Leukoplakia Patients Research Articles

Differences in the landscape of colonized microorganisms in different oral potentially malignant disorders and squamous cell carcinoma: a multi-group comparative study

BackgroundThe role of microbes in diseases, especially cancer, has garnered significant attention. However, research on the oral microbiota in oral potentially malignant disorders (OPMDs) remains limited. Our study investigates microbial communities in OPMDs.Materials and methodsOral biopsies from19 oral leukoplakia (OLK) patients, 19 proliferative verrucous leukoplakia (PVL) patients, 19 oral lichen planus (OLP) patients, and 19 oral lichenoid lesions (OLL) patients were obtained. 15 SCC specimens were also collected from PVL patients. Healthy individuals served as controls, and DNA was extracted from their paraffin-embedded tissues. 2bRAD-M sequencing generated taxonomic profiles. Alpha and beta diversity analyses, along with Linear Discriminant Analysis effect size analysis, were conducted.ResultsOur results showed the microbial richness and diversity were significantly different among groups, with PVL-SCC resembling controls, while OLK exhibited the highest richness. Each disease group displayed unique microbial compositions, with distinct dominant bacterial species. Noteworthy alterations during PVL-SCC progression included a decline in Fusobacterium periodonticum and an elevation in Prevotella oris.ConclusionsDifferent disease groups exhibited distinct dominant bacterial species and microbial compositions. These findings offer promise in elucidating the underlying mechanisms of this disease.

Prognostic significance of neutrophil to lymphocyte ratio (NLR) in oral squamous cell carcinoma

BackgroundTumor microenvironment is one of the evading research in the progression of cancers. Circulating neutrophils reflect the system's inflammatory response and study the growth of tumor cells thereby inducing cancer progression. Aim and objectivesThe current study aims to evaluate the NLR ratio in oral squamous cell carcinoma group and to compare it with that of normal control and Premalignant group. Material and methodsThis study is retrospective, the medical records of 48 OSCC patients, 48 Leukoplakia patients, and 48 normal healthy controls were taken. Association between NLR and clinicopathological variables were assessed. Tukey's post-hoc test was done to compare the variables. Kaplan Meier analysis was done for the analysis of overall survival. ResultsThe NLR ratio was assessed and compared between the three groups. The mean NLR ratio was found to be 3.79 in Oral squamous cell carcinoma patients, 2.62 in leukoplakia patients, and 2.07 in the normal control group. ConclusionAn increase in NLR levels was seen in OSCC patients. Increased NLR was also seen associated with advanced tumor staging and grading.

To Estimate and Compare the Level of Serum Immunoglobulin A in Patients of Oral Submucous Fibrosis, Leukoplakia, and Control Group.

To estimate and compare the level of serum immunoglobulin A in patients of oral submucous fibrosis (OSMF), leukoplakia, and control group. OSMF is a well-recognized potentially malignant condition in the oral cavity, and a transformation rate as high as 7.6% over a period of 10 years has been reported from India. Leukoplakia has evolved as a clinico-pathologic concept over many years, with the current clinical designation being accepted worldwide. Quantia IgA is a turbidometric immunoassay for the detection of IgA in human serum and is based on the principle of agglutination reaction. The test specimen is mixed with the activation buffer (R1) and then with antihuman IgA reagent (R2) and allowed to react. For the estimation of serum IgA, 500 μl of Quantia IgA activation buffer R1 is taken in a clean test tube. Serum sample is diluted in the ratio of 1:5 with normal saline. Ten microliters of diluted serum sample are added to R1. After incubation for 10 min, 50 μl of R2 is added, which is Quantia IgA antihuman IgA reagent to the sample, and the reading is recorded at wavelength 340 nm at 37°C. The level of IgA will be estimated by a semi-automatic biochemical analyzer machine and then data analysis will be done. The immunologic alterations observed by us in OSMF and leukoplakia are almost similar or nearer to the alterations observed in oral cancer, so it is reasonable to assume that OSMF and leukoplakia can be an intermediate stage in the transformation process of a normal cell to oral malignancy. Prevention and early detection is the best way to reduce the incidence and mortality of premalignant disorders and oral cancer. There are many ways and methods of early detection of premalignant disorders and oral cancer, and the immunological method is one of them.

Dermatoglyphic Pattens in OSMF and Leukoplakia - A Comparative Study.

The term dermatoglyphics was first used in 1926 by anatomist Cummins. Dermatoglyphic is a term that refers to "a skin carving." The unique carve present on the skin surface of humans and other living beings is referred to as dermatoglyphics. This research was conducted to understand the variations in patterns and assess the usefulness of dermatoglyphics among individuals diagnosed with potentially malignant diseases and mouth cancer. Three hundred subjects were selected. The study consisted of three groups. Patients were divided into the following groups: Group I: oral submucous fibrosis (OSMF) (100), Group II: ORAL LEUKOPLAKIA (100), and Group III: Healthy Individuals (100). Our research study indicates a difference in dermatoglyphic patterns between patients and healthy individuals. Specifically, there is a lower percentage of loop patterns and an increase in arch patterns in OSMF and leukoplakia patients. These findings suggest that dermatoglyphics may have relevance in understanding or identifying certain health conditions.

A Study to Assess the Role of Psychological Stress in the Severity of Oral Lichen Planus, OSMF, and Leukoplakia and its Correlation with Serum Cortisol Levels.

The most prevalent oral mucosal conditions in humans are leukoplakia, oral lichen planus (OLP), and oral submucous fibrosis (OSMF). These conditions should be studied as psychosomatic conditions, but little research has been done on how these conditions relate to stress. A total of 280 patients of both genders with ages 35-60 years were divided into four groups: leukoplakia, OSMF, OLP, and control. Whole blood samples were collected from all the participants and analyzed for serum cortisol by using ELISA with a commercial kit. Detailed case history and DASS-21 questionnaire were recorded for psychological assessment. To assess the variables, the student t test and Chi-square test were used. P values of <0.05 were regarded as significant. In this study, it was found that higher levels of serum cortisol were found in OLP patients, with advanced erosive lesions showing the highest levels of serum cortisol, followed by OSMF and leukoplakia. Patients with Oral Lichen Planus showed higher levels of stress, anxiety and depression than patients of OSMF and leukoplakia. This study emphasized stress and anxiety as a predisposing factor in the occurrence and progression of OSMF apart from re-establishing the role of stress and anxiety in OLP with consideration to depression in triggering role for OLP. Furthermore, it demonstrated that serum cortisol levels play a significant role as an indicator of psychological stress.

A study to evaluate the role of antioxidants and iron as diagnostic markers in oral cancer

Oral cavity (OC) and oropharyngeal (OP) cancers afflict about 500,000 people globally. Dental health care practitioners need to understand the causes, diagnosis, and treatment of oral cavities and oropharyngeal cancer. The constant and direct exposure of oral mucosal cells to tobacco product chemical carcinogens such as polynuclear aromatic hydrocarbons and nitrosamines causes the generation of free radicals/reactive oxygen species. Changes in salivary antioxidant and micronutrient levels may prove to be an important factor in the pathogenesis of oral premalignant and malignant disorders. The present study aimed to evaluate the levels of Vitamin C, MDA and iron levels in oral cancer, pre-malignant group and the oral cancer group. 15 oral cancer patients, 30 premalignant condition patients, and 15 normal controls were analyzed in the study. Among the 30 patients of the premalignant conditions- group (Group S1) 12 were diagnosed with oral submucous fibrosis, and 18 of them were diagnosed with oral leukoplakia. The oral cancer patients (Group S2) were segregated based on the TNM system from stages I to IV. The oral submucous fibrosis patients were subdivided into four grades. The oral leukoplakia patients were categorized into homogenous and speckled types. A detailed history was obtained from the patients. After obtaining informed consent, saliva samples will be collected from patients who have not consumed any food or any form of tobacco 2 hours before collection. Following a thorough mouth rinse using distilled water, saliva is allowed to accumulate in his or her mouth for 5 minutes. Accumulated saliva is collected by the spit method. 5ml of blood sample will also be collected from these patients, Serum was separated by Centrifugation. Collected serum and saliva were stored at -80℃ until further analysis. The serum and salivary samples were then analyzed for Vitamin C, iron, and malonaldehyde (lipid peroxidation) according to the standardized protocol. In the present study, a total of 60 samples were screened for Vitamin C, Iron, and MDA in both serum and saliva of normal control, premalignant, and oral cancer patients. We found that there was a decreased level of Vitamin C and iron in both serum and saliva of premalignant and oral cancer patients. MDA level was higher in premalignant, and oral cancer patients and there was a significant increase in the level of MDA in oral cancer patients. Understanding antioxidants can help reduce the incidence of oral cancer in its early stages. More study is needed before this supplement may be recommended as an adjuvant treatment.

Expression of salivary levels of S100A7 in oral submucous fibrosis and oral leukoplakia.

The aim of the study is to evaluate the expression of S100A7 levels in saliva of oral sub-mucous fibrosis, oral leukoplakia patients, and healthy control. The study comprised of saliva samples from 15 patients each with clinically diagnosed oral sub-mucous fibrosis, oral leukoplakia, and healthy control. Salivary S100A7 levels were estimated using Enzyme-Linked Immunosorbent Assay. Statistical analysis was performed using SPSS. The significance level is fixed at 5% (α = 0.05). To compare the mean values of concentration between the disease group oral leukoplakia (OL) and oral submucous fibrosis (OSMF) and control, one-way analysis of variance was used followed by a post hoc test for multiple pairwise comparisons. The results of the study indicated a statistically significant increase in the salivary S100A7 level among the OSMF and OL when compared with the control group. When a pairwise comparison was done between OSMF with a control group and leukoplakia with a control group, a statistically significant difference was observed, subsequently while comparing OSMF with leukoplakia, and no statistically significant difference was observed. Results from this study demonstrated increased S100A7 levels in OSMF and OL when compared with control group. This indicated that salivary S100A7 can be used as an adjunctive marker to identify patients at risk of progression into oral squamous cell carcinoma (OSCC).

Architectural and cytological features of epithelial dysplasia associated with transformation risk.

This study explored associations between histological features of dysplasia and malignant transformation, as well as genomic copy number alterations. Overall, 201 samples were collected from patients of oral leukoplakia. The associations of dysplastic features with malignant transformation and copy number alterations were investigated by Cox proportional hazards regression analysis and the Mann-Whitney U-test. Eight individual histological features, such as irregular epithelial stratification (p = 0.001), mitoses high in epithelium (p = 0.033), extension of changes along minor gland ducts (p < 0.001), etc., were associated with greater risk of malignant transformation. A model including histological features and age showed good performance for predicting malignant transformation (area under receiver operating characteristic curve: 0.806). Irregular epithelial stratification (p = 0.007), abnormal nuclear shape (p = 0.005), abnormal cell size (p = 0.004), etc. were associated with greater genomic instability. A Cox proportional hazards model using eight histological features and patient age reliably predicted the malignant potential of oral epithelial dysplasia. Identification of these histological features closely related to malignant transformation may aid the management of oral potentially malignant disorders and early detection of oral squamous cell carcinoma.

Dual Sec62/Ki67 immunocytochemistry of liquid-based cytological preparations represents a highly valid biomarker for non-invasive detection of head and neck squamous cell carcinomas.

Head and neck squamous cell carcinomas (HNSCC) are frequently diagnosed in advanced stages, which limits therapeutic options and results in persistently poor patient outcomes. The aim of this study was to use liquid-based swab cytology (LBC) in combination with dual immunocytochemical detection of migration and proliferation markers Sec62 and Ki67 in order to allow non-invasive early detection of HNSCC as well as to analyse the diagnostic validity of this method for predicting the malignancy of suspicious oral lesions. 104 HNSCC patients and 28 control patients, including healthy patients (n = 17), papilloma (n = 1) and leukoplakia patients (n = 10), were included in this study. For all patients, an LBC swab followed by simultaneous immunocytochemical detection of Sec62 and Ki67 was performed. Immunocytochemical as well as cytopathological results were correlated with histological diagnoses and clinical findings. All HNSCC patients (100%) showed dual Sec62/Ki67 positivity, and all control patients except for the papilloma patient were negative for Sec62/Ki67 (96.4%), resulting in a 100% sensitivity and 96.4% specificity of Sec62/Ki67 dual stain for non-invasive detection of HNSCC. The positive predictive value was 99% and the negative predictive value was 100%. Sec62 expression levels showed a positive correlation with tumour de-differentiation (p = 0.0489). Simultaneous immunocytochemical detection of Sec62/Ki67 using LBC represents a promising non-invasive and easy-to-apply tool for the early detection of HNSCC in routine clinical practice. This novel technique can help to avoid incisional biopsies and reduce the frequency with which general anaesthesia is used in diagnostic procedures in patients with suspicious oral lesions.

The role of differentiated dysplasia in the prediction of malignant transformation of oral leukoplakia.

Oral leukoplakia is the most common oral potentially malignant disorder. Malignant transformation of oral leukoplakia occurs at an annual rate of 1%-7%. WHO-defined classic epithelial dysplasia is an important predictor of malignant transformation of oral leukoplakia, but we have previously shown in a proof of concept study that prediction improves by incorporation of an architectural pattern of dysplasia, also coined as differentiated dysplasia. We aimed to analyze this finding in a larger cohort of patients. For this retrospective study 176 oral leukoplakia patients were included. Biopsies for all patients were assessed for the presence of dysplasia and analyzed for cytokeratin 13 and 17 expression. Moreover, the inter-observer agreement for the diagnosis of differentiated dysplasia was determined. In total, 33 of 176 patients developed oral squamous cell carcinoma during follow-up. Presence of classic epithelial dysplasia increased cancer risk two-fold (HR = 2.18, p = 0.026). Lesions without classic epithelial dysplasia could be further risk-stratified by the presence of differentiated dysplasia (HR = 7.36, p < 0.001). Combined classic epithelial and differentiated dysplasia imparted a seven-fold increased risk of malignant transformation (7.34, p = 0.001). Inter-observer agreement for the diagnosis of dysplasia, including differentiated dysplasia, was moderate (κ = 0.56, p < 0.001). This study emphasizes the importance of the recognition of the architectural pattern of differentiated dysplasia as a separate entity for risk prediction of malignant transformation of oral leukoplakia. Presence of any pattern of dysplasia results in accurate prediction of malignant transformation risk of oral leukoplakia.

Treatment experience with and clinicopathological analysis of vocal fold leukoplakia per appearance classification guidance: a cohort of 1442 patients.

To analyse the comparative clinical outcomes and clinicopathological significance of vocal fold leukoplakia lesions treated by appearance classification and traditional methods. A total of 1442 vocal fold leukoplakia patients were enrolled. Group A patients were treated according to appearance classification and Group B patients were treated according to traditional methods. In Group A, 24.4, 14.9 and 60.6 per cent of patients had grade I, II and III dysplasia, respectively. Grade I dysplasia (63.4 per cent) was more than twice as frequent in Group B patients than in Group A patients, while grade II dysplasia (20.4 per cent) and grade III dysplasia (16.2 per cent) were significantly less frequent in Group B patients than in Group A patients (p = 0.000). There was a significant correlation between vocal fold leukoplakia appearance and the degree of dysplasia (p = 0.000). The recurrence and malignant transformation rates (17.6 and 31 per cent, respectively) in Group B were significantly greater than those in Group A (10.8 and 25.9 per cent, respectively) (p = 0.000). Vocal fold leukoplakia appearance classification is useful for guiding treatment decision-making and could help to improve therapeutic accuracy.

Clinical Significance of Pathogenicity of Somatic Mutations in Oral Leukoplakia: a Prospective Observational Study

Background. The vast majority of malignant neoplasms of the oral mucosa refer to squamous cell carcinomas. The development of squamous cell carcinoma of the oral mucosa is often promoted by previous potentially malignant diseases, with oral leukoplakia dominating among them.Objective. To determine the clinical significance of the pathogenicity of somatic mutations in oral mucosal leukoplakia.Methods. The study material included 24 samples of abnormal epithelium of the oral mucosa from leukoplakia patients. QIAamp DNA FFPE Tissue Kit (Qiagen, Germany) was used for deoxyribonucleic acid (DNA) extraction from the samples. DNA sequencing was performed using IlluminaNextSeq 550 sequencer and TruSight™ Oncology 500 DNA Kit For Use with NextSeq (Illumina, USA). All DNA extractions from biological samples, preparation and sequencing of DNA libraries were performed step-by-step in strict accordance with the guidelines provided with the respective reagent kits. Bioinformatics analysis was carried out using specific software Illumina Base Space (Illumina, USA) and Galaxy Project (The Galaxy Community, a non-profit international project) according to current guidelines. The desired power of the study accounted for 90%. Two Proportions Z test was performed by means of The Sample Size Calculation of Statistica 12 (StatSoft, Inc.) with the set option “one-tailed hypothesis”, because it was initially assumed that pathogenic (oncogenic) genetic variants occur in the tissue of oral leukoplakia much more frequently than in the human reference genome used for sequence alignment.Results. The pathogenic somatic mutations in the TP53, KRAS, APC, NRAs and BRAF genes, identified in this study, alone or in combination, are highly likely (hazard ratio 3000-11000) to be associated with the development of oral mucosal leukoplakia and low-grade epithelial dysplasia. The multiplicity of pathogenic and likely pathogenic genetic variants associated with epithelial dysplasia, as well as the fact that a number of variants do not occur in all patients, suggests that the same histotype of oral mucosal dysplasia may develop under the influence of different mutations.Conclusion. The pathogenic and likely pathogenic variants of the TP53, KRAS, APC, NRAS and BRAF genes, identified in this study, alone or in combination, are highly likely (hazard ratio 3000–11000) to be associated with the development of leukoplakia and low-grade epithelial dysplasia.

Evaluation of salivary Chemerin in oral leukoplakia, oral squamous cell carcinoma and healthy controls

Introduction: Chemerin is a multifunctional chemoattractant cytokine, encoded by RARRES2 gene. Its role in oral carcinogenesis is largely unexplored. Study was undertaken to evaluate the role of salivary chemerin as a potential biomarker in the early diagnosis of oral cancer and to elucidate the clinicopathologic and prognostic significance of chemerin in different grades of Oral Squamous Cell Carcinoma (OSCC) and oral leukoplakia. Materials and Methods: The study comprised 96 subjects, divided into 3 groups, each consisting of 32 subjects. These groups were clinically and histopathologically diagnosed with OSCC, oral leukoplakia, and healthy controls, respectively. The whole unstimulated saliva (1.5 mL) was collected from the subjects using passive drool technique and immediately centrifuged at 3000 rpm for 10 minutes. The salivary chemerin level was estimated using the competitive sandwich enzyme-linked immunosorbent assay method. The statistical analysis was performed using R software EZR version 1.32. Independent t-tests, analysis of variance tests, and Scheffe Multiple Comparisons were carried out to compare quantitative parameters. A receiver operating characteristic (ROC) curve was plotted to assess diagnostic accuracy of salivary chemerin. Results: Significantly higher levels of salivary chemerin were observed in OSCC and oral leukoplakia patients compared to healthy controls (P &lt; 0.05). The study further established an increase in salivary chemerin concentration in different histological grades of OSCC and oral leukoplakia. ROC curve analysis suggested that salivary chemerin is a highly sensitive and specific indicator for early screening and detection of oral leukoplakia and OSCC. Conclusion: Salivary chemerin can be considered as a reliable and specific indicator for the early screening and detection of oral leukoplakia and OSCC and a highly sensitive biomarker which may aid in detection of malignant transformation of oral leukoplakia.

A study to estimate the serum IgA and salivary IgA levels in patients with oral leukoplakia and oral squamous cell carcinoma.

The increasing death rate because of oral cancer is mainly due to its late diagnosis. Tumour markers are often detected in abnormal amounts in blood, urine or saliva of patients with certain types of cancer. Diagnosing cancer through human saliva has advantages such as low invasiveness, minimum cost and easy sample collection. We have used serum immunoglobulin A (IgA) and salivary IgA for our present study. The aim of present study was to estimate serum and salivary IgA levels in oral leukoplakia and oral squamous cell carcinoma (OSCC) patients. The study included 40 patients; 10 in the control group, 15 cases with oral leukoplakia and 15 cases with OSCC. The blood samples and saliva were taken from clinically diagnosed oral leukoplakia and OSCC patients and were tested for IgA levels. The data were analysed using SPSS 16.0. The mean values were compared between the groups by using analysis of variance (ANOVA) followed by post-hoc test for group-wise comparison. P value ≤ 0.05 was considered significant. It was observed that the comparison of levels of serum IgA in control and leukoplakia group; control and OSCC group; leukoplakia and OSCC group were found to be statistically significant. Also, comparison between the levels of salivary IgA in control and OSCC group was found to be statistically significant. It is suggested that the serum and salivary IgA levels could be a better adjuvant diagnostic marker along with routine markers in patients with premalignant and malignant lesions.

Clinicopathological correlation of microRNA 21 expression in oral cancer patients with tobacco chewing habits

Introduction: Oral cancer is one of the most common malignancies and has a significant association with tobacco chewing. Micro RNA 21 (mi-RNA 21) has been identified as oncogenic RNA, implicated in etio-pathogenesis of various neoplastic lesions. Objective: (i) To assess the microRNA 21 expression in oral cancer patients with history of tobacco chewing habits and (ii) to carry out its clinicopathological correlation. Material and Method: The present study was carried out as a case-control study, for a two year period in which a total of 20 biopsy tissue specimen obtained from oral cancer patients having tobacco chewing habits were enrolled as cases and an equal number of biopsy tissue specimen obtained from age, and sex-matched patients of oral leukoplakia were enrolled as controls. Clinical details, personal habits were noted and histopathological assessment was done. miRNA21 was done using Taq Man microRNA assay (ABI) using Polymerase Chain Reaction (PCR).Real time PCR results was analyzed and expressed as relative miRNA expression of the threshold cycle (CT) values. Mean values were compared using independent samples ‘t’-test and ANOVA using SPSS 21.0 software. p-value less than 0.05 indicated a statistically significant association. Results: Mean age of cases was 52.65±7.82 years, majority of cases were males (85%). Among cases, majority (70%) had clinical stage I and were histo-pathologically well differentiated (75%). There were 3 (15%) cases with metastasis. Mean relative miRNA 21 expression were significantly higher in cases (10.53±3.52) as compared to that in controls (8.76±1.16) (p=0.029). Relative miRNA21 values showed a significant increasing trend with increasing clinical stage, histopathological grade and metastasis. Conclusion: miRNA21 expression was able to differentiate between leukoplakia and oral cancer cases. miRNA21 also showed a significant correlation with clinicopathological factors.

Predicting oral cancer risk in patients with oral leukoplakia and oral lichenoid mucositis using machine learning

Oral cancer may arise from oral leukoplakia and oral lichenoid mucositis (oral lichen planus and oral lichenoid lesions) subtypes of oral potentially malignant disorders. As not all patients will develop oral cancer in their lifetime, the availability of malignant transformation predictive platforms would assist in the individualized treatment planning and formulation of optimal follow-up regimens for these patients. Therefore, this study aims to compare and select optimal machine learning (ML)-based models for stratifying the malignant transformation status of patients with oral leukoplakia and oral lichenoid mucositis. One thousand one hundred and eighty-seven patients with oral leukoplakia and oral lichenoid mucositis treated at three tertiary health institutions in Hong Kong, Newcastle UK, and Lagos Nigeria were included in the study. Demographic, clinical, pathological, and treatment-based factors obtained at diagnosis and during follow-up were used to populate and compare forty-six machine learning-based models. These were implemented as a set of twenty-six predictors for centers with substantial data quantity and fifteen predictors for centers with insufficient data. Two best models were selected according to the number of variables. We found that the optimal ML-based risk models with twenty-six and fifteen predictors achieved an accuracy of 97% and 94% respectively following model testing. Upon external validation, both models achieved a sensitivity, specificity, and F1-score of 1, 0.88, and 0.67 on consecutive patients treated after the construction of the models. Furthermore, the 15-predictor ML model for centers with reduced data achieved a higher sensitivity for identifying oral leukoplakia and oral lichenoid mucositis patients that developed malignancies in other treatment settings compared to the binary oral epithelial dysplasia system for risk stratification (0.96 vs 0.82). These findings suggest that machine learning-based models could be useful potentially to stratify patients with oral leukoplakia and oral lichenoid mucositis according to their risk of malignant transformation in different settings.

MiRNAs as non-invasive biomarkers in the serum of Oral Squamous Cell Carcinoma (OSCC) and Oral Potentially Malignant Disorder (OPMD) patients

ObjectiveCell-free microRNAs have shown differential levels in the serum of individuals under disease conditions suggesting its potential to act as biomarkers. A population specific miRNA signature in oral cancer is reported in different studies. We aim to identify a set of serum specific miRNAs that may differentiate oral cancer, oral pre-malignant conditions from the healthy individuals. DesignWe investigated the levels of 24 miRNAs in the serum of 47 Oral squamous cell carcinoma (OSCC) patients, 20 patients with Oral potentially malignant disorders (OPMD) and 42 healthy controls from Eastern India. Small RNAs were isolated from serum samples followed by cDNA synthesis. Levels of miRNAs were determined using qRT-PCR. The sources of serum specific miRNAs were evaluated using GTEx-RNAseq and TCGA-HNSCC database. ResultsFive miRNAs, miR-483–5p, miR-31–5p, Let-7b-5p, miR-486–5p and miR-30e-5p showed significant elevation in OSCC patients. An Elastic-Net model with 4 miRNAs classified OSCC from healthy controls with 80 % sensitivity, 64.3 % specificity, and 72.4 % accuracy. Mir-483–5p and miR-31–5p was significantly overexpressed in OSCC tissues as well as significantly higher in the serum of Leukoplakia and Verrucous carcinoma patients suggesting their potential as early disease markers. MiR-483–5p showed a consistent elevated level in the serum/plasma of oral cancer patients across different population and was found to be tumour specific while, the rest of the miRNAs showed variable results across different studies. ConclusionsOur study suggested that the serum miRNAs in oral cancer and pre-malignant disorder conditions can be used as a non-invasive marker for screening of these oral conditions.

Correlation of serum trace elements (Fe, Cu, and Zn) in the blood samples of Indian patients with leukoplakia, oral squamous cell carcinoma, and normal subjects.

Trace elements (iron, copper, and zinc), leukoplakia, and oral squamous cell carcinoma. To study and correlate serum trace element (iron, copper, and zinc) levels in leukoplakia, oral squamous cell carcinoma, and normal subjects. The present study comprised a total of 80 patients, which included 30 patients of leukoplakia, 30 patients of oral squamous cell carcinoma, and the normal control group comprising 20 healthy individuals who were not having any relevant medical, dental, and habit history. Peripheral blood samples of a volume of 10 ml each will be collected by anti-cubical vein puncture from control groups and from patients suffering from leukoplakia and oral squamous cell carcinoma. The blood will be collected in a plain red top vein puncture tube without additives or anti-coagulants and allowed to stand undisturbed to clot at room temperature, and serum will be separated from the cells by centrifugation at 4°C at a speed of 3000 rev/min, the separated sera will be kept at -20°C until analysis is performed. The estimation of serum zinc (Zn) and copper (Cu) levels is performed by atomic absorption spectrometry (AAS). In the present study, the estimation of copper and zinc levels was conducted by using an atomic absorption spectrophotometer (model no: AA-6300 SHIMADZU, Japan). Serum iron estimation is performed using the kit RANDOX (siedel, 1984). Statistical analysis is performed by the paired and Scheffe tests. From the results, it was concluded that there was a decrease in serum iron and zinc levels and copper had increased serum levels. It was concluded that evaluation of serum trace elements can be a cost-effective and non-invasive alternative for screening, diagnosis, and monitoring of pre-malignant lesions such as leukoplakia and malignant lesions such as oral squamous cell carcinoma. Thus, these parameters can be used as biomarkers that provide important tools in formulating an adequate diagnosis, treatment plan, and prognosis for oral squamous cell carcinoma.

A comparative study to evaluate the therapeutic effects of nutraceuticals in oral leukoplakia:- A randomized clinical trail.

Oral leukoplakia, usually white changes in the oral mucosa, is one of the most common conditions affecting the oral cavity. Oral leukoplakia can occur anywhere in the mouth and is usually asymptomatic. Clinical diagnosis is reliant on visual inspection and manual palpation. It has a global prevalence of 2.6% and a malignant transformation rate of 0.13-34%. In India, OL has a higher prevalence (0.2-5.2%) but a lower a malignant transformation rate (0.13-10%). It was a randomized control trial in which study was conducted on clinically diagnosed 300 oral leukoplakia patients. All patients were randomly categorized in three groups of 100 each. Group-A: Patients were given commercially available curcumin 500 mg. daily orally. Group-B: Patients were given 4 mg of oral lycopene daily. Group-C: Patients were treated with 4 mg of lycopene + 500 mg curcumin daily by oral route. After recording the pre-treatment clinical findings, all the participants were evaluated regularly after 30 days, 60 days and 90 days of active treatment and once in a month for another 3 months of post-treatment follow-up and to evaluate concomitant medication, lesion(s), compliance, and adverse events. The clinical response was evaluated by bi-dimensional measurement of the lesions and color photography. Safety assessment measures: Physical examination and laboratory tests were performed at baseline, and every 30 days for 3 months after randomization. Number of participants cured after treatment with oral curcumin was 51%. Participants took lycopene tablets showed 63% cure rate and 72% participants cured after treatment with combination curcumin and lycopene. Results showed that curcumin, lycopene, and a combination of the two are effective in the treatment of oral leukoplakia. When compared, we found that lycopene is a better nutraceutical as compared to curcumin. When both nutraceuticals were given to the participants, they showed better results than single nutraceuticals when the data were analyzed after 90 days of treatment. There is a significant difference in the response of curcumin and combinations of both nutraceuticals, although the difference between lycopene and combinations of curcumin and lycopene is insignificant.

Elucidating the Genetic Landscape of Oral Leukoplakia to Predict Malignant Transformation.

Oral leukoplakia is the most common oral potentially malignant disorder with an annual malignant transformation rate of 1% to 5%. Consequently, oral leukoplakia patients have a 30% to 50% lifetime risk to develop oral squamous cell carcinoma. Although risk factors for malignant transformation of oral leukoplakia have been investigated, no definitive risk stratification model has been proposed. Next-generation sequencing can elucidate the genetic landscape of oral leukoplakia, which may be used to predict the risk for malignant transformation. We investigated a retrospective cohort of 89 oral leukoplakia patients, and analyzed their oral leukoplakia lesions for the presence of genomic copy-number alterations and mutations in genes associated with oral squamous cell carcinoma. In 25 of 89 (28%) patients, oral squamous cell carcinoma developed during follow-up. Seventy-nine of 89 (89%) oral leukoplakias harbored at least one genetic event. Copy-number alterations were present in 61 of 89 (69%) oral leukoplakias, most commonly gains of chromosome regions 8q24 (46%) and 20p11 (20%) and loss of 13q12 (19%). Mutations were present in 59 of 89 (66%) oral leukoplakias, most commonly in TP53 (28%), FAT1 (20%), and NOTCH1 (13%). Genetic data were combined with the presence of dysplasia to generate a prediction model, identifying three groups with a distinct risk for malignant transformation. We provide an extensive description of genetic alterations in oral leukoplakia and its relation to malignant transformation. On the basis of our data we provide a model for the prediction of malignant transformation of oral leukoplakia using dysplasia and genetic markers.