Background: Recurrent aphthous stomatitis (RAS) is one of the most frequent inflammatory disorders of the oral mucosa. Cytokines, which play an important role in RAS pathogenesis, participate directly or indirectly in normal, immunological and inflammatory processes and are secreted from cells belonging to innate and adaptive immunity as a consequence of microbial and antigenic stimuli. Gene polymorphisms in specific cytokines may predispose to RAS development. The aim of this study was the investigation and association of IL-10 and TGF-beta 1 gene polymorphisms with RAS.Material and Methods: Study's cohort consisted of 60 Greek patients diagnosed with RAS, including 40 patients with minor, 10 patients with major and 10 with herpetiform aphthous ulcers. Forty age-and sex-matched control subjects were included in this study. DNA was extracted from whole blood samples of all patients and sequence -specific primers (SSP)-based polymerase chain reaction (PCR) was used for genotyping. Gene polymorphisms for cytokines IL-10 at loci-592 and-819 and for TGF-beta 1 at codon 10 were detected. Results: Significant differences between patients with minor RAS and healthy controls were recorded for IL-10 genotypes distribution at position-592 (p=0.042) and-819 (p=0.045) with predominance of C/A and C/T geno-types in RAS patients, respectively. Also, in patients with minor and herpetiform aphthous ulcerations, heterozygous TGF-beta 1 genotype C/T at codon 10 was associated with increased risk of RAS (p= 0.044 andp= 0.020, respectively). Conclusions: These data provide evidence that genetic predisposition for RAS and possibly its specific clinical vari-ants is related with the presence of gene polymorphisms for specific cytokines, including IL-10 and TGF-beta 1, which, in turn, may vary according to geographic origin and genetic background.

Background: In the current study the presence of extracellular IL-1B, IL-8, OAZ and SAT mRNAs in the saliva was evaluated as a tool in the early detection of oral squamous cell carcinoma.Methods: 34 patients with primary oral squamous cell carcinoma stage T1N0M0/T2N0M0, 20 patients with oral leukoplakia and dysplasia (15 patients with mild dysplasia and 5 with severe dysplasia/in situ carcinoma) and 31 matched healthy-control subjects were included in the study. The presence of IL-1B, IL-8, OAZ and SAT mRNA was evaluated in extracellular RNA isolated from saliva samples using sequence-specific primers and real-time RT-PCR. ROC curve analysis was used to estimate the ability of the biomarkers to detect oral squamous cell carcinoma patients.Results: The data reveal that the combination of these four biomarkers provides a good predictive probability of up to 80% (AUC = 0.799, p = 0.002) for patients with oral squamous cell carcinoma but not patients suffering from oral leukoplakia with dysplasia. Moreover, the combination of only the two biomarkers (SAT and IL-8) also raises a high predictive ability of 75.5% (AUC = 0.755, p = 0.007) approximately equal to the four biomarkers suggesting the use of the two biomarkers only in the prediction model for oral squamous cell carcinoma patients limiting the economic and health cost in half.Conclusion: SAT and IL-8 mRNAs are present in the saliva in high quality and quantity, with a good discriminatory ability for oral squamous cell carcinoma patients only but not for patients with oral leukoplakia and dysplasia an oral potentially malignant disorder. (C) 2016 Published by Elsevier Ltd.

Several topical and systemic factors have been reported to influence the eruption of teeth. Some of the local lesions include eruption cysts, eruption sequestra, fibrous developmental malformations and dentigerous cysts. The systemic factors include Down’s syndrome, cleidocranial dysostosis, hypothyroidism, hypopituitarism and achondroplastic dwarfism. All these lesions and factors generally influence the eruption of the primary, as well as the permanent dentition. The purpose of this review article is to present up-to-date aspects of these conditions.

Objective: To describe an unusual case of bilateral masseter and pterygoid muscle hypertrophy. Clinical Presentation and Intervention: A 53-year-old female patient presented with a bilateral, painless swelling at the parotid areas without improvement after using antibiotics/systemic corticosteroids/nonsteroidal anti-inflammatory agents. Her medical history included thyroid nodules, but no dental/occlusal disorders were observed. The initial differential diagnosis included salivary gland/jaw bone/masseter pathology, but the CT/MRI revealed only an increase in the size of the masseter and pterygoid muscles. The patient was informed of the benign nature of the swelling and was advised to discontinue the use of nonsteroidal anti-inflammatory agents. Conclusion: The bilateral hypertrophy of masseter muscles should be considered in differential diagnosis in cases of unilateral or bilateral swelling of the parotid or lateral mandible area.

Oral squamous cell carcinomas (OSCCs) are the most frequent malignancy of the oral cavity, representing 90%-95% malignancies in the oral region. The development of OSCC is a multi-step process requiring the accumulation of multiple genetic and epigenetic alterations, influenced by the patient's genetic predisposition and environmental factors. Cell-cycle regulation is crucial for tumorigenesis. This control in eukaryotic cells involves the sequential activation of cyclins, cyclin-dependent kinases (CDKs), and two great CDK inhibitors (CDK-Is): the INK4 family, comprised by inhibitors p16, p15, p18, and p19, and the Cip/Kip family, comprised by p27, p57, and p21. The inactivation of p16(INK4a) and p21(Waf1/CIP1) has been widely associated with this type of tumors. The OSCC is a characteristic locally aggressive tumor that presents large areas of tumoral necrosis, in which the levels of acidity and hypoxia are very high, causing low response to chemotherapy. Hypoxia-inducible factors (HIFs) include hypoxia-inducible factor 1 alpha (HIF-1 alpha), hypoxia-inducible factor 1 beta (HIF-1 beta), hypoxia-inducible factor 2 alpha (HIF-2 alpha), and hypoxia-inducible factor 3 alpha (HIF-3 alpha). HIF-1 alpha overexpression has been associated with tumor cell growth and survival of these in head and neck tumors. Carbonic anhydrases (CAs), mainly CA-IX expression, have been thoroughly described in different tumors, including cervical carcinoma, lung, bladder, breast, esophagus, and colorectal cancers; however, this has not been so for head and neck carcinomas. Tumor development and progression are complex processes involving oncogenes, tumor suppressor genes, and tumoral microenvironment, through their contact with malignant cells, surrounding stromal cells (fibroblasts, endothelial cells, and inflammatory cells), and the extracellular matrix (ECM), formed by structural proteins (such as collagen and elastin), specialized proteins (such as fibrillin, fibronectin, and laminin), and proteoglycans. There are several proteases responsible for remodeling the ECM that enable dissemination and metastasis of tumor cells, such as serine proteases, cysteine cathepsins, and matrix metalloproteinases (MMPs). There is a group of specific and strong MMPs called endogenous tissue inhibitors of metalloproteinases (TIMPs), and although the expression of MMPs has been largely described in almost all the tumors of the body, the expression of TIMPs has been studied in a lesser degree. Altered mRNA transcripts are also a promising field for OSCC detection. Therefore, the goal of this chapter is to describe the expression of p16(INK4a); p21(Waf1/CIP1); HIFs; CA-IX; TIMPs; IL-8 and IL-1B; DUSP1; H3 histone, family 3A (H3F3A); ornithine decarboxylase antizyme 1 (OAZ1); S100 calcium binding protein P (S100P); and spermidine/spermine N1-acetyltransferase1 (SAT1) in OSCC, determining their relation with clinical, histological, and prognostic factors regarding the different stages of the evolution of the tumors, looking for their detection through noninvasive techniques as exfoliative cytology.

Background: Angiokeratoma is an asymptomatic, hyperkeratotic, capillary disorder of the skin present as solitary or multiple, keratotic papules or plaques, which may also be related to Fabry disease. Oral involvement may be observed in cases of widespread muco-cutaneous angiokeratomas, whereas solitary buccal angiokeratoma without systemic/ cutaneous involvement is extremely rare. Case Report: A 45-year-old woman was referred with a 3-month, painless, bluish lesion, located on left buccal mucosa. The medical record of the patient was free of any systemic disease or medication. After a careful clinical oral, mucosal as well as skin examination, an excisional biopsy was taken. A mutine haematoxylin-eosin staining and additional immunohistochemistry were performed. Differential diagnosis included haemangioma, haematoma or lesions of melanocytic origin. Clinical examination showed a solid, lobulated bluish lesion, located on left buccal mucosa without other skin or mucosal involvement. The microscopic findings revealed dilated vascular spaces covered by normal endothelium without atypia, extending into the epithelium, indicating the diagnosis of angiokeratoma. Conclusions: Despite its rare occurrence, solitary angiokeratoma of oral mucosa should be included in the differential diagnosis of black-bluish lesions. Further investigation for other similar lesions throughout skin or mucosa is needed to avoid complications as haemorrhage.

Usually increased presence of Heat Shock Proteins (HSPs) is considered to mediate inflammation process in Oral Lichen Planus (OLP) lesions, but in contrast HSP-70 expression found to be stable or even decreased in those lesions. The purpose of this study was to detect the serum titres of the HSP-70 in patients with OLP compared to healthy individuals, which may indicate an alternative, systemic role. Serum levels of HSP- 70 were detected by sandwich-ELISA in 45 patients with reticular (n=28) and erosive (n=17) OLP, respectively. A group of 35 healthy individuals was used as control. HSP-70 was detected in significantly increased levels in OLP (p<0.05) compared to controls. The increase was prominent in reticular-OLP (p<0.05), whereas no difference was observed between serum HSP-70 in erosive OLP compared to controls. These results indicate a systemic initiation of the immune response in the pathogenesis and process of OLP. The higher titres of HSP-70 in reticular but not erosive form of OLP indicate rather an immunoregulatory role in chronicity than in the acute inflammatory process of OLP. Consequently, the evaluation of serum imbalances of HSP-70 in OLP using ELISA may be a useful marker for disease's monitoring and/or efficacy of systemic treatment.

AIMTo investigate the role of matrix-degrading metalloproteinases 9, 12 (MMPs), as mediators of functional connective tissue damage in actinic cheilitis.METHODSThirty five formalin-fixed, paraffin embedded specimens of actinic cheilitis, and twelve specimens of normal lower lip vermillion, which were obtained by the archives of the Department of Oral Medicine and Maxillofacial Pathology, were examined. From each block, 5 μm thick sections were cut and routinely stained with Hematoxylin and Eosin. Immunohistochemical studies were performed on 4-μm thick sections of formalin-fixed paraffin embedded actinic cheilitis lesions and of normal lower lip vermillion, for MMP-9 and MMP-12 in serial sections of our specimens. Appropriate positive and negative controls were performed to confirm the specificity of the staining reaction. MMP immunohistochemistry was evaluated using a semiquantitative immunoreactive score.RESULTSHaematoxylin and eosin staining revealed in actinic cheilitis lesions atrophic stratified squamous cell epithelium, or focally and irregularly hyperplastic of variable thickness, in some areas was observed marked keratin production. Varying degrees of epithelial dysplasia were noticed with a wide spectrum of change within the same specimen. Characteristic was the appearance of chronic inflammatory infiltration, and a band of amorphous acellular, basophilic change like solar elastosis (elastin replacement of collagen). In normal lower lip specimens weak and scanty positive expression of MMP-9 and MMP-12 was observed. Anti-MMP-9 antibody showed a weak reaction, in actinic cheilitis lesions, focal in the elastotic material, in chronic inflammatory cells and mostly in macrophages and neutrophils. Strong and in some cases diffused immunohistochemical expression of MMP-12 was detected in actinic cheilitis lesions in the areas of the fragmented, distorted and thickened elastic fibers. MMP-12 was also expressed in chronic inflammatory cells and mostly macrophages. MMP-12 was significantly higher in actinic cheilitis specimens compared with the normal lower lip specimens (P = 0.0029).CONCLUSIONOur results suggest that especially MMP-12 may play an important role in remodeling events occurring in the connective tissue during long-term exposure to sunlight in the actinic cheilitis lesions.

Objectives Cells with stem/progenitor properties have been detected in major salivary glands, but no data are available on their presence within minor salivary glands (MSGs). This study aimed to isolate and characterize potential stem/progenitor cells from human MSGs. Materials and methods MSGs of the lower lip were surgically obtained during biopsy for Sjogren’s syndrome investigation that finally proved to be histologically normal. The established MSG cultures were assessed for morphology, proliferation, colony-forming-unit efficiency, multipotentiality, and immunophenotypic characteristics. Results A mixed population of fibroblast-like and a few flat-shaped epithelial-like cells was obtained. These cells were capable for osteogenic, adipogenic, and neurogenic differentiation. Evidence for strong stem cell potency was observed by the detection of early stem cell markers, like Nanog, Oct-3/4, and SSEA-3. These cells also expressed characteristic mesenchymal stem cell markers, including CD90-Thy1, CD105, CD49f, CD81, nestin, CD146, and Stro-1, but were negative for CD117/C-KIT, CD45, and CD271/NFG. In addition, positivity for keratins 7/8 in part of the population was indicative of an epithelial phenotype, whereas these cells were negative for aquaporin-1 expressed in acinar/myoepithelial cells during development. Conclusions Based on these data, a cell population with stem/progenitor characteristics was primarily isolated from labial MSGs. The morphologic and immunophenotypic features indicated that this population is mixed with mesenchymal (mainly) and epithelial characteristics. Clinical relevance Due to their large number and superficial distribution in labial mucosa, MSGs may be proposed as a potential easily accessible source of adult stem/progenitor cells for regenerative therapies of glandular organs with parenchymal pathology.

Purpose Primary oral melanoma is an uncommon malignant tumor of melanocytic origin, with potentially aggressive capacity. Several immunohistochemical markers influencing invasiveness and metastatic dissemination of melanoma have been identified and used previously. Our aim was to analyze the immunohistochemical expression for cathepsin-D, c-kit, E-cadherin, cyclin D1, and MUM-1 in oral malignant melanomas. Material and Methods Immunoperoxidase staining was performed on formalin-fixed tissue specimens from oral melanoma specimens and oral melanocytic nevi lesions which were used as control group. The percentages of immunoreactive melanocytes were estimated and qualitatively abnormal immunoreactivity patterns were also tabulated. Results Cathepsin-D, c-kit, cyclin D1, and MUM-1 were highly expressed in oral melanomas, whereas negative or very weak expression of the above mentioned markers was recorded in oral melanocytic nevi lesions. E-cadherin was expressed uniform throughout the entire oral melanocytic nevi, whereas oral melanomas expressed heterogeneous E-cadherin staining. Furthermore oral melanomas showed qualitatively and quantitavely abnormal E-cadherin immunoreactivity, with gradual loss of staining and increasing lesional depth. The results indicate that cathepsin-D, c-kit, E-cadherin, cyclin D1, and MUM-1 may be involved in the development of oral melanomas, and eventually they may be useful in the differential diagnosis of oral malignant melanomas and oral melanocytic nevi lesions. Conclusions The above mentioned markers seems to correlate with oral melanoma development and have an important functional role in tumor development or progression. Melanoma has a wide spectrum of histologic features which mimic epithelial, hematologic, mesenchymal, and neural tumors. Immunohistochemistry has been the primary tool to distinguish melanomas from these other tumors. Use of immunohistochemical markers should therefore give additional information which cannot be determined by routine histopathology.

Oral cancer is one of the most common cancers and it constitutes a major health problem particularly in developing countries. Oral squamous cell carcinoma (OSCC) represents the most frequent of all oral neoplasms. Several risk factors have been well characterized to be associated with OSCC with substantial evidences. While tobacco and alcohol are the primary risk factors for OSCC development, many epidemiological studies report a strong association with human papillomavirus (HPV) in a subset of OSCC. This article presents our current knowledge on the relationship between HPV and development of OSCC. HPVs are DNA viruses that specifically target the basal cells of the epithelial mucosa. Most experimental data are consistent with the hypothesis that HPV plays a causal role in oral carcinogenesis. Genotypes, such as HPV1 infect epidermal cells, whereas HPV6, 11, 16 and 18 infect epithelial cells of the oral cavity and other mucosal surfaces. Several studies have shown that there is an increased risk of head and neck cancer in the two major HPV 16 oncogenes E6 and E7 -positive patients. The presence of antibodies to HPV E6 and E7 proteins was found to be more associated with tumors of the oro-pharynx than of the oral cavity. However, HPV alone appears to be insufficient as the cause of OSCC but requires other co-factors. Although a viral association within a subset of OSCC has been shown, the molecular and histopathological characteristics of these tumors have yet to be clearly defined.

Vascular endothelial growth factor (VEGF) is an angiogenic cytokine and mast cells play a role in neoangiogenesis in various malignancies. The aim of the present study was to elucidate the role of VEGF and mast cells in the early stages of tumorigenesis in oral squamous cell carcinoma (OSCC). Immunohistochemistry was conducted to study VEGF expression and microvessel density (MVD) in 49 tissue samples, 31 OSCCs, 13 leukoplakias (8 with and 5 without dysplasia) and 5 samples from normal oral tissue. Counterstaining with tolouidine blue was conducted to reveal mast cells. The number of microvessels and mast cells were counted at the same optical field. A gradually increased VEGF expression was observed from normal oral epithelium to leukoplakia and OSCC. MVD was found to increase significantly between normal oral tissue and OSCC (p=0.000). The number of mast cells was found to increase significantly between normal oral tissue, dysplasia (p=0.012) and OSCC (p=0.000). In the early stages of tumorigenesis in OSCC, VEGF, which is secreted by the epithelium, is gradually increased immediately affecting the population of mast cells, which are then related to the increase of microvessels.

Oral squamous cell carcinoma is the most common malignant epithelial neoplasm affecting the oral cavity. This article overviews the essential points of oral squamous cell carcinoma, highlighting its risk and genomic factors, the potential malignant disorders and the therapeutic approaches. It also emphasizes the importance of the early diagnosis.

Objective: To detect immunohistochemically the N-cadherin expression in different types of benign and malignant salivary gland tumors in an attempt to note any possible correlation to their development, stage and invasive properties. Materials and Methods: N-cadherin expression was examined in tissue specimens from 49 salivary gland tumors including: pleomorphic adenomas (4), Warthin’s tumors (10), and myoepitheliomas (4) (benign tumors), as well as adenoid cystic carcinomas (14), mucoepidermoid carcinomas (4), polymorphous low-grade adenocarcinomas (6), and adenocarcinomas not otherwise specified (5) (malignant tumors). Twelve specimens of normal salivary glands were used as control. The perineural invasion and stage of malignant salivary gland tumors were evaluated. Immunohistochemical procedure was performed automatically using the Bond Polymer Refine Detection Kit. Results: N-cadherin expression was not found in normal salivary glands. In benign salivary gland tumors, N-cadherin along membranes of neoplastic cells as well as in centrocytes of lymphoid germinal centers was seen in 1 and 4 cases of Warthin’s tumors, respectively. Varied degree of N-cadherin expression was found in 13 (45%) cases of malignant salivary gland tumors. N-cadherin expression was significantly correlated with perineural invasion (χ2 = 11.7, p < 0.0001), but not with stage of malignant salivary gland tumors. Conclusion: N-cadherin expression was observed in malignant salivary gland tumors and could be an indicator of potentially aggressive behavior. N-cadherin expression by tumor cells could be attributed to perineural invasion.

Although osteonecrosis of the jaw is a well-known adverse reaction of bisphosphonates (BPs), random cases of oral mucosal ulceration after per os administration of BP-aledronate have been attributed to prolonged mucosal irritation. This report, for the first time, describes the mucosal ulceration related to intravenous use of zoledronic acid (ZA). A 52-year-old female patient presented with painful ulcers on both cutaneous/mucosal surfaces of the lower lip and a 2-month history of osteonecrosis of the mandible beside the right lower canine. Her medical record included intravenous administration of ZA for 10 months for primary breast cancer metastatic to bone. Examination of the peripheral blood showed severe anemia and a slightly increased white blood cell count, due to urinary tract infection by E. coli, but no evidence of a viral infection. The treatment of anemia and E. coli infection did not improve the labial ulcers. Biopsy from the mucosal lesion revealed a non-specific ulceration with moderate inflammatory infiltration. There was no evidence of infection or malignancy. ZA administration was discontinued and within 3 months the lesions were resolved after treatment with systemic antibiotics (amoxicillin), vitamins A and E, chlorexidine and H2O2 (hydrogen peroxide) solutions and local pantothenic acid/vitamin A creams. Recurrence was detected a month after ZA re-administration. Nevertheless, after new treatment, the patient was free of oral/skin lesions 18 months later. This case, which is the first report of ulceration associated with intravenous administration of bisphosphonates, suggests that systemic mechanisms may be implicated in BP-induced oral mucosal ulceration. Furthermore, ZA appears to cause the same oral mucosal manifestations as alendronate. This emphasizes the need for oral examination in all cases of BP therapy, whether per os or intravenously administrated.

Tenascin-C is an extracellular matrix glycoprotein that has been implicated in the development of fibrosis in certain chronic inflammatory/sclerosing conditions. This study was undertaken to expand our understanding of the processes involved in fibrosis that occurs in chronic sclerosing sialadenitis (CSS) by investigating the distribution of tenascin-C. Fifteen specimens of CSS with varying degrees of fibrosis and five normal submandibular glands were retrospectively examined immunohistochemically for the distribution of TNC. Linear deposition of TNC was found around collecting ducts in normal glands and around collecting ducts without surrounding fibrotic tissue in CSS; percentage incidences were not statistically different. In contrast, broader, band-like deposition of TNC was found in the fibrous tissue around collecting ducts in CSS with widespread degree of fibrosis compared to little or no fibrosis; the percentage incidence was statistically different. In addition, deposition of TNC was found around duct-like structures and extremely atrophic acini but, interestingly, however, was not found in fibrotic interlobular septa. The results of this investigation suggest that TNC is likely involved in the fibrosis that occurs around collecting ducts in CSS.

The presence of extracellular circulating or cell-free RNA in biological fluids is becoming a promising diagnostic tool for non invasive and cost effective cancer detection. Extracellular RNA or miRNA as biological marker could be used either for the early detection and diagnosis of the disease or as a marker of recurrence patterns and surveillance. In this review article, we refer to the origin of the circulating extracellular RNA, we summarise the data on the biological fluids (serum/plasma, saliva, urine, cerebrospinal fluid and bronchial lavage fluid) of patients suffering from various types of malignancies reported to contain a substantial amount of circulating extracellular (or cell-free) RNAs and we discuss the appropriate reagents and methodologies needed to be employed in order to obtain RNA material of high quality and integrity for the majority of the experimental methods used in RNA expression analysis. Furthermore, we discuss the advantages and disadvantages of the RT-PCR or microarray methodology which are the methods more often employed in procedures of extracellular RNA analysis.