tumoral invasion process is the result of a complex interaction between the tumor cells and microenvironment which plays an important role in modulating the growth and invasion of the cancer. The myoepithelial cells, present in glandular organs such as the breast and salivary glands, seem to exert paracrine effects on the glandular epithelium, acting as natural tumor suppressors. To verify the influence of the benign myoepithelial cells in the invasion of malignant cells, simulating an in situ carcinoma ex pleomorphic adenoma, we have cultured three different high-potential invasive malignant tumors (breast ductal adenocarcinoma, melanoma and oral squamous cell carcinoma) in conditioned medium of myoepithelial cells from salivary gland pleomorphic adenomas using transwell chambers with 8-µm pores membrane coated with matrigel. After 96 h, quantitative analyses of the results were performed by calculating the invasion index (number of cells that invaded in relation to the total number of cells). The results showed that there was a reduction of the invasion index mean for the three different malignant tumors. This study supports a tumoral suppressor function of the myoepithelial cells from pleomorphic adenoma in in vitro invasion process.

Histological and mast cells evaluation after application of botulinum toxin type a in masseter muscles of rats.

Background: Botulinum Toxin Type A (BTX-A) has been largely used to reduce muscle strength of masseter and temporal muscles by producing a temporary weakening of their activity. This study aimed to evaluate the histological changes and the number of mast cells after the injection of BTX-A. Material and Methods: In the masseter muscle of rats in the periods of 1, 7, 15, and 30 days. These muscles were stained with hematoxylin and eosin (H&E) and toluidine blue (TBO). The presence or absence of an inflammatory process and necrosis were analyzed by H&E in all area of the slide at 10X magnification. The number of mast cells was evaluated by counting 10 "hotspots" in the intra-muscular region on TBO-stained slides, 400X magnification. Statistical analysis was performed through two-way analysis of variance and Tukey's test. Results: As a result, the inflammatory process and necrosis were not observed in any periods studied in both groups Regarding mast cells, there was no statistically significant increase in their quantity in the study group when compared to the control group in the evaluation periods of 7 days and 15 days. However, these mast cells increased significantly during the periods of 1 and 30 days. Conclusions: This study showed that even in the absence of an inflammatory process, there was an increase in the number of mast cells in the first 24 hours after the application of BTX-A, with a subsequent balance between the numbers of mast cells at 7 and 15 days, and again an increase after 30 days. Key words:Botulinum toxins type A, mast cells, masseter muscle.

Increased mucin 1 expression in recurrence and malignant transformation of salivary gland pleomorphic adenoma.

AIMS: Pleomorphic adenoma (PA) is the most common salivary gland tumour with a tendency to recur (RPA) and a risk of malignant transformation. Mucin 1 (MUC-1) plays a role in the progression of many tumours and may be a marker to predict RPA. The aim of this study was to evaluate MUC-1 expression in different phases of the adenoma to carcinoma sequence. METHODS AND RESULTS: Twenty-one cases of PA, 18 cases of RPA, three cases of RPA with focal transformation (TRPA) and 11 cases of carcinoma ex-pleomorphic adenoma (CXPA) were analysed immunohistochemically for MUC1 expression using an antibody to MUC1/DF3. MUC1 reactivity in RPA was stronger than that observed in PA and, in all the different carcinoma groups, MUC-1 expression was significantly higher in carcinoma than in RPA and PA. CONCLUSION: This study has confirmed that MUC-1 is related to the recurrence of PA and that this molecule is associated with malignant transformation of PA with carcinoma cells overexpressing MUC-1.

Study of histopathological, morphological and immunohistochemical features of recurrent pleomorphic adenoma: an attempt to predict recurrence of pleomorphic adenoma.

BACKGROUND: Recurrent pleomorphic adenoma (RPA) is an uncommon and challenging disease. The aim of this study was to review the clinical information obtained by examining RPA patients, to determine its histomorphological features and to verify the expression of the Mcm-2 markers of cell proliferation and the expression of p-53 in pleomorphic adenoma (PA), RPA, and RPA with malignant transformation (TRPA). METHODS: A total of 10 cases of PA and 29 cases of RPA were examined in detail for the presence of nodules and their histomorphological features. Cell proliferation and expression of p-53 were detected by the immunohistochemical technique using the antibodies against Mcm-2 and p-53. RESULTS: Histopathologically, RPA is very similar to PA; the only difference found was that all the cases of RPA were multinodular. When comparing primary and recurrent tumor, no significant difference was found in terms of cell proliferation and the expression of p-53; however, in the RPA with areas of malignant transformation there was an increased expression of these proteins. CONCLUSION: This study showed that recurrences were multinodular, with nodules varying in numbers and sizes. No significant difference in histological features was found between RPA and PA. Moreover, the immunohistochemical study showed a low expression of p-53 and Mcm-2 in PA and RPA and an increased expression of these proteins in the RPA with areas of malignant transformation.

Recurrent oral pyogenic granuloma in port-wine stain.

Pyogenic granuloma (PG) is a benign inflammatory lesion, nonneoplastic in nature, which occurs in the oral cavity and skin. This lesion arises in response to various stimuli such as low-grade local irritations, traumatic injury, or hormonal factors. Recently, in some cases, the occurrence of recurrent PGs in skin associated with vascular lesions, such as port-wine stains, has been described. It has been postulated that this association is promoted by arteriovenous anastomoses in the vascular lesions, leading to the development of PG. The authors discuss 2 cases of recurrent PG in patients with a port-wine stain, and the treatment options adopted.

Salivary immunoglobulin A responses in professional top-level futsal players.

The purpose of this study was to investigate the responses of salivary immunoglobulin A (SIgA) in 10 professional top-level Brazilian futsal players after 2 highly competitive games separated by 7 days. Unstimulated saliva was collected over a 5-minute period at PRE- and POST-match. The SIgA was measured by an enzyme-linked immunosorbent assay and expressed as the absolute concentration (SIgAabs) and secretion rate of IgA (SIgArate). Rate of perceived exertion and heart rate were used to monitor the exercise intensity. A 2-way analysis of variance with repeated measures showed nonsignificant differences between matches to SIgAabs, SIgArate, and saliva flow rate (p > 0.05). However, significant time differences were observed for all these parameters. In summary, we showed that a competitive training match induced a decrease in SIgA levels in top-level futsal players, which suggests an increment of the vulnerability to infections meditated by the training stimulus. This decrease suggests that the athletes were at an increased risk of developing an upper respiratory tract infection, and therefore, it could be necessary to take protective actions to minimize contact with cold viruses or even reduce the training load for athletes.