Assessment of pharyngeal airway space with MRI In oral submucous fibrosis: A cross-sectional observational study.

Introduction: Oral submucous fibrosis (OSMF) alters the pharynx, which may affect airway size. MRI will be useful for diagnosing pharyngeal abnormalities. MRI is used to evaluate pharyngeal airway and soft palate changes in OSMF patients. Materials and methods: This study is a cross-sectional observational study that included a sample size of 42 patients. Group A consisted of 21 patients with OSMF, while Group B consisted of 21 volunteers without OSMF, who served as the control group. The patients with OSMF were classified into Stages I, II, and III according to the categorization established by Pindborg JJ in 1989, Stop-Bang questionnaire was employed to assess obstructive sleep apnoea. Magnetic Resonance Imaging (MRI) was utilized to acquire evaluations of the pharyngeal airway, encompassing measurements in the midsagittal, cross-sectional width, length planes, and cross-sectional area with volume, for all participants. The Shapiro-Wilk test determines distribution normality. We utilized one-way ANOVA to compare the means between groups. Results: The average age of OSMF patients was 45.9 ± 8.16, while the control group was 39.19 ± 4.21. Stage I of OSMF had the highest mean Stop Bang questionnaire score (2.75), followed by stage III (2.22), and stage II (1.75). Statistically significant differences (p < 0.001) were seen in volume, linear midsagittal planes, cross-sectional width and length planes, cross-sectional area, and soft palate breadth and length between OSMF and control groups. Conclusion: MRI can effectively examine early changes in the pharyngeal airway of patients with OSMF thereby serving as a constructive diagnostic and motivational tool.

Estimation of the Salivary Copper Levels in Oral Submucous Fibrosis Condition: An In Vivo Study.

AIM: The aim of the present study was to estimate the salivary copper levels in oral submucous fibrosis (OSMF) condition. MATERIALS AND METHODS: A total of 60 individuals of which an equal number of 30 each in normal healthy control group as well as in clinically and histopathologically (the biopsy was done once after the clinical confirmation of OSMF in the patient) confirmed patients of OSMF were included in the study group. Total of 51 males and 9 females were considered for the present study and the age distribution of these groups ranged from a minimum of 21 years to a maximum of 74 years. Unstimulated whole saliva was collected from the patient followed by the conventional biopsy practice. The collected saliva was then subjected for the analysis of copper levels. Trace element copper was estimated by using Digital Semiautomatic Analyzer with the help of copper kit. The clinical mouth opening of OSMF was estimated. Analysis of variance (ANOVA) and Tukey HSD post hoc test was used analyze the data wherein the participants were grouped into age ranges of 20-30, 31-40, 41-50, and >60 years. RESULTS: The mean salivary copper level among OSMF and control groups with respect to age in 20-30 years was 55.98 ± 15.50 and 30.87 ± 7.70, in 31-40 years was 63.96 ± 21.13 and 32.95 ± 4.56, in 41-50 years was 50.11 ± 6.83 and 30.46 ± 3.28, and >60 years was 45.65 and 13.67 µg/dL, respectively. The mean salivary copper levels among OSMF and Control groups with respect to males were 55.60 ± 15.27 and 31.18 ± 6.97 and among females were 67.0 ± 24.25 and 30.06 ± 5.77 µg/dL, respectively. The mean salivary copper levels with histopathological grades in very early stage was 47.18 ± 5.73, in early stage was 49.22 ± 7.65, in moderately advanced was 73.53 ± 10.62 and in OSMF with mild dysplasia was 79.98 ± 16.27 µg/dL, respectively. The mean salivary copper levels in individuals with clinical mouth opening more than 35 mm was 45.65 ± 6.57, in 25-35 mm was 48.94 ± 21.60, in 15-25 mm was 70.54 ± 3.52 and in less than 15 mm was 81.50 ± 16.66, respectively. CONCLUSION: The present study concluded that salivary trace element levels could be used as potential diagnostic and prognostic markers in patients with OSMF. CLINICAL SIGNIFICANCE: Trace elements are involved in many different physiological and metabolic processes in humans, either directly or indirectly. Copper is involved in vital biochemical activities like different redox and free radical formation and in maintaining cellular proton homeostasis. It is also associated with the processing of oxygen and a component of arecanut in all forms, which is implicated in the etiology of OSMF. How to cite this article: Gupta R, Jayanti I, Das A, et al. Estimation of the Salivary Copper Levels in Oral Submucous Fibrosis Condition: An In Vivo Study. J Contemp Dent Pract 2024;25(5):498-502.

Outcome of a Novel Drug Combination in the Medical Management of Oral Submucous Fibrosis Patients: A Retrospective Cohort Study.

Aim: To evaluate the efficacy of a new drug combination in the medical management of oral submucous fibrosis (OSMF) patients. Materials and Methods: This retrospective cohort study included 89 patients who were clinically diagnosed of having OSMF (grade I, II, III). These patients had been administered the new drug combination for 6 months and clinical parameters such as burning sensation score, mouth opening and tongue protrusion were evaluated at 1 month, 3 months, 6 months and 12 months post-medication retrospectively. Results: Out of the total 89 patients included in the study, 18 patients had Grade-I, 50 patients had Grade-II and 21 patients had Grade-III OSMF. A significant improvement was observed in the mouth opening and tongue protrusion in all the three grades of OSMF from baseline to 1 month, 3 months, 6 months and 12 months after treatment (p < 0.0001) on intra- and intergroup comparison. A significant reduction in the burning sensation score from baseline to 1 month, 3 months, 6 months and 12 months after treatment was observed (p < 0.0001) in all the three grades on intra group comparison. Conclusion: This retrospective study showed that this novel drug combination is significantly effective in improving burning sensation score, mouth opening and tongue protrusion in OSMF grade I, II and III patients with results consistent after 1 year of follow-up.

Hindi Translation of Oral Health-Related Quality of Life (OHRQoL) Questionnaire and its Validation for Oral Submucous Fibrosis Patients: A Pilot Study.

Background: Oral Submucous fibrosis has a substantial burden on the Indian population; thus there is a substantial need for a validated questionnaire assessing QoL in Hindi. Thus, a prospective cross-sectional study was designed to translate and validate the English version of the OHR QoL-OSF questionnaire into Hindi. Methodology: We followed the guidelines suggested by the European Organisation for Research and Treatment of Cancer (EORTC) for the translation procedure and subsequently validated the translated questionnaire with reliability and validity. Results: A total of 50 patients were included in this pilot study. Cronbach's alpha for 17 items was 0.87 signifying high reliability. All questions had an I-CVI of 1.00, demonstrating complete agreement among the content experts regarding the relevance and clarity of items in the questionnaire. Convergent validity was adequately established for all factors except for factor 3 for which it was around 0.43. The intraclass coefficient was 0.803 showing good agreement between the test-retest results. Conclusion: The Hindi version of the OHR QoL questionnaire is a reliable instrument having good reliability, good content, good acceptability, and good responsiveness, and thus appears to be a valid instrument. Large studies with large sample sizes can be conducted in the future. Supplementary Information: The online version contains supplementary material available at 10.1007/s12663-023-01927-2.

MIAT promotes myofibroblastic activities and transformation in oral submucous fibrosis through sponging the miR-342-3p/SOX6 axis.

Oral submucous fibrosis (OSF) is an oral potentially malignant disorder that is closely related to the habit of areca nut chewing. Long non-coding RNA (lncRNA) myocardial infarction-associated transcript (MIAT) has been identified as an essential regulator in the fibrosis progression. However, the role of MIAT in the development of OSF remains unknown. The transcriptomic profile showed that MIAT is significantly overexpressed in the OSF cohort, with a positive correlation to fibrotic markers. The silencing of MIAT expression in primary buccal mucosal fibroblasts (BMFs) markedly inhibited arecoline-induced myofibroblast transformation. Mechanistically, MIAT functioned as a miR-342-3p sponge and suppressed the inhibitory effect of miR-342-3p on SOX6 mRNA, thereby reinstating SOX6 expression. Subsequent RNA expression rescue experiments confirmed that MIAT enhanced resistance to apoptosis and facilitated myofibroblastic properties such as cell mobility and collagen gel contraction by regulating the miR-342-3p/SOX6 axis. Taken together, these results suggest that the abnormal upregulation of MIAT is important in contributing persistent activation of myofibroblasts in fibrotic tissue, which may result from prolonged exposure to the constituents of areca nut. Furthermore, our findings demonstrated that therapeutic avenues that target the MIAT/miR-342-3p/SOX6 axis may be a promising approach for OSF treatments.

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

Aim: To estimate and compare the level of serum immunoglobulin A in patients of oral submucous fibrosis (OSMF), leukoplakia, and control group. Introduction: 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. Materials and Methods: 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. Results: 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. Conclusion: 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.

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.

Introduction: 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. Materials and Methods: 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. Results: 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. Conclusion: 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.

Targeting microRNA-190a halts the persistent myofibroblast activation and oxidative stress accumulation through upregulation of Krüppel-like factor 15 in oral submucous fibrosis.

Background/purpose: Oral submucous fibrosis (OSF) is a condition characterized by inflammation and excessive collagen deposition, which has been identified as a potentially malignant disorder. Recently, several microRNAs (miRNAs) have been shown to be implicated in various disorders associated with fibrosis. However, how these miRNAs modulate OSF development is poorly understood. Therefore, the study aimed to identify the specific miRNAs that contribute to the progression of OSF and to investigate their molecular mechanisms in promoting fibrosis. Materials and methods: The expression and clinical significance of potential pro-fibrosis miRNA in the OSF cohort and primary buccal mucosal fibroblasts were confirmed through RNA sequencing and qRT-PCR. Luciferase reporter activity assay, miRNA mimic or inhibitor, and short-hairpin RNA silencing were used to elucidate the molecular mechanism of miRNA. Transwell migration, collagen contraction, and reactive oxygen species (ROS) generation detection were used to investigate the effects of this mechanism on the myofibroblast phenotype and cellular pro-fibrosis capacity. Results: This study demonstrated that miR-190a was overexpressed in fibrotic buccal mucosal fibroblasts (fBMFs). Transfecting fBMFs with miR-190a inhibitor resulted in reduced cell migration, collagen gel contraction, ROS generation, and expression of fibrotic markers. Furthermore, miR-190a exerted this pro-fibrosis property by direct binding to its target, Krüppel-like factor 15 (KLF15). The results also indicated that the aberrant upregulation of miR-190a, in turn, downregulated the expression of KLF15, which resulted in the activation of myofibroblast. Conclusion: Our findings demonstrated that miR-190a was involved in myofibroblast activation, suggesting that targeting the miR-190a/KLF15 axis may be a feasible approach in the therapy of OSF.

Effects of lactobacillus pentosus postbiotics on fibrotic response in arecoline-induced oral fibrogenesis.

Background/purpose: Oral submucous fibrosis (OSF), characterized by excessive collagen deposition by myofibroblasts, is often linked to Areca nuts consumption. Probiotics consumption has shown protective effects against fibrotic diseases, and recently, their metabolic byproducts, known as postbiotics, have demonstrated superior advantages over probiotics. However, studies on the therapeutic impact of postbiotics on OSF have been scarce. Therefore, this study aims to examine the effect of PostBio GK4, a postbiotic derived from Lactobacillus pentosus GK4, on OSF and explore its underlying mechanisms. Materials and methods: The cytotoxicity of GK4 in normal buccal mucosal fibroblasts (BMFs) and fibrotic BMFs (fBMFs) were assessed. Following this, we evaluated the effects of GK4 on collagen contraction, migratory, and wound healing capacities in arecoline-induced fibrotic BMFs. Next, Western blotting and ELISA were employed to assess GK4's impact on fibrosis-related proteins such as COL1A1, and α-SMA, as well as on TGF-ß and Smad2/3 signaling pathway. Results: Arecoline was shown to stimulate cell migratory, contractile and wound healing abilities as well as the expression of α-SMA and COL1A1 in BMFs. Treatment with GK4 reduced all arecoline-induced phenomena in BMFs. Moreover, GK4 diminished the increased expression of TGF-ß and Smad2/3. Conclusion: Our findings proposed that GK4 may exert a suppressive effect on arecoline-induced myofibroblast activities via the inhibition of TGF-ß and Smad2/3 signaling pathway. Therefore, GK4 holds promise as an adjunct therapeutic approach for intervening in OSF. Further in-vivo and clinical studies are warranted to validate these observations.

Comprehensive analysis of clinicopathological characteristics in oral squamous cell carcinoma arising from oral submucous fibrosis: a retrospective study.

BACKGROUND: Oral squamous cell carcinoma arising from oral submucous fibrosis is recognized as a clinicopathologically distinct disease. This study aims to summarize and compare the population and clinicopathologic characteristics of OSCC-OSF with other OSCC in Hunan Province, China. The objective is to formulate treatment strategies more in line with regional characteristics. METHODS: A retrospective review of OSCC cases recruited from the Second Xiangya Hospital from 2010 to 2020 was conducted. A total of 1,413 OSCC patients were selected, including 481 patients with OSCC-OSF and 932 were other OSCC. Population characteristics, risk factors and clinicopathological manifestations were explored in OSCC-OSF and other OSCC, as well as thinprep cytologic test and DNA quantification. RESULTS: We found that OSCC-OSF patients were younger than those with other OSCC. Both types of disease were predominantly observed in males compared to females. Tumor biopsy analysis indicated that tumor cells within OSCC-OSF patients were more likely to be well differentiated and showed a higher frequency of lymph node metastases. Clinicopathological factors, such as the chewing betel nuts habit and smoking, were more prevalent in OSCC-OSF patients in contrast to other OSCC. DNA quantification revealed that the number of DNA aneuploid cells was higher in OSCC-OSF compared to other OSCCs. CONCLUSION: In this study, OSCC-OSF is considered a clinicopathologically distinct disease. Compared with other OSCC, OSCC-OSF patients have a higher incidence of nodal metastases, an early clinical TNM stage, and a lower three-year survival rate. Therefore, early diagnosis and treatment are crucial to improving the prognosis of OSCC-OSF.

Prevalence of oral submucous fibrosis linking with Areca Nut usage among Indians.

The prevalence of oral submucous fibrosis amongst population of Southern Rajasthan is of interest to dentists. Hence, a cross sectional study had been conducted on 3548 patients from 18-60 years age group who visited the Department of the Oral Medicine and Radiology, Pacific Dental College and Research Centre, Bedla, Udaipur was completed. They were subjected to thorough case history related to quid habit in arecanut (Areca catechu L.) form and to diagnose OSMF clinically. 1645 processed form of arecanut users were identified. The prevalence of arecanut chewers in study population has been reported to be 46.36%. The prevalence of OSMF in study population has been reported to be 458 (12.9%). Exact component associated and the mechanism involved in the occurrence of OSMF is still not available in literature and research is going on regarding unveiling this mysterious lesion.

The role of M1 (CD11c) and M2 (CD163) interplay in the pathogenesis of oral submucous fibrosis and its malignant transformation: An immunohistochemical analysis.

OBJECTIVES: The M1/M2 macrophage framework is crucial in organ fibrosis and its progression to malignancy. This study investigated the possible role of M1/M2 macrophage interplay in the pathogenesis of oral submucous fibrosis (OSF) and its malignant transformation by analysing immunohistochemical expression of CD11c (M1) and CD163 (M2) markers. METHODS: Immunohistochemistry was performed using primary antibodies against CD11c and CD163 on ten formalin-fixed paraffin-embedded tissue blocks for each group: (i) Stage 1 OSF, (ii) Stage 2 OSF, (iii) Stage 3 OSF, (iv) Stage 4 OSF, (v) well-differentiated squamous cell carcinoma (WDSCC) with OSF, and (vi) WDSCC without OSF. Ten cases of healthy buccal mucosa (NOM) served as controls. RESULTS: Epithelial quick scores of M1 (CD11c) in NOM, Stages 1-4 OSF, and WDSCC with and without OSF were 0, 1.8, 2.9, 0.4, 0, 0, and 0, while connective tissue scores were 0, 3.2, 4.3, 2.7, 0.5, 1.2, and 2.4, respectively. Epithelial scores for M2 (CD163) were 0, 0.8, 0.8, 2.1, 0.6, 0.8, and 0.2, and connective tissue scores were 0, 1.8, 2.6, 3.9, 2.2, 5, and 4.4, respectively. Stages 3 and 4 OSF, WDSCC with and without OSF exhibited higher M2/M1 ratios compared to NOM and Stages 1-2 OSF. CONCLUSION: The interaction between M1 (CD11c) and M2 (CD163) macrophages, leading to M2 polarisation, plays a crucial role in the pathogenesis of OSF and its potential malignant transformation.

Carvacrol inhibits the progression of oral submucous fibrosis via downregulation of PVT1/miR-20a-5p-mediated pyroptosis.

Oral submucous fibrosis (OSF) is a precancerous condition in the oral cavity, which is closely related to the myofibroblast conversion of buccal mucosal fibroblasts (BMFs) after chronic consumption of areca nut. Emerging evidence suggests pyroptosis, a form of programmed cell death that is mediated by inflammasome, is implicated in persistent myofibroblast activation and fibrosis. Besides, numerous studies have demonstrated the effects of non-coding RNAs on pyroptosis and myofibroblast activities. Herein, we aimed to target key long non-coding RNA PVT1 with natural compound, carvacrol, to alleviate pyroptosis and myofibroblast activation in OSF. We first identified PVT1 was downregulated in the carvacrol-treated fBMFs and then demonstrated that myofibroblast features and expression of pyroptosis makers were all reduced in response to carvacrol treatment. Subsequently, we analysed the expression of PVT1 and found that PVT1 was aberrantly upregulated in OSF specimens and positively correlated with several fibrosis markers. After revealing the suppressive effects of carvacrol on myofibroblast characterisitcs and pyroptosis were mediated by repression of PVT1, we then explored the potential mechanisms. Our data showed that PVT1 may serve as a sponge of microRNA(miR)-20a to mitigate the myofibroblast activation and pyroptosis. Altogether, these findings indicated that the anti-fibrosis effects of carvacrol merit consideration and may be due to the attenuation of pyroptosis and myofibroblast activation by targeting the PVT1/miR-20a axis.

Emerging roles of the renin-angiotensin system in select oral diseases.

OBJECTIVES: The renin-angiotensin system (RAS) plays essential roles in cardiovascular and renal function regulation. Recent studies have shown that the RAS components are widely expressed in oral tissues, but their roles in oral diseases remain underexplored. This review aims to summarize the effects of the RAS in select oral diseases including oral squamous cells carcinoma (OSCC), periodontitis, oral submucous fibrosis (OSF), and ageusia/dysgeusia. SUBJECTS AND METHODS: Data searches were performed using PubMed, Web of Science and Scopus through July 2024. A narrative overview of current literature was undertaken to synthesize the contexts with elaboration and summary. RESULTS: In OSCC, ACE/Ang II/AT1R promotes OSCC by inducing angiogenesis, cell proliferation and invasiveness. Conversely, ACE/Ang II/AT2R and ACE2/Ang (1-7)/MasR inhibit OSCC progressions. In periodontitis, ACE/Ang II/AT1R upregulates inflammatory cytokines and promotes osteoclast differentiation factor RANKL, whereas ACE2/Ang (1-7)/MasR exerts opposite effects by preventing inflammation and alveolar bone loss. In OSF, Ang (1-7) counters the profibrotic and proinflammatory action of Ang II. In dysgeusia, Ang II suppresses salt taste responses and enhances sweet taste sensitivities, while Ang (1-7) exhibits opposite effects. CONCLUSIONS: The RAS cascade plays crucial roles in OSCC, periodontitis, OSF and ageusia/dysgeusia. The imbalanced RAS may aggravate the progression of these diseases.

Quantitative Assessment of Receptors of Advanced Glycation End Products Expression in Tissue Samples from Patients with oral Submucous Fibrosis, Leukoplakia, and Oral Squamous Cell Carcinoma.

Background and Aim: Oxidative stress markers have been firmly established as elevated in oral squamous cell carcinomas (OSCCs). These markers play a crucial role in the pathogenic mechanism underlying the accumulation of advanced glycation end products (AGEs) and their respective receptors. The primary objective of this study is to discern and compare the levels of receptors of AGEs (RAGEs) within tissue samples from patients diagnosed with oral submucous fibrosis (OSMF) at varying stages, oral leukoplakia at various stages, and OSCC. Materials and Methods: A cross-sectional investigation was conducted, enrolling a total of 49 patients, distributed across three distinct groups. Tissue samples were meticulously collected from the aforementioned patient groups. Subsequently, these samples underwent a process of homogenization and centrifugation. The supernatant obtained was subjected to enzyme-linked immunosorbent assay analysis to precisely determine the concentration of RAGE. Results: The concentration of RAGEs was found to be significantly higher at various stages of OSMF when compared to the reference group of OSCC (P < 0.05). This difference was statistically significant, indicating a substantial association. In contrast, the levels of RAGE in patients with hyperkeratosis accompanied by epithelial dysplasia at various stages were observed to be lower than those in the OSCC group, with the difference in concentration being statistically insignificant (P > 0.05). Conclusion: This comprehensive study has provided compelling evidence demonstrating the heightened levels of RAGE in OSMF when compared to OSCC. These findings collectively suggest the potential utility of anti-RAGE interventions as a promising avenue for novel therapeutic strategies in potentially malignant disorders such as OSMF.

Novel role of CD34+ stromal cells/telocytes in oral submucous fibrosis development and its malignant transformation.

BACKGROUND: Telocytes are interstitial cells widely distributed in the extracellular matrix of numerous tissues distinguished by their long, thin, and moniliform projections. Telocytes have a role in the stimulation of angiogenesis and contribute to the development and progression of fibrosis. AIM: The current study aimed to assess and compare the telocyte distribution in normal mucosa, oral submucous fibrosis (OSF), and OSCC associated with OSF (OSCCOSF). MATERIALS AND METHODS: Formalin-fixed and paraffin-embedded tissue blocks of 30 OSF cases, 15 OSCCOSF cases, and 15 normal oral mucosae were obtained. Immunohistochemical staining was done with antibodies to CD34 to assess the vasculature and telocytes. The mean vascular density (MVD) and mean telocyte density were compared between the groups using the Kruskal-Walli test. RESULTS: A statistically significant high MVD (3.4 ± 1.22) and mean telocyte density (3.8 ± 1.35) was observed in OSCCOSF cases while it was lowest in advanced OSF cases. MVD was higher in early OSF cases than in normal mucosa. CONCLUSION: This study showed a decrease in CD34-positive telocytes in OSF, indicating that telocyte loss promotes the development of fibrosis.Increased angiogenesis coexisted with an increase in telocytes in OSCCOSF.

Arecoline promotes fibroblast activation and M2-macrophage polarization by up-regulating the expression of IL-4.

OBJECTIVE: To determine the biological effects of arecoline on oral submucous fibrosis (OSF). DESIGN: The differential genes between OSF tissue and normal oral tissue were collected form GSE64216 dataset, analyzed by Gene Expression Omnibus (GEO) database. Real-time PCR and immunohistochemistry were used to analyze the expression of IL-4 gene and protein in oral tissue. Enzyme-linked immunosorbent assay (ELISA) was used to analyze the expression of exocrine IL-4 protein in human oral fibroblasts (HOF) pre-treated by arecoline. Cell Counting Kit-8 (CCK-8) and transwell assays were used to analyze the proliferation and migration of HOF cells, respectively. After IL-4 was knocked down by short hairpin (sh) plasmid, the proliferation and migration of HOF cells were detected. Flow cytometry was used to analyze the proportion of M2-macrophages. Real-time PCR and immunohistochemistry were used to verify the expression of biomarker proteins of macrophages in OSF tissues. RESULTS: The expression of IL-4 gene and protein were both up-regulated in OSF tissue. Arecoline could enhance the expression of IL-4 gene and exocrine protein in HOF cells, and promote the proliferation and migration of HOF cells. While knockdown of IL-4 could inhibit arecoline-induced proliferation and migration in HOF cells. The results of flow cytometry showed that recombinant human IL-4 (rhIL-4) protein could increase the proportion of M2-macrophages. Similarly, the results of real-time PCR and immunohistochemistry showed the expression of ARG1 (Biomarker proteins of M2-macrophage) was up-regulated in OSF tissues. CONCLUSION: Arecoline promotes activation of fibroblasts and polarization of M2-macrophages by up-regulating the expression of IL-4.

Alteration of salivary LPO, MDA, LDH, glutathione, GPx, SOD and vitamins in oral submucous fibrosis: A three-level meta-analysis study.

This study aims to investigate the alteration of salivary biomarker profiling in the development of oral submucous fibrosis (OSMF) and to explore the influence of saliva in the diagnosis of OSMF. A systematic search of published articles using the PRISMA guidelines was conducted to identify relevant studies on OSMF and saliva. All eligible studies, including case-control, cross-sectional studies, cohort, and pilot studies, contained the evaluation of salivary biomarker profiling in patients with OSMF. Salivary biomarker data from 28 selected articles were categorized into nine groups, and their mean values were determined. A three-step meta-analysis was performed by grouping salivary biomarker profiling into more heterogeneous categories based on OSMF classification, considering functional, histological, and clinical grading. The salivary biomarker profiling analysis revealed significant alterations in all markers, indicating their efficacy in OSMF diagnosis. Subgroup analyses highlighted significant associations in oxidative stress and protein with increased mean values, particularly emphasizing lipid peroxidase (LPO), malondialdehyde (MDA), and lactate dehydrogenase (LDH). Conversely, decreased mean values were observed in glutathione, glutathione peroxidase (GPx), superoxide dismutase (SOD), and vitamins. Notably, OSMF grading analysis demonstrated a significant difference in weighted effect sizes for histological grading, particularly in stage IV. The study underscores the alteration of specific salivary biomarkers, particularly those associated with LPO, MDA, LDH, glutathione, GPx, SOD, and vitamins, in diagnosing and grading OSMF.

TUSC3, p53 and p21 genetic association with development of oral submucous fibrosis and oral squamous cell carcinoma among addictive tobacco chewers of Pakistan.

BACKGROUND: This study delves into the intricate landscape of oral cancer, a global concern with a high incidence in Asian countries. We focus on oral squamous cell carcinoma (OSCC), primarily driven by the consumption of betel nut and its derivatives. OSCC often arises from premalignant lesions like oral submucous fibrosis (OSF). In Pakistan, OSCC is prevalent among men due to various addictive substances, including smokeless tobacco and chewing materials. Mutations in tumor suppressor genes, such as TP53 and p21, play crucial roles in this malignancy's development. We also explore the involvement of TUSC3 gene deletion in OSCC and OSF. METHODS: In this study we investigated demographics, TUSC3 gene expression, deletion analysis, and TP53 and p21 genetic alterations in OSCC and OSF patients (blood and tissue of 50 samples in each condition) who had tobacco derivates usage history. The association analysis was carried out mainly through PCR based genotyping. RESULTS: The study's patient cohort (OSCC and OSF) displayed a wide age range from 13 to 65 years (Mean = 32.96 years). Both conditions were more prevalent in males, with a male-female ratio of approximately 2.5:1. Chewing habits analysis revealed high frequencies of gutka use in both OSF and OSCC patients. TUSC3 expression analysis in OSCC cell lines indicated significant downregulation. Genotyping showed no TUSC3 deletion in OSF cases, but a deletion rate of over 22% in OSCC tissue samples. Analysis supported a significant association of TUSC3 deletion with OSCC development but not with OSF. Polymorphism in p53 exon 4 and p21 (rs1801270) were significantly associated with both OSCC and OSF, adding to their pathogenesis. Our findings further revealed a strong correlation between TUSC3 deletion and the excessive use of tobacco and related products, shedding light on the genetic underpinnings of OSCC development. CONCLUSIONS: Notably, our study provides a crucial insight into genetic aspects underlying OSCC and OSF in response of addictive consumption of areca nut, betel quid, and tobacco derivatives. A significant association between TUSC3 deletion and OSCC development, along with polymorphisms in TP53 and p21, underscores the importance of further research into the molecular mechanisms driving oral cancer progression for improved diagnosis and treatment outcomes.

Targeting MetaLnc9/miR-143/FSCN1 axis inhibits oxidative stress and myofibroblast transdifferentiation in oral submucous fibrosis.

Background/purpose: Persistent activation of myofibroblasts is attributed to various dysregulated biological events conferring multiple types of fibrosis diseases, including oral submucous fibrosis (OSF). Although the significance of non-coding RNAs (ncRNAs) in the occurrence of fibrosis has been appreciated, the detailed mechanisms still have not been fully elucidated. The aim of this study was to identify key dysregulated ncRNAs and elucidate their pro-fibrotic mechanisms in promoting myofibroblast activation and the pathological development of OSF. Materials and methods: Expression of non-coding RNAs and mRNAs in OSF cohort was determined using RNA sequencing and qRT-PCR. The molecular axis of pro-fibrotic ncRNAs were exploited via luciferase reporter activity assay and RNA expression rescue experiments. Functional assays, including collagen gel contraction, wound healing ability, cell migration, and reactive oxygen species (ROS) production, were conducted to assess the changes in the myofibroblastic phenotypes of primary human buccal mucosal fibroblasts. Results: Herein, we found that long non-coding RNA MetaLnc9 was upregulated in OSF specimens and positively associated with several fibrosis markers. Silencing of MetaLnc9 diminished the features of activated myofibroblasts and the production of ROS. We not only showed that MetaLnc9 functioned as a competitive endogenous RNA of microRNA (miR)-143, but also demonstrated that the pro-fibrosis effect of MetaLnc9 on myofibroblast activities was mediated by suppression of miR-143. Moreover, our data showed that fascin actin-bundling protein 1 (FSCN1) was a direct target of miR-143 and positively related to MetaLnc9. Conclusion: Upregulation of MetaLnc9 may enhance the activation of myofibroblasts by sponging miR-143 and titrating its inhibitory property on FSCN1.