IL-10 and TGF-beta1 gene polymorphismsin Greek patients with recurrent aphthous stomatitis

Background: Recurrent aphthous stomatitis (RAS) is one of the most frequent inflammatory disorders of theoral mucosa. Cytokines, which play an important role in RAS pathogenesis, participate directly or indirectly innormal, immunological and inflammatory processes and are secreted from cells belonging to innate and adaptiveimmunity as a consequence of microbial and antigenic stimuli. Gene polymorphisms in specific cytokines maypredispose to RAS development. The aim of this study was the investigation and association of IL-10 and TGF-β1gene polymorphisms with RAS.Material and Methods: Study’s cohort consisted of 60 Greek patients diagnosed with RAS, including 40 patientswith minor, 10 patients with major and 10 with herpetiform aphthous ulcers. Forty age- and sex-matched controlsubjects were included in this study. DNA was extracted from whole blood samples of all patients and sequencespecific primers (SSP)-based polymerase chain reaction (PCR) was used for genotyping. Gene polymorphisms forcytokines IL-10 at loci -592 and -819 and for TGF-β1 at codon 10 were detected.Results: Significant differences between patients with minor RAS and healthy controls were recorded for IL-10genotypes distribution at position -592 (p=0.042) and -819 (p=0.045) with predominance of C/A and C/T genotypes in RAS patients, respectively. Also, in patients with minor and herpetiform aphthous ulcerations, heterozygousTGF-β1 genotype C/T at codon 10 was associated with increased risk of RAS (p=0.044 and p=0.020, respectively).Conclusions: These data provide evidence that genetic predisposition for RAS and possibly its specific clinical variants is related with the presence of gene polymorphisms for specific cytokines, including IL-10 and TGF-β1, which,in turn, may vary according to geographic origin and genetic background. (AU)

Salivary mRNA markers having the potential to detect oral squamous cell carcinoma segregated from oral leukoplakia with dysplasia.

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.

Bilateral masseter and internal pterygoid muscle hypertrophy: a diagnostic challenge.

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.

Expression of matrix metalloproteinases 9 and 12 in actinic cheilitis.

AIM: To investigate the role of matrix-degrading metalloproteinases 9, 12 (MMPs), as mediators of functional connective tissue damage in actinic cheilitis. METHODS: Thirty 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. RESULTS: Haematoxylin 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). CONCLUSION: Our 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.

Current aspects on oral squamous cell carcinoma.

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.

Role of human papillomavirus in the pathogenesis of oral squamous cell carcinoma.

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.

Lip ulceration associated with intravenous administration of zoledronic acid: report of a case.

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 H(2)O(2) (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 in chronic sclerosing sialadenitis.

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.

Recovering circulating extracellular or cell-free RNA from bodily fluids.

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.

Salivary markers for oral cancer detection.

Oral cancer refers to all malignancies that arise in the oral cavity, lips and pharynx, with 90% of all oral cancers being oral squamous cell carcinoma. Despite the recent treatment advances, oral cancer is reported as having one of the highest mortality ratios amongst other malignancies and this can much be attributed to the late diagnosis of the disease. Saliva has long been tested as a valuable tool for drug monitoring and the diagnosis systemic diseases among which oral cancer. The new emerging technologies in molecular biology have enabled the discovery of new molecular markers (DNA, RNA and protein markers) for oral cancer diagnosis and surveillance which are discussed in the current review.

Heat shock protein 70 membrane expression in oral cancer: a possible new target in antineoplastic therapy?

BACKGROUND: Heat shock proteins (HSPs) are a group of highly immunogenic proteins with an exceptional degree of conservation. Cells or tissues from a wide range of tumors have been shown to express atypical levels of 1 or more HSPs. Experimental evidence suggests that HSPs may promote tumorigenesis by suppressing apoptosis. AIM: The aim of this investigation was to study the expression and the localization of HSP70 in oral squamous cell carcinoma (OSCC) and to compare HSP70 expression between neoplastic, dysplastic lesions and normal oral tissues. METHODS: Immunohistochemistry staining was conducted to study HSP70 expression in 50 paraffinized tissue samples: 30 OSCCs, 10 leukoplakias with dysplasia and 10 samples from normal oral tissue. RESULTS: Our results showed that all slides with OSCC intensely expressed HSP70. Dysplastic lesions were positive to a lesser extent for HSP70. Samples from normal oral tissue were all negative for HSP70. CONCLUSIONS: It is concluded that increased HSP70 immunoexpression could be a marker for the presence of epithelial dysplasia or epithelial malignant transformation.

Detection, enumeration and characterization of T helper cells secreting type 1 and type 2 cytokines in patients with recurrent aphthous stomatitis.

One of the factors involved in the pathogenesis of recurrent aphthous stomatitis (RAS) is a cell-mediated immune response in which several cytokines seem to play a major role. The aim of this study was to detect, enumerate and characterize T helper cells which are secreting type 1 cytokines (interleukin [IL]-2, IL-12, interferon [IFN]-gamma, and tumor necrosis factor [TNF]-alpha) and type 2 cytokines (IL-4, IL-5, IL-6, and IL-10) in the peripheral blood of patients with RAS. Thirty-two patients in the active phase of RAS (14 men and 18 women) and 40 healthy individuals participated in the study. T helper (T) cells were detected and characterized using Elispot assay. T cells secreting IL-2, IL-12 or IFN-gamma were increased in patients with RAS compared with the controls (p < 0.05, p < 0.001 and p < 0.001, respectively). T cells secreting TNF-alpha in RAS patients and controls were not statistically different (p > 0.05). T cells secreting IL-10 were increased in patients with RAS compared with the controls (p < 0.05). T cells secreting IL-4 were decreased in patients with RAS compared with the controls (p < 0.001), No statistical difference was observed between T cells secreting IL-5 or IL-6 in patients with RAS and controls. Our findings suggest that the increased numbers of T cells secreting type 1 cytokines may influence the immune response against RAS. Whether this action is of etiological importance or epigenetic phenomenon is a question that needs to be answered.

Epidemiologic survey of pemphigus vulgaris with oral manifestations in northern Greece: retrospective study of 129 patients.

OBJECTIVE: To evaluate the epidemiology of pemphigus vulgaris (PV) in a Greek population and to compare it with other countries. MATERIALS AND METHODS: A retrospective study was conducted based on the records of 129 patients (41 males and 88 females) with PV who visited the Department of the Oral Medicine and Maxillofacial Pathology, Aristotle University of Thessaloniki, Greece and the State Hospital for Skin and Venereal Diseases of Thessaloniki, Greece, between 1985 and 2004. A group of 73 individuals was used as controls. RESULTS: The average annual incidence was found to be eight patients per year. The male to female ratio was 1 : 2.25. The difference in the age of onset between the two genders was statistically significant in marginal levels (P = 0.05). In addition, 86.1% of the patients showed oral lesions only, 13.3% oral and skin lesions and 1.3% manifested oral, skin, and ocular lesions. Twenty-eight of the 88 females were in the premenopausal period-of-life. Additionally, 19 males were farmers who had daily contact with organophosphoric pesticides. Co-existing pathologic conditions were present in 75 of the 129 patients, and of these 75 patients six (8%) were diabetics, 15 (20%) presented with hypertension, two (2.6%) faced problems from their thyroid gland, and 10 (13.3%) of the patients complained of allergic reactions. CONCLUSIONS: The results of this study demonstrated a relatively high incidence of PV in northern Greece compared with that in other countries. The disease most frequently occurred in the sixth decade-of-life and the majority of the patients manifested oral lesions. Further epidemiological studies are needed to elucidate whether this region is constituted from population groups with high susceptibility to PV.

Pemphigus vegetans of the oral cavity.

BACKGROUND: Pemphigus vegetans, a variant of pemphigus vulgaris, constitutes a rare form of all pemphigus cases, and oral involvement is common. Two clinical subtypes of pemphigus vegetans exist, characterized initially by flaccid bullae and erosions (Neumann) or pustules (Hallopeau). Both subtypes subsequently develop into hyperpigmented vegetative plaques with pustules and hypertrophic granulation tissue at the periphery. METHODS: We report three cases of pemphigus vegetans with oral manifestations exclusively. Two patients were male aged 30 and 45 years old, respectively, while one was a 51-year-old female. CONCLUSION: Oral lesions in all cases consisted of erosions and whitish, vegetating plaques. The histopathological characteristics were in all cases identical. The spinous cell layer was characterized by intense acanthosis and by the presence of vesicles between the spinous and basal cell layers. Inside the vesicles exudative elements were observed consisting mainly of eosinophils. In the upper lamina propria severe inflammatory reaction was observed. Streptavidin-biotin immunoperoxidase technique showed in all cases intercellular epithelial deposition of IgG and C3.

Ulcerative uremic stomatitis associated with untreated chronic renal failure: report of a case and review of the literature.

Uremic stomatitis represents a relatively uncommon intraoral complication seen, mostly, in cases of end-stage renal disease or undiagnosed/untreated chronic renal failure. Its incidence has decreased due to the advent of renal dialysis. Clinically uremic stomatitis is characterized by the presence of painful plaques and crusts that are usually distributed on the buccal mucosa, dorsal or ventral surface of the tongue, gingiva, lips, and floor of the mouth. Treatment consists of improvement of urea blood concentration and the underlying renal failure, supported by increased oral hygiene with antiseptic mouthwashes and antimicrobial/antifungal agents if necessary. Although uremic stomatitis occurs in patients with end-stage renal disease, we report a case of a patient who exhibited an ulcerative form of uremic stomatitis related to the sudden relapse of uremia, although not in an advanced stage of her renal disease. A description of the clinical and microscopic appearance is given along with our hypothesis for the pathogenesis of the disease.