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Objectives Frequent consumption of acidic beverages and dietary preservatives in younger generation, diet-conscious (celebrities), and obese individuals have a rapid impact on demineralization of the teeth. An attempt was made to analyze the erosive potential of various acidic beverages. Materials and Methods One hundred and ninety extracted human permanent teeth were sectioned longitudinally, pre-weighed, randomly grouped, and placed in nine acidic beverages (200 mL) with predetermined pH, i.e., three commercially available fruit juices, three carbonated drinks, and three dietary preservatives. Statistical Analysis The sectioned specimens (n = 10) were analyzed at time intervals of 12, 24, 48, and 96 days. Mean weight loss was calculated, and surface changes were assessed under a stereomicroscope. The demineralization pattern and microscopic changes were observed under a compound and polarizing microscope. One-way analysis of variance test followed by Tukey's post-hoc analysis was employed. Results Overall the maximum demineralizing effect was caused by vinegar and apple cider. In the fruit juices category, lemon juice induced significant changes, while in the carbonated drinks category Coca-Cola induced the maximum changes and in the category of food preservatives vinegar induced the maximum changes. Severe discoloration was seen with respect to Coca-Cola followed by Mountain Dew (carbonated drink). Conclusion The present study is unique as three different types of microscopes have been employed and both dentin and enamel of permanent teeth have been analyzed. In addition, the effect of dietary preservatives on hard tissues was evaluated. Oral health educators can reinforce important practices such as decreasing the frequency of consumption and time duration of beverage contact with the teeth. Also, the use of mouth rinses and buffering agents after the consumption of dietary beverages can be advocated along with regular fluoride application for those who are regular consumers.

The impact of SARS-CoV-2 infection on heart transplant recipients is unknown. Literature is limited to case reports and series. The purpose of this study is to identify the clinical features, outcomes, and immunosuppression strategies of heart transplant recipients with COVID-19 infection. A systematic review was conducted using the search term “Coronavirus” or COVID,” “SARS-CoV-2,” “cardiac transplantation,” and “heart transplant.” Case reports and retrospective studies were gathered by searching Medline/PubMed, Google Scholar, CINAHL, Cochrane CENTRAL, and Web of Science. Thirty-three articles were selected for review. We identified 74 cases of SARS-CoV-2 infection in heart transplant and heart-kidney transplant recipients. The mean age was 60.5 ± 15.8 years, and 82.4% were males with median time from transplant of 6.5 years. Commonest symptoms were fever, cough, and dyspnea, but new left ventricular (LV) dysfunction was rare. Leukocytosis, lymphopenia, elevated inflammatory markers, and bilateral ground-glass opacities were common. Mortality was high, with particularly poor survival in patients who required intensive care unit (ICU) admission and older patients. Immunosuppression involved discontinuation of antimetabolites and steroids. COVID-19 infection in heart transplant (HT) recipients presents similarly to the general population, but new onset of LV dysfunction is uncommon. Immunosuppression strategies include increase in corticosteroids and discontinuation of antimetabolites.

Aim The assessment of hyalinization to determine aggressive behavior in oral pathological lesions is a scarcely researched field that requires further exploration. The current study aims to predict the biological behavior of oral hyalinizing extraosseous lesions (OHEOL) by employing four differential stains with clinicopathologic correlation. Materials and methods The study was performed on retrospectively diagnosed formalin-fixed paraffin-embedded cases of salivary gland tumors (SGTs) (n = 13), benign soft tissue (BST) lesions (n = 24), and oral submucous fibrosis (OSMF) (n = 53). The hematoxylin and eosin-stained sections were analyzed for the severity of hyalinization (SOH). Differential stains periodic acid Schiff (PAS), Alcian blue, safranin O, and picrosirius red with polarizing microscopy were used to assess the components of hyalinized tissue. The SOH was correlated with differential staining characteristics and clinicopathologic features to analyze possible correlation with aggressive potential in BST, advancement of disease in OSMF, and recurrence in SGT. Results Intensity of picrosirius red stain significantly correlated with SOH of SGTs (p = 0.044). The intensity of PAS stain (p = 0.040), picrosirius red polarizing greenish-yellow color (p = 0.002), and pattern of distribution of picrosirius red (p = 0.023) significantly correlated with recurrence of SGTs. The intensity of differential stains increased with the SOH in BST lesions indicating their correlation with SOH. The intensity (p = 0.008) and pattern (p = 0.010) of Alcian blue staining and intensity of safranin O stain (p = 0.003) significantly correlated with SOH in OSMF. Picrosirius red polarizing color reddish and yellowish red (p = 0.002) significantly correlated with SOH distinguishing early and advanced OSMF. Conclusion Picrosirius red and PAS stains are reliable indicators of SOH and recurrence potential in SGT. Alcian blue, safranin O, and picrosirius red polarizing colors enable detection of SOH and accurately distinguish early from advanced OSMF. Clinical significance SOH can be considered as a histological predictor of aggressive biologic behavior in OHEOL. These findings will result in appropriate management protocols.

Aim Odontogenic tumors (OTs) and bone lesions of the oral cavity present diverse histological features and varying clinical behavior that makes predicting their biologic behavior difficult. The research undertaken in the current study aims to predict the biological behavior of oral hyalinizing odontogenic and bone lesions (OHO-BL) for the first time by employing four differential stains with clinicopathologic correlation. Materials and methods The study was performed on retrospectively diagnosed formalin-fixed paraffin-embedded cases of OTs (n = 53) and bone lesions (n = 10). The severity of hyalinization (SOH) was assessed from stained tissue sections. Polarizing microscopy was used to analyze hyalinization in tissues stained with differential special stains, namely periodic acid-Schiff (PAS), Safranin-O, Alcian Blue, and Picrosirius red. SOH was also analyzed for possible correlation with recurrence and clinicopathologic correlation in OHO-BL. Results Intense staining was observed with PAS, Alcian Blue, and Safranin-O in OTs with increased SOH with a statistical significance. Polarizing greenish yellow color correlated significantly with the recurrence potential of the OT group. Recurrence in individual lesions of the OT group showed a statistically significant association with SOH. Such individual correlation was not observed in bone diseases. Conclusion PAS, Alcian Blue, Safranin-O, and Picrosirius red are reliable stains to assess hyalinization in OHO-BL. Picrosirius red-polarizing microscopy is a dependable tool for identifying recurrent odontogenic lesions. Clinical significance SOH can be considered a histological predictor of aggressive biologic behavior in oral hyalinizing odontogenic lesions that can enable the surgeon to arrive at an appropriate management protocol.

Oral cancer progression is a multi-step process in which adhesion molecules play a pivotal role in the development of recurrent, invasive, and distant metastasis. The aim of this study was to adopt prognostic biomarkers to assess the lymph node metastasis of OSCC that will facilitate in deciding the treatment modality by the surgeons. The objectives of the study were to assess the biological behaviour of OSCC by correlating the expression levels of P-Cadherin and WNT5A immunohistochemically. A total of 60 selected OSCCs cases (lymph node metastasis n = 30, non-metastatic n = 30) and 10 normal healthy controls were quantitatively and qualitatively analysed by immunohistochemistry for P-Cadherin and WNT5A. A survival analysis was also performed. The expression levels of P-Cadherin and WNT5A in OSCC groups were statistically significant between metastatic and non-metastatic groups (p

Background Hyalinization is a process of conversion of stromal connective tissue into a homogeneous, acellular translucent material. Nevertheless, hyalinization could provide insights into the biologic behaviour and prognosis of pathological lesions. Few studies with limited sample size have intended to assess the correlation of hyalinization and biologic behaviour in oral lesions. Aim The current review aims to comprehensively appraise the mechanism of hyalinization in pathological oral hyalinizing lesions (OHL) and its clinical implications with emphasis on differential stains employed. Methods An electronic search was performed in the PubMed database (from year 2000–2020) using the keywords “special stains in oral hyalinizing lesions”, “significance of hyalinization in oral lesions” and “hyalinization and biologic behaviour”. Original research articles analyzing the effect of hyalinization on biologic characteristics of the lesion were evaluated in this review. Narrative review articles that provided insights into the mechanism of hyalinization and maturity of collagen fibers were also considered for analysis. Conclusion The presence of hyalinization does seem to have a significant effect on the biologic behaviour of pathological lesions. There is substantial scope to further investigate the process of hyalinization on larger samples and its correlation with the aggressive behaviour of OHLs. Special stains and advanced investigations such as immunohistochemistry for stromal markers would define the nature of hyalinized material and validate the correlation. Clinical significance The prediction of the biologic behaviour of a lesion established through assessment of hyalinization would prevent unwanted over or under treatment leading to a better prognosis.

(1) Background: Odontogenic keratocysts (OKCs) are enigmatic developmental cysts that deserve special attention due to their heterogeneous appearance in histopathological characteristics and high recurrence rate. Despite several nomenclatures for classification, clinicians still confront challenges in its diagnosis and predicting its recurrence. This paper proposes an ensemble deep-learning-based prognostic and prediction algorithm, for the recurrence of sporadic odontogenic keratocysts, on hematoxylin and eosin stained pathological images of incisional biopsies before treatment. (2) Materials and Methods: In this study, we applied a deep-learning algorithm to an ensemble approach integrated with DenseNet-121, Inception-V3, and Inception-Resnet-V3 classifiers. Around 1660 hematoxylin and eosin stained pathologically annotated digital images of OKC-diagnosed (60) patients were supplied to train and predict recurrent OKCs. (3) Results: The presence of SEH (p = 0.004), an incomplete epithelial lining, (p = 0.023), and a corrugated surface (p = 0.049) were the most significant histological parameters distinguishing recurrent and non-recurrent OKCs. Amongst the classifiers, DenseNet-121 showed 93% accuracy in predicting recurrent OKCs. Furthermore, integrating and training the traditional ensemble model showed an accuracy of 95% and an AUC of 0.9872, with an execution time of 192.9 s. In comparison, our proposed model showed 97% accuracy with an execution time of 154.6 s. (4) Conclusions: Considering the outcome of our novel ensemble model, based on accuracy and execution time, the presented design could be embedded into a computer-aided design system for automation of risk stratification of odontogenic keratocysts.

The histologic properties of tumors seem to affect their biological behavior, and the same holds good for solid multicystic ameloblastoma (SMA), a benign, locally destructive lesion. Hyalinization is one such histological factor that has been demonstrated to correlate with the biological behavior of neoplasms. The present study aimed to analyze the correlation between the severity of hyalinization (SOH) and the recurrence potential of SMAs. The study was performed on formalin-fixed, paraffin-embedded (FFPE) diagnosed archival cases of SMA, follicular SMA (n = 35) and plexiform SMA (n = 25). The cases were evaluated for SOH and scored from 0–3, and the correlation between SOH and recurrence was analyzed for statistical significance. The clinical parameters of the lesion were analyzed for statistical correlation with recurrence. The SOH significantly correlated with the recurrence of SMA (p = 0.001). The histologic type did not influence the biological behavior of SMA. The location of SMA in the body of the mandible (p = 0.036), multilocular radiolucency (p = 0.001) and root resorption (p = 0.002) also showed strong statistical correlation with recurrence. It is evident from the present study that hyalinization strongly correlates with the biological behavior of SMA. Future studies with advanced investigations could validate the presence of hyalinization and identify the origin of the hyalinized product in SMAs.

The present study evaluated the histopathological changes in oral tissues induced by pesticide poisoning.This was a cross-sectional pilot study. The sample consisted of oral tissues obtained from deceased patients during autopsy. The study samples were obtained from 10 cases of ingested pesticide poisoning, and the control samples were obtained from road traffic accident cases. All the obtained samples were subjected to histopathological examinations. The changes observed in poisoning cases were compared to those in the road traffic accident cases.Significant degenerative changes were observed in the epithelial cells and connective tissue components, such as collagen, muscles, nerves, vasculature, adipose tissue, and salivary acini and ducts, in the poisoning cases. The oral tissues of the road traffic accident cases did not show any significant degenerative changes.The degenerative changes in the study samples can be attributed to the direct contact of the pesticide with the autopsied oral tissues when the poison was consumed. There are instances in which the entire body may not be recovered or may not be in an examinable state. In such cases, an oral autopsy could provide additional evidence for determining the cause of death in suspected poison cases.

The inductive effect of hyalinisation and its influence on the biologic behaviour of ameloblastoma variants represent a scarcely researched domain of oral pathology. The complexity of the induction effects within the odontogenic apparatus, with the involvement of both ectodermal and mesodermal tissues, is responsible for diverse histopathological characteristics, hyalinisation being the major feature. The present study aims to deduce for the first time the correlation between the severity of hyalinisation (SOH) and recurrence in three unicystic ameloblastoma (UA) variants, namely, intra-luminal (UA-IL), luminal (UA-L) and mural (UA-M). Retrospectively diagnosed archival cases of UA-IL (n = 08), UA-L (n = 22) and UA-M (n = 30) were assessed for SOH and its correlation with recurrence. A subgroup comparison (between UA-IL/UA-L and UA-M) was also performed. The clinical parameters of the patients were also analysed from files for clinicopathological correlation with recurrence. Results: sub-epithelial hyalinisation (SEH) significantly correlated with the recurrence of UA-L and UA-M (p = 0.001). When the histologic types (UA-L and UA-IL vs. UA-M) were grouped and the correlation of SOH with recurrence was checked, it was observed that both groups (p = 0.001) showed strong statistical correlation. UA-M lesions with multilocular radiolucency (p = 0.001) also showed significant correlation with recurrence. SOH can be a reliable histological predictor of recurrence and of aggressive biologic behaviour in UA. The present study shows a significant association of hyalinisation with the biologic behaviour of UA. Further studies with immunohistochemical investigations could validate the presence of hyalinisation and identify the origin of the hyalinised product in UAs.

Background The presence and grading of oral epithelial dysplasia (OED) are considered the gold standard for predicting the malignant risk of oral potentially malignant disorders. However, inter-observer and intra-observer agreement in the context of reporting on OED grading has been reputedly considered unreliable. Methods We undertook a multi-centre study of six Indian oral pathologists to assess variations in reporting OED using the World Health Organization (WHO; 2005) system and also the recently introduced binary system. The observer variability was assessed with the use of kappa statistics. Results The weighted kappa intra-observer agreement scores improved (κw = 0.5012) on grouping by two grades as no and mild dysplasia versus moderate and severe dysplasia compared to binary grading system (κ = 0.1563) and WHO grading system (κw = 0.4297). Poor to fair inter-observer agreement scores were seen between the principal investigator (PI) and the other five observers using the WHO grading system (κ = 0.051-0.231; κw = 0.145 to 0.361; 35% to 46%) and binary grading system (κ = 0.049 to 0.326; 50 to 65%). Conclusions There is considerable room for improvement in the assessment of OED using either system to help in standardised reporting. The professional pathology organisations in India should take steps to provide external quality assessment in reporting OED among oral and general pathologists who are engaged in routine reporting of head and neck specimens.

Recent scientific advances have presented substantial evidence that there is a multifaceted relationship between the microbiome and cancer. Humans are hosts to multifarious microbial communities, and these resident microbes contribute to both health and disease. Circulating toxic metabolites from these resident microbes may contribute to the development and progression of cancer. The aim of this systematic review was to evaluate microbiome and microbial shift contribution to the development and progression of cancer. This systematic review provides an analytical presentation of the evidence linking various parts of the microbiota to cancer. Searches were performed in databases such as PubMed, Google Scholar, Scopus, EBSCO, E-Journals and Science Direct from the time of their establishment until May 2018 with the following search terms: cancer or human microbe or cancer and human microbiome AND shift in microbes in cancer. The merged data were assessed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Cochrane’s Risk of Bias Tool was used to assess the bias. Initially, 2,691 articles were identified, out of which 60 full-text articles were screened and re-evaluated. Among them, 14 were excluded based on inclusion/exclusion criteria; eventually, 46 articles were included in the systematic review. The reports of 46 articles revealed that microbial shift involving Candida species, Fusobacterium nucleatum, Porphyromonas gingivalis, Helicobacter pylori and Human papilloma virus (HPV) 16 & 18 were most commonly involved in various human cancers. In particular, organisms, such as Candida albicans, Fusobacterium nucleatum, Porphyromonas gingivalis and HPV-16 were found to be more prevalent in oral cancer. The present systematic review emphasizes that the role and diverse contributions of the microbiome in carcinogenesis will provide opportunities for the development of effective diagnostic and preventive methods.

Aim: This in vivo study compared clinical, histological, and radiological differences in bone formation in human extraction sockets grafted with demineralized freeze-dried bone allograft (DFDBA) and platelet-rich fibrin (PRF), with nongrafted sockets and bone–implant contact (BIC) at 3 and 6 months after implant placement. Settings and Design: Randomised controlled trial. Materials and Methods: The study comprised thirty posterior teeth sockets in either arch in patients ranging from 25 to 60 years. The patients were divided into two equal groups – Group I: control group wherein no graft was placed and the extraction socket was left to heal normally and Group II: test group in which DFDBA and PRF were placed after extraction. 12–16 weeks after extraction, a trephine biopsy was done just prior to implant placement, followed by implant placement. Cone-beam computed tomography (CBCT) at 3 and 6 months after implant placement was done to assess BIC. Statistical Analysis Used: Descriptive and Inferential statistical analysis was done. Parametric test: Independent t-test was used for intergroup analysis and dependent t-test for intra-group analysis. Results: Lower buccal bone levels were seen in the control group versus test group at all intervals though moderately significant. Lingual bone levels significantly reduced at all the three intervals for the control group as compared to the test group. Ridge width in both groups reduced in a time span of 6–7 months without any significant difference. Better bone conversion was noted in the preserved sockets. The preserved sockets also showed better BIC 3 months after implant placement and loading. Conclusion: Indigenously developed DFDBA material shows promising results as an osteoinductive material.

Autophagy, first described by the Belgian biochemist Christian De Duve in 1963, is derived from the Greek word "autóphagos," which means "self-devouring." It is a cellular homeostatic process in which the body rids itself of flawed or damaged cells and other defective cellular organelles.

Background: The goal of the study was to create a histopathology image classification automation system that could identify odontogenic keratocysts in hematoxylin and eosin-stained jaw cyst sections. Methods: From 54 odontogenic keratocysts, 23 dentigerous cysts, and 20 radicular cysts, about 2657 microscopic pictures with 400× magnification were obtained. The images were annotated by a pathologist and categorized into epithelium, cystic lumen, and stroma of keratocysts and non-keratocysts. Preprocessing was performed in two steps; the first is data augmentation, as the Deep Learning techniques (DLT) improve their performance with increased data size. Secondly, the epithelial region was selected as the region of interest. Results: Four experiments were conducted using the DLT. In the first, a pre-trained VGG16 was employed to classify after-image augmentation. In the second, DenseNet-169 was implemented for image classification on the augmented images. In the third, DenseNet-169 was trained on the two-step preprocessed images. In the last experiment, two and three results were averaged to obtain an accuracy of 93% on OKC and non-OKC images. Conclusions: The proposed algorithm may fit into the automation system of OKC and non-OKC diagnosis. Utmost care was taken in the manual process of image acquisition (minimum 28–30 images/slide at 40× magnification covering the entire stretch of epithelium and stromal component). Further, there is scope to improve the accuracy rate and make it human bias free by using a whole slide imaging scanner for image acquisition from slides.

Targeting cancer stem cell (CSC) subpopulation within the tumor remains an obstacle for specific therapy in head-and-neck squamous cell carcinoma (HNSCC). Few studies in the literature describe a panel of stem cell makers, however a distinct panel has not been put forth. This systematic review aims to enhance the knowledge of additional markers to accurately relate their expression to tumorigenesis, metastasis, and therapy resistance. Databases, including PubMed, Google Scholar, Ebsco, and Science Direct, were searched from 2010 to 2017 using various combinations of the following keywords: "Stem cell markers in HNSCC" and "chemoresistance and radioresistence in HNSCC." Original experimental studies (both in vitro and in vivo) published in English considering stem cell markers in HNSCC, were considered and included. We excluded articles on tumors other than HNSCC, reviews, editorial letters, book chapters, opinions, and abstracts from the analyses. Forty-two articles were included, in which 13 types of stem cell markers were identified. The most commonly expressed CSC markers were CD44, aldehyde dehydrogenase, and CD133, which were responsible for tumorigenesis, self-renewal, and therapy resistance, whereas NANOG, SOX-2, and OCT-4 were involved in metastasis and invasion. Identification of an accurate panel of CSC markers is the need of the hour as nonspecificity of the current markers poses a problem. Further studies with a large sample size would help validate the role of these CSC markers in HNSCC. These CSC proteins can be developed as therapeutic targets for HNSCC therapy, making future treatment modality more specific and effective.

Odontogenic tumors (OTs) and bone lesions of the oral cavity present diverse histological features and varying clinical behavior that makes predicting their biologic behavior difficult. The research undertaken in the current study aims to predict the biological behavior of oral hyalinizing odontogenic and bone lesions (OHO-BL) for the first time by employing four differential stains with clinicopathologic correlation.The study was performed on retrospectively diagnosed formalin-fixed paraffin-embedded cases of OTs (n = 53) and bone lesions (n = 10). The severity of hyalinization (SOH) was assessed from stained tissue sections. Polarizing microscopy was used to analyze hyalinization in tissues stained with differential special stains, namely periodic acid-Schiff (PAS), Safranin-O, Alcian Blue, and Picrosirius red. SOH was also analyzed for possible correlation with recurrence and clinicopathologic correlation in OHO-BL.Intense staining was observed with PAS, Alcian Blue, and Safranin-O in OTs with increased SOH with a statistical significance. Polarizing greenish yellow color correlated significantly with the recurrence potential of the OT group. Recurrence in individual lesions of the OT group showed a statistically significant association with SOH. Such individual correlation was not observed in bone diseases.PAS, Alcian Blue, Safranin-O, and Picrosirius red are reliable stains to assess hyalinization in OHO-BL. Picrosirius red-polarizing microscopy is a dependable tool for identifying recurrent odontogenic lesions.SOH can be considered a histological predictor of aggressive biologic behavior in oral hyalinizing odontogenic lesions that can enable the surgeon to arrive at an appropriate management protocol.

Human telomerase reverse transcriptase enzyme, the catalytic subunit of telomerase are seen to be frequently reactivated in cancers including Oral squamous cell carcinoma (OSCC). Increased hTERT expression have been seen in potentially malignant conditions including Oral submucous fibrosis (OSMF). The aim of the study was to evaluate the expression levels in OSMF, OSCC in the background of OSMF and OSCC using immunohistochemistry and also to correlate hTERT expression with clinicopathologic parameters. A total of 50 histopathologically diagnosed cases of 20 OSMF, 20 OSCC wherein 5 were OSCC in the background of OSMF and 10 Normal oral mucosae were retrieved from the departmental archives and subjected to immunohistochemical analysis of hTERT. The expression of hTERT increased from normal, OSMF, to OSCC with statistically significant differences in mean labelling score (LS). We also found a shift in cellular localization of stain where, normal mucosal tissues showed a nuclear stain unlike OSMF, where combined nuclear and cytoplasmic staining as noted. The tumor cells in OSCC showed predominant cytoplasmic staining. There was no correlation between hTERT expression and clinicopathological parameters of OSMF. However, a significant increase of hTERT expression was seen with increasing histological grading of OSCC. These results suggest the role of hTERT in the early event of malignant transformation of OSMF. Telomerase could be used as a potent diagnostic marker to identify high-risk group of OSMF.

Background Researchers have struggled to understand the natural history of lesions presenting with both lichenoid features and epithelial dysplasia. Thus the present study was designed to differentiate between OLP, OLP with dysplasia, epithelial dysplasia and epithelial dysplasia with lichenoid features based on the expressions of ki-67, p53, COX-2, and α-SMA. Materials and methods Formalin-fixed paraffin-embedded archival specimens of OLP, OLP with dysplasia, epithelial dysplasia and epithelial dysplasia with lichenoid features were subjected to immunohistochemical staining with ki-67, p53, COX-2, and α-SMA. Results Ki-67 exhibited strong positivity in 100% (6/6) of epithelial dysplasia cases, 71.4% (5/7) of lichenoid dysplasia cases, 57.1% (4/7) of OLP cases and 60% (3/5) of OLP with dysplasia cases. Strong p53 staining was evident in more cases of lichenoid dysplasia [42.8% (3/7)], while moderate staining was more frequent in OLP cases [42.8% (3/7)] and OLP with dysplasia cases [42.8% (3/7)] and mild intensity was more frequent in epithelial dysplasia cases [50% (3/6)] followed by lichenoid dysplasia cases [42.8% (3/7)], OLP cases [28.5 (2/7)] and OLP with dysplasia cases [40% (2/5)]. COX-2 strong positivity was more frequent in cases of epithelial dysplasia cases [57.1% (4/7)] and OLP [50% (3/6)]. Strong α- SMA staining was noted more frequently in lichenoid dysplasia cases [71.4 (5/7)], followed by OLP cases [42.8% (3/7)] and OLP with dysplasia cases [60% (3/5)]. Conclusions Ki-67, p53, α-SMA and COX-2 expression do not differentiate between OLP, LP with dysplasia and epithelial dysplasia with lichenoid features.

Graphical abstract, Highlights • Coelomic fluid of 3 earthworms exhibited cytotoxicity on oral cancer cells. • Coelomic fluid induced DNA strand breakage in SCC-9 cells. • Coelomic fluid of Perionyx excavatus showed highest percentage of G2M cycle arrest. • Coelomic fluid of Eudrilus eugeniae caused a 37-fold rise in apoptosis percentage., The current protocol of cancer management includes surgery, radiotherapy and chemotherapy. However, these modalities have significant adverse effects and affect the quality of life. Further intensification of treatment is hindered as maximal toxicity levels are reached impeding improvement. Hence researchers are in the quest for adjunctive naturally available therapies that can alter tumor proliferation without causing significant adverse reactions. The present study aims to explore the cytotoxic potential of earthworm coelomic fluid (ECF) of Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) on oral cancer cell line SCC-9. The effect of ECF on cell cycle analysis and mechanism of cell death have also been investigated. All experiments reported in this paper were performed as 3 replicates per experiment. The results indicated that ECF of EE, EF and PE have potent variable cytotoxic effect on SCC-9 cells demonstrated through LDH, clonogenic and comet assay. An effective cell cycle arrest was observed at the G2M phase of cell cycle with apoptotic induction that was observed through an Annexin V – FITC/PI assay. ECF of EE was found to be superior in its cytotoxic action closely followed by ECF of PE. The present findings provide evidence for the first time that ECF of EE, EF and PE have potent cytotoxic effect on oral cancer cells in vitro. They significantly induce G2M cell cycle arrest and promote apoptosis in SCC-9 cell line. Gene expression studies have been planned to ascertain the pathways of cell death.