Bioimpedance a Diagnostic Tool for Tobacco-induced Oral Lesions – A Mixed Model Cross-sectional Study

Salivary Lactate Dehydrogenase (sLDH) levels seem to be higher in patients with Oral Squamous Cell Carcinoma (OSCC) and Oral Potentially Malignant Disorders (OPMD) than a control group (CG). Case-control study. Patients with OPMD [oral leukoplakia (OL) and oral lichen planus (OLP)] and OSCC who attended two services in Spain were selected. sLDH in saliva was measured. Epidemiological, periodontal and specific variables related to OPMD and OSCC were collected. A total of 92 patients were included: 12 with OSCC, 51 with OPMD (17 OL and 34 OLP), and 29 controls. sLDH values were higher in the OSCC, followed by the OPMD and CG groups, although no significant differences were observed. In the OSCC group, larger tumor size was associated with higher sLDH levels. In the OLP group, sLDH values were higher in patients with symptomatic lesions than in patients with only white lesions, but not significantly. No associations were observed between sLDH and the type of OL (homogeneous vs. non-homogeneous) and the degree of dysplasia. When analyzing periodontal variables among OSCC, OPMD and CG, periodontal probing depth (PPD) and bleeding on probing were significantly higher in the OSCC group, while the plaque index was higher in OPMD patients. The linear regression model for sLDH in the total group identified age and PPD as significant predictors of sLDH levels. Although sLDH values were higher in OSCC and OPMD patients than in a CG, the results do not support the use of sLDH as a reliable prognostic biomarker of malignancy. Future studies need to consider other factors that may influence sLDH levels, such as age and periodontal status.

Introduction:Oral squamous cell carcinoma (OSCC) is sixth most common cancer in Indian subcontinent with a high morbidity and mortality rate. It is frequently preceded by oral potentially malignant disorders (OPMD). Early diagnosis of OSCC might halt the poor prognosis. Therefore, present study was aimed to evaluate the diagnostic significance of preoperative hematological parameters and serum lipid indices in OSCCs and OPMDs.Material and Methods:Study included 75 cases each of OSCC, OPMD, and controls. Mean and standard deviation of various hematological and lipid parameters were evaluated, including: a) RBC metrics: RBC count, hemoglobin (Hb%), hematocrit, MCV, MCH, RDW. b) WBC metrics: WBC count, neutrophils, lymphocytes, monocytes, atypical lymphocytes, immature leukocytes count, NLR. c) Platelet metrics: Platelet count, MPV, PDWSD, PCT, PLCR. d) Lipid metrics: Total Cholesterol, Triglycerides, HDL, LDL, VLDL, Non-HDL. The parameters were statistically analyzed using one-way ANOVA.Result:Mean values of neutrophil count, immature leucocytic cells, NLR, MPV, PDWSD, PLCR, and VLDL were found to be significantly increased (P < 0.00), while, mean values of RBC count, Hb%, atypical lymphocytes, platelet count and HDL were observed to be significantly decreased (P < 0.05) in OPMD and OSCC groups as compared to control. However, the remaining indices were found to be non-significant.Conclusion:Present study findings corroborate the role of hematological and serum lipid profile parameters in the etiology and progression of OPMDs and OSCCs. This highlights their potential as early diagnostic predictive markers to forecast the occurrence of high-risk OPMD and OSCC cases and also guiding personalized treatment strategies.

Objective:Objectives of this study were to compare expression of Programmed Death-Ligand 1(PD-L1) protein in oral squamous cell carcinoma (OSCC) and oral potentially malignant disorder (OPMD) cases; and to compare the PD-L1 protein expression in histological grades of OSCC and also in OPMD’s with Dysplasia and without Dysplasia. Materials & Methods:In this study, 25 cases of Oral squamous cell carcinoms, 25 cases of Oral Potentially Malignant Disorders and 10 cases of non-neoplastic oral mucosa (control) cases were included. FFPE blocks of OSCC and OPMD cases were contributed by Department of Pathology, Histopathology Division,Pakistan Institute of Medical Sciences, Islamabad. Immunohistochemical staining of cases with PD-L1 monoclonal antibody (1:100; Dako) was carried out at Histopathology division , PMC Labs, Peshawar Medical College,Peshawar, Riphah International University, Islamabad . Epithelial cells (membranous and cytoplasmic) positivity was observed for PD-L1 Antibody. Data was analyzed in SPSS version20. For qualitative variables frequencies and percentages were calculated whereas for quantitative variables means and standard deviations were recorded. The Chi-square test was applied to evaluate the significant difference in categorical variables . p-value of ≤0.05 was taken as significant.Results:PD-L1 expression in OSCC cases turned out to be 48% (n=12/25) as compared to 8% of OPMD cases (n=2/25) with a significant p value of 0.002 and all non-neoplastic oral mucosa cases were negative. PD-L1 expression in high grade OSCC cases was quite high (61% n=11/18) as compared to low grade OSCC (14% n=1/7) cases with a significant p value of 0.035. Conclusion:A statistically significant increased PD-L1 expression was noted in OSCC as compared to OPMD. Expression of PD-L1 was more intense in high grade OSCC cases. The relation of PD-L1 expression to age ,gender or location of OSCC and OPMD cases , and presence of dysplasia in OPMD cases was statistically not significant.

Introduction: Oral squamous cell carcinomas (OSCCs) are malignancies which develop predominantly from preexisting potentially malignant disorders. Immunoglobulin A (IgA) in saliva and serum is responsible for local immune response. The present study evaluates the role of serum and salivary IgA in OSCC and oral potentially malignant disorders (OPMDs). Materials and Methods: Ninety individuals were recruited into the study in three groups of thirty each. The participants were categorized as control group, study Group I consisting of OPMDs, and study Group II consisting of OSCC. Serum and salivary IgA was analyzed using nephelometry. Results: Serum IgA showed statistically significant increase between controls and study Group II and between study Group I and study Group II (P < 0.05). Salivary IgA showed statistically significant difference between controls and study Group I and between controls and study Group II (P < 0.05). Conclusion: The results show increased serum IgA in OSCC group as compared to controls while salivary IgA increased in both OPMD and OSCC groups. Thus, the present study indicates a role for serum IgA in OSCC while salivary IgA has potential as a prognostic indicator in OPMDs and OSCCs.

Oxidative stress has been implicated in tumourigenesis by affecting the actions of various protein kinases and transcription factors, and resulting in damage to DNA structure by the deamination of nucleotides and the generation of free radicals. Derangements in the antioxidant defence mechanism have been considered in the pathogenesis of oral cancer. To evaluate and compare the serum levels of nitric oxide and vitamin C in cases of Oral Potentially Malignant Disorders (OPMDs) and Oral Squamous Cell Carcinoma (OSCC) and healthy controls. Serum from 20 patients with OSCC, 20 patients with OPMDs and 10 healthy subjects were analyzed for levels of nitric oxide and vitamin C using UV spectrophotometry. Antioxidant-Oxidant Index (AOI) was calculated by calculating the ratio between the levels of nitric oxide and vitamin C. The mean nitric oxide levels were elevated in OSCC group, as compared to OPMD and control group, whereas, mean serum vitamin C levels were reduced in OSCC group as compared to OPMDs and control group (statistically significant, p<0.05). There was significant increase in AOI from control group (0.023), OPMDs (0.167) and OSCC group (0.279) (p<0.05). Assessment of levels of antioxidant oxidant markers are useful tools in monitoring the progression of OPMDs to OSCC. AOI is an objective tool to assess the oxidative stress in these patients.

Background: Bacterial and virus infection may cause pre-malignant and malignant disorder such as hepatocellular carcinoma and cervical cancer. Recent studies reported that oral bacterium is associated with gastric and colon cancer. However, it is not clear yet if the oral microbiome affects the development of pre-malignant disorder in oral cavity and oral cancer. Thus, this study aims to identify the periodontal pathogen candidates involved in the onset and progression of oral squamous cell carcinoma (OSCC). Materials and Methods: Saliva was collected from 112 patients without oral mucosal diseases (OMDs) as control, 36 patients with oral potentially malignant disorders (OPMDs), and 104 patients with OSCC, and DNA was extracted in each sample. Periodontal examinations were performed on all patients. Endpoint-PCR was performed on seven species of oral pathogens using the extracted DNA. 16S ribosomal RNA (rRNA) was utilized for analysis of oral microbiome with 20 samples in each group. 4-nitroqquinoline-1-oxide (4-NQO)-induced tongue tumor model in mice was analyzed with or without endotoxin of one of the oral bacteria, Porphyromonas gingivalis (P. gingivalis). Results: From the statistical analysis of the clinical parameters, periodontal disease tended to worsen in the OMPD and OSCC groups compared to the control group. The number of oral bacteria was significantly higher in the OSCC group than those in other groups. From PCR results, the detection rates of P. gingivalis, Aggregatibacter actinomycetemcomitans, and Treponema denticola were significantly higher in the OSCC group than those in the control group. From 16S rRNA analysis, the relative abundance of Prevotella buccae and intermedia was significantly higher in OSCC groups than in the control group. Furthermore, the relative abundance of P. gingivalis was higher in the OSCC group compared to other groups, regardless of age and the number of teeth. In mouse model, mice treated with P. gingivalis-derived endotoxin developed oral leukoplakia at earlier time points and tumors significantly higher lesions compared to control group. Conclusion: These results suggest that specific periodontal pathogens are present in the oral cavity of OPMDs and OSCC patients and those changes in the bacterial flora due to their presence may contribute to the onset and progression of OMDs. Citation Format: Mitomu Kioi, Hiroshi Isono, Shintaro Nakajima. Specific oral microbiome is closely associated with oral potentially malignant disorders and oral squamous cell carcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4238.

The incidence of oral squamous cell carcinoma (OSCC) of the tongue is increasing in the younger population without traditional risk habits that lead researchers to find other related factors such as diet and viruses, especially human papillomavirus (HPV). It is noteworthy that many OSCCs develop from oral potentially malignant disorders (OPMDs). Correct diagnosis and timely management of OPMDs may help to prevent malignant transformation, and therefore it is worth seeing the involvement of HPV in OPMDs and oral cancers, as the preventive and curative measures in HPV-induced cancer types are different from the conventional types of OPMDs and OSCCs. Therefore, the main objective of this study was to identify a relationship between HPV and p16 in OPMDs and compare it with OSCC. This study was conducted on 83 cases of known OSCCs and OPMDs (oral submucous fibrosis, leukoplakia, and oral lichen planus). Assays, such as polymerized chain reaction (PCR) and reverse transcription-PCR, were carried out for HPV and p16. The results were compared with clinical information and with the literature. The results were analyzed using SPSS 16.0 for windows. P16 expression was mostly seen in males than in female patients. Out of 21 cases of keratosis with dysplasia, 19% expressed p16. Of 26 oral lichen planus patients, 29% showed the p16 gene with immunohistochemistry. Interestingly, a high percentage of OSF cases expressed p16 (48.27%). Minimal expression was observed in OSCC (6.25%). HPV DNA was detected in 2.4% of the total sample. Both p16 and HPV were detected in a single case of OSCC. OPMDs expressed a significant amount of the p16 gene by immunohistochemistry and reverse transcription-PCR technique when compared with malignant lesions, suggesting a possible inactivation of the p16 gene. HPV and p16 are mostly negative in our OSCC sample, exhibiting low prevalence. OPMDs expressed a significant amount of the p16 gene when compared with malignant lesions, suggesting a possible inactivation of the p16 gene. Although OSF expressed p16, HPV was not detected, suggesting that over-expression could be independent of HPV. OSCC shows low HPV prevalence.

Introduction:Oral squamous cell carcinoma (OSCC) accounts for 95% among all head and neck cancers. Literature reveals saliva as a predictive, diagnostic and prognostic tool in carcinoma, inflammatory and genetic disorders. Expression of salivary interleukin-6 (IL-6) has been reported in patients with OSCC and in oral potentially malignant disorders (OPMDs). This study aims at the following objectives: • To evaluate the changes in the salivary levels of IL-6 in healthy individuals and those with chronic periodontitis (CP), OPMD and OSCC. • To compare the estimated levels of salivary IL-6 individually in healthy individuals and those with CP, OPMD and OSCC. • To assess the estimated levels of salivary IL-6 individually within histopathologically differentiated OSCC. • To analyse salivary IL-6 as a reliable biomarker in the diagnosis of OSCC.Materials and Methods:Totally, 60 patients were divided into four groups consisting of 15 patients in each group. Salivary samples were collected by simple drooling method. The concentration of IL-6 is to be determined by using Quantitative sandwich ELISA technique. All analyses were carried out using Statistical Package for Social Sciences (SPSS).Results:The concentration values of IL-6 were found to be more in OSCC group in comparison with the other three groups and the concentration values of OPMD were found to be more than in the CP and control group and was statistically significant.Discussion:We attempted a study to evaluate the salivary IL-6 in patients with OSCC, OPMDs and CP in comparison with the healthy controls. We achieved a pragmatic result showed that salivary IL-6 can be a reliable biomarker in the detection of OSCC. Saliva, due its wide array of functional characteristics, is an upcoming diagnostic fluid in the field of medicine and salivary IL-6 can be one such biomarker in the diagnosis of OSCC.

Oral Squamous Cell Carcinoma (OSCC) is the 12th most common cancer in the world. 90-95% of oral cancer is OSCC and it can be preceded by oral potentially malignant disorder (OPMD). Diabetes mellitus (DM) is a major global health issue and various epidemiological studies have implicated diabetes mellitus as a risk factor for the development of oral squamous cell carcinoma and other oral potentially malignant disorders. The study aimed to evaluate the correlation of diabetes mellitus in OSCC, leukoplakia and oral lichen planus patients and also evaluate age and gender predilection in OSCC and OPMD cases with diabetes mellitus. This retrospective study included a sample size of n=226 [156-OPMD, 70-OSCC]. Data regarding the sample cases were selected after the review and analysis of records of about 86000 patients who visited our institution between June 2019 and March 2020. From n=226, 30% of OSCC cases showed the positive diabetic history and 15% of OLP and 8.3% of leukoplakia were associated with DM. A statistically significant association found between diabetes in OSCC, Leukoplakia, OLP cases with p-value (0.001). 90.48% of OSCC and 75% of leukoplakia cases with a positive diabetic history were males. Among OLP, 77.8% of cases with diabetes mellitus were females. The association of prevalence of diabetes mellitus with gender in Oral squamous cell carcinoma(OSCC), Leukoplakia and Oral lichen planus(OLP) was found to be statistically significant (p=0.001). The most prevalent age range for both OSCC and OPMD patients with a history of diabetes mellitus was found to be 50-60 years, followed by above 60 years. In conclusion, there found to be a significant prevalence of DM in OSCC and OPMD cases. The DM status of oral cancer and OPMD patients may have a significant impact on their disease progression and treatment. DM could be considered as a prognostic factor for OSCC and OPMD.

Oral squamous cell carcinoma (OSCC), a growing global health concern, is often preceded by oral potentially malignant disorders (OPMDs). Emerging evidence suggests mitochondrial dysfunction is crucial in carcinogenesis, affecting malignancy's aggressiveness. This study uses Janus Green B (JGB) staining to investigate mitochondrial dysfunction in OPMDs and OSCC and examines its relationship with tobacco exposure to better understand its etiopathogenesis. Utilizing the exfoliative cytology technique, thirty buccal smears of epithelial cells from normal, OPMD and OSCC samples were collected and subjected to JGB staining. The intensity of staining and the total percentage of positively stained cells were quantified and subsequently correlated with clinicopathological parameters. The mean values of varying grades of staining intensity including mild, moderate, and high as well as the total percentage of positive cells, demonstrated statistically increased significant differences among control, OPMD, and OSCC cases. Additionally, moderate, high-grade staining intensity and the total positive stained cells exhibited significant correlations with tobacco use frequency and duration. A cut-off value of 44.31 and 51.32 of JGB-positive cells was found to categorize the cases into high-risk OPMDs and OSCC transformation. The progressive increase in staining intensity and the number of positive cells from OPMDs to increasing histopathological grades of OSCC highlights mitochondrial dysfunction as a key factor in oral carcinogenesis and a reliable indicator for assessing malignant transformation in OPMDs and aggressive potential in OSCC. Furthermore, the study underscores tobacco as a primary cause of mitochondrial dysfunction in both OPMDs and OSCC.

Background: Oral squamous cell carcinoma (OSCC) accounts for nearly 90% of oral malignancies and represents a major global health care problem. It is often preceded by oral potentially malignant disorders (OPMD). Although regular clinical examination forms the backbone for oral cancer screening, subtle lesions go unnoticed and there is a need for more sensitive and specific molecular biomarkers in mass screening of population. Salivary proteomics offer an attractive alternative to serum and tissue testing. Aims: To find the diagnostic utility of salivary interleukin-6 (IL-6) in the differential diagnosis of OSCC, OPMD from healthy controls. Study Design: In vivo study. Methods: After approval from the Institutional Review Board, unstimulated whole saliva was collected from 90 subjects, 30 in each group of OSCC, OPMD and controls after ethical clearance. Salivary IL-6 was measured by ELISA, and the results were statistically analysed. Results: Significant difference in salivary IL-6 was seen between OSCC, OPMD and controls. Receiver operating characteristic curve analysis showed the highest area under a curve of 0.982 in distinguishing OSCC from controls. It showed a sensitivity of 71% and specificity of 100% at a cut-off value of 33.4 pg/mL (P = 0.000). Moderately differentiated OSCC (MDSCC) showed a significant increase in salivary IL-6 concentration compared to well-differentiated OSCC (WDSCC). Conclusion: Results of the present study showed strong predictive power of salivary IL-6 in distinguishing OSCC from controls. Its levels also increased with tumor aggressiveness from WDSCC to MDSCC. Thus, salivary IL-6 could have a diagnostic and/or prognostic significance in identifying high-risk groups in mass screening of the population.

Geriatric Emergency Medicine

Tumour heterogeneity in oral cancer is attributed to the presence of cancer stem cells (CSCs). CSCs are the most migratory and metastatic cellular subpopulation within tumours. Assessment of CSC markers as significant predictors of lymph node metastasis may prove valuable in the clinical setting. Furthermore, analysis of this panel of putative stem cell markers in oral dysplasia may additionally inform of the likelihood for oral potentially malignant disorders (OPMDs) to progress to oral squamous cell carcinoma (OSCC). The present study aims to assess the significance of CSC markers in the progression of OPMDs to OSCC and assessment of lymph node metastasis in OSCC. CD44 and ALDH1 were assessed immunohistochemically in 25 normal, 30 OPMDs, and 24 OSCCs. CD44 is a membranous marker and ALDH1 is a cytoplasmic marker. The immunohistochemical expression of these markers were compared between OPMDs with and without dysplasia, as well as between low-risk and high-risk dysplasias. Similarly, expression was compared between OSCC with and without lymph node metastasis and among grades of OSCC. Positive CD44 expression was seen in all normal mucosal tissues. The expression decreased from normal epithelium to OPMDs but increased in OSCC. CD44 expression was positive in 21 cases of OSCC (87.5%) and reduced from well-differentiated to poorly differentiated OSCC. CD44 staining index was higher in OSCC without lymph node metastasis (3.59) when compared with OSCC with lymph node metastasis (1.33). There was a statistically significant difference observed in the ALDH1 staining index among three groups (p < 0.05), with highest expression seen in OSCC. Within OPMDs, the ALDH1 staining index was statistically higher in OPMDs with dysplasia as compared to OPMDs without dysplasia. Furthermore, the expression was higher in OPMDs with high-risk dysplasia when compared with low-risk dysplasia, but this was not statistically significant (p = 0.82). In conclusion, The CD44 positive population possesses properties of CSCs in head and neck carcinoma, and continuous shedding could be found after CD44 down-regulation. The present study reports differences in ALDH1 expression between OPMDs with and without dysplasia, dysplastic and non-dysplastic epithelia, and low-risk and high-risk dysplasia. These findings may suggest ALDH1 as a specific marker for dysplasia. CD44 demonstrated a difference in staining index in OSCC without lymph node metastasis versus OSCC with lymph node metastasis. These findings may suggest CD44 as a marker for lymph node metastasis. Both proteins may play key roles in the tumorigenicity of CSCs in OPMDs and OSCC.

Objectives:to determine salivary electrolyte concentration of oral potentially malignant disorders (OPMD) and oral squamous cell carcinoma (OSCC) patients. A related systematic review was performed. Methods:Observational study. Unstimulated saliva from 18 patients with OSCC, 18 with OPMD, and 18 without oral lesions was collected. A biochemical analysis was performed to evaluate the salivary concentrations of potassium (K), phosphorus (P), sodium (Na), calcium (Ca), magnesium (Mg), zinc (Zn), copper (Cu), and iron (Fe). Kruskal–Wallis test was performed, and p < 0.05 was interpreted as statistically significant. The literature search for the systematic review retrieved 9 studies that associated salivary electrolyte levels with presence and progression of OSCC. Results:A highly significant increase was found in the salivary Mg levels in the OPMD group (5.41 µg/mL) in comparison with the OSCC (3.71µg/mL) and control group (3.51 µg/mL) (p = 0.041). No differences were observed in other salivary levels elements. The results of the systematic review revealed that one article indicated a decrease, and three papers reported an increase in salivary Na levels in patients with OPMD and OSCC. Two articles indicated a decrease in salivary K levels in OSCC, and the other two reported high Mg levels in OPMD and OSCC. Conclusion:High salivary Mg levels can be a potential biomarker indicating the presence of OPMD, however, the evidence is still contradictory and more studies are required.

Comparative evaluation of salivary and serum CA 15-3 levels in tobacco users with and without potentially malignant oral disorders (PMOD) and oral squamous cell carcinoma (OSCC)