A Comparative Evaluation of Collagen in Ameloblastoma and Oral Squamous Cell Carcinoma using Picrosirius Red Staining with Polarizing Microscopy and CD44v6 Immunoreactivity.

Background: Solid multicystic ameloblastoma (SMA) is a locally aggressive, benign odontogenic tumor of odontogenic origin with greater rate of recurrence. Epithelial-mesenchymal interaction plays an important role in tooth morphogenesis that shows complete differentiation of epithelial and ectomesenchymal components to the level of tooth formation. Tumor stroma in ameloblastoma is normal mature collagen that prevents differentiation to the level of tooth formation. Current study evaluates the role of stromal elements in aggressive behavior of SMA using picrosirius red staining with polarizing microscopy and CD44v6 immunohistochemistry (IHC). Objectives: To compare nature of collagen using picrosirius red staining under polarized microscope and IHC expression of CD44v6 marker in SMA and oral squamous cell carcinoma (OSCC). Methods: Thirty blocks were retrieved from departmental archives and subjected to picrosirius red staining and CD44v6 IHC staining. Slides stained with picrosirius red were observed under polarized microscope to report the birefringence pattern. IHC slides were annotated for intensity of staining of tumor cells. Results: In contrast to OSCC's 40% red, 40% yellowish-red, and 20% greenish-yellow birefringence, SMA displayed 87% red, 13% yellowish-red, and 0% greenish-yellow. Compared to OSCC, which had tumor cells stained 9% strongly, 64% moderately, 27% mildly, and 0% negatively, SMA revealed 0% strong, 10% moderate, 60% weak, and 30% negative staining. Conclusion: As opposed to OSCC, which exhibited a greater quantity of greenish-yellow birefringence of immature collagen, SMA showed predominantly red birefringence, which is suggestive of mature collagen with a lack of metastasis. Comparing SMA to OSCC, the lack of significant CD44v6 positivity suggests that there has not been perineural invasion or regional metastases in SMA.

Enhancement of Odontoblastic Differentiation of Stem Cells from Exfoliated Deciduous Tooth Using N-acetylcysteine-An In Vitro Study.

AIM AND OBJECTIVE: The study was conducted to evaluate the effects of N-acetylcysteine (NAC) on the propagation and differentiation of stem cells from human exfoliated deciduous teeth(SHED). MATERIALS AND METHODS: SHEDs were isolated by explant culture method and characterized for stem cell properties using flow cytometry method. MTT assay and Cell Counting Kit-8 (CCK-8) assay were used to examine the viability and proliferation of the SHEDs. The effects of NAC-induced osteo/odontoblastic differentiation of SHEDs were determined by functional staining for mineralization, and the gene expression of osteo/odontoblastic transcription factors and proteins was evaluated by real-time quantitative reverse transcription-polymerase chain reaction(qRT-PCR) analyses. Protein levels of collagen type 1 (COL1), dentin sialophosphoprotein (DSPP), and dentin matrix acidic phosphoprotein 1(DMP-1) were calculated by the Western blot method to assess the osteo/odontogenic differentiation. RESULTS: SHEDs presented mesenchymal stem cell (MSC)-like characteristics on flow cytometric analysis. The cell viability and metabolic activity of SHEDs were increased with an increase in the concentrations of NAC from 0.5 to 10 nM. However, the concentrations of NAC from 0.5 to 2.5 mM did not affect cell proliferation. NAC incorporated at a concentration of 2.5 mM showed higher mineralization and considerably increased gene expression levels of runt-related transcription factor 2 (RUNX2), COL1A1, DSPP, and DMP-1. It significantly increased the protein expression of odontoblast-related matrix proteins like COL1, DSPP, and DMP-1. CONCLUSION: NAC regulates the healthy propagation of dental stem cells in vitro. Its effects on the differentiation of dental pulp SHEDs remain unidentified. This study explores that NAC can encourage the mineralization of SHEDs and differentiate them into the odontoblastic lineage. CLINICAL SIGNIFICANCE: The results propose that NAC could have a significant pharmacological role in activating and enhancing odontogenic differentiation of dental stem cells and possibly a prospect in regenerative dentistry.

Evaluation of Antibacterial Properties of Organic Gutta-percha Solvents and Synthetic Solvents Against Enterococcus faecalis.

BACKGROUND: The use of solvents is necessary to remove remnants of filling materials within dentinal tubules to allow penetration of irrigating solution within the tubules. AIM AND OBJECTIVES: We aimed at determining the antibacterial effect of various gutta-percha (GP) solvents against Enterococcus faecalis (E. faecalis). MATERIALS AND METHODS: An in vitro study was conducted by measuring the zone of inhibition using the disk diffusion method. The test organism used for the study was E. faecalis, and the solvents were divided into five groups: eucalyptus oil, chloroform, turpentine oil, xylene, and orange oil. About 500 µL of the suspension was spread over the agar plates, and the empty sterilized disks were impregnated with 10 µL of pure essential oils (EOs). The inoculated plates were incubated at 37°C for 18 to 24 h. The antimicrobial activity of each solvent was expressed and measured in terms of the mean diameter of the zone of inhibition (in mm) produced by each solvent at the end of the incubation period. ANOVA was used for intergroup comparison. The P-value of <0.05 was considered significant. RESULTS: The mean zone of inhibition for E. faecalis was 24.00 ± 1.21 for eucalyptus oil, 16.30 ± 0.92 for chloroform, 26.50 ± 1.24 for turpentine oil, 13.70 ± 1.26 for xylene, and 19.80 ± 1.32 for orange oil. The difference between the groups was statistically significant (P < 0.001). CONCLUSION: This study demonstrated that the use of turpentine oil during endodontic retreatment significantly reduced the levels of E. faecalis as compared with other solvents.

Staining the Ground Section of the Tooth Using an Innovative Plant Stain Found in the Pravara Region, India.

AIMS: To assess the staining efficacy of the innovative "Pravara" stain on ground sections of human teeth under a stereomicroscope. In this study, the ground sections were stained with the innovative stain. The stain is obtained from a leafless tree called Terminalia chebula. Terminalia chebula, commonly known as Harad, is found in South Asia from India. Few species are found in the Pravara region of Maharashtra, India. MATERIALS AND METHODS: A total of 40 extracted teeth were collected and ground sections of each tooth were prepared. The samples were randomly divided into two groups: group I-control (without stain), group II-study group (the samples in this group were stained with the help of innovative "Pravara" stain). The sections were graded from I to III according to the differentiation and contrast of tissues. RESULTS: The ground sections in group II stained with Pravara stain demonstrated better contrast for structures within sections under a microscope in comparison with group I (control group). CONCLUSION: The study demonstrates the efficacy of a new innovative Pravara stain, which is a simple, economical, quick, and effective staining method for the ground sections of the teeth. CLINICAL SIGNIFICANCE: This innovative Pravara stain is cost-effective, saves time, ecofriendly, and highlights the anatomical structures more effectively and does not fade easily with time.

Study of the Role of Salivary Lactate Dehydrogenase in Habitual Tobacco Chewers, Oral Submucous Fibrosis and Oral Cancer as a Biomarker.

AIM: To evaluate the salivary lactate dehydrogenase (LDH) levels in clinico-pathologically confirmed oral submucous fibrosis (OSMF), oral cancer and clinically diagnosed tobacco pouch keratosis patients. MATERIALS AND METHODS: A prospective, comparative study was carried out in a tertiary healthcare centre located in Loni from October 2013 to January 2014. A total of 120 patients were separated into 4 groups depending upon the clinical diagnosis as follows. Group I: healthy control (with no addictions and diseases). Group II: oral cancer. Group III: oral submucous fibrosis. Group IV: habitual tobacco chewers (tobacco addiction without any disease). Substantiation was done using biopsy. The samples were inspected for salivary LDH levels by the technique in line with the recommendations of the International Federation of Clinical Chemistry with the help of Erba Chem semi auto analyser. RESULTS: The mean salivary LDH levels in the control, oral cancer OSMF and habitual tobacco chewer group were 86.12 ± 7.05 IU/L, 592.09 ± 28.57 IU/L, 350.43 ± 5.90 IU/L and 125.19 ± 13.42 IU/L, respectively. Out of 4 groups, LDH activity was increased in saliva of patients with tobacco pouch keratosis, OSMF, and oral cancer consistently. Notable difference was found in the mean salivary levels of the above groups. Results were subjected to appropriate statistical analysis: one-way ANOVA, Student's unpaired t test for group-wise comparison followed by post hoc Tukey's test. CONCLUSION: We observed congruous higher levels of salivary LDH in oral precancer and cancer, and hence it could be a future marker.

Oral Manifestation in Patients diagnosed with Dermatological Diseases.

AIM: This study aimed to estimate the frequency of oral lesions in dermatological diseased patients attending the outpatient department of Pravara Rural Medical and Dental College and Hospital, Pravara Institute of Medical Sciences (Deemed University), Loni, Ahmednagar, Maharashtra, India, and the Department of Dermatology at Medical College, Ahmednagar, Maharashtra, India. MATERIALS AND METHODS: A cross-sectional hospital-based study was conducted in Loni from October 2013 to January 2014. A total of 310 patients (mean age 37.2 ± 16 years, 61.2% males) completed an oral examination and a personal interview. Oral lesions were recorded. Biopsy and smear were used as adjuvant techniques for confirmation. Data were analyzed using Statistical Package for the Social Sciences (SPSS) (version 15.0.1). RESULTS: Of 310 cases (n = 310) observed for skin lesions, 99 cases were psoriasis (31.93%) and 68 cases were lichen planus (LP; 21.9%), followed by herpes zoster in 44 cases (14.1%), herpes simplex in 13 cases (4.1%), pemphigus vulgaris (PV) in 15 cases (4.8%), erythema multiforme (EM) in 8 cases (2.5%), bullous pemphigoid (BP) in 4 cases (1.2%), chicken pox in 3 cases (0.9%), eczema in 52 cases (16.7%), and nutritional deficiency and candidiasis in 2 cases (Table 1). CONCLUSION: The dermatologic diseases studied frequently occur in the oral cavity. Among them, psoriasis was the most common dermatological disease, and LP frequently showed. CLINICAL SIGNIFICANCE: This study also depicts that diagnosis and management of these oral lesions should also be carried out by oral clinicians so as to improve the oral health functioning during the course of the disease. The intraoral examination should be incorporated to the routine of dermatologic assistance as the oral manifestations can represent preliminary signs or can coexist with the diseases.

Study to assess Activity and Concentration of Gammaglutamyl Transpeptidase in Precancerous and Cancer Patients and Its Comparison with Controls.

OBJECTIVE: The purpose of the study is to highlight the use of gamma-glutamyl transpeptidase (GGT) as salivary enzyme tumor marker and assess the activity and concentration of GGT in precancerous and cancer patients and compare it with the normal control. MATERIALS AND METHODS: Individuals in the age group of above 20 years were included in the study. In this study, salivary GGT was analyzed in 75 cases. The selected patients were divided into three main groups as group I (controls with normal health), group II (patients with precancerous lesions and conditions), and group III (patients with oral cancer lesions). All the selected individuals were analyzed for salivary GGT. RESULTS: A significant difference was observed between control and precancerous groups with GGT values at 5% level of significance. The mean GGT value is significantly higher in precancerous group as compared with control group (p < 0.05). A significant difference was observed between control and cancerous groups with GGT values at 5% level of significance. The mean GGT value is significantly higher in cancerous group as compared with control group (p < 0.05). A significant difference was observed between precancerous and cancerous groups with GGT values at 5% level of significance. The mean GGT value is significantly higher in cancerous group as compared with precancerous group (p < 0.05). CONCLUSION: There is a remarkable increase in salivary GGT activity in both precancerous and cancerous conditions. The increased activity was more marked in cancerous conditions than in precancerous conditions. The GGT levels were two- to threefold increased in precancerous conditions as compared with control group. This finding was statistically significant and also suggested the strong correlation between GGT levels and presence of precancerous conditions. CLINICAL SIGNIFICANCE: Integration of all these observation strengthens our thinking that elevation of salivary GTT in pre-cancerous and cancerous conditions can be a reliable biomarker in early detection and prevention of oral malignancy.