Giovanni Giannatempo

RNA-Mediated Gene Silencing of Nicotinamide N-Methyltransferase Is Associated with Decreased Tumorigenicity in Human Oral Carcinoma Cells

Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer. Despite progress in the treatment of OSCC, overall survival has not improved substantially in the last three decades. Therefore, identification of reliable biomarkers becomes essential to develop effective anti-cancer therapy. In this study, we focused on the enzyme Nicotinamide N-methyltransferase (NNMT), which plays a fundamental role in the biotransformation of many xenobiotics. Although several tumors have been associated with abnormal NNMT expression, its role in cancer cell metabolism remains largely unknown. In this report, 7 human oral cancer cell lines were examined for NNMT expression by Real-Time PCR, Western blot and HPLC-based catalytic assay. Subsequently, we evaluated the in vitro effect of shRNA-mediated silencing of NNMT on cell proliferation. In vivo tumorigenicity of oral cancer cells with stable knockdown of NNMT was assayed by using xenograft models. High expression levels of NNMT were found in PE/CA PJ-15 cells, in keeping with the results of Western blot and catalytic activity assay. PE/CA PJ-15 cell line was stably transfected with shRNA plasmids against NNMT and analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and soft agar Assays. Transfected and control cells were injected into athymic mice in order to evaluate the effect of NNMT silencing on tumor growth. NNMT downregulation resulted in decreased cell proliferation and colony formation ability on soft agar. In athymic mice, NNMT silencing induced a marked reduction in tumour volume. Our results show that the downregulation of NNMT expression in human oral carcinoma cells significantly inhibits cell growth in vitro and tumorigenicity in vivo. All these experimental data seem to suggest that NNMT plays a critical role in the proliferation and tumorigenic capacity of oral cancer cells, and its inhibition could represent a potential molecular approach to the treatment of oral carcinoma.

Orofacial Granulomatosis: Clinical Signs of Different Pathologies

Orofacial granulomatosis (OFG) is an uncommon disease characterized by persistent or recurrent soft tissue enlargement, oral ulceration and a variety of other orofacial features. It could be an oral manifestation of a systemic disease. For a correct differential diagnosis, local and systemic conditions characterized by granulomatous inflammation should be excluded using appropriate clinical and laboratory investigations. In fact, the diagnosis of OFG may be confirmed only by histopathological identification of noncaseating granulomas. The literature from 1943 to 2014 was reviewed with emphasis on the etiology of OFG and on clinical manifestations of systemic pathologies associated with OFG. The precise cause of OFG is still unknown, although several theories have been suggested, such as infection, hereditary factors and allergy. OFG is a disease that has a wide spectrum of presentation, which may include the oral manifestation of a systemic condition such as Crohns disease, sarcoidosis, granulomatosis with polyangiitis and Melkersson-Rosenthal syndrome.

Adverse drug reactions in the oral cavity

Several drugs may have a number of adverse reactions (ADRs) involving the oro-facial region. The dose of the drug and the time required for the reaction to take place are relevant parameters; nonetheless, ADRs mechanisms are not always known and ADRs are not always predictable since aspects other than drug pharmacodynamics and/or pharmacokinetics, as well as various interacting variables contribute to the final outcome. All tissues and many functions of the oral cavity can be affected. In particular, salivary function is frequently involved and hypo-salivation is the main manifestation; several mucosal lesions with different morphology (ulcerations, vesiculobullous lesions, white lesions, pigmentations, swelling) are also possible. Taste, sensation and trigeminal function alterations have been reported and the recent evidence regarding the occurrence of jawbones osteonecrosis, especially in bisphosphonates treated patients, is increasing. Clinical management may be quite difficult due to the multiplicity of involved classes of drugs and substances (dental materials, foods), the variety of affected tissues and functions, the type of produced lesions and disturbances, the complexity of related pathogenetic mechanisms (if known), the difficulties in assessing causality and managing drug withdrawal and/or dose adjustment, as well as in establishing specific treatments, if any. In this paper the most common and significant oral ADRs, their related aspects and importance (including medico-legal implications) for health care providers will be discussed.

Expression of salivary biomarkers in patients with oral mucositis: evaluation by SELDI-TOF/MS

OBJECTIVE This study aims to evaluate changes in proteomic salivary profile of patients with oral mucositis after adjuvant cancer treatments. MATERIALS AND METHODS Samples were collected from patients after adjuvant cancer therapies, and were analyzed by means of SELDI/TOF. Patients were separated in two groups: patients affected by mucositis (MUCOSITIS) and patient without mucositis (NO MUCOSITIS). All patients were divided in function of the anticancer treatment: patients who had radiotherapy (MUCOSITIS RADIO), had not radiotherapy (MUCOSITIS NO RADIO), had chemotherapy (MUCOSITIS CHEMO), and those who had not chemotherapy (MUCOSITIS NO CHEMO). Statistical evaluation PCA (Principal Component Analysis) was conducted with the software BIO-RAD Data Manager(™) (Version 3.5). RESULTS We found the increased peaks of 3443, 3487, and 4135 m/z in MUCOSITIS group, while 6237 m/z was reduced. These same peaks would the same modifications in MUCOSITIS RADIO, while in MUCOSITIS CHEMIO are increased 3443 and 6237 m/z but 3487, 4135 m/z are reduced. These data were confirmed by the PCA. CONCLUSION Anticancer therapy influenced the level expression of many salivary biomarkers in mucositis with a good significance. Therefore, 3443, 3487, 4135, and 6237 m/z are good biomarker candidates of oral mucositis.

Oral manifestations in chronic uremia patients.

Abstract The incidence of chronic renal failure (CRF) is approximately 200 cases per million people in different Western countries. Recent data indicate that the incidences of these pathologies are increasing. Ninety percent of patients with CRF report oral signs and symptoms that affect both the bone and soft tissues. A broad range of lesions may be observed in chronic uratemia patients, including the following: gingival hyperplasia, enamel hypoplasia, petechiae, gingival bleeding, and others lesions. These patients require various types of treatment ranging from dietary and lifestyle changes to dialysis and kidney transplantation. CRF often leads to multiple oral manifestations that are difficult for dentists to manage. The present study examined the characteristics of this disease, the existing therapeutic options and the relevant considerations for dental professionals.

Mandibular Class II Furcation Defect Treatment: Effects of the Addition of Platelet Concentrates to Open Flap: A Systematic Review and Meta-Analysis of Randomized Clinical Trials

BACKGROUND To improve the efficacy and outcomes of regenerative therapy for furcation defects, the use of platelet concentrates (PCs) in addition to open flap debridement (OFD) has been investigated. The aim of this systematic review is to evaluate whether mandibular Class II furcation defects treated with the addition of PC to OFD heal with a significant improvement in the following: 1) horizontal clinical attachment level (HCAL); 2) vertical clinical attachment level (VCAL); 3) probing depth (PD); and 4) level of gingival margin (LGM). METHODS Electronic databases (MEDLINE, EBSCO Library, Cochrane Database of Systematic Reviews, and SCOPUS) were searched for randomized clinical trials to address the use of PCs in combination with OFD compared with a control group without PCs for the treatment of mandibular Class II furcation defects in humans and to provide data on the above reported outcome measures. The results of selected studies were converted to mean difference and standard error and interpolated using the inverse of variance test. Heterogeneity was investigated using both the Higgins index and the Q test. RESULTS Of 254 articles screened, only 11 were read in full text, and three of these were included in the meta-analysis. The addition of PCs to OFD revealed a small improvement in the following: 1) HCAL (mean difference, 1.36 mm; 95% confidence interval [CI] = 1.07 to 1.65); 2) VCAL (mean difference, 1.54 mm; 95% CI = 1.23 to 1.85); and 3) PD (mean difference, 1.83 mm; 95% CI = 1.36 to 2.29). No differences were found for the LGM. Heterogeneity across the studies was high, and all the three included studies were performed in one country. CONCLUSIONS Adding PCs to OFD for the treatment of mandibular Class II furcation defects may lead to slight improvements in clinical parameters. Nonetheless, because of the high heterogeneity of the very small number of reported studies and the small effect size, no definitive conclusion can be achieved about the clinical application of such a treatment option.

Salivary proteomic signatures of oral squamous cell carcinoma

Delay in diagnosing oral squamous cell carcinoma (OSCC) can be still identified as a major cause of its high morbidity and mortality. To date, the early diagnosis for OSCC is mainly based on clinical oral examination and there is an urgent need for reliable markers; thus, advancements in molecular technologies has set the stage for investigating new markers, as well as new diagnostic matrices. The aim of the present study is to investigate the presence of proteomic signatures of OSCC in saliva and their use as potential biomarkers for early and non-invasive diagnosis. Saliva from 45 OSCC patients and 30 healthy controls was analysed by SELDI-TOF mass spectrometry and ProteinChip® technology. A supervised multivariate statistical analysis (Classification and Regression Tree - CART) was used to build models for discriminating between OSCC and controls, and between early (ES-OSCC) and late stage (LS-OSCC) cancers. The peptide with 8041 Da mass was 22-fold more expressed in OSCC, thus being a suitable potential biomarker. Classification and regression analysis allowed to build a model that was capable of correctly classifying all cancers and controls in an independent testing set, using the 8041 m/z peak as splitter. Eleven peaks were also differently expressed between ES-OSCCand LS-OSCC, but, basing on these differences, it was not possible to build an algorithm to predict tumour staging. These findings confirm that saliva proteome in OSCC patients is different from healthy controls and these variations might reflect different stages of disease progression and are worthy of further validation as diagnostic and prognostic biomarkers.

Sinus Lift Augmentation by Using Calcium Sulphate. A Retrospective 12 Months Radiographic Evaluation Over 25 Treated Italian Patients

Purpose: The aim of this investigation was to assess bone healing of sinus lift procedure in which the augmentation has been performed by using calcium sulphate like bone substitutes. The methods of this investigation how the use of Cone Beam Computed Tomography (CBCT) may be a valid instrument to support reconstructive surgery of the jaws. Patients and Methodology: 25 Patients presented large bone defects after tooth extractions located in the upper jaw posterior area. Vertical bone volume was assessed by CBCT examinations before and about six months after sinus lift surgery. Results: Examined defects treated with sinus lift surgery and evaluated by CBCT showed a strong increasing on the bone volume at 6 months follow up control. Conclusion: Calcium sulphate application in sinus lift surgery represent a safe and predictable option in the place of autologous bone. Therefore the application of CBCT investigation may give the clinicians the opportunity of evaluating with high precision value, the consistence of the bone defects before the surgery.

Nuclear Survivin as a Prognostic Factor in Squamous-Cell Carcinoma of the Oral Cavity.

Oral squamous-cell carcinoma (OSCC) and most human tumors are characterized by an imbalance of regulatory mechanisms controlling cell processes such as apoptosis. Survivin, a member of the inhibitor of apoptosis family, is overexpressed in most solid and hematological malignancies and correlates with a reduced overall survival rate. Thus, the aim of this study was to find a correlation between nuclear Survivin expression and clinicopathologic data and the prognosis in OSCC patients. A total of 152 OSCC samples were investigated by immunohistochemistry for nuclear Survivin expression. Then, Survivin was scored semiquantitatively using an immunoreactivity score (IRS), calculated by multiplying the percentage of positive cells with the staining intensity. Using a digital image analysis software, OSCC patients were stratified into 4 groups. Results showed that patients with a lower IRS score displayed better survival rates than patients with a higher IRS score, reaching statistical significance. As the expression of Survivin at the nuclear level seems to suggest a poor prognosis in OSCC patients, the evaluation of nuclear Survivin IRS may be a useful tool to identify patients with more aggressive and disseminated disease, influencing follow-up and therapeutic protocols.

Ectopic manifestation of a cholesterol granuloma in the nasopalatine duct

The nasopalatine duct is an anatomical structure that connects nose to oral cavity. This anatomic area may be affected by a large group of pathologies. In this paper, we reported a very rare case of cholesterol granuloma located in the nasopalatine duct. No other case of nasopalatine localization of a cholesterol granuloma has been reported in literature up to the present time. The Cholesterol Granuloma (CG) is a foreign body reaction due to the deposition of cholesterol crystals. It is very common in the middle ear, although ectopic manifestations can be vary. In the nasopalatine duct CG may simulate other pathological conditions with similar clinical and radiological features, for this reason a careful histological examination is necessary to avoid mistakes and wrong diagnosis.