Formulation and Development of a Herbal Antifungal Gel Containing Origanum vulgare and Syzygium aromaticum Essential Oils Against Oral Candida albicans.

Background Oral candidiasis is the most prevalent oral fungal infection, and existing antifungal agents have side effects such as drug intolerance, resistance, and toxicity. Herbal essential oils are emerging as an alternative therapeutic approach for treating fungal infections. Origanum vulgare (O. vulgare), commonly known as oregano, and Syzygium aromaticum (S. aromaticum), commonly known as clove, are known to have antifungal properties and are effective against fluconazole-resistant strains. A combination of essential oils has a synergistic effect and aids in achieving effective antifungal activity at sufficiently low concentrations, which could lead to reduced side effects and resistance. Aim of the study This study aimed to formulate and develop an herbal antifungal gel containing O. vulgare and S. aromaticum and evaluate its synergistic antifungal efficacy against oral Candida albicans (C. albicans). Methodology Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) determinations of O. vulgare and S. aromaticum essential oils were performed individually and in combination to assess the antifungal activity against C. albicans. Based on the obtained MIC and MFC of essential oils in combination, an herbal antifungal gel was formulated. Further, to determine the biocompatible nature of the gel, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed. Results We found that a combination of O. vulgare and S. aromaticum essential oils showed antifungal activity at a lesser concentration, with a MIC of 0.19 µl/ml and MFC of 0.39 µl/ml when compared to their individual concentrations. Based on our results, an antifungal herbal gel comprising a concentration of 0.6 µl/ml of both essential oils was developed to achieve synergistic antifungal activity against oral C. albicans. The MTT assay of the herbal gel did not show any cytotoxicity. Conclusion The novel herbal antifungal gel containing O. vulgare and S. aromaticum is biocompatible in nature and provides an alternative therapeutic approach for treating oral candidiasis.

Evaluation of Immunoexpression of AJUBA Protein in Normal Oral Mucosa and Oral Squamous Cell Carcinoma.

AJUBA is multifunctional scaffold protein which belongs to Zyxin family of proteins. It is known to have dual role in cancer as a tumor promoter and tumor suppressor. AJUBA has a key role in systemic malignancies like esophageal squamous cell carcinoma, colorectal cancer, cervical, breast, prostate cancer, etc. But there is very sparse literature available regarding its expression profile in oral squamous cell carcinoma (OSCC) and moreover its expression has not been observed in normal oral mucosa (NOM). Thus, the aim of this research is to explore the expression profile of AJUBA by immunohistochemical method in NOM and OSCC. Furthermore, we also evaluated the association of AJUBA expression with clinicopathologic parameters. A total of 84 samples of formalin fixed paraffin embedded tissue blocks comprising of 42 cases each of NOM and OSCC were subjected to detect immunoexpression of AJUBA. We found enhanced intense immune-expression of AJUBA in OSCC cases than compared with NOM and found to be statistically significant. The parameters specific to histologic tumor grade and inflammatory response in OSCC also found to have statistically significant with AJUBA expression. Our study is first of its kind to reveal AJUBA expression in basal and suprabasal layer of NOM suggestive of its definitive role in differentiation and stratification process. We also observed its intense expression in peripheral cell of tumor islands of OSCC cases, which can suggest its possible role in tumor growth and progression.

Isolation, culture and characterization of primary cell lines of human buccal mucosal fibroblasts: A combination of explant enzamytic technique.

BACKGROUND: The cell culture technique has become a routine and a popular method for its wide applications in the field of cell biology and biotechnology and in medical research. Isolation of primary cells over the cancer cells is an essential component of cell culture technology as they are the reliable source to understand normal physiological, morphological and molecular process of human cells. As fibroblasts are the prominent cells of the connective tissue of oral mucosa, many disease entities and histogenesis are linked to fibroblasts. Culture of oral fibroblast cells helps the oral biologists and researchers to study the morphological and molecular process in the oral diseases. AIM: The aim of our experiment is to isolate and culture the human buccal mucosal fibroblast cells from healthy individuals using a combination of explant-enzymatic method and characterization of the cells by short tandem repeat (STR) profiling. MATERIALS AND METHODS: The tissue samples were collected from healthy individuals undergoing routine impacted third molar extraction. A combination of explant-enzymatic technique was used for the isolation from the tissue samples. The cells were further subcultured, maintained and stored as per the standard protocols. Thus, to confirm the oral fibroblasts of human origin and its uniqueness, they were characterized using STR profiling. RESULTS AND CONCLUSION: Using the combination technique, we were successful in isolating the cells at a faster rate by detachment of cells on day 3 and confluency on day 10. The morphological assessment and STR profiling further confirmed that the isolated cell lines resemble human fibroblast cells.

Roles of the matricellular protein Tenascin-C in T-lymphocyte trafficking and etiopathogenesis of Oral Lichen Planus.

OBJECTIVE: This study was aimed at examining the role of Tenascin-C in T cell trafficking in Oral Lichen Planus (OLP). DESIGN: For the in vivo immunohistochemical analyses, 115 OLP samples were collected from patients and immunostaining was performed. The intensity and distribution of TN-C expression were quantified and correlated with histological analyses of basement membrane integrity and presence of inflammatory infiltrate. For the in vitro study, TN-C and collagen were coated on culture plates and migration of T lymphocytes was assessed. RESULTS: TN-C immunoexpression was increased in terms of both distribution and intensity along the basement membrane zone. These changes were significantly associated with basement membrane duplication (distribution p < 0.002 and intensity p < 0.001) and bands of inflammation (distribution p < 0.002 and intensity p < 0.001) assessed by Chi-square test. T lymphocytes demonstrated significant migration towards TN-C as compared to collagen (n = 3, p < 0.05). CONCLUSIONS: These findings indicate TN-C may have a key role in promoting T cell migration at the epithelial-mesenchymal junction in OLP. These observations suggest TN-C could be a good target for therapeutic intervention, either in itself or synergistically with anti-inflammatory directed strategies in this chronic disease management.

Stature estimation from regression analysis of facial anthropometry in Indian population.

BACKGROUND: Stature or body height is one of the most important and useful anthropometric parameters which determines the physical identity of an individual. As craniofacial structures have the advantage of being composed largely of hard tissue which is relatively indestructible, the careful study of these can enable reliable determination of stature of the person in life. Studies pertaining to stature estimation from facial measurements are limited in an Indian population. The present investigation attempts to estimate stature from anthropometric dimensions of face. MATERIALS AND METHODS: The material for the present study comprises 361 Indian students (151 males and 210 females) in the age range of 21-45 years. Stature and six facial measurements were taken on each participant following standard methods and techniques. Karl Pearson's correlation coefficient and linear regression were done to estimate stature. RESULTS: The results indicate that facial measurements are strongly and positively correlated (P < 0.001) with stature. The accuracy of the computed equations was further tested on 50 randomly selected study participants of each group, which shows close approximation of actual and estimated stature. CONCLUSION: Within the limits of this study, we conclude that facial dimensions can be used as a supplementary approach for the estimation of stature but with caution, as these are population-specific approach.

Immunohistochemical expression of extracellular matrix metalloproteinase inducer (EMMPRIN) in normal oral mucosa, oral epithelial dysplasia and oral squamous cell carcinoma.

BACKGROUND: In oral squamous cell carcinoma (OSCC), extracellular matrix metalloproteinase inducer (EMMPRIN) expression has been noted in the cell membrane throughout the epithelium of the lesion, suggesting its increased expression. OBJECTIVES: The present study was conducted to evaluate and compare the expression of EMMPRIN in the normal oral mucosa (NOM), different histological grades of oral epithelial dysplasia (OED) and OSCC. MATERIALS AND METHODS: A total of 100 formalin-fixed paraffin-embedded tissue blocks of NOM (n = 10), 20 cases each of mild, moderate and severe (OED) (n = 60), and 10 cases each of well differentiated, moderately differentiated and poorly differentiated carcinomas (n = 30) were included in the study. The tissues sections were immunohistochemically stained and were evaluated for intensity and area of expression in different groups. RESULTS: Out of 60 cases of OED, 29 (48%) cases showed intense dark brown staining in the epithelium. The stroma in 38 (63%) cases showed positive immunoexpression. The expression of EMMPRIN in OSCC revealed intense dark brown staining in 9 (90%) cases of well differentiated, and a decent thereon in 8 (80%) cases of moderately differentiated and 4 (40%) cases of poorly differentiated carcinomas. CONCLUSION: The role of EMMPRIN in precarcinogenesis and early carcinogenesis needs to be studied on considerable sample size. This can enable oncologists to detect cancer at an early stage before it progresses to malignancy.

Evaluation of cannibalistic cells: a novel entity in prediction of aggressive nature of oral squamous cell carcinoma.

OBJECTIVE: Cellular cannibalism is a distinctive morphologic feature exclusively seen in aggressive malignancies and is defined as a large cell enclosing a slightly smaller one within its cytoplasm. This phenomenon has been illustrated in several malignancies and is correlated well with the degree of anaplasia, invasive and metastatic potential of tumor cells. However, this marker has not been extensively studied and is often unnoticed during the routine histopathological assessment of Oral squamous cell carcinoma (OSCC). Thus, the aim of this research was to evaluate the presence of cannibalistic cells (CC) and to find if there exists any association with the aggressive nature of OSCC. MATERIALS AND METHODS: In total, 20 neck dissection cases of OSCC with follow-up data were included in the study. The cannibalistic cells were evaluated in the lesion tissues. Cellularity of cannibalism is graded asGrade I: < 5 cells, Grade II: 6-15 cells and Grade III:> 16 cells. The data was analyzed using Fischer Exact test. RESULTS: Out of 20 cases, all the cases showed presence of CC, Grade I in five cases, Grade II in eight cases and Grade III in seven cases. A statistically significant relation between advanced grade of cellular cannibalism and lymph node positive status (p≤ 0.001) was obtained. CONCLUSION: Interestingly the cases with positive lymph node metastasis demonstrated Grade 3 CC.Hence, during routine histopathological examination, the search of CC can be considered as one of the important parameters to note the aggressive nature of OSCC.

Expression of cytokeratin subtypes: MMP-9, p53, and αSMA to differentiate basaloid squamous cell carcinoma from other basaloid tumors of the oral cavity.

Basaloid squamous cell carcinoma (BSCC) is rare variant of oral squamous cell carcinoma (OSCC) with predilection for upper aerodigestive tract. Although it is characterized by distinct histologic features it is often confused with conventional OSCC and other basaloid tumors. The study aims to establish differentiating features of BSCC with oral basaloid tumors using immunohistochemical (IHC) markers. This retrospective study included 34 cases, including BSCC, OSCC, and basaloid tumors. IHC staining was performed with primary antibodies against cytokeratin (CK) 19, 14, 8/18, α-smooth muscle actin (αSMA), p53, and MMP-9. A prominent CK 19, 14, and 8/18 expression was observed in BSCC as compared with basaloid tumors suggesting of basal cell origin with undifferentiated type of tumor cells. Expression of αSMA was intense in tumor cells of myoepithelial differentiation but lacked in BSCC. The intense expression of p53 and MMP-9 was noted in all basaloid malignancies. Considering standard histologic criteria in diagnosing BSCC, when in confusion with other basaloid tumors, IHC markers gain importance. Hence, enhanced expression of CK 19, 8/18, and 14 and coexistence of p53 and MMP-9 expression and negativity for αSMA suggest an accurate diagnosis of BSCC.

Expression of transforming growth factor ß and its correlation with lipodystrophy in oral submucous fibrosis: an immunohistochemical study.

OBJECTIVE: In our routine review of Oral Submucous Fibrosis (OSMF) biopsies, we observed decreased adipose tissue even though most are from buccal mucosa. Pathogenesis of OSMF has demonstrated the role of Transforming Growth Factor ß (TGF ß), in causing fibrosis. This study aims to correlate the role of TGF ß with loss of adipose tissue in OSMF. STUDY DESIGN: From our archives, 84 OSMF cases (24 early and 60 advanced OSMF) were screened for adipose tissue. Immunoexpression of TGF ß in these cases were investigated. RESULTS: Adipose tissue was seen in 67% of early OSMF and in 13% of advanced cases. Early cases showed more intense TGF ß staining of epithelium, fibroblast, macrophages and inflammatory cells than the advanced cases. CONCLUSION: These findings suggest that TGF ß plays a key role in causing lipodystrophy in OSMF and is secreted more during early course of the disease than in advanced stage.

Establishment of field change by expression of cytokeratins 8/18, 19, and MMP-9 in an apparently normal oral mucosa adjacent to squamous cell carcinoma: A immunohistochemical study.

BACKGROUND: Squamous cell carcinoma is the most frequent malignancy of the oral cavity. The survival rate of this malignancy has not improved from past two decades. The major factors responsible for this could be due to loco regional and distant metastatic spread. However, the other important prognostic factor is concomitant occurrence and recurrence of multiple primary carcinomas in the head and neck region, which is explained as the concept of field cancerization. The evidence to support the field change in normal mucosa of Oral squamous cell carcinoma (OSCC) through biological markers using immunohistochemistry has always been challenging. AIM: Hence, the aim of the present research is to identify changes in the expression of CK 8/18, 19, and MMP-9 to visualize field changes in the clinically normal mucosa adjacent to OSCC and compare with non neoplastic normal oral mucosa. MATERIALS AND METHODS: 20 cases of OSCC with radical resection specimens were included in the study. Lesional tissue and adjacent normal looking mucosa were taken during grossing. Ten cases of non-neoplastic normal oral mucosa are also included in the study. Markers such as CK 8/18, CK 19, and MMP-9 are used by the immunohistochemical method in this present study. RESULT AND CONCLUSION: The enhance expression of CK 8/18 (80%), CK 19 (70%), and MMP 9 (90%) in ANM was noted and furthermore in six ANM showing severe dysplasia with enhance expression of CK 8/18, CK 19, and MMP 9 in the apparently normal oral mucosa can suggest a field cancerization.

Intraosseous fibrosarcoma of maxilla in an HIV patient.

Fibrosarcoma is a malignant mesenchymal neoplasm of fibroblasts that rarely affects the oral cavity and can cause local recurrences or metastasis. Fibrosarcomas account for 15% of all soft tissue sarcomas, which represent only 1% of all malignant tumors of the head and neck region. The clinical behavior of the fibrosarcoma is characterized by a high local recurrence rate, and low incidence of loco regional lymph node and/or distant hematogenous metastasis. The etiology for fibrosarcoma has no definite cause but is thought to occur from preexisting lesions or in previously irradiated areas of bone lesions. Immunosuppression associated with HIV infection and acquired immune deficiency syndrome (AIDS) has been consistently linked to various cancers, including Kaposi's sarcoma, non-Hodgkin's lymphoma, and invasive cervical cancer. Rare neoplasms like Hodgkin's disease, anal cancer, leukemia, basal cell carcinoma, and squamous cell carcinoma have also been demonstrated. This paper presents one such a rare incidence of an intraosseous fibrosarcoma occurring in an HIV-positive patient.

Immunohistochemical expression of Tenascin in embryogenesis, tumorigenesis and inflammatory oral mucosa.

OBJECTIVE: Tenascin is a large extracellular matrix glycoprotein that plays specific role in cell matrix interaction. This protein is mainly attracted because of its oncofetal predominance expression at epithelial-mesenchymal interaction and also been associated with inflammatory response. Thus the aim was to study the expression of Tenascin within the oral cavity in a developing tooth, normal oral mucosa, squamous cell carcinoma and inflammatory mucosa and further to compare its expression in inflammatory mucosa with that of squamous cell carcinoma. DESIGN: A total numbers of 92 cases were included, with 22 being all morphological stages of developing tooth, 10 cases of normal oral mucosa, 30 cases each of inflammatory gingival hyperplasia and oral squamous cell carcinoma. The intensity and pattern of expression was assessed immunohistochemically using anti-human mouse monoclonal Tenascin antibody. RESULTS AND CONCLUSION: Tenascin expression in developing tooth was seen mainly at epithelial-mesenchymal junctions, but temporally reduced at cap stage. In normal mucosa TN expression was restricted only at basement membrane zone. Inflammatory gingival hyperplasia intensity of expression was enhanced at the juxtraepithelial stroma and showed reticular pattern of expression. In oral squamous cell carcinoma, intensity of expression was seen in superficial front of the stroma and also around tumour islands with intraepithelial expression and predominantly showed fibrillar pattern of expression. Furthermore, Tenascin expression was noticed around neovascularization. Hence, there is a regulatory system in Tenascin expression and plays a vital role in embryogenesis, tumerogenesis and inflammation in remodelling the stroma for cell migration and also for healing.

Oral pulse granuloma associated with keratocystic odontogenic tumor: Report of a case and review on etiopathogenesis.

Pulse granuloma is a distinct oral entity characterized as a foreign body reaction occurring either centrally or peripherally. It is usually seen in the periapical or in the sulcus area. Occasionally the lesions occur in the wall of the cyst, commonest being the inflammatory odontogenic cyst. Histologically, they present as eosinophilic hyaline mass with giant cell inclusions and inflammatory cells. They may show different histological characteristics, possibly related to the length of time in the tissue. Adequate recognition is important to avoid misdiagnosis. Many authors suggest that pulse granuloma results due to implantation of food particles of plant or vegetable origin into the tissue following tooth extraction. This paper aims to report a case of pulse granuloma associated with keratocystic odontogenic tumor with its histochemical and polarizing microscopic features and discuss on etiopathogenesis of pulse granuloma.

Anthropometric and anthroposcopic analysis of different shapes of faces in group of Indian population: a pilot study.

Focus of facial identification research is to isolate features that can be considered a factor of individualization. Anthropometry and Anthroposcopy have been the most important research tools in the field of forensic anthropology. These two methods of observation and data collection are made both on the living and on skeletonized human remains. Anthropometry is a systematic study of human measurements. Morphological analysis of unmeasurable features usually is the area of anthroposcopy. Hence, this study aims to assess different human faces using anthropometry and anthroposcopy. A total of 100 subjects were included in the study, prosopic index given by Martin and Saller was used to classify facial types. An anthropometric measurement was used to assess the facial proportions and anthroposcopy for facial morphology. A significant number of subjects were of hyperleptoprosopic facial type, and with a significant sexual dimorphism observed among the Indian population. Anthroposopic analysis showed common morphological features in each prosopic indices. The findings in our study may throw light on anthropometric and anthroposcopic features of Indians and help significantly in the field of forensic anthropology.

Trichilemmal carcinoma associated with xeroderma pigmentosa: report of a rare case.

Trichilemmal carcinoma is a rare malignant skin adnexal tumor derived from the outer root sheath epithelium of the hair follicle. We describe a rare case of trichilemmal carcinoma of the upper lip associated with xeroderma pigmentosa in a young patient. We demonstrate that this tumor, as well as the more frequently reported squamous cell carcinoma and basal cell carcinoma can occur in association with xeroderma pigmentosa. Making this diagnosis is imperative as the biological behavior of Trichilemmal carcinoma is less aggressive than that of other epithelial malignancies, requiring a different approach to treatment planning.