Anticancer Effects of Carica papaya L. and Benzyl Isothiocyanate on an Oral Squamous Cell Carcinoma Cell Line: An In Vitro Study.

AIM: The study aimed to assess the anticancer effects of leaves of the male and female plant and seeds Carica papaya L. extract and the active compound benzyl isothiocyanate on oral squamous cell carcinoma (OSCC) cell line. MATERIALS AND METHODS: Extracts of CO2 strain C. papaya L. seeds were prepared using water, ethanol, and ethanol:water by maceration, and benzyl isothiocyanate was quantified. Alkaloid fractions of leaves of male and female plants of C. papaya L. were prepared and quantified. The anticancer effects of the test substances on the SCC-25 cell line were assessed by MTT, apoptosis assay, cell cycle analysis, and determination of mitochondrial membrane potential. RESULTS: The ethanol:water extract of C. papaya L. (seeds) demonstrated the highest quantity of benzyl isothiocyanate. Male plant leaves demonstrated greater alkaloid content. The leaves of the male plant exhibited apoptosis induction and S-phase arrest, whereas the leaves of the female plant and seeds of C. papaya L. demonstrated G2M-phase arrest and apoptosis induction. CONCLUSION: C. papaya L. and benzyl isothiocyanate demonstrated anticancer effects. There was a difference in the anticancer effects of leaves of male and female plants of C. papaya L. CLINICAL SIGNIFICANCE: The anticancer effects of papaya leaves and seeds could be further explored to develop an adjunct therapy for oral cancer to improve prognosis and reduce recurrence rates.

In Vitro Anticancer Effects of Cinnamomum verum J. Presl, Cinnamaldehyde, 4 Hydroxycinnamic Acid and Eugenol on an Oral Squamous Cell Carcinoma Cell Line.

AIM AND OBJECTIVE: The present study was conducted to assess the in vitro anticancer effects of Cinnamomum verum J. Presl extract and its active constituents, such as cinnamaldehyde, 4 hydroxycinnamic acid, and eugenol on oral squamous cell carcinoma cell line. MATERIALS AND METHODS: Aqueous, ethanolic, and hydroalcoholic extracts of C. verum J. Presl (bark) were prepared using standardized protocols. Cinnamaldehyde, 4 hydroxycinnamic acid, and eugenol were quantified in the extracts. Total saponins, tannins, and polyphenols were quantified in the selected extracts. A commercially available SCC-25 cell line was cultured according to standard protocol. The anticancer effects of the extract, active compounds, and standard cisplatin were assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cytotoxicity, acridine orange/ethidium bromide staining, DNA, fragmentation assay, cell cycle analysis by flow cytometry, and JC-1 staining (5,5',6,6'-tetrachloro1,1',3,3'tetraethylbenzimidazolylcarbocyanine iodide). RESULTS: The hydroalcoholic extracts demonstrated a higher quantity of the active ingredients cinnamaldehyde, 4 hydroxycinnamic acid, and eugenol. The selected extract and active compounds demonstrated anticancer effects via apoptosis induction and S-phase arrest. Apoptosis induction was exerted by the extract via alteration in mitochondrial membrane potential. CONCLUSION: Cinnamomum verum J. Presl and its active compounds exhibited in vitro anticancer effects on oral squamous cell carcinoma. Further studies in animal models have to be carried out to assess toxicity and in vivo effects. CLINICAL SIGNIFICANCE: The anticancer properties of Cinnamomum verum J. Presl could be explored further for prevention and management of oral squamous cell carcinoma.

Overexpression of deubiquitinase (usp 36) in oral squamous cell carcinoma.

Aim: Oral cancer is one of the top three types of cancer and is of significant public health importance in India. A common post-translational modification in cells is ubiquitination/deubiquitination, and its dysregulation is closely associated with the development of cancer. Studies on the role of ubiquitination in oral squamous cell carcinoma (OSCC) are lacking. Increased expression of usp36 has been observed in various types of cancer, and this study aimed to check the gene expression of usp36 in OSCC patients. In this study, we analyzed the expression of ubiquitin-specific proteases (USPs) 36 in OSCC. Materials and Methods: A total of 15 OSCC patients at different stages of tumor differentiation and age- and sex-matched controls were recruited for the study. The patients were categorized based on their differentiation patterns. RNA was extracted from the tissues, and usp36 gene expression was checked in these samples using a quantitative real-time PCR technique. Results: Our study showed increased expression of usp36 gene in OSCC patients. The usp36 mRNA was 231.8 ± 137.94 folds higher in well-differentiated squamous cell carcinoma patients, 38.18 ± 3.77 folds higher in moderately differentiated squamous cell carcinoma patients, and 25.49 ± 7.30 folds higher in poorly differentiated squamous cell carcinoma patients compared to control tissues. Conclusion: Our study reports, for the first time, an increased gene expression of usp36 in OSCC tissues.

Lipoid proteinosis presenting with an unusual nonsense Q32X mutation in exon 2 of the extracellular matrix protein 1 gene.

Lipoid proteinosis (LP) is a rare disorder characterized by extensive hyaline-like deposits on the skin, mucous membranes and various internal organs with varying clinical manifestations. The disorder has been recently shown to result from loss-of-function mutations in the extracellular matrix protein 1 gene (ECM1) on 1Q21. The two cases reported here had typical clinical and histological features consistent with LP. Direct sequencing of amplified DNA from the second patient showed a single nucleotide substitution (C > T) at nucleotide 94 within exon 2 of the ECM1 gene, nonsense mutation Q32X. This is the second case reported of LP with involvement of exon 2 of ECM1.

Centromere Protein F (CENPF): A novel marker for salivary gland pathology.

Background: Salivary gland tumours are relatively uncommon, and there exists considerable diagnostic difficulty. This is due to individual lesions having diverse histopathological features, presence of number of types and variants, and overlapping histological features in different tumour entities. Aim: The current study aimed at assessing the expression of centromere protein F (CENPF) in benign and malignant salivary gland tumours and to evaluate the efficacy of CENPF as a proliferative marker to aid in the diagnosis of malignancy so that it will help in surgical pathology practice. Materials and Methods: The study group involved 20 cases of benign salivary gland tumours, 20 cases of malignant salivary gland tumours, and 10 normal salivary gland tissues. All the cases were subjected to immunohistochemical analysis for CENPF expression and were assessed by two independent observers and further taken up for evaluation. Statistical Analysis: The results were analysed statistically among different groups using analysis of variance (ANOVA) or Kruskal-Wallis test with Chi-squared test using IBM's Statistical Package for the Social Sciences (SPSS) version 17.0. Results: CENPF expression in normal salivary gland was negative with gradual increase in expression from benign salivary gland tumours to malignant salivary gland tumours. CENPF expression was high in malignant salivary gland tumours. Conclusion: Findings of the study suggest that CENPF can be regarded as a new cell proliferation marker for malignant salivary gland tumours.

Evaluation of the genotoxic effects of fixed appliances on oral mucosal cells and the relationship to nickel and chromium concentrations: an in-vivo study.

INTRODUCTION: The release of metal ions from fixed orthodontic appliances is a source of concern. The aim of this study was to evaluate genotoxic damage in the oral mucosal cells of patients wearing fixed appliance, and the nickel and chromium ion contents in these cells. METHODS: Twenty patients undergoing orthodontic treatment formed the experimental group, and 20 untreated subjects comprised the control group. Oral mucosal smears were collected at 2 times: at debonding and 30 days after debonding. The smears were stained with Papanicolaou stain and studied under a light microscope to evaluate the presence of micronuclei. Inductively coupled plasma-mass spectrometry was used to quantify the presence of metal ions. The data were subjected to the Mann-Whitney U test and the Spearman rank correlation test. RESULTS: The mean micronuclei frequency was significantly higher in the treated group than in the control group at debonding; the difference was smaller and not statistically significant 30 days after debonding. The nickel and chromium ion contents in the experimental group were not significantly higher than in the control group. No correlation could be established between micronuclei frequency and metal ion content. CONCLUSIONS: Nickel and chromium alloys of orthodontic appliances emit metal ions in sufficient quantities to induce localized genotoxic effects, but these changes revert on removal of the appliances.

In vitro anticancer activity of a pentacyclic triterpenoid via the mitochondrial pathway in bone-invasive oral squamous cell carcinoma.

CONTEXT: Oral cancer is the most dreadful cancer worldwide with a 5-year survival rate of approximately 50%. Anticancer therapies such as chemotherapy and radiotherapy result in severe side effects. AIM: We aimed to evaluate the in vitro anticancer activity of Asiatic acid (AA) on bone-invasive oral squamous cell carcinoma (BHY) cell line. SETTINGS AND DESIGN: This was an in vitro laboratory setting. MATERIALS AND METHODS: BHY cell lines were used for the experiment. Confocal microscopy was used to observe cellular alterations. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the IC50 concentration of AA and flow cytometry to analyze the percentage of cells in each phase of the cell cycle post treatment. Immunoblot assays and semiquantitative reverse transcriptase-polymerase chain reaction (rt-PCR) were used to study the expression level of genes involved. STATISTICAL ANALYSIS USED: Student's t-test and one-way analysis of variance were used for statistical analysis. RESULTS: IC50 concentration of AA was 15.6 µM. On flow cytometry analysis, treatment with 15.6 µM and 31.25 µM of AA for 24 h increased the percentage of cells in the G2/M phase to 45.63% and 53.12%, respectively, compared to 9.62% in control group. Immunoblot analysis and semiquantitative rt-PCR demonstrated an upregulation of p53, cyclin-dependent kinase inhibitors (p21 and p27), caspase-3, caspase-9, cytochrome c and Bax in a time-dependent manner and downregulation of cyclins and anti-apoptotic protein Bcl-2 (**P < 0.05, ***P < 0.001 versus control) post AA treatment. CONCLUSION: AA induces apoptosis via the mitochondrial-dependent pathway and causes cell cycle arrest at the G2/M phase in BHY cell line.

Pleomorphic Sarcoma of the Oral Cavity: A Rare Case Entity.

Pleomorphic sarcomas are extremely rare in the oral cavity, representing only 5% of all soft tissue sarcomas in the extremities of adults. They form a group of highly undifferentiated pleomorphic malignant tumors. A 36-year-old male reported to the OP with a chief complaint of a tooth ache in the right lower quadrant for the past 6 months. The patient gave a history of extraction of a mobile tooth in that region. A radiograph of the region was advised. An incisional biopsy was also done and sent for histopathological examination. The microscopy revealed the presence of spindle-shaped cells showing dysplastic features. A panel of immunohistochemical markers were performed to identify the tissue of origin of the lesion. The article details the clinical, radiographic, and microscopic features of a pleomorphic soft tissue lesion along with the array of immunohistochemical markers, leading to the final diagnosis of such enigmatic lesions.

Oral Lichen Planus in a 7-year-old Child: A Rare Case Report.

Lichen planus is a chronic inflammatory mucocutaneous disease reported most frequently in adults and relatively rare in children with the prevalence being 0.03%. This article reports a case of oral lichen planus (OLP) in a 7-year-old girl without any attendant skin lesions, diagnostic workup, and management protocol for the same. This article also emphasizes the need to consider OLP as a differential diagnosis for white lesions of oral mucosa in children. How to cite this article: Chinnasamy NK, Venugopal DC, Sankarapandian S, et al. Oral Lichen Planus in a 7-year-old Child: A Rare Case Report. Int J Clin Pediatr Dent 2020;13(1):91-93.

Prevalence of Common Clinically Manifested Developmental Anomalies of the Oral Cavity Among Adults - An Epidemiological Study in a South Indian Population.

Introduction Developmental anomalies are malformation which arises due to the disturbances during the development of the organs. Although there have been many studies that have described the prevalence of these anomalies in the oral cavity, none have specified the prevalence of clinically manifested anomalies and their distribution between gender. Materials and methods In this study, 500 patients aged 18 to 50 years were screened for clinically manifested developmental anomalies. These were then divided based on age, sex, and jaws, which were then analyzed using a chi-square test and tabulated. Results We detected anomalies in 12.2% of the study population. Supernumerary teeth were the most prevalent anomaly detected (4.25%). The frequency of developmental anomalies was higher in men (57.1%). Conclusions Supernumerary teeth were the most widely recognized anomaly. Other anomalies identified were related to the shape and size of teeth. These anomalies can lead to severe orofacial problems. Therefore, proper care of these anomalies should be taken.

Evaluation of Oral Health Status Using the Geriatric Oral Health Assessment Index Among the Geriatric Population in India: A Pilot Study.

Introduction and aim Clinical indicators alone are insufficient for evaluating oral health. In addition to health and disease, oral health includes socio-dental indicators of physical, psychological, and social aspects of well-being. The adaptive capacity of an individual influences the perception of oral health-related quality of life (OHRQoL). Indices such as the Oral Health Impact Profile, Oral Impacts on Daily Performances, and the Geriatric Oral Health Assessment Index (GOHAI) have been used to measure OHRQoL. This study was designed to assess OHRQoL in older individuals using the GOHAI. Methods Subjects aged older than 65 years who visited our institution from January to March 2016 were included. Subjects with cognitive behavior disorders were excluded. Subjects were assigned into three groups based on age: 65-69 years, 70-74 years, and 75 years or older. The participants were asked 12 questions, and their responses were assessed by age group. Our Institutional Ethics Committee approved the study protocol. Results The 219 subjects recruited included 126 (57.5%) patients aged 65-69 years, 57 (26.0%) patients aged 70-74 years, and 36 (16.4%) patients aged 75 years or older. Several physical, physiological, and psychological aspects of the GOHAI differed significantly among these three groups, with overall OHRQoL decreasing with age. Conclusion Although oral healthcare problems were widespread in the geriatric population, they were not a primary concern. Attitudes toward dentistry require improvement. However, further studies in larger populations are required to assess geriatric oral health.

Oral medullary plasmacytoma: Rare case reports.

Medullary plasmacytoma (MP) or osseous or solitary bone plasmacytoma is a specializing hematopathology lesion typically present with local symptoms, such as pain, paresthesia and pathologic bone fractures as a result of proliferation of plasma cells. The most often involved sites are active hematopoietic long bones and the vertebrae. The clinical course of disease is identical to spectrum of other plasma cell dyscrasias. The diagnostic criteria include punched-out radiolucencies, monoclonal plasma cells and M protein. This lesion should be considered for the differential diagnosis of bone tumors. It is highly radiosensitive although combination modalities of radiation, surgery and chemotherapy have been used in the treatment. The long-term follow-up is essential. We report two rare cases of oral MP with unusual clinical presentation.

A Rare Case of Intraoral Psoriasis.

Psoriasis is a chronic inflammatory mucocutaneous disease predominantly affecting the skin. While oral manifestations are common in many papulosquamous conditions, the occurrence of oral lesions in psoriasis is relatively rare. This case report highlights the presence of oral lesions in cutaneous psoriasis and the need for early identification of such lesions for timely management and better symptomatic improvement.

Characterization of oral fibroblasts: An in vitro model for oral fibrosis.

BACKGROUND: Oral submucous fibrosis (OSMF) is a chronic debilitating condition of the oral mucosa that has been classified as a potentially malignant disorder with a malignant transformation rate of 2%-8%. Several in vitro and in vivo experiments have been performed to formulate a treatment modality for OSMF, yet no ideal in vitro primary oral fibroblast model has been developed. AIM: To establish an in vitro primary oral fibroblast model. SETTING AND DESIGN: In vitro laboratory setting. MATERIALS AND METHODOLOGY: Primary cell culture protocol was performed after obtaining normal oral tissue. Karyotyping was performed to rule out chromosomal abnormalities. Immunofluorescence staining was carried with a panel of fibroblast-specific markers (vimentin, phalloidin, transforming growth factor-ß receptor 1 [TGFßR1] and s100a4) and Masson trichrome staining (MTS) to demonstrate the presence of extracellular matrix (ECM) qualitatively. RESULTS: A monolayer of oral fibroblasts was observed on the 9th-day postseeding. No chromosomal abnormality was observed in the patient samples. Positive staining was observed with vimentin, phalloidin, TGFßR1 and s100a4, thereby confirming the cell type. MTS revealed fibroblasts with spindle morphology and scanty ECM. CONCLUSION: The present study lays down a protocol to design and characterize primary buccal fibroblast cell culture model that would aid researchers in performing in vitro preliminary experiments in areas concerning fibrosis.

Toxicity of xylene in occupationally exposed workers: A high-performance liquid chromatography analysis.

BACKGROUND: Xylene is one of the most commonly used solvents in industrial and medical technologies. Several health hazards of xylene have been documented in literature. Workers in certain forces appear to have the greatest potential for exposure to high concentrations of xylene - histopathology technicians and painters are two such groups. This study was undertaken with the aim to determine the level of xylene exposure and the various systemic health effects among these groups. METHODOLOGY: The study was performed by analyzing the urine samples of the participants for methylhippuric acid, the established biomarker of xylene with the aid of high-performance liquid chromatography. RESULTS AND CONCLUSION: The work hours per week of the occupationally exposed participants were statistically analyzed with that of the excretory values of the metabolites of xylene, and the P value was found to be highly significant. Various side effects of xylene including respiratory, dermatological, neurological and gastrointestinal symptoms were observed among the study groups.

Immunohistochemical quantification of mast cells in inflamed and noninflamed pulp tissue.

BACKGROUND: Dental pulp inflammation is a very complex process due to its situation in low compliance (confined environment) surrounded by mineralized dentin. Mast cells are one of the mediators of inflammation. Immunohistochemical localization of mast cells by anti-tryptase antibodies in formalin-fixed paraffin-embedded sections has been shown to be highly specific. The aim of the present study is to quantify the mast cells in inflamed and noninflamed human pulp tissue using immunohistochemical analysis. MATERIALS AND METHODS: Immunohistochemical localization of mast cells by anti-tryptase antibody was done in 15 inflamed and 15 noninflamed formalin-fixed paraffin-embedded pulp tissue. A number of mast cell per five high-power fields were performed using three observers and the mean was calculated. Statistical analysis was performed using t-test. RESULTS: The results of the present study showed an increased number of mast cells in inflamed pulp in comparison with noninflamed pulp. CONCLUSION: Thus, the presence of mast cells in pulp inflammation could be used as a diagnostic marker. It can also aid in the management of pulpitis as mast cell stabilizers and antihistaminic agents could be used to control pulpal pain and inflammation.

Estimation of time since death based on light microscopic, electron microscopic, and electrolyte analysis in the gingival tissue.

BACKGROUND: Estimation of time since death is an important parameter in forensic science. Although there are various methods available, precise estimation is still to be established. AIM: The present study aimed to evaluate the histological and ultrastructural changes in the gingival tissue along with the changes in electrolyte levels (sodium, potassium, calcium, and magnesium) among the three groups which included normal, 2, and 4 h since death. MATERIALS AND METHODS: For light microscopic examination and electrolyte analysis, five normal gingival tissue samples were collected from patient following impaction procedure and five gingival tissue samples were obtained from postmortem specimen at 2 and 4 h since death. Each sample was divided into two parts. The first part was fixed in 10% formalin solution for the light microscopic analysis, and microscopic changes were observed between the groups. The second part was snap frozen at -80°C, until measurement of electrolyte using inductively coupled plasma-optical emission spectrometer, and the values were compared among the groups using Kruskal-Wallis test. For electron microscopic examination 2 and 4 h postmortem, gingival tissue samples were collected from the same individual and immediately fixed in 2.5% buffered glutaraldehyde, and the ultrastructural changes were compared with the normal gingival tissue. RESULTS: The light microscopic changes were observed as early as 2 h since death, but there was no significant difference observed between 2 and 4 h postmortem samples whereas ultrastructurally significant difference in morphology was observed between 2 and 4 h postmortem gingival tissue. Our results can confirm histomorphological changes within 2 and 4 h since death.

Intraoral neurilemmoma of mental nerve mimicking as traumatic fibroma: An unusual presentation - A case report and literature review.

Neurilemmoma, neurinoma or schwannoma is a benign tumor of neurogenic origin, arising from the myelin sheath of a neuron. Head and neck region accounts for 25%-48% of these tumors reported till date. On the contrary, intraoral sites form only 1% of the cases accounted. Although a few cases of intraoral neurilemoma of mental nerve has been reported in English literature, none has been reported to arise from the terminal fibers of the mental nerve. In this particular article, we present a case were the tumor was noted in the lower labial mucosa.

Antifibrotic effect of Centella asiatica Linn and asiatic acid on arecoline-induced fibrosis in human buccal fibroblasts.

AIM: The aim of the present study was to investigate the in vitro antifibrogenic effects of Centella asiatica Linn (CA) and its bioactive triterpene aglycone asiatic acid (AA) on arecoline-induced fibrosis in primary human buccal fibroblasts (HBF). METHODS: An ethanolic extract of CA was prepared, and AA was purchased commercially. High-performance thin-layer chromatography (HPTLC) was performed to quantify AA in the CA extract; colorimetric assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) was performed to determine an half-maximal inhibitory concentration. HBF were cultured and stimulated with arecoline. The inhibitory effects of CA and AA at different concentrations were assessed using gene-expression studies on fibrosis-related markers: transforming growth factor-ß1, collagen 1 type 2, and collagen 3 type 1. The stimulatory effect of arecoline and the inhibitory effect of AA on fibroblast morphology and extracellular matrix were assessed qualitatively using Masson trichrome stain. RESULTS: The HPTLC analysis determined 1.2% AA per 100 g of CA extract. Arecoline produced a concentration-dependent increase in the fibrotic markers, treatment with CA significantly downregulated fibrotic markers at higher concentrations, and AA downregulated at lower concentrations. Arecoline altered fibroblast morphology and stained strongly positive for collagen, and AA treatment regained fibroblast morphology with faint collagen staining. CONCLUSION: CA and AA can be used as antifibrotic agents.

The effect of fluorosis on human teeth under light microscopy: A cross-sectional study.

BACKGROUND: Fluoride is needed for the normal development of bone and teeth; in high levels, it affects developing teeth and bone. Dental fluorosis (DF) is caused by ingestion of excess fluoride mainly through drinking water. AIM: The present study aims to observe and understand the histological changes of fluorosed teeth under light microscope (LM). MATERIALS AND METHODS: Teeth which were indicated for extractions for orthodontic or periodontal problems were selected. Thirty extracted teeth were selected with varying degrees of DF based on modified Dean's fluorosis index. Ground sections of these teeth were prepared and the sections were studied under binocular LM. Photomicrographs were taken under high power objective using 15 megapixels Nikon camera. RESULTS AND CONCLUSION: Qualitative histologic changes in different grades of fluorosed teeth were evaluated in enamel, dentin, cementum and between their junctions. Fluoride interacts with enamel in both mineral phases and organic macromolecules by strong ionic and hydrogen bonds resulting in incomplete crystal growth at prism peripheries. This presents as hypomineralization of enamel and dentin, increased interglobular dentin, increased secondary curvatures and changes in cementum such as diffuse cementodentinal junction and increased thickness of Tomes' granular layer. Changes in the structure of the teeth with Dean's index below 2 and teeth with Dean's index of 2 and above were compared using Chi-square test. P value was found to be highly significant being 0.00047. Many of the features of dental fluorosis seen in the present study under light microscope are comparable to those results studied under specialized microscopes.