Advancement in therapeutic strategies for immune-mediated oral diseases.

BACKGROUND: Immune-mediated diseases are a diverse group of conditions characterized by alteration of cellular homeostasis and inflammation triggered by dysregulation of the normal immune response. Several immune-mediated diseases exhibit oral signs and symptoms. Traditionally, these conditions are treated with corticosteroids or immunosuppressive agents, including azathioprine, cyclophosphamide, and thalidomide. Recent research into the developmental pathways of these diseases has led to the exploration of novel approaches in treatment. This review examines newer treatment modalities for the management of immune-mediated diseases with oral presentations. Topical calcineurin inhibitors (TCIs) such as tacrolimus and pimecrolimus have been employed successfully in managing oral lichen planus and pemphigus vulgaris. Biologic agents, comprising monoclonal antibodies, fusion proteins, and recombinant cytokines, can provide targeted therapy with fewer adverse effects. Neutraceutical agents comprising aloe vera, curcumin, and honey are commonly used in traditional medicine and offer a holistic approach. They may have a place as adjuvants to current standard therapeutic protocols. Photodynamic therapy (PDT) and low-level laser therapy (LLLT) utilize a specific wavelength of light to achieve desired cellular change. While the use of PDT in immune-mediated diseases is contentious, LLLT has shown positive results. Newer therapeutic modalities involve kinase inhibitors, S1P1 receptor modulators, MSCs, and iRNA providing targeted treatment of specific diseases.

Hypertonic Saline Use and Traditional Nasal Irrigation: A Possible Preventative Measure against COVID-19.

Saline nasal irrigation (SNI) is a clinically established treatment that has been used to manage upper respiratory infections and allergies; there is also some indication that it may be effective in the setting of coronavirus disease-2019 (COVID-19). The possible advantages of SNI include the following benefits due to the well-known antiviral impact of sodium chloride (NaCl) and the mechanical cleansing effect arising from the irrigation method. First, there will most likely be a decrease in COVID-19 infection rates; second, illness severity will be lowered; and third, community transmission will be mitigated. Despite the need for more concentrated research into these aspects, public health organizations should emphasize alternate infection mitigation measures such as SNI in light of the ongoing COVID-19 problem, low global vaccine supply, and the rapid introduction of SARS-CoV-2 variants. Keywords: Community transmission, COVID-19, Pandemic, Prevention, Saline, SARS-CoV-2.

Effect of Supplementation of Vitamin D in Patients with Periodontitis Evaluated before and after Nonsurgical Therapy.

Background: Vitamin D has anti-inflammatory properties and the potential to increase the generation of antimicrobial peptides like cathelicidin and defensins that may have a good impact on oral health. Higher vitamin D consumption has also been linked to a reduced risk of periodontal disease progression. Hence, the primary objective of this study was to evaluate and compare the clinical and laboratory parameters of oral supplementation of vitamin D as an adjuvant to scaling and root planing and to assess the bone mineral density via qualitative ultrasound bone density scanner in chronic periodontitis patients. Methodology. This study included 40 patients with periodontitis categorized into 2 groups with twenty patients each, Group I comprising scaling and root planing (SRP) alone and Group II comprising SRP along with vitamin D supplementation. Plaque index, gingival index, probing pocket depth, and clinical attachment loss was measured as clinical parameters. Serum vitamin D levels were assessed before and after SRP at both baseline and 6 weeks. Results: The intergroup comparison of clinical parameters (PI, GI, PPD, and CAL) at 6 weeks for both the groups showed statistical significance. Intragroup comparison of clinical parameters from baseline to 6 weeks showed a statistically significant reduction in both groups. The mean bone mineral density level in both the control and test groups demonstrated a mean T score of -1.3 and -1.21, respectively. The mean vitamin D levels were 27.8460 and 28.1020 for the test and control groups, respectively, which was statistically insignificant (p = 0.705) and those at six-week intervals improved to 31.3650 and 28.0240 which were statistically significant (p ≤ 0.001). Conclusion: It could be stated that a positive relationship exists between periodontitis and osteopenia which could aggravate periodontal destruction. All periodontitis cases should thus be evaluated for BMD and supplemented with vitamin D3 in an appropriate dosage and time frame to treat both these diseases.

Efficacy of Spirulina 500 mg vs Triamcinolone Acetonide 0.1% for the Treatment of Oral Lichen Planus: A Randomized Clinical Trial.

AIM: The present study aimed at evaluating the efficacy of spirulina 500 mg in reducing the burning sensation and lesion size in oral lichen planus (OLP). MATERIALS AND METHODS: A total of 60 subjects who attended the oral medicine specialty clinic with histopathologically confirmed OLP and having symptoms of burning sensation were recruited for the study. They were randomly divided into two groups: group A (30) subjects were prescribed Spirulina 500 mg twice daily along with only a week application of topical triamcinolone acetonide 0.1% thrice daily; group B subjects were prescribed topical triamcinolone acetonide 0.1% alone thrice daily for 8 weeks. Both the groups were followed up posttreatment monthly for three consecutive months. RESULTS: Data were recorded, and statistical analysis by using ANOVA one-way test, and Chi-square test were performed, which showed statistically significant p-value (<0.005) for the parameters "burning sensation" and "size of the lesion". When compared between groups, group A showed a favorable outcome of the intervention. CONCLUSION: Spirulina 500 mg supplementation twice daily could be effective adjunct therapy with steroids to treat OLP. CLINICAL SIGNIFICANCE: This research allowed us to delve into spirulina as one of the treatment modalities for OLP. Further studies are needed as it is a rich source of proteins and vitamins and demonstrates potent anti-inflammatory, immunomodulatory, and antioxidant actions.

Severe Acute Respiratory Syndrome Coronavirus 2 Nosode: A Potential Alternative Vaccine Platform.

Besides conventional vaccinations, viable alternatives are needed to elicit an immune response against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We propose and highlight the value of a homeopathic approach known as the "nosode" for the prevention of coronavirus disease-2019 (COVID-19). Nosode is an extract prepared from disease-affected tissues which is subsequently processed and administered as an antidote for the same medical condition. This concept might be a crucial therapeutic approach for viral infections since infected tissues contain a wide range of important viral antigens that could induce a functional host response via immunological sensitization. Thereby, nosode preparation produced from SARS-CoV-2-affected tissues may provide protection against COVID-19. "SARS-CoV-2 nosode" warrants more scientific investigation as a viable alternative vaccination platform. Keywords: COVID-19, Nosode, SARS-CoV-2, Variolation.

Multi-target potential of Indian phytochemicals against SARS-CoV-2: A docking, molecular dynamics and MM-GBSA approach extended to Omicron B.1.1.529.

BACKGROUND: SARS-CoV-2, an emerged strain of corona virus family became almost serious health concern worldwide. Despite vaccines availability, reports suggest the occurrence of SARS-CoV-2 infection even in a vaccinated population. With frequent evolution and expected multiple COVID-19 waves, improved preventive, diagnostic, and treatment measures are required. In recent times, phytochemicals have gained attention due to their therapeutic characteristics and are suggested as alternative and complementary treatments for infectious diseases. This present study aimed to identify potential inhibitors against reported protein targets of SARS-CoV-2. METHODOLOGY: We computationally investigated potential SARS-CoV-2 protein targets from the literature and collected druggable phytochemicals from Indian Medicinal Plants, Phytochemistry and Therapeutics (IMPPAT) database. Further, we implemented a systematic workflow of molecular docking, dynamic simulations and generalized born surface area free-energy calculations (MM-GBSA). RESULTS: Extensive literature search and assessment of 1508 articles identifies 13 potential SARS-CoV-2 protein targets. We screened 501 druggable phytochemicals with proven biological activities. Analysis of 6513(501 *13) docked phytochemicals complex, 26 were efficient against SARS-CoV-2. Amongst, 4,8-dihydroxysesamin and arboreal from Gmelina arborea were ranked potential against most of the targets with binding energy ranging between - 10.7 to - 8.2 kcal/mol. Additionally, comparative docking with known drugs such as arbidol (-6.6 to -5.1 kcal/mol), favipiravir (-5.5 to -4.5 kcal/mol), hydroxychloroquine (-6.5 to -5.1 kcal/mol), and remedesivir (-8.0 to -5.3 kcal/mol) revealed equal/less affinity than 4,8-dihydroxysesamin and arboreal. Interestingly, the nucleocapsid target was found commonly inhibited by 4,8-dihydroxysesamin and arboreal. Molecular dynamic simulation and Molecular mechanics generalized born surface area (MM-GBSA)calculations reflect that both the compounds possess high inhibiting potential against SARS-CoV-2 including the recently emerged Omicron variant (B.1.1.529). CONCLUSION: Overall our study imparts the usage of phytochemicals as antiviral agents for SARS-CoV-2 infection. Additional in vitro and in vivo testing of these phytochemicals is required to confirm their potency.

A Comparison in Patient Comfort Using Conventional Syringe and Needleless Jet Anesthesia Technique in Periodontal Surgery-A Split-Mouth Randomized Clinical Trial.

Background and Objectives: Periodontal surgery requires local anesthetic coverage to alleviate patient discomfort. Needles and injections can engender feelings of fear and anxiety in individuals. This study aimed to assess the level of comfort and anxiety in patients during the administration of local anesthesia using needleless jet anesthesia (JA) when compared to a conventional syringe (CS) in periodontal surgery. Method and Materials: 60 sites were designated for injection in a split-mouth design in 30 subjects who required periodontal surgery. Local anesthesia was administered in two appointments scheduled one week apart using either a JA system or a CS. The Visual Analogue Scale (VAS), Verbal Rating Scale (VRS), and Beck's anxiety inventory were used to report the pain and anxiety levels while injecting local anesthesia. Statistical analysis of the results was performed using the Shapiro-Wilks test and Paired t-test. Results: Patients reported greater comfort with JA. The VAS and VRS values were statistically significant-(p = 0.003) and (p = 0.001), respectively. Patients showed fear and were nervous about receiving a local anesthetic using a CS. A few subjects experienced lingering pain with the CS, whereas greater comfort and no lingering soreness were reported post-operatively at the site of JA administration. Conclusions: This study provides the first comprehensive assessment of using JA for periodontal surgical procedures. Lower pain scores were consistently observed with the use of jet injectors. Patients were at ease and reported lesser anxiety and greater comfort with jet injectors, making it ideally suited for providing local anesthesia in periodontal surgery.

Assessing the therapeutic potential of agomelatine, ramelteon, and melatonin against SARS-CoV-2.

BACKGROUND: The SARS-Cov-2(severe acute respiratory syndrome coronavirus 2) infection affecting human populations worldwide is now a very concerning issue considering the morbidity and mortality rates. Despite several measures followed by the medical fraternity and general public, there is no resolution. Therapeutic measures to tackle the infection have been based on researching new designer drug molecules that could prevent viral entry into the human host. Melatonin has been tried as an adjuvant in the management of COVID 19(coronavirus disease) illness but its specific antiviral role has not been investigated. Objectives: The objectives of the present study were to conduct an in-silico analysis to investigate if melatonin and related drugs namely ramelteon and agomelatine could be used as antiviral agents in SARS-CoV-2 infection based on their binding to the SARS-CoV-2 receptor binding site (RBD) and Angiotensin-converting enzyme 2 (ACE 2). METHODS: For docking studies (Pdb Id 1M0J), the SARS-CoV-2 spike protein receptor-binding domain (RBD) crystal structure which was ACE2 cell receptor bounded was employed. From the PubChem database, the three-dimensional configuration of the ligands melatonin, ramelteon, and agomelatine was retrieved, and conceptual density functional theory (CDFT) was performed to determine molecular descriptors. Charges were added and optimized with the universal force field to prepare the ligands for the process of docking. For facilitation of readability by the AutoDock software conversion to PDBQT(Protein Data Bank, Partial Charge (Q), & Atom Type (T)) format was performed. AutoDock version 4.2.6 docking program and AutoDock Tools (ADT) version 1.5.6 were used for molecular docking. Desmond, a Package of Schrödinger LLC was used to simulate molecular dynamics for hundred nanoseconds using. RESULTS: Data from the present study reveal that melatonin, ramelteon, and agomelatine demonstrate significant binding with SARS-CoV-2 RBD and ACE 2 demonstrating the fact that they can strongly prevent viral entry into the host cells through dual binding effects. However, Ramelteon was found to be the most superior amongst the 3 drugs analyzed in its antiviral properties against SARS-CoV-2. CONCLUSION: Results advocate further research in exploring the potential therapeutic applications of melatonin, ramelteon, and agomelatine for the management of SARS-CoV-2 infection.

Pharmacological Means of Pain Control during Separator Placement: A Systematic Review.

AIM: To assess the effectiveness of adjuvant analgesics/anesthetics in pain control after separator placement compared with no medication. BACKGROUND: Separator placement to create space for cementing bands is the first clinical procedure done in orthodontics. Pain in this stage can negatively affect patient compliance and trust in the clinician. To date, there is no universally accepted regimen for pain control. MATERIALS AND METHODS: Electronic databases of PubMed, Scopus, and Web of Science were searched. One hundred and thirty-two potentially relevant studies were found. A total of eight randomized clinical trials including 642 subjects were selected. Data were extracted into customized forms, and selected studies were assessed for risk of bias using the Joanna Briggs Institute. RESULTS: Results showed the use of analgesics led to lower reported pain scores at almost all time intervals. NSAIDs resulted in a statistically significant reduction in pain compared to a control group. CONCLUSION: According to the available literature, the use of analgesics is effective in controlling orthodontic pain due to separators. Acetaminophen and ibuprofen show a stable analgesic effect. CLINICAL IMPLICATION: Acetaminophen 650 mg or ibuprofen 400 mg taken 1 hour prior to separator placement can reduce pain associated with the procedure.

Efficacy of vitamin C supplementation as an adjunct in the non-surgical management of periodontitis: a systematic review.

BACKGROUND AND OBJECTIVE: The antioxidant potential of vitamin C is useful in reducing oxidative stress, free radicals, and reactive oxygen species, which may assist in the improved outcomes of periodontal therapy. This systematic review was aimed to evaluate the effectiveness of vitamin C supplementation as an adjunct to non-surgical periodontal therapy, in the management of periodontitis. DATA SOURCES: PubMed, EMBASE, Cochrane Library, and Web of Science. STUDY ELIGIBILITY CRITERIA: Randomized controlled trials published between January 1990 and March 2020. PARTICIPANTS: People 18 years and older with periodontitis. STUDY APPRAISAL AND SYNTHESIS METHODS: The Critical Appraisal Skills Programme (CASP) quality appraisal tool. RESULTS: The initial search yielded 441 articles out of which six studies fulfilled the inclusion criteria. Vitamin C supplementation helped improve bleeding indices in gingivitis but did not significantly lead to reduction of probing depths or clinical attachment gain for periodontitis. CONCLUSION: Administration of vitamin C as an adjunct to non-surgical periodontal therapy did not result in clinically significant improvements in pocket probing depths at 3 months in periodontitis patients. With the limited evidence available, no recommendation can be made for supplementation of vitamin C in conjunction with initial periodontal therapy for subjects with periodontitis to improve primary treatment outcome measures.

Moringa oleifera: Antioxidant, Anticancer, Anti-inflammatory, and Related Properties of Extracts in Cell Lines: A Review of Medicinal Effects, Phytochemistry, and Applications.

Moringa oleifera (MO), or the common drumstick possesses antioxidant properties, and its pods, seeds, leaves, and bark have been used for the treatment of inflammatory and cancerous conditions. This systematic review attempts to synthesize original studies of MO extracts in cell lines to determine their specific antiproliferative, antioxidant, anti-inflammatory, and related effects. The literature was obtained from PubMed central, the Cochrane registry, and other sources including Google Scholar, and Embase. Studies fulfilling the inclusion criteria were selected. Custom data collection forms were employed and two independent evaluators compiled the relevant information. Eighteen studies were selected after applying inclusion and exclusion criteria. In most studies, MO leaves had more potent properties compared to other parts of the plant. Ethyl acetate and ethanolic extractions improved the potency of the extract. Effects were selective (different for normal cells and cancer cells) and dose-dependent. Anticancer and antioxidant activities were consistently reported, with effects exerted at the genetic and molecular levels. MO extracts potentially could be employed for therapeutic applications. The optimal sources, preparation protocols, and dosages have been researched, though further scrutiny is needed for a comprehensive formulation. Keywords: Anticancer, Anti-inflammatory, Antioxidant, Moringa oleifera.

Signaling alterations in oral keratinocytes in response to shisha and crude tobacco extract.

BACKGROUND: Tobacco consumption in smoking and non-smoking forms has been consequential in the rise of oral cancer cases. Among different forms, epidemiological studies from Middle Eastern countries and rural parts of northern India have reported increasing association of oral cancer with waterpipe (hookah) smoking. However, molecular mechanisms and role played by waterpipe smoking in the onset of oral carcinogenesis remains unexplored. METHODS: In this study, immortalized normal human oral keratinocytes were chronically treated with extracts of two varieties of waterpipe tobacco-crude tobacco and processed shisha. Phenotypic changes and molecular aberrations were examined using cell culture-based assays and mass spectrometry-based quantitative proteomic analysis, respectively. Bioinformatics analysis was utilized to analyze proteomics data and identify dysregulated pathways. RESULTS: Our data indicate that chronic treatment with waterpipe tobacco extracts increased proliferation, invasion, migration, and significant dysregulation of protein expression in oral keratinocytes. Altered expression of proteins involved in interferon signaling pathway were observed with both varieties of tobacco. Overexpression of cholesterol metabolism and vesicle-mediated transport proteins were identified exclusively in cells treated with crude tobacco extract. Bioinformatics analyses revealed different oncogenic response in oral cells based on the type of waterpipe tobacco used. CONCLUSIONS: This study may serve as a useful resource in understanding the early onset of oral cancer attributed to waterpipe smoking.

Enteromorpha compressa extract induces anticancer activity through apoptosis and autophagy in oral cancer.

Marine algae are an auspicious source of innovative bioactive compounds containing possible therapeutic agents against mammalian cancers. However, the mechanism by which bioactive algal compounds exhibit anticancer activity against oral squamous cell carcinoma (OSCC) is scant. The main objective of the current study was to explore the properties of the Enteromorpha compressa solvent extracts that induced autophagy and apoptosis with reference to their potent phytochemical and antioxidant properties. The presence of bioactive compounds were confirmed by UV and FT-IR spectroscopy. The free radical scavenging activity were analyzed by evaluating H2O2, DPPH, superoxide and hydroxyl activity. The anticancer activities of the extracts were investigated by employing clonogenic and scratch assay. The apoptosis potential was evaluated by DAPI and MMP by Rh123 fluorescence assay. Moreover, the CAT, SOD, GPX, APX, and GR activities were measured. The autophagy potential was evaluated by LC3 puncta formation, acridine orange in addition to LysoTracker staining. The present investigation revealed that the methanolic extract of E. compressa elicited robust free radical scavenging activity that discerns its antiproliferative potency. Moreover, the methanolic algal extract boosted intrinsic apoptosis against OSCC by downregulating protective antioxidant enzymes. Furthermore, it also revealed induction of autophagy to promote cell death in oral cancer cells. The presence of novel bioactive compounds in E. compressa has uncovered possible therapeutic value against OSCC by modulating antioxidant defense system, apoptosis and autophagy that could be used to explore very competent algal candidates for the development of potential alternative anticancer drugs.

EMMPRIN/BASIGIN as a biological modulator of oral cancer and COVID-19 interaction: Novel propositions.

Extracellular matrix metalloproteinase inducer (EMMPRIN), which is also called BASIGIN/CD147, is a cell surface glycoprotein that belongs to the immunoglobulin superfamily and plays a significant role in intercellular recognition in immunology, cellular differentiation and development. Apart from ACE-2, recently EMMPRIN, has been regarded as a target for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) attachment and entry into the host cell. Since one of the routes of entry for the virus is the oral cavity, it becomes imperative to percept oral comorbidities such oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMDs) in terms of EMMPRIN as a target for SARS-CoV-2. In the present paper, it is proposed that OSCC, by the virtue of upregulation of EMMPRIN expression, increases the susceptibility to coronavirus disease (COVID-19). In turn, COVID-19 in OSCC patients causes exhaustion of EMMPRIN receptor due to binding with 'S' receptor leading to a downregulation of related carcinogenesis events. We proposed that in the ACE-2 depleted situation in OSCC, EMMPRIN receptor might get high jacked by the COVID-19 virus for the entry into the host cells. Apart from the anti-monoclonal antibody, it is recommended to explore the use of grape seed and skin containing mouthwash as an adjunct, which could also have anti EMMPRIN effects in patients with OSCC and OPMDs.

Meditation and Yoga-assisted Alteration of Dietary Behavior as a Useful Strategy for Limiting Zoonotic Pandemics: A Novel Insight.

Vegetarian diets are known to have significant positive effects on personal and planetary health and are likely to curb zoonotic infection transmission. We propose that minimizing meat consumption should become an essential dietary shift in the post-COVID-19 era. To date, however, there is limited knowledge concerning suitable methods that could catalyze this change on a global scale. Meditation and Yoga are practical and easy to implement psychomodulatory strategies that can naturally trigger vegetarianism and related eating behaviors, lowering our reliance on animal meat. Decreasing dependence on animal meat reduces the need for animal markets and may substantially minimize the likelihood of spillover (passage of viruses from animal reservoirs into human populations). Global implementation of these strategies, in our opinion, can add to spiritual wisdom, compassion, and cooperative human behavior, thus reducing the encroachment of wild-life reserves and animal exploitation. The application of these ancient Indian approaches represents a novel and focused strategy toward curbing zoonotic pandemics. Keywords: Coronavirus disease-19, Meat-eating, Meditation, Pandemics, Viral spillover, Yoga.

Terminalia bellirica extract induces anticancer activity through modulation of apoptosis and autophagy in oral squamous cell carcinoma.

Terminalia bellirica (TB) has been used in traditional Indian medical system, Ayurveda. However, the mechanism underlying the efficacy of the TB extract against oral squamous cell carcinoma (OSCC) is yet to be explored. The present study established a connecting link between the TB extract induced apoptosis and autophagy in relation to reactive oxygen species (ROS). Our study revealed, that gallic acid in the TB extract possess a strong free radical scavenging capacity contributing towards the selective anti-proliferative activity. Furthermore, TB extract markedly enhanced the accumulation of ROS that facilitated mitochondrial apoptosis through DNA damage, indicating ROS as the vital component in regulation of apoptosis. This effect was effectively reversed by the use of a ROS scavenger, N-acetyl cysteine (NAC). Moreover, it was observed to induce autophagy; however, it attenuated the autophagosome-lysosome fusion in Cal33 cells without altering the lysosomal activity. Pharmacological inhibitors of autophagy, namely, 3-methyladenine and chloroquine, were demonstarated to regulate the stage-specific progression of autophagy post treatment with the TB extract, favouring subsequent activation of apoptosis. These findings revealed, presence of gallic acid in TB extract below NOAEL value causes oxidative upset in oral cancer cells and promote programmed cell death which has a potential therapeutic value against oral squamous cell carcinoma.

A Holistic Intervention for Oral Lichen Planus.

Oral lichen planus (OLP) is a chronic immune-mediated, inflammatory, and psychosomatic condition that frequently affects the oral mucosa in a typical bilateral pattern, often noticed in middle-aged females.1 It involves the buccal, lingual, and/or gingival mucosa and is also an oral potentially malignant disorder (OPMD). OLP is associated with etiological factors ranging from viruses like hepatitis C, systemic factors like thyroid disorders,2 local factors like dental plaque, calculus and cuspal trauma, to deep rooted, often underestimated, psychological triggers. OLP is routinely encountered in oral medicine clinics and is sometimes accompanied by the simultaneous involvement of skin, nails, genitals, and/or mucous membranes. Although OLP is multifactorial in origin, a large share of these presentations can be attributed chiefly to psychological factors like stress, anxiety, and depression. OLP is even more striking as its genesis, progression, and exacerbation closely mirror the dynamics of emotional disturbance.

Chronic Exposure to Chewing Tobacco Induces Metabolic Reprogramming and Cancer Stem Cell-Like Properties in Esophageal Epithelial Cells.

Tobacco in its smoke and smokeless form are major risk factors for esophageal squamous cell carcinoma (ESCC). However, molecular alterations associated with smokeless tobacco exposure are poorly understood. In the Indian subcontinent, tobacco is predominantly consumed in chewing form. An understanding of molecular alterations associated with chewing tobacco exposure is vital for identifying molecular markers and potential targets. We developed an in vitro cellular model by exposing non-transformed esophageal epithelial cells to chewing tobacco over an eight-month period. Chronic exposure to chewing tobacco led to increase in cell proliferation, invasive ability and anchorage independent growth, indicating cell transformation. Molecular alterations associated with chewing tobacco exposure were characterized by carrying out exome sequencing and quantitative proteomic profiling of parental cells and chewing tobacco exposed cells. Quantitative proteomic analysis revealed increased expression of cancer stem cell markers in tobacco treated cells. In addition, tobacco exposed cells showed the Oxidative Phosphorylation (OXPHOS) phenotype with decreased expression of enzymes associated with glycolytic pathway and increased expression of a large number of mitochondrial proteins involved in electron transport chain as well as enzymes of the tricarboxylic acid (TCA) cycle. Electron micrographs revealed increase in number and size of mitochondria. Based on these observations, we propose that chronic exposure of esophageal epithelial cells to tobacco leads to cancer stem cell-like phenotype. These cells show the characteristic OXPHOS phenotype, which can be potentially targeted as a therapeutic strategy.

Secretome analysis of oral keratinocytes chronically exposed to shisha.

BACKGROUND: Shisha smoking has been associated with multiple diseases including oral cancer. However, a mechanistic study to investigate alteration of secreted proteins in oral cells due to shisha smoking is lacking. OBJECTIVES: Elucidation of differentially secreted proteins by immortalized human normal oral keratinocytes (OKF6/TERT1) upon chronic exposure to shisha. METHODS: OKF6/TERT1 was chronically treated with 0.5% shisha extract for 8 months. Conditioned media from shisha treated (OKF6/TERT1-Shisha) and untreated (OKF6/TERT1-Parental) cells were subjected to TMT-based quantitative proteomic analysis. Bioinformatics analysis of differentially secreted proteins was carried out using SignalP, SecretomeP and TMHMM. Immunoblot validation of selected proteins was carried out to confirm the proteomics results. RESULTS: Proteomic analysis of OKF6/TERT1-Parental and OKF6/TERT1-Shisha secretome resulted in the identification of 1,598 proteins, of which 218 proteins were found to be differentially secreted (⩾ 1.5-fold; p-value ⩽ 0.05) in shisha treated cells. Bioinformatics analysis using prediction tools showed secretory potential of differentially secreted proteins identified in OKF6/TERT1-Shisha. Western blotting validated the expression of AKR1C2, HSPH1 and MMP9 in OKF6/TERT1-Shisha secretome in agreement with proteomic data. CONCLUSION: This study serves as a useful resource to understand the effect of chronic shisha smoking on the milieu of secreted proteins of oral cells. In vivo studies are warranted to supplement our in vitro data to elucidate the role of these proteins as early diagnostic biomarkers for oral carcinogenesis among shisha smokers.

New Directions for Oral Submucous Fibrosis Research: Whole Evaluation for Holistic Rehabilitation!

Oral submucous fibrosis (OSF) is an oral potentially malignant disorder (OPMD) associated with prolonged betel-quid (BQ) use. It affects both physiological and psychological dimensions, leading to a significant morbidity. Some challenges in OSF research include: Assessment and correction of: (i) Quality of life, (ii) burning sensation, and (iii) nutritional deficiency. There are numerous medical treatments for OSF, but most of them fail to offer complete rehabilitation. This is because some underpinnings of this condition like "mental factors" that contribute to habit continuation at root level, "poor quality of life," and "nutritional deficiencies" are still uncorrected. We believe that OSF must be managed at the "mind and body" level to accomplish holistic rehabilitation. Herein, we briefly describe the general condition of OSF patients, and highlight the possibility of a novel telomere-based assessment of psychosocial stress, and support use of meditation to counter excessive "telomere shortening" which is linked to oral cancer. We also expand the current view on "burning sensation" in OSF from an entire physical concept to a possibility of mental factors, and finally recommend rich, micronutrient supplementation (through diet) to counter free radical production within the hypoxic oral epithelium.