Effect of short-term fasting on the cisplatin activity in human oral squamous cell carcinoma cell line HN5 and chemotherapy side effects.

BACKGROUND: Ketogenic interventions like short-term fasting show potential as complementary therapies to enhance the effectiveness of chemotherapy for cancer. However, the specific effects of fasting on head and neck squamous cell carcinoma (HNSCC) cells and healthy oral mucosa cells during these treatments are not well understood. This study investigates whether short-term fasting can differentially impact HNSCC cell survival and viability compared to healthy keratinocytes while undergoing standard chemotherapy regimens. METHODS: This study investigated the effects of fasting on cell viability in HN5 cell line and healthy oral keratinocyte cells. The HN5 cell line, derived from human tongue squamous cell carcinoma, and primary human keratinocytes isolated from the basal layer of gingival epithelium were divided into three groups: (1) control, (2) treated with the standard chemotherapeutic agent cisplatin, and (3) treated with cisplatin under fasting conditions achieved through 48-hour glucose restriction mimicking the blood glucose levels of fasted individuals. Cell proliferation was assessed at 48 and 72 h using the MTT assay, a colorimetric method based on mitochondrial dehydrogenase activity. Flow cytometry analysis with specific apoptosis and necrosis markers distinguished between early and late apoptotic, necrotic, and viable cells. RESULTS: Cell viability in HN5 and healthy keratinocyte cells decreased in cisplatin with low glucose groups compared to cisplatin and control groups. The same results were observed for healthy keratinocyte cells; only a decrease in cell viability in cisplatin groups compared to control groups was observed, which was not statistically significant. Cell apoptosis in HN5 and healthy keratinocyte cells increased in cisplatin with low glucose groups compared to cisplatin and control groups. In healthy keratinocyte cells, the cisplatin with low glucose group showed an impressive increase in necrosis, late apoptosis, and early apoptosis and a significant decrease in live cells compared with other groups. CONCLUSION: This study revealed that short-term fasting chemotherapy significantly improved HNSCC cell line apoptosis and necrosis.

Identification of common salivary miRNA in oral lichen planus and oral squamous cell carcinoma: systematic review and meta-analysis.

BACKGROUND: Oral lichen planus (OLP) is a chronic inflammatory condition that can impact patients' quality of life. While its exact etiology remains unclear, it is associated with an increased risk of malignant transformation. Currently, the diagnosis of OLP relies on clinical examination and histopathological analysis, which can be invasive. Therefore, there is an urgent need for non-invasive and accurate diagnostic biomarkers. This systematic review and meta-analysis aims to investigate the potential of salivary microRNAs as promising candidates for OLP diagnosis. This meta-analysis seeks to identify specific microRNAs that are differentially expressed and could serve as reliable biomarkers for OLP diagnosis. METHODS: Our strategy involved searching for pertinent keywords in multiple academic databases including Cochrane Library, Embase, LIVIVO, MEDLINE, Ovid, ProQuest, Scopus, Web of Science, Espacenet, and Google Scholar search engine. Upon identification, articles were screened and data extracted from the eligible studies. Split component synthesis method was utilized to assess specificity, sensitivity, likelihood and diagnostic odds ratios. The random-effects meta-analysis approach was used to combine study findings and develop pooled diagnostic performance metrics. Hierarchical summary receiver operating characteristic (ROC) plots were generated to determine area under the curve. Subgroup analyses concerning the type of saliva and control groups were also performed. RESULTS: Among the fourteen studies included in our systematic review, five were eligible for meta-analysis. Salivary microRNAs showed the pooled sensitivity of 0.80 (95% Confidence Interval (95% CI): 0.68-0.88), specificity of 0.89 (95% CI: 0.82-0.94), diagnostic odds ratio of 28.45 (95% CI: 10.40-77.80), and area under the curve (AUC) of 0.93 for OLP diagnosis. Unstimulated saliva had higher sensitivity and specificity than oral swirl samples as the biomarker medium for OLP diagnosis. Meta-analysis uncovered that miR-27a, miR-137, miR-1290, miR-27b, miR-4484, miR-142, and miR-1246 had the highest diagnostic odds ratio for OLP. CONCLUSIONS: Our systematic review and meta-analysis demonstrate that salivary microRNAs can serve as valuable biomarkers for the diagnosis of OLP. The findings highlight the exceptional accuracy of salivary microRNAs in differentiating OLP patients from healthy controls and assessing the risk of malignant transformation.

Transdifferentiation of periodontal ligament stem cells into acinar cells using an indirect co-culture system.

Serous Acinar Cells (ACs) are mature and functional secretory epithelial cells that develop and complete through other stem cells at the end of the ductal system. So, the regeneration of the salivary gland damaged by radiation does not occur without cell therapy. Todays, an accessible tissue like the Periodontal Ligament (PDL) of the tooth was considered to easily extract the Mesenchymal Stem Cells (MSCs). In-vitro differentiation of stem cells before transplantation to damaged tissue reduces the risk of tumorigenesis. This study was conducted to evaluate the feasibility of differentiation of PDLSCs into salivary acinar cells by a co-culture system. PDLSCs were isolated from adult human PDL tissue and co-cultured with rat parotid ACs using an indirect co-culture system. The transdifferentiation of PDLSCs was evaluated by PCR of Aquaporin 5 (AQP5) and Carbonic anhydrase 6 (CA6) genes, then quantitative real-time PCR was used to measure the gene expression levels. The data were analyzed by ANOVA. Specific bond with the correct size on 6% acrylamide gel and TBE5X buffer showed the expression of AQP5 and CA6 in PDLSCs co-cultured with acinar cells. RT-PCR revealed co-cultured PDLSCs with or without KGF (Keratinocyte Growth Factor) showed significantly increased expression of AQP5 genes in compared to the initial PDLSCs. Expression of AQP5 and CA6, indicating successful transdifferentiation of PDLSCs into ACs, in co-culture system for 3 weeks.

Evaluation of CSTB and DMBT1 expression in saliva of gastric cancer patients and controls.

BACKGROUND: Gastric cancer (GC) is the fifth most common cancer and the third cause of cancer deaths globally, with late diagnosis, low survival rate, and poor prognosis. This case-control study aimed to evaluate the expression of cystatin B (CSTB) and deleted in malignant brain tumor 1 (DMBT1) in the saliva of GC patients with healthy individuals to construct diagnostic algorithms using statistical analysis and machine learning methods. METHODS: Demographic data, clinical characteristics, and food intake habits of the case and control group were gathered through a standard checklist. Unstimulated whole saliva samples were taken from 31 healthy individuals and 31 GC patients. Through ELISA test and statistical analysis, the expression of salivary CSTB and DMBT1 proteins was evaluated. To construct diagnostic algorithms, we used the machine learning method. RESULTS: The mean salivary expression of CSTB in GC patients was significantly lower (115.55 ± 7.06, p = 0.001), and the mean salivary expression of DMBT1 in GC patients was significantly higher (171.88 ± 39.67, p = 0.002) than the control. Multiple linear regression analysis demonstrated that GC was significantly correlated with high levels of DMBT1 after controlling the effects of age of participants (R2 = 0.20, p < 0.001). Considering salivary CSTB greater than 119.06 ng/mL as an optimal cut-off value, the sensitivity and specificity of CSTB in the diagnosis of GC were 83.87 and 70.97%, respectively. The area under the ROC curve was calculated as 0.728. The optimal cut-off value of DMBT1 for differentiating GC patients from controls was greater than 146.33 ng/mL (sensitivity = 80.65% and specificity = 64.52%). The area under the ROC curve was up to 0.741. As a result of the machine learning method, the area under the receiver-operating characteristic curve for the diagnostic ability of CSTB, DMBT1, demographic data, clinical characteristics, and food intake habits was 0.95. The machine learning model's sensitivity, specificity, and accuracy were 100, 70.8, and 80.5%, respectively. CONCLUSION: Salivary levels of DMBT1 and CSTB may be accurate in diagnosing GCs. Machine learning analyses using salivary biomarkers, demographic, clinical, and nutrition habits data simultaneously could provide affordability models with acceptable accuracy for differentiation of GC by a cost-effective and non-invasive method.

Effect of synbiotic mouthwash on oral mucositis induced by radiotherapy in oral cancer patients: a double-blind randomized clinical trial.

PURPOSE: Considering the complex pathobiology of oral mucositis, especially in oral cancer patients, the prevention and treatment of oral mucositis in patients undergoing radiotherapy remains an essential and clinically crucial unmet need. The present study aims to investigate and compare the effects of synbiotic mouthwash with normal saline mouthwash on the prevention and control of radiotherapy-induced oral mucositis in oral cancer patients. METHODS: Double-blind, randomized clinical trial (RCT) performed on 64 oral cancer patients who underwent radiotherapy (IRCT20201106049288N1, registration date: 2020-12-23). Patients were divided randomly into the case (32 subjects) and control (32 subjects) groups. All patients underwent intensity-modulated radiotherapy and received 6000 cGY of radiotherapy in 34 fractions. All patients received the usual treatment for mucositis, but in the case group, synbiotic mouthwash was prescribed and in the control group, normal saline mouthwash was prescribed from a day before the start to the end of radiotherapy treatment. Patients were monitored every session for 6 weeks to check the progression, oral involvement severity, and mucositis grade. RESULTS: The case group showed a significant reduction in the oral mucositis severity. The mucositis grade in the case group from the 7th session of oral examination was significantly lower than the control (p < 0.05), and this significant difference persisted until the last session of oral examination. Incidence rates of severe oral mucositis (grade 3) during the treatment period were 11.59% in the case and 36.45% in control (p < 0.001). CONCLUSION: Synbiotic mouthwash significantly reduces and prevents oral mucositis intensity in oral cancer patients undergoing radiotherapy.

Salivary cystatin S levels in children with early childhood caries in comparison with caries-free children; statistical analysis and machine learning.

BACKGROUND: Early childhood caries is the most common infectious disease in childhood, with a high prevalence in developing countries. The assessment of the variables that influence early childhood caries as well as its pathophysiology leads to improved control of this disease. Cystatin S, as one of the salivary proteins, has an essential role in pellicle formation, tooth re-mineralization, and protection. The present study aims to assess salivary cystatin S levels and demographic data in early childhood caries in comparison with caries-free ones using statistical analysis and machine learning methods. METHODS: A cross-sectional, case-control study was undertaken on 20 cases of early childhood caries and 20 caries-free children as a control. Unstimulated whole saliva samples were collected by suction. Cystatin S concentrations in samples were determined using human cystatin S ELISA kit. The checklist was collected from participants about demographic characteristics, oral health status, and dietary habits by interviewing parents. Regression and receiver operating characteristic (ROC) curve analysis were done to evaluate the potential role of cystatin S salivary level and demographic using statistical analysis and machine learning. RESULTS: The mean value of salivary cystatin S concentration in the early childhood caries group was 191.55 ± 81.90 (ng/ml) and in the caries-free group was 370.06 ± 128.87 (ng/ml). T-test analysis showed a statistically significant difference between early childhood caries and caries-free groups in salivary cystatin S levels (p = 0.032). Investigation of the area under the curve (AUC) and accuracy of the ROC curve revealed that the logistic regression model based on salivary cystatin S levels and birth weight had the most and acceptable potential for discriminating of early childhood caries from caries-free controls. Furthermore, using salivary cystatin S levels enhanced the capability of machine learning methods to differentiate early childhood caries from caries-free controls. CONCLUSION: Salivary cystatin S levels in caries-free children were higher than the children with early childhood caries. Results of the present study suggest that considering clinical examination, demographic and socioeconomic factors, along with the salivary cystatin S levels, could be usefull for early diagnosis ofearly childhood caries in high-risk children; furthermore, cystatin S is a protective factor against dental caries.

Three-dimensional simulation of human teeth and its application in dental education and research.

Background: A comprehensive database, comprising geometry and properties of human teeth, is needed for dentistry education and dental research. The aim of this study was to create a three-dimensional model of human teeth to improve the dental E-learning and dental research. Methods: In this study, a cross-section picture of the three-dimensional model of the teeth was used. CT-Scan images were used in the first method. The space between the cross- sectional images was about 200 to 500 micrometers. Hard tissue margin was detected in each image by Matlab (R2009b), as image processing software. The images were transferred to Solidworks 2015 software. Tooth border curve was fitted on B-spline curves, using the least square-curve fitting algorithm. After transferring all curves for each tooth to Solidworks, the surface was created based on the surface fitting technique. This surface was meshed in Meshlab-v132 software, and the optimization of the surface was done based on the remeshing technique. The mechanical properties of the teeth were applied to the dental model. Results: This study presented a methodology for communication between CT-Scan images and the finite element and training software through which modeling and simulation of the teeth were performed. In this study, cross-sectional images were used for modeling. According to the findings, the cost and time were reduced compared to other studies. Conclusion: The three-dimensional model method presented in this study facilitated the learning of the dental students and dentists. Based on the three-dimensional model proposed in this study, designing and manufacturing the implants and dental prosthesis are possible.

Comparison of salivary nitric oxide and oral health in diabetic patients with and without xerostomia.

BACKGROUND AND OBJECTIVE: Diabetes mellitus (DM) is a chronic multi-systemic metabolic disorder; diabetic patients are more prone to xerostomia and oral health problems than others. There are evidences that nitric oxide (NO) plays an important role in healthy salivary gland function, prevention of insulin resistance and progression of diabetes mellitus. The aim of this study was to compare the salivary NO level between type 2 diabetic (T2DM) patients with and without xerostomia. METHODS: In this case control study, 70 patients with T2DM, which were matched according to age, sex, type of disease control, were enrolled conveniently. The subjects based on abeslang test were allocated to the two groups; 35 patients with xerostomia and 35 patients without xerostomia. Unstimulated whole saliva was collected by spitting method. NO levels was measured by ELISA method using Griess reaction. Data was analyzed using t-test, ANOVA and logistic regression analysis to examine the association of salivary NO and xerostomia. RESULTS: The mean and standard deviation of salivary NO in the diabetic subjects with xerostomia was significantly lower than diabetic subjects without xerostomia (138 ± 94.58 µmol/L vs. 356.61 ± 302.81 µmol/L (P-value = 0.001). In logistic regression analysis, salivary NO level was associated with 0.994 fold decreased risk of xerostomia in diabetic subjects after adjustment for age, gender, FBS and HbA1c. CONCLUSIONS: The present study indicates salivary nitric oxide level was a predictor of xerostomia in diabetic patients. More longitudinal studies are necessary to understand the association of salivary NO level with diabetes-induced xerostomia.

Er,Cr:YSGG laser surface treatment of gamma titanium aluminide: Scanning electron microscopy-energy-dispersive X-ray spectrometer analysis, wettability and Eikenella corrodens and Aggregatibacter actinomycetemcomitans bacteria count-in vitro study.

Dental implants play an important role in oral health. Titanium dental implants must endure the complex microflora environment of the oral cavity. Moreover, bacterial infections have been considered as one of the most important factors of implant failure. The issue of dental improvement through modification of chemical composition and surface treatment has received considerable critical attention. γ-TiAl as a novo biocompatible material revealed a slower corrosion rate in biological media rather Ti-6Al-4V. The objective of this study is to investigate the effect of Er,Cr:YSGG laser on γ-TiAl in comparison with sandblasted and acid-etched samples as the control groups and machined samples.Wettability, surface roughness, surface topography, scanning electron microscopy-energy dispersive X-ray spectrometer analysis of surface and subsurface of samples were investigated and bacteria counts of two periodontal bacterial strains (Aggregatibacter actinomycetemcomitans and Eikenella corrodens) were evaluated on the Er,Cr:YSGG laser surface-treated sandblasted and acid-etched and machined samples.The results of this investigation show that Er,Cr:YSGG laser surface treatment affects surface roughness, surface topography, wettability, chemical composition of the surface and bacteria count. Scanning electron microscopy-energy dispersive X-ray spectrometer analysis of the sample revealed the increment of titanium and oxygen content and reduction of aluminum content in the surface and subsurface layer. A. actinomycetemcomitans and E. corrodens count were found from the lowest level to highest in the sandblasted and acid-etched samples, laser samples and machined samples, respectively.Using controlled parameters of Er,Cr:YSGG laser ensured no significant adverse alteration. The findings to emerge from this study revealed the significant correlation between microbial count and wettability. Furthermore, the contact angle strongly correlated with surface roughness.

Comparison of the effect of regular and probiotic cake (Bacillus coagulans) on salivary pH and Streptococcus mutans count.

BACKGROUND: Dental caries is considered the most common infectious disease in humans worldwide. Cariogenesis is the outcome of a complex interaction between the host's oral flora and diet. The consumption of snacks such as cake, which have the potential to promote dental caries, has increased. OBJECTIVES: The aim of this study was to investigate the effect of including probiotic bacteria (Bacillus coagulans - B. coagulans) in consumed snack cake on the Streptococcus mutans (S. mutans) count and salivary pH. MATERIAL AND METHODS: We conducted a randomized, double-blind, cross-sectional cohort study on 40 healthy volunteers. The subjects were divided into 2 groups. In the 1st group, the subjects consumed probiotic cake as breakfast for 1 week and then, following a 4-week wash-out period, consumed regular cake as breakfast for 1 week. In the other group, the administration of probiotic and regular cake was reversed. For both groups, samples of at least 5 mL of non-stimulated saliva were collected using the spitting technique before and after the 1st and the 6th week. A colony counter was used to determine the number of S. mutans colonies. Salivary pH was measured before eating (8-9 a.m.). RESULTS: We detected no statistically significant difference in the S. mutans count before and after the consumption of probiotic cake, but noted a statistically significant difference in the count before and after the consumption of regular cake. We did not detect a significant difference in salivary pH with respect to the consumption of probiotic and regular cake, although the consumption of both foods caused a drop in salivary pH. CONCLUSIONS: The addition of probiotic bacteria to sweet snack cake caused a minimal increase in the salivary count of S. mutans, a bacterial species with a definite role in cariogenesis, but did not impact salivary pH. Since probiotic cake has a slight impact on the S. mutans count, it is preferred over regular cake as a snack food.

Lineage and phylogenetic analysis of HPV-16, -18 in saliva of HNSCC patients

Aim: Head and Neck Squamous Cell Carcinoma (HNSCC) is a global health problem whose incidence varies by geographic region and race according to risk factors. Human papillomavirus (HPV) infection is a significant risk factor for HNSCC. HPV-16 and HPV-18 are two forms of HPV that are carcinogenic. HNSCCs that are HPV positive have a better prognosis rather than HPV negative. The purpose of this research was to characterize HPV-16, -18 variations in the saliva of HNSCC patients by examining the genetic diversity of HPV-16, -18 utilizing the full E6, E7, and L1 genes. Methods:The case-control research included 15 patients with HNSCC and 15 healthy volunteers. Unstimulated entire saliva samples were obtained from the case and control groups by spitting method. Genomic DNA was isolated from all saliva samples. A PCR reaction was used to determine the presence of HPV in saliva. HPV-positive samples were genotyped and data were analyzed. We conducted a variant study on the HPV-16, -18 E6, and E7 genes. Results: Three patients with HNSCC were HPV-positive for two HPV genotypes out of 30 people diagnosed with HPV-DNA. HPV-16 and -18 were the most common genotypes. The HPV-16, -18 E6, and E7 genes were sequenced and compared to the HPV-16, -18 (E6, E7) prototype sequence. In all, HPV-16 lineages A1 and HPV-18 lineages A3 were discovered. Conclusion: Regarding the variation of HPV found in Iranian HNSCC patients, the need for further studies in HPV genotyping was seen. Sequencing HPV genes in HNSCC may help answer questions about HPV genotyping in the Iranian population. HPV genotype analysis aids in the development of vaccinations against HNSCC, halting disease progression and preventing HPV-associated HNSCC