Prognostic value of histamine H1 receptor expression in oral squamous cell carcinoma

Objectives

Overexpression of the histamine H1 receptor (H1R) has been described in a variety of tumor models, but experience in oral squamous cell carcinomas (OSCC) is not available. Current adjuvant treatment options for OSCC can be improved by the identification of new targets of therapy. Herein, we evaluated H1R expression in a large patient cohort of OSCC.

Materials and methods

H1R immunoexpression was evaluated in 191 cases of OSCC and two OSCC cell lines BICR56 and BICR3. Scanned images were digitally analyzed using ImageJ and the immunomembrane plug-in. The combined score of computer-assisted semiquantitative analysis was correlated with manually counted percentages of tumor cells by Kendall’s tau (т) correlation coefficient. Disease-free survival times were estimated using the Kaplan–Meier method and were compared by using the log-rank test. Multivariate analyses were performed using the Cox proportional hazards model.

Results

H1R was rarely expressed in OSCC but significantly related with advanced tumor stages (n?=?21/191, mean expression 63.5 % of cancer cells in positive tumor samples, 95 % confidence interval of the mean 53.5 to 73.6 %, p?=?0.006). Following univariate analysis, patients with H1R expression showed a significant poorer prognosis (p?=?0.0004). Multivariate analysis revealed H1R expression as an independent prognostic factor (p?=?0.0164). Expression of H1R in cancer cell lines was confirmed by specific staining of OSCC cell lines BICR56 and BICR3.

Conclusion

This is the first study focusing on H1R expression showing a significant poorer DFS rate in the H1R+ patient cohort. Based on these data, H1R activation may promote carcinogenesis in OSCC.

Clinical relevance

Investigation of H1R regulation and its antagonists shows a clear rationale for future supportive anticancer therapies in OSCCs.     

Cyclin D1 expression and its possible regulation in chewing tobacco mediated oral squamous cell carcinoma progression

Objective

Proto-oncogene cyclin D1 is a G1 phase specific cell cycle regulator and known for its role in various cancers. The aim of the study was to understand oral cancer progression by observing the mRNA and protein expression of cyclin D1.

Methods

Different oral tissue samples were selected as a model to study oral cancer progression. Those include healthy oral mucosa, premalignant lesions (Leukoplakia, Erythroplakia, Oral SubMucous Fibrosis) and oral cancer (OSCC) samples. Cyclin D1 mRNA and protein expression were detected by slot-blot and by immunohistochemical methods, respectively.

Results

Premalignant lesions (PML) showed average 3-fold increase in the mRNA expression than normal oral mucosa (p = 0.001) whereas only 1.3-fold increase in mRNA has been observed in OSCC samples over the PML. On the other hand OSCC showed average 4-fold increase in mRNA expression than normal oral mucosa (p < 0.001). Cyclin D1 protein accumulation has been observed in 31.3% (16/51) of the OSCC samples whereas the normal oral mucosa and the PML showed no immunoreactivity. Oral cancer samples showing positive cyclin D1 immunoreactivity has increased from 15.0% (3/20) well differentiated SCC to 31.2% (5/16) moderately differentiated SCC to 53.3% (8/15) poorly differentiated SCC, found statistically significant (p = 0.05).

Conclusion

By observing the expression of cyclin D1 in different stages, we have noticed two major transitions that occur in normal oral mucosa that leads to oral cancer. The first transitional event transforms the normal oral mucosa to PML whereas the second transition drives the PML to OSCC. These findings give evidence that the first transition induces cyclin D1 mRNA with no detectable cyclin D1 protein. The induction of mRNA is maintained with increased cyclin D1 protein accumulation in the second transition.     

Levels of potential oral cancer salivary mRNA biomarkers in oral cancer patients in remission and oral lichen planus patients

Objectives

To gather preliminary data concerning the feasibility of using seven salivary mRNAs—IL-8; IL-1β; dual specificity phosphatase 1 (DUSP1); H3 histone family 3A (H3F3A); ornithin decarboxylase antizyme 1 (OAZ1); S100 calcium-binding protein P (S100P); and spermidine/spermine N1-acetyltransferase 1 (SAT1)—for detecting development of oral squamous cell carcinoma (OSCC) in oral lichen planus (OLP) patients and OSCC patients whose disease was in remission.

Materials and methods

Saliva samples were collected from five study groups (25 subjects/group): newly diagnosed OSCC, OSCC-in-remission, disease-active OLP, disease-inactive OLP, and normal controls. The salivary mRNA levels were determined by a pre-amplification RT-qPCR approach with nested gene-specific primers. Mean fold changes between each pair of study groups were analyzed by the Mann–Whitney U test.

Results

Salivary levels of OAZ1, S100P, and DUSP1 mRNAs were significantly higher in newly diagnosed OSCC patients, compared to: (1) normal controls (p?=?0.003; p?=?0.003; and p?<?0.001, respectively); (2) OSCC-in-remission (p?<?0.001; p?=?0.001; and p?<?0.001, respectively); (3) disease-active OLP (p?<?0.001; p?=?0.016; and p?<?0.001, respectively); and (4) disease-inactive OLP (p?=?0.043; p?<?0.001; and p?<?0.001, respectively). No significant differences were found in the levels of salivary IL-8, IL-1β, H3F3A, and SAT1 mRNAs between newly diagnosed OSCC patients and the normal controls (p?=?0.093, 0.327, 0.764, and 0.560, respectively).

Conclusion

Salivary OAZ1, S100P, and DUSP1 mRNAs are candidate biomarkers for detecting OSCC development in OSCC patients in remission and in OLP patients.

Clinical relevance

The results of this study serve as the basis for a further large-scale study which may lead to a non-invasive screening method for early detection of OSCC.     

Cyclin D1 and Ki-67 expression correlates to tumor staging in tongue squamous cell carcinoma

Background

The immunohistochemical expression of Cyclin D1 and Ki-67 were analyzed in tongue squamous cell carcinomas (SCC), relating them to the clinical and morphological exhibition of these tumors.

Material and Methods

Twenty-nine patients fulfilled the inclusion criteria; clinical data included gender, age, ethnicity and use of licit drugs such as alcohol and tobacco. The TNM staging and histopathological differentiation grading was assessed for each case. In addition, T1 patients were gathered with T2 patients; and T3 patients were gathered with T4 patients to assemble two distinct groups: (T1/T2) and (T3/T4).

Results

The mean follow-up time was 24 months and 30% of the patients died as a consequence of the disease, while 23.3% lived with the disease and 46.7% lived lesion-free. T1 and T2 tumors showed statistically lesser Ki-67 and Cyclin D1 staining when compared to T3 and T4 tumors.

Conclusions

Ki-67 and Cyclin D1 pose as auxiliary tools when determining the progression of tongue SCC at the time of diagnosis. Key words:Carcinoma, squamous cell, cyclin D, immunohistochemistry, Ki-67 antigen, prognosis.     

Investigation of trefoil factor expression in saliva and oral mucosal tissues of patients with oral squamous cell carcinoma

Objectives

The aims of our study were to determine levels of trefoil factor (TFF) peptides in saliva and oral mucosal tissues from patients with oral squamous cell carcinoma (OSCC), and to evaluate whether individual members of TFFs (TFF1, TFF2, and TFF3) might act as biomarkers of disease.

Materials and methods

Saliva samples were from 23 healthy subjects and 23 OSCC patients. Tissue samples were collected from 32 normal oral mucosa (NOM) and 32 OSCC biopsy specimens. ELISA and immunohistochemical methods were used to evaluate the expression of TFF1, TFF2, and TFF3 in saliva and oral mucosal tissues, respectively.

Results

Expression of TFF2 and TFF3 in oral mucosal tissues of OSCC patients was strongly downregulated when compared to healthy subjects (p?<?0.001 and p?=?0.002, respectively). However, there were no differences in levels of salivary TFF concentrations between OSCC patients and healthy subjects.

Conclusions

The present study extends previous observations, demonstrating the reduction of TFF2 and TFF3 expression in oral mucosal tissues of OSCC patients.

Clinical relevance

These findings suggest the clinical significance of TFF2 and TFF3 molecules as negative markers of tumor progression in OSCC. Quantification of TFF levels in saliva may not be optimal in terms of diagnostic or predictive value for OSCC derived from oral mucosa.     

Daxx and TCF4 interaction links to oral squamous cell carcinoma growth by promoting cell cycle progression via induction of cyclin D1 expression

Objectives

Death domain-associated protein (Daxx) has been recently implicated as a positive factor in ovarian cancer and prostate cancer, but the role of Daxx in oral squamous cell carcinoma (OSCC) has never been addressed. Herein, we investigate the expression and function of Daxx in OSCC.

Materials and methods

RT-quantitative PCR, Western blotting, and immunohistochemistry were used to evaluation of the expression of Daxx in human OSCC cell lines and clinical surgical specimens. Short hairpin RNA targeting Daxx was transduced by lentivirus infection to knockdown the expression of Daxx in SAS and SCC25 cell lines, and the influence of this knockdown was evaluated by analyzing the growth and the cell cycle in transduced cells. Immunoprecipitation and sequential chromatin immunoprecipitation-quantitative PCR were used to analyze the associations between Daxx, TCF4, and cyclin D1 promoter. Xenograft tumor model was used to evaluate the in vivo tumorigenicity of Daxx in OSCC.

Results

Daxx mRNA and protein expression are elevated in several OSCC cell lines and human OSCC samples in comparison to those in normal tissue. We further find that depletion of Daxx decreases OSCC cell growth activity through G1 cell cycle arrest. Daxx silencing reduces cyclin D1 expression via a Daxx-TCF4 interaction, whereas the Daxx depletion-mediated G1 arrest can be relieved by ectopic expression of cyclin D1. Moreover, we show that in OSCC clinical samples, the expression of Daxx is significantly correlated with that of cyclin D1.

Conclusion

Our data demonstrate the importance of Daxx in regulation of cyclin D1 expression and provide the first evidence that Daxx exhibits tumor-promoting activity in OSCC.

Clinical relevance

Daxx plays an important role in malignant transformation of OSCC and may serves as a target for cancer prevention and treatment.

     

Eosinophils may predict occult lymph node metastasis in early oral cancer

Objective

The aim of this study was to investigate whether tumor-associated tissue eosinophilia (TATE) in early oral squamous cell carcinoma (OSCC) would aid in predicting occult lymph node metastasis.

Patients and methods

Seventy-one patients undergoing elective neck dissection for T1 and T2 OSCC were evaluated for clinical features, prognosis, and TATE. The degree of TATE in OSCC was statistically analyzed in relation to the clinicopathological features, tumor invasion, occult lymph node metastasis, and survival using χ ² test and Kaplan–Meier method.

Results

Statistical analysis revealed that intense TATE was a significant feature (p?=?0.004) to predict occult lymph node metastasis in patients with early OSCC. All regional recurrences of the OSCC occurred in patients showing intense TATE.

Conclusions

These results suggest that intense TATE can be clinically used as a predictive factor for occult lymph node metastasis.

Clinical relevance

The presence of intense TATE is an adjunctive histopathological marker to reinforce the indication of elective neck dissection of the patients with early OSCC.     

Salivary level of interleukin-8 in oral precancer and oral squamous cell carcinoma

Background

Interleukin 8 (IL-8) is a pro-angiogenic, pro-inflammatory mediator that belongs to the family of chemokines. Due to its pro-angiogenic characteristic, it may play a vital role in tumour angiogenesis and progression.

Objectives

This study was designed to estimate the levels of salivary IL-8 in oral precancer and oral squamous cell carcinoma (OSCC) patients and compare them with healthy controls. The aim was to evaluate its efficacy as a potential biomarker for these diseases.

Materials and methods

Each group comprised 25 individuals. The salivary IL-8 levels were determined by enzyme-linked immunosorbent assay.

Results

The levels of salivary IL-8 were found to be significantly elevated in patients with OSCC as compared to the precancer group (p?<?0.0001) and healthy controls (p?<?0.0001). However, the difference in salivary IL-8 concentrations among the precancer group and controls was statistically non-significant (p?=?0.738).

Conclusions

Our results suggested that salivary IL-8 can be utilised as a potential biomarker for OSCC. Salivary IL-8 was found to be non-conclusive for oral premalignancy in this preliminary study. Hence, its possible role in transition from premalignancy to malignancy needs further research with larger sample sizes.

Clinical relevance

Saliva as a diagnostic biofluid offers a number of advantages over blood-based testing. The role of IL-8 in oral cancer if validated further by future research can provide an easy diagnostic test as well as a prognostic indicator for patients undergoing treatment. Therefore, if it’s role in tumourigenesis can be sufficiently assessed, it could open up new avenues to find out novel treatment modalities for oral cancer.     

Increased immunoexpression of trefoil factors in salivary gland tumors

Objective

Very little is known about the role of trefoil factors (TFFs) in salivary gland tumors, and TFF immunoexpression has never been investigated in such tumors. The aim of this study was to evaluate TFF immunoexpression in benign and malignant salivary gland tumors.

Materials and methods

Benign (n?=?25) and malignant (n?=?25) salivary gland tumor specimens were included in this study, using mucocele (n?=?25) specimens as a control group. Immunohistochemical staining was performed to evaluate the expression of TFFs (TFF1, TFF2, and TFF3) by semiquantitative means.

Results

Expression of TFF1, TFF2, and TFF3 was significantly increased in benign (p?=?0.001, p?=?0.005, p?<?0.001, respectively) and malignant (p?<?0.001, p?<?0.001, p?<?0.001, respectively) groups as compared with the control group. Patterns of co-expression between TFF1/TFF2, TFF2/TFF3, and TFF1/TFF3 were different among the three groups.

Conclusions

The present study provided new information showing that all TFFs were significantly increased in benign and malignant salivary gland tumors, and overexpression of TFFs could be associated with neoplastic transformation in salivary gland tissues.

Clinical relevance

Overexpression of TFFs may be useful as biomarkers in terms of differential diagnosis between salivary gland tumors and other oral neoplasms for which clinical manifestations are indistinguishable.     

An update on the implications of cyclin D1 in oral carcinogenesis

Cyclin D1 promotes cell cycle progression during G1 phase, a key event in G1‐S transition. The protein is encoded by gene CCND1, located in chromosomal band 11q13. Cyclin D1 plays key roles in cell biology, including cell proliferation and growth regulation, mitochondrial activity modulation, DNA repair, and cell migration control. CCND1 gene and its protein cyclin D1 are frequently altered by different molecular mechanisms, including amplification, chromosomal translocations, mutations, and activation of the pathways involved in cyclin D1 expression, alterations which appear to be essential in the development of human cancers, including oral carcinoma. This is the first published review of the specific features of cyclin D1 overexpression in oral oncogenesis. Starting with the physiological regulation of cyclin D1, there is an evaluation of its functions, overexpression mechanisms, and the implications of the oncogenic activation of CCND1/cyclin D1 in oral squamous cell carcinoma . The potential diagnostic and prognostic value of cyclin D1 is reviewed. The influence of CCND1/cyclin D1 on tumor size and clinical stage is reported, and an update is provided on the utilization of cyclin D1 as therapeutic target and on the combination of cyclin D1 inhibitors with cytotoxic agents. Future research lines in this field are also proposed.     

Prognostic and predictive value of serum vascular endothelial growth factor (VEGF) in squamous cell carcinoma of the head and neck

Introduction

This study aimed to investigate the prognostic and predictive value of serum vascular endothelial growth factor (VEGF) in head and neck squamous cell carcinoma (HNSCC).

Method

Preoperative and 6-month postoperative serum VEGF levels were measured using a quantitative sandwich enzyme immunoassay technique in 55 consecutive patients with HNSCC and two control groups. The first control group included normal, healthy, age- and sex-matched individuals (n?=?20), while the second control group included the patients who had history of HNSCC and were free of disease for at least 5 years (n?=?25).

Results

The mean baseline serum VEGF concentrations of the 55 patients with HNSCC and the first and the second control groups were 437.86, 42.56, and 48.03 pg/ml, respectively (P?<?0.001). After a median follow-up of 75 months, 15 patients of the study group developed recurrent disease and 40 patients remained free of disease. The mean preoperative and 6-month postoperative serum VEGF levels for the 40 patients who did not have recurrent disease were respectively 327.69 and 153.50 pg/ml compared to 731.72 and 692.96 pg/ml for the 15 patients with recurrent disease (P?<?0.001). High (≥540 pg/ml) serum VEGF level was associated with poor overall survival (P?<?0.001). Moreover, multivariate analysis showed node stage (P?<?0.001) and preoperative serum VEGF level (P?=?0.020) as significant, independent prognostic factors for overall survival.

Conclusion

Preoperative or postoperative elevated serum levels of VEGF are highly predictive for disease recurrence and are associated with poor disease-free and overall survival of patients with HNSCC.     

Spin electron paramagnetic resonance of albumin for diagnosis of oral squamous cell carcinoma (OSCC)

Objectives

Albumin has a known capability to modulate free serum concentrations of proteins produced by tumour cells. The technique of spin probe labelling of albumin followed by electron paramagnetic resonance (EPR) spectroscopy may allow identification of these structural and functional changes, which regularly occur as consequence of binding tumour metabolites as ligands. The aim of the present study was a proof of principle evaluation of EPR-analysis of peripheral blood samples as possible predictor for oral squamous cell carcinoma (OSCC).

Material and methods

The present study is designed as gender-matched cohort. EPR was tested after retrieval of peripheral blood samples. The study group is represented by 32 patients with OSCC, and the control group consisted of 30 healthy patients.

Results

Overall analysis exhibited a diagnostic sensitivity of 72% (23/32 OSCC group) and a specificity of 80% (24/30 control group). Subgroup analysis revealed ten patients with elevated leukocytes (>10,000/μl; n?=?9 [OSCC group] and n?=?1 [control group]). After exclusion of patients with elevated white blood cell count, sensitivity considerably increased to 87% and specificity to 83%.

Conclusion

EPR analysis of peripheral blood samples might be appropriate to support the clinician in primary and follow-up diagnosis of potential tumours such as OSCC. Unfortunately, subgroup analysis characterises the method vulnerable to inflammation.

Clinical relevance

Nevertheless, our preliminary results are intriguing, as diagnosis of OSCC appears possible by simple peripheral blood examination. Thus, further appraisal of this novel method with inclusion of different tumour entities, systemic conditions and inflammation in a larger study population appears highly valuable.     

Changes in mastication after an immediate loading implantation with complete fixed rehabilitation

Objective

This study aimed to measure modifications of mastication after immediate loading full-arch prosthesis (ILFAP) rehabilitation.

Materials and method

Fourteen patients were observed before and 6 months after ILFAP rehabilitation when masticating two natural, standardized foods (peanut and carrot) and three model foods with increasing hardness. The granulometry of the expectorated boluses from carrot and peanuts was characterized by median particle size (D50), determined at the natural point of swallowing. Chewing time (CT), number of chewing cycles (CC), and chewing frequency (CF) were video recorded. A self-assessment questionnaire for oral health-related quality of life [Geriatric Oral Health Assessment Index (GOHAI)] was also used.

Results

After ILFAP rehabilitation, the mean D50 values for carrot and peanuts were smaller [Repeated Model Procedures (RMP), F?=?41, p?<?0.001]. Mean CT and CC values recorded with the three model foods decreased, while CF increased regardless of the model food hardness (RMP, F?=?14, F?=?10, and F?=?11, respectively, p?<?0.001). The GOHAI score increased from 43?±?9 to 56?±?3 (t test, p?<?0.001).

Conclusion

ILFAP rehabilitation improves the ability to reduce the bolus particle size and the ability to discriminate between different food hardnesses in the 6 months post-surgery period.

Clinical relevance

This study encourages the clinical development of immediate loading implantation with a fixed full-arch prosthesis protocol.     

P53 immunohistochemical expression does not correlate with clinical features in 207 carcinomas of the oral cavity and in the head and neck region

Objectives

The present study aims to investigate the relevance of immunohistochemical p53 expression in carcinomas of the oral cavity and of the head and neck region. Long-term clinical and histopathological follow-up findings as well as HPV status are correlated with the results of this examination.

Materials and methods

Sections made from two tissue arrays composed of 222 oral squamous cell carcinomas and 427 squamous cell carcinomas of the head and neck region, respectively, were examined for p53 expression and Ki-67 index by means of immunohistochemistry. Correlation of long-term clinical findings of the patients and pathological features of tumours with laboratory results were examined statistically.

Results

No significant correlation was found between the p53 immunohistochemical expression in the 207 oral carcinomas and features of the tumours and patient outcomes. There was no significant association between the Ki-67 labelling index and the p53 expression.

Discussion

Our failure in detecting any association of the p53 immunohistochemical expression regardless of HPV status with clinical features of these tumours suggests it lacks a prognostic value for squamous cell carcinomas of the oral cavity.

Clinical relevance

The prognostic value of p53 immunostaining in oral squamous carcinoma is not clarified yet. In the present study, there is no impact on any prognostical item nor even a correlation with cell proliferation (Ki-67) regardless of HPV status.     

Dentofacial characteristics of patients with rheumatoid arthritis

Objectives

The aims of this study are to evaluate the dentofacial morphology of patients with rheumatoid arthritis (RA) and to compare the morphological data with those of healthy age- and sex-matched control subjects.

Methods

Twenty-seven RA patients (mean age, 45.77?±?8.64 years) and 25 healthy subjects (mean age, 44.80?±?8.24 years) participated in this prospective study. Clinical and functional evaluations of the RA patients were assessed. The erythrocyte sedimentation rate, C-reactive protein level, rheumatoid factor level, and anti-citrullinated peptide antibodies (ACPA) titers of RA patients were determined, and DAS28 scores were calculated. Linear and angular measurements were performed on cephalometric tracings and condylar erosion was evaluated on lateral panoramic radiographs. Statistical comparison of the two groups was performed with an independent samples t test. Pearson correlation analysis was used to assess the relationship between the clinical and laboratory parameters.

Results

Based on DAS28 scores, no patient with RA was in the remission period, 3 patients had low, 23 had medium, and 1 had high disease activity. Sixteen (59.26 %) patients with RA had positive ACPA titers. Lateral cephalometric radiographs revealed statistically significant difference between the two groups for the measurement of U1–NA (millimeter; p?=?0.047), U1–NA (degrees; p?=?0.031), L1–NB (degrees; p?=?0.030), IMPA (L1–MP; p?=?0.001), interincisal angle (U1–L1; degrees; p?=?0.022) and midface length (Co–A; millimeter; p?=?0.033). A significant positive linear correlation was found between disease duration time and DAS28 scores (r?=?0.066, p?=?0.040).

Conclusions

Dentoalveolar effects of RA on dentofacial morphology are more significant than the skeletal effects. Future studies with larger sample sizes are required to evaluate the exact effects of RA on dentofacial morphology.

Clinical relevance

Clinicians should consider the fact that RA-associated dentoalveolar changes can be observed and may affect the orthodontic treatment process.     

Diagnostic accuracy of ultrasound, 18F-FDG-PET/CT,and fused 18F-FDG-PET-MR images with DWI for the detection of cervical lymph node metastases of HNSCC

Objective

This study aimed to compare ¹⁸F-fluorodesoxyglucose positron emission tomography/MRI (¹⁸F-FDG-PET-MRI) fusion images, including diffusion-weighted imaging (DWI), ¹⁸F-FDG-PET/CT, and ultrasound (US) regarding their performance in nodal staging of patients with head and neck squamous cell carcinoma (HNSCC).

Materials and methods

Eighteen patients prospectively underwent ultrasound examination, ¹⁸F-FDG- PET/CT, and MRI before oral tumor resection and bilateral neck dissection. PET data sets were fused with contrast-enhanced T1-weighted MR images. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy for nodal detection were calculated for all the imaging modalities. Furthermore, the accuracy of the correct N-staging was calculated for all methods. Detailed histopathology served as the standard of reference.

Results

The sensitivity, specificity, PPV, NPV, and accuracy for detection of lymph node metastases were 63, 99, 86, 96, and 95 % for ultrasound; 30, 97, 56, 92, and 90 % for ¹⁸F-FDG-PET/CT; 52, 96, 59, 94, and 91 % for ¹⁸F-FDG-PET-MRI; and 53, 97, 67, 95, and 92 % for ¹⁸F-FDG-PET-MRI plus DWI, respectively. There was no significant difference in the diagnostic accuracy for lymph node metastasis detection between ¹⁸F-FDG-PET-MRI and ¹⁸F-FDG-PET/CT (p?=?0.839) and between ¹⁸F-FDG-PET-MRI plus DWI and ¹⁸F-FDG-PET/CT (p?=?0.286), respectively. US was significantly more accurate than ¹⁸F-FDG-PET/CT (p?=?0.009), whereas no significant difference was seen between ¹⁸F-FDG-PET-MRI and US (p?=?0.223) or ¹⁸F-FDG-PET-MRI plus DWI and US (p?=?0.115). The nodal stage was correctly rated by ¹⁸F-FDG-PET-MRI in eight patients, ¹⁸F-FDG-PET-MRI plus DWI in nine patients, US in 12 patients, and ¹⁸F-FDG-PET/CT in five out of 18 patients.

Conclusion

Software-based fusion of ¹⁸F-FDG-PET-MRI and ¹⁸F-FDG-PET-MRI plus DWI may not increase nodal detection and N-staging performance in patients with oral malignancies compared to US and ¹⁸F-FDG-PET/CT.

Clinical relevance

Surgical staging of cervical lymph nodes will not be replaced even by advanced imaging modalities in the near future.     

Salivary and serum interleukin-6 levels in proliferative verrucous leukoplakia

Background

Cytokines and chemokines have been analysed in patients with oral squamous cell carcinoma and potentially malignant disorders. We selected interleukin-6 (IL-6) because it is a multifunctional interleukin reported to be altered in potentially malignant oral disorders and in malignant lesions. To date, this has not been evaluated or tested in proliferative verrucous leukoplakia (PVL), however.

Objectives

This study aimed to analyse the differences in serum and saliva IL-6 levels among patients with PVL, oral squamous cell carcinoma (OSCC) and healthy controls and to examine the relationship between salivary IL-6 levels and the extent of the verrucous area.

Methods

Using an enzyme-linked immunosorbent assay, we determined the serum and saliva IL-6 levels in three groups: 20 patients with PVL, 20 with OSCC and 20 healthy controls.

Results

There were significant (p?<?0.01) differences in the serum and saliva IL-6 levels among the three groups and among the three grades of extent of the verrucous areas (p?=?0.01). In the OSCC group, there was a significant difference in the saliva IL-6 levels between patients with and without lymph node metastasis at diagnosis (p?=?0.02).

Conclusions

We found that patients with OSCC had the highest salivary and serum IL-6 levels, while PVL had lower values than OSCC, but higher than the controls, and these altered levels were associated with the extent of the verrucous areas.

Clinical relevance

Salivary and plasma IL-6 are altered in patients with PVL, with more extensive verrucous areas being associated to the highest IL-6 levels. This could be a significant tool for monitoring patients with PVL, their progression to more advances stages and their recurrences.

     

Adverse effects of the amlodipine on bone healing of the mandibular fracture: an experimental study in rats

Background

Calcium channel blockers are effective antihypertensive agents, but they may affect many metabolic processes, including bone metabolism.

Purpose

The aim of this study was to evaluate by radiographic, histologic, and biochemical analyses the effects of amlodipine on bone healing of a defect simulating a fracture in mandibular ramus of rats.

Methods

Fifty male Wistar rats were submitted to the same unilateral surgical procedure simulating a mandibular fracture. Experimental group received oral doses of amlodipine 0.04?mg/rat/day starting 12?days before procedure, while control group received water. Animals were sacrificed at 1, 7, 14, 30, and 90?days postoperatively. Numerical values were submitted to statistical analyses.

Results

Radiographic analysis showed larger radiolucent area into bone defect to the experimental group at the periods of 14 (p?=?0.016), 30 (p?=?0.009), and 90 (p?=?0.028)?days. In the histological analysis, the experimental group had a slight delay in the chronology of the repair process. In the histomorphometric analysis, the experimental group presented significant lowering of newly formed bone volume at 7 and 14?days periods (p?=?0.049). There was a significant decrease of alkaline phosphatase levels in experimental group in the initial periods (p?=?0.049).

Conclusions

It was concluded that chronic use of amlodipine compromised bone neoformation in the repairing process of surgical defect in the mandibular ramus of rats, but no precluded occurrence of fracture consolidation.     

Role of amniotic fluid mesenchymal cells engineered on MgHA/collagen-based scaffold allotransplanted on an experimental animal study of sinus augmentation

Objectives

The present research has been performed to evaluate whether a commercial magnesium-enriched hydroxyapatite (MgHA)/collagen-based scaffold engineered with ovine amniotic fluid mesenchymal cells (oAFMC) could improve bone regeneration process in vivo.

Materials and methods

Bilateral sinus augmentation was performed on eight adult sheep in order to compare the tissue regeneration process at 45 and 90 days after implantation of the oAFMC-engineered scaffold (Test Group) or of the scaffold alone (Ctr Group). The process of tissue remodeling was analyzed through histological, immunohistochemical, and morphometric analyses by calculating the proliferation index (PI) of oAFMC loaded on the scaffold, the total vascular area (VA), and vascular endothelial growth factor (VEGF) expression levels within the grafted area.

Results

MgHA/collagen-based scaffold showed high biocompatibility preserving the survival of oAFMC for 90 days in grafted sinuses. The use of oAFMC increased bone deposition and stimulated a more rapid angiogenic reaction, thus probably supporting the higher cell PI recorded in cell-treated sinuses. A significantly higher VEGF expression (Test vs. Ctr Group; p?=?0.0004) and a larger total VA (p?=?0.0006) were detected in the Test Group at 45 days after surgery. The PI was significantly higher (p?=?0.027) at 45 days and became significantly lower at 90 days (p?=?0.0007) in the Test Group sinuses, while the PI recorded in the Ctr Group continued to increase resulting to a significantly higher PI at day 90 (CTR day 45 vs. CTR day 90; p?=?0.022).

Conclusions

The osteoinductive effect of a biomimetic commercial scaffold may be significantly improved by the presence of oAFMC.

Clinical relevance

The amniotic fluid mesenchymal cell (AFMC) may represent a novel, largely and easily accessible source of mesenchymal stem cells to develop cell-based therapy for maxillofacial surgery.