E-cadherin-mediated impairment increases anti-apoptotic mechanism through upregulation of Bcl-2: An immunohistochemical study in various patterns of invasion of oral squamous cell carcinoma.

BACKGROUND: Bcl-2 and E-cadherin proteins are known to be involved in the control of apoptotic cell death and invasive potential, respectively, which is an important hallmark of tumor regulation that influences their biologic behavior. AIM: This study investigates the relationship of Bcl-2 and E-cadherin immunoexpression in various Bryne's patterns of invasion. MATERIAL AND METHODS: Immunohistochemical analyses for Bcl-2 and E-cadherin were performed on paraffin-embedded tissue sections on 40 cases (32 cases of Oral squamous cell carcinoma and eight cases of controls) and were scored using qualitative and quantitative (percentage positive) analysis. STATISTICAL ANALYSIS: The resulting data were analyzed using SPSS software version 19. Correlation between patterns of invasion and qualitative scores of Bcl-2 and E-cadherin was calculated using Spearman rho correlation. Difference of mean percentage of positive cells of Bcl-2 and E-cadherin in different patterns of invasion was tested by ANOVA followed by Tukey HSD test. RESULTS: Bcl-2 and E-cadherin immunoreactivity was positively correlated with Bryne's pattern of invasion (P value<.05). An inverse relation was found between Bcl-2 and E-cadherin expression with Bryne's patterns 1-5 of invasion. CONCLUSIONS: The results pointed to the antagonistic role of E-cadherin and Bcl-2 and thus provide the opportunity for cell survival along with increased invasive potential.

Role of p63 in Determining the Histogenesis of Low-Grade Neoplasms versus Cystic Lesion.

The biological nature of salivary gland neoplasms and the overlapping characteristics that result from the heterogeneity of the cells of origin make diagnosis difficult. Hence, we intend to present a case of low grade mucoepidermoid carcinoma (MEC) on the palate and to understand the importance of biomarker such as p63 in the early diagnosis of tumor as it also has a role in its histogenesis. A 53-year-old female reported with a unilateral swelling for 3 months on posterolateral palatal region of the right side. Clinical differentials for such palatal swellings include a varied spectrum of lesions such as reactive, benign, and malignant lesions. Based on the incisional and excisional biopsy, histopathological findings and immunohistochemical examination with p63 the case were diagnosed with low grade MEC. The tumor cell differentiation in MEC could be the result of multiplicity of differentiation pathways leading to the formation of various histological patterns. This case report highlights the complexity of salivary gland pathology diagnosis and role of specific tumor marker such as p63 as an early marker for differentiation of salivary gland tumor such as low grade MEC from other cystic lesions occurring on the palate.

Proliferative potential of impacted tooth lesions using Ki67 labelling index-A clinicopathological insight.

Tooth impaction is a frequent phenomenon, and the prevalence and distribution of this entity in different regions of the jaws may vary considerably. The third molars, maxillary canines, maxillary and mandibular premolars, and maxillary central incisors are the most commonly affected teeth. Impacted teeth in children and adolescents are rarely associated with pathological changes, but the prevalence of problems has been found to increase in later decades. Impacted teeth are commonly asymptomatic and not associated with any pathologic lesions for years. Proliferative potential of various odontogenic lesions were calculated using Ki-67 labeling index calculation, with the highest index of Unicystic Ameloblastoma followed by Adenomatoid odontogenic tumor, Unicystic Ameloblastoma, followed by the dental follicle. Ki-67 is a marker of cell proliferation, used as an important diagnostic marker in the pathologic differentiation of various lesions. It is always better to orthodontically treat or extract asymptomatic impacted teeth to avoid or to restrict the proliferative capacity of the dental follicle. Treatment decisions about the third molar have important clinical and cost implications.

Neuroectodermal influence in odontogenic cyst and tumor: evidence through CD56 immunoexpression.

BACKGROUND AND OBJECTIVES: CD56, associated with neuroectodermal differentiation of the embryonal cells, is often considered a marker of neural lineage. Odontogenic keratocysts (OKCs) are of particular interest because of their characteristic histopathologic features, high recurrence rate, and aggressive behavior. CD56 immunoreactivity in these lesions has been reported with very high frequency. The present study analyzes the immunoexpression of CD56 in ameloblastoma (AM) and OKC to infer neuroectodermal influence in the pathogenesis of odontogenic lesions and its correlation with clinicopathologic parameters. MATERIALS AND METHODS: Fifty histopathologically confirmed cases of OKC and AM, 25 from tooth-bearing (TB) and molar-ramus (MR) regions each, and 5 dental follicular tissues as control were collected from the department archives and immunohistochemical analysis with CD56 was carried out. RESULTS: CD56 immunopositivity was seen in 64% AM and 36% OKC cases. The majority of AM cases showed cytoplasmic expression in the peripheral cells of odontogenic islands; similarly, OKC cases showed continuous and uniform cytoplasmic expression in the basal and parabasal cells of the cystic lining. CD56 immunopositivity was found in more AM cases as compared to OKC cases in both the TB and MR regions. INTERPRETATION AND CONCLUSIONS: The assessment of CD56 immunoexpression in odontogenic cyst and tumor (AM) may aid in understanding the role of neuroectodermal influence in the etiopathogenetic pathways and a possible influence of CD56 on the clinical behavior and aggressiveness of the odontogenic lesions. A correlation of CD56 expression with the clinical outcome of the disease (site, perforation, root resorption, and tooth displacement) can help envisage possible prognostic assessment for these lesions.

Assessment of changes in Streptococcus pyogenes levels using N-acetylgalactosamine-6-sulfatase marker and pharyngeal airway space with appliance therapy in mouth breathers - An ELISA-based study.

Background: The frequency of adenotonsillar hypertrophy in mouth-breathing children when compared to the average found in the general population is considered to be higher. Mouth breathing is considered as one of the causative factors for tonsillitis in children. Through continuous irritation on tonsillar wall, tonsils swell up and inflammation develops. Purpose: The purpose of the study is to evaluate Streptococcus pyogenes count using colony-forming units (CFUs) and N-acetylgalactosamine-6-sulfatase side chain marker on ELISA (enzyme linked immunosorbent assay) in mouth breathers and to establish its correlation with pharyngeal airway space pre- and post-oral screen appliance therapy. Materials and Methods: A total number of 24 (n) mouth breathers aged between 5 and 12 years were included in the study and given oral screen appliance therapy. The subjects were evaluated for the various parameters before the delivery of a habit-breaking appliance and then reevaluated for the same parameters (presence of S. pyogenes and its counts, size of tonsils, and pharyngeal airway space dimensions) after 6 months of appliance usage. Results: A statistically significant difference was seen in levels of S. pyogenes using ELISA and CFUs. Furthermore, statistically significant difference was observed in Friedman tonsil scoring and pharyngeal airway space and pre- and post-oral screen appliance therapy. Conclusion: Oral screen appliance therapy reduced the frequency of occurrence of tonsillitis in mouth breathers by decreasing the counts of S. pyogenes bacteria. Upper and lower pharyngeal airway space dimensions were increased after 6 months of appliance therapy in mouth breathers.

Case series of head-and-neck adenosquamous carcinoma: Role of histogenetic model in immunotherapy as a future perspective for nonconventional squamous cell carcinoma.

Adenosquamous carcinoma (ASC) is not only a rare variant of squamous cell carcinoma (SCC) but it is also an aggressive and highly infiltrating neoplasm with distinct histomorphology. Histogenetic concept in the pathogenesis of the tumor would eradicate controversies regarding the resemblance of ASC with mucoepidermoid carcinoma and adenoid SCC. Hence, we intend to present a case series of four cases of ASC in the head-and-neck region reported in a single institution in the past 10 years. Head-and-neck ASC lesions have been reported in the thyroid gland, nasal cavity, oral cavity, pharynx, and larynx. Intraoral lesions are mostly seen in the tongue and floor of the mouth, but in our case series, the most common site was the maxillary alveolus. Therapeutic approach toward nonconventional malignancies of epithelial origin should include proper analysis of the biological behavior of disease, the anatomical site, sensitivity of lesion to radiation, and systemic treatment modality. Hence, for better understanding of the behavior of lesions such as ASC, immunohistochemical analysis plays a significant role as it provides an insight into the origin and also accentuates the chances of enhanced therapeutic model for treating any such variant of SCC.

Emerging insights of NK cells immunosurveillance in histomorphologic prognostic indicators of oral squamous cell carcinoma.

Background: IFN-gamma and natural killer (NK) cells have been considered the most effective cells in the combat of cancer, contributing to better prognosis and longer survival. The aim of the study was to analyze and correlate the CD 57 immunopositive NK cell-mediated Interferon-γ pathway in regulating immune mechanisms in Oral Squamous Cell Carcinoma. Materials and Methodology: The study sample was composed of a total of 40 cases of histopathologically confirmed cases of Oral Squamous cell carcinoma (OSCC). Clinical data such as age, gender, habit history, signs and symptoms, and TNM staging were obtained for each case. The biopsy specimens of the cases obtained were fixed with 10% neutral buffered formalin and processed and embedded in paraffin wax. 3-4 µ thick sections were taken for hematoxylin and eosin staining and immunohistochemistry procedure. A saliva sample was collected from each patient and stored at 20 degree Celsius for estimation of salivary interferon-gamma levels using the sandwich ELISA technique. Results: CD 57 NK cells quantitative assessment was significantly associated with tumor budding, cell nest size, the pattern of invasion, lymphocytic host response, NK cell morphology, Depth of invasion, and Tumor thickness. The ratio of CD 57 immunopositive NK cells to salivary IFN-γ levels showed a significant association with histopathological grades, tumor size, and lymph node status. Conclusion: Adoptive cellular transfer therapy with NK cells has been advocated in both experimental models and clinical trials in treating hematopoietic malignancies. The strategy is based on reviving the patient innate immune surveillance and control of tumor invasion by the infusion of activated NK cells. The IFN-gamma and NK cell infiltration in oral squamous cell carcinoma might show a distinctive tumor microenvironment with a favorable local cytotoxic immune response against neoplastic cells.

Potentiated action on the progression of OSMF by hypoxia mediated signaling pathway by the epithelial mesenchymal transition and angiogenic apparatus.

Background: Epithelial-mesenchymal transition (EMT) is a complex process, in which epithelial cells acquire the characteristics of invasive mesenchymal cells. EMT has been implicated in cancer progression and metastasis as well as the formation of many tissues and organs during development. Aim: The aim of the study was to ascertain the role of hypoxia-mediated signaling pathways influencing EMT and angiogenesis in progression of oral submucous fibrosis (OSMF). Materials and Methods: Evaluation of the immunoexpression of alpha-smooth muscle actin (α-SMA), E-cadherin, vimentin, and factor VIII receptor antigen in OSMF and oral squamous cell carcinoma (OSCC) arising from OSMF was done. Differences between the different variables were analyzed using ANOVA test and Pearson's Chi-square test, and Mann-Whitney test was also calculated. Results: The mean α-SMA positive myofibroblasts increased from Group 1 (OSMF) to Group 2 (OSCC), especially those in the deeper connective tissue stroma. The mean labeling index of vimentin and mean vessel density immunoexpression was more in Group 2 (OSCC) as compared to Group 1 (OSMF). Mean α-SMA correlated negatively with E-cadherin expression and positively with vimentin and factor VIII immunoexpression. E-cadherin expression correlated negatively with factor VIII and positively with Vimentin expression. Conclusions: The molecular mechanisms responsible for the development of OSCC in patients with OSMF require unification of multiple progressive pathogenetic mechanisms involved in the progression of the disease.

A comparative assessment of antibacterial properties of neem oil coated sutures: An in vitro study.

Background: When it comes to surgical site infection (SSI), it has been shown that two-thirds of wound infections starts around the incision line and the number is even greater in the presence of sutures. Therefore, a number of compounds have been used to coat the suture materials which tend to decrease the SSI some of which include ciprofloxacin, Aloe vera, and chlorhexidine. Objective: The objective of this study was to assess the antibacterial properties of neem oil-coated sutures in anaerobic culture media when compared to triclosan-coated sutures and uncoated sutures. Materials and Methods: The phosphate-buffered saline solution containing the plaque samples of the patients was transferred evenly onto the sterile agar media. Equal segments of 100% neem oil coated (Group 1), 50% neem oil coated (Group 2), triclosan-coated (Group 3), and uncoated (Group 4) sutures were placed on the agar plate. These were then incubated at 37°C for 24 h. The inhibition zone was calculated in mm by measuring of the zone of inhibition (ZOI) in terms of length, breadth, and area. Results: The results were calculated following the incubation. The mean length for 100% neem oil-coated suture was 2.61 mm ± 0.2 and for 50% neem oil-coated suture was 2.49 mm ± 0.24. The mean breadth for 100% neem oil-coated suture was 1.5 mm ± 0.41 and for 50% neem oil-coated suture the mean was 0.95 mm ± 0.58. The mean area for 100% neem oil-coated suture was 77 mm ± 32.9 and for 50% neem oil-coated suture was 16.8 mm ± 11.14. Triclosan and uncoated sutures did not show any ZOI. Conclusion: The 100% neem oil-coated sutures had the largest length, breadth, and area of ZOI in the anaerobic culture media, followed by 50% neem oil-coated sutures. Triclosan-coated and uncoated sutures did not show any ZOI. Therefore, neem oil-coated suture can be used in the overall healing and prevention of postoperative discomfort after oral surgical procedures.

Pathogenetic model of survivin-dependent molecular signalling pathways in tumorigenesis of oral cancer and precursor lesions.

Background: p53 tumour suppressor gene limits unchecked cellular growth in response to DNA damage, by causing G1 arrest and the activation of apoptosis. Inhibitors of apoptosis include survivin which acts by inhibition of caspases. Survivin has a significant role as a cell cycle modulator and is only minimally present in mature tissues. Aberrant expression of p53 and survivin has been evaluated in various carcinomas. Thus, the objective of this research was to elucidate the co-expression of p53 and survivin in tissue samples of Oral Potentially Malignant Disorders (OPMDs) and Oral Squamous Cell Carcinoma (OSCCs). Method: Thirty tissue samples of OPMDs and 30 tissue samples of OSCCs taken from department archives were used in the study. Expression of p53 and survivin was analyzed in the study groups by the help of immunohistochemistry. Also, co-expression of both the markers was evaluated. Results: The expression of p53 and survivin in the oral epithelium of patients with OSCCs was significantly higher than that in patients with OPMDs (P value ≤0.05). Conclusion: Our results provide insights into the altered survivin and p53 co-expression with significant immunoexpression within the study groups. Therefore, survivin and p53 could be better markers for identifying cell proliferation and apoptotic pathway. Also, malignant transformation rate of OPMD increases with increased expression of these markers.

Evaluation of EGFR in prognostic characterization in histopathological variants of oral squamous cell carcinoma.

AIM OF THE STUDY: Epidermal growth factor receptor (EGFR) is a tyrosine kinase receptor of the Erb B family contributing to proliferation, invasion, and metastasis. EGFR overexpression is frequently associated with poor clinical outcome in malignant neoplasms.• To evaluate and compare immunoexpression of EGFR in histopathological variants of oral squamous cell carcinoma (OSCC).• To hypothesize the role of EGFR in determining biological behavior and prognostic course of histopathological variants of OSCC. MATERIALS AND METHODS: The study comprised a total of 40 cases including 10 cases each of Squamous cell carcinoma, Verrucous carcinoma, Adenosquamous cell carcinoma, and Adenoid squamous cell carcinoma. EGFR immunoexpression was observed qualitatively as low (1), moderate (2) and strong (3) and quantitatively as score 1 for <10%, 2 for 10%-50%, and 3 for >50% positive cells. The resulting data were analyzed using SPSS software version 19. Data have been expressed as mean and standard deviation. Differences between the different variables were analyzed using ANOVA, and Pearson's Chi-square. (p ≤ 0.05). RESULT: The study results revealed that the EFGR immunoexpression was highest in adenosquamous cell carcinoma followed by adenoid squamous cell carcinoma then conventional squamous cell carcinoma followed by lowest immunoexpression in verrucous carcinoma. The results were statistically significant. (p ≤ 0.05). CONCLUSION: Expression of EGFR could be established as a valuable biomarker with significant association in predicting aggressive potential and treatment response in various histopathological variants of OSCC. Further studies where EGFR could be linked to predictive indicators and tumor prognosis could be undertaken.

Immunolocalization of epithelial cell adhesion molecule and matrix metalloproteinase-9 in oral epithelial dysplasia and oral squamous cell carcinoma.

INTRODUCTION: Cancers are complex tissues composed of multiple distinct cell types that participate in heterotypic interactions with one another. Physiologically cell-to-cell contacts formed by dense populations of normal cells operate to suppress further cell proliferation. OBJECTIVES: The objective of the study is to evaluate and compare the immunoexpression of matrix metalloproteinase-9 (MMP-9) and epithelial cell adhesion molecule (EpCAM) in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC) and to hypothesize their role in the progression in varying grades of these lesions. MATERIALS AND METHODS: A total of 60 samples comprising of 30 cases each of OED and OSCC. Three micrometers thin sections were taken and subjected for hematoxylin and eosin stain and immunohistochemical procedure. The sections were incubated with monoclonal anti-EpCAM anti-MMP-9 antibody. The data were analyzed using SPSS software version 19. RESULTS: The results of the study show EpCAM immunoexpression decreased in OSCC when compared to OED. MMP-9 immunoexpression increased in OSCC when compared to OED (statistically significant, P ≤ 0.05). CONCLUSION: Correlation between EpCAM and MMP-9 may help to unravel the signaling cascades involved in the carcinomatous changes, tumor cell invasion, and progression of OSCCs.

Evaluation of Bacterial Reduction at Various Stages of Endodontic Retreatment After Use of Different Disinfection Regimens: An In Vivo Study.

OBJECTIVE: The present study was conducted to evaluate the presence of aerobic bacteria, anaerobic bacteria, E. faecalis, F. nucleatum, Propionibacteria sp., Actinomyces sp., and their reduction at various stages of endodontic retreatment with the use of conventional protocol (5.25 % Sodium Hypochlorite (NaOCl) as the irrigant along with Calcium Hydroxide (Ca (OH)2) as intracanal medicament and advocated protocol (SmearOFF as the irrigant along with 2% Chlorhexidine (CHX) gel as intracanal medicament). METHODS: Twenty eight patients fulfilling the eligibility criteria were selected for root canal retreatment and randomly allocated into two groups. Group 1: Final irrigant as SmearOFF+Chlorhexidine 2% gelas intracanal medicament (n=14). Group 2: Final irrigant as 5.25% NaOCl+Ca(OH)2 as intracanal medicament (n=14). With aseptic environment, access opening was performed followed by Gutta Percha (GP) removal and sample S1 was collected for bacterial analysis. The biomechanical preparation was done by using Reciproc system with additional finishing with XP-Endo Finisher R. Sample S2 was then collected for bacterial analysis after the final irrigation protocol in the respective groups. Intracanal medicaments were placed for one week and sample S3 was collected. All the samples were subjected to qualitative analysis using PCR and quantification was done by Colony Forming Unit (CFU) analysis. RESULTS: Aerobic [28/28], Anaerobic [28/28], Propionibacterium sp. [20/28] and F. nucleatum [24/28] were the most frequently isolated in S1 sample followed by Actinomyces sp. [16/28] and E. faecalis sp. [19/28]. Chemico-mechanical preparation followed by irrigation (S2 sample) resulted in significant reduction of all types of bacteria in both groups. Group-1 (SmearOFF as the final irrigant) had significantly superior efficacy against aerobic bacteria, E. faecalis and F. nucleatum (P<0.05) as compared to Group-2 (NaOCl). After medicament placement, significant differences between the groups were noted only for the E. Faecalis group. For the S3 samples, the mean bacterial reduction was significant in Aerobic and F. nucleatum in S3 samples for Group 1 and Group 2. CONCLUSION: Chemico-mechanical preparation followed by irrigation resulted in significant reduction in bacterial load irrespective of the final irrigant. SmearOFF was significantly better than NaOCl in minimizing bacterial load of E. faecalis and F. nucleatum. 2% Chlorhexidine gel has superior antimicrobial efficacy against E. faecalis and may be recommended in secondary endodontic treatment.

Effect of second-hand smoke on systemic, oral and psychological health of primary school children in Western Uttar Pradesh: a biochemical analysis.

BACKGROUND: Secondhand smoke (SHS) exposure causes several systemic problems like asthma, dental problems like caries and effects overall wellbeing. This study was conducted to study the impact of SHS on primary school children by associating it with dental caries, cotinine level, asthma severity and overall wellbeing. METHODS: A case control study was conducted amongst 200 children (9-15 years old) that came to the Dental College in Muradnagar-Ghaziabad, India. Children who lived in smoking households were identified as SHS subjects-100 children and those who lived in nonsmoking households were identified as control group-100 children. Parents completed questionnaire regarding their smoking status. In children's questionnaire severity of asthma was determined by using expert panel report (EPR3), dental caries was recorded by dmft Index and psychological assessment of children was done using WHO-5 Wellbeing Index. Collection of unstimulated saliva of children was done in sterile plastic tubes and cotinine level was measured using Enzyme-Linked-Immunosorbent Assay (ELISA). Students' t-test and One-Way Analysis of Variance (ANOVA) were used to analyze data. RESULTS: Mean number of children suffering from asthma was higher in SHS subjects in comparison to control group i.e. 348.9±166.509 and 247.3±15.86 respectively. dmft and WHO wellbeing scores increased as level of cotinine increased. Mean cotinine level was higher in SHS subjects than control group i.e. 348.9±166.509 and 247.3±15.86 respectively. CONCLUSIONS: SHS has an adverse impact on young children reflected by poor dmft scores and psychological wellbeing, increased cotinine levels and asthma severity in comparison to control group.

Effect of dental acid etchant-mediated photodynamic therapy on bacterial reduction and microshear bond strength of composite to dentin - An in vitro study.

OBJECTIVE: The objective of this study was to assess the effect of dental acid etchant (DAE)-mediated photodynamic therapy on bacterial reduction and microshear bond strength of composite to dentin. MATERIALS AND METHODS: Eighty permanent third molars after sample preparation were exposed to a cariogenic challenge with Streptococcus mutans. After incubation, specimens were randomly divided into four groups (n = 20): Group I - DAE, Group II - low-level laser (LLL), Group III - diode laser + methylene blue (MB + L), and Group IV - diode laser + DAE (DAE + L). Half of the specimens from each group were selected for bacterial reduction assessment and the other half for microshear bond strength. All the samples for assessment of bacterial reduction (before and after intervention) were seeded onto the surface of mitis-salivarius-bacitracin medium. After incubation, the viable bacterial count was determined in colony-forming unit/mL. For microshear bond strength assessment, samples were subjected to various treatment modalities and then bonding and debonding procedure was performed for blocks of composite and values were recorded. RESULTS: Significant reductions in S. mutans were observed in all the groups - Group I (DAE) 68.50%, Group II (LLL) 55.90%, Group III (MB + L) 88.60%, and Group IV (DAE + L) 87% with comparable bacterial reduction between Group III (MB + L) and Group IV (DAE + L). Furthermore, a significant difference in bond strength values was seen in Group III (MB + L) 10.99 MPa and Group IV (DAE + L) 17.99 MPa whereas an insignificant difference was found between Group I (DAE) 20.74 MPa, Group II (LLL) 18.27 MPa, and Group IV (DAE + L). CONCLUSION: DAE caused a comparable reduction in bacterial count to MB-assisted PDT and also there was no adverse effect on bond strength values. PDT can be performed while acid etchant containing MB dye is being applied in the cavity, thus reducing operative time and enhancing cavity disinfection.

"Neuroectodermal influence of CD 99 immunoexpression correlates with the clinical behavior of odontogenic cysts and tumors".

Background: Odontogenic tumors show a variety of characteristic features that are dependent on the tumor cell origin and the stage of tumor cell differentiation. Odontogenic cysts arise from the enamel organ or remnants of dental epithelium which influences their pathogenetic model and further clinical behavior of these lesions. Aim: The study aims at assessment of CD 99 immunoexpression in odontogenic keratocyst (OKC) and ameloblastoma (in tooth bearing [anterior to third molar] and nontooth bearing areas [molar ramus area]) to postulate neural influence in their pathogenesis and the clinical behavior. Materials and Methods: Immunohistochemical analysis for CD 99 was performed on paraffin-embedded tissue sections on 50 histopathologically confirmed cases of OKC and ameloblastoma (25 each) arising within the oral cavity and were scored qualitatively, topographically, and according to cellular localization. Statistical Analysis: The resulting data were analyzed using the SPSS software version 20.0. The significance of the parameters was tested by the Pearson's Chi-square test (P ≤ 0.05 as statistically significant). Results: CD99 immunoreactivity was distributed in both tooth bearing and nontooth bearing groups of OKC and ameloblastoma with an increased immunoexpression in basal and suprabasal layers of OKC in nontooth bearing area and in peripheral cells of ameloblastoma in nontooth bearing area confined to the cell membrane. Conclusions: The results point toward the role of CD99 in the pathogenesis and aggressive behavior of such odontogenic lesions and it can be used as a promising therapeutic target.

Effect of antimicrobial agents on the oral microflora in patients undergoing fixed orthodontic therapy-An ex vivo comparative analysis.

AIM: To evaluate and compare the efficacy of Ozonated Olive Oil Gel, Chlorhexidine gel, and Amflor (Fluoridated) mouthwash on reducing the count of Streptococcus mutans and Lactobacillus in patients undergoing fixed orthodontic therapy evaluated at different time intervals. METHODS: Sixty patients undergoing orthodontic treatment were randomly divided into three groups (n = 20) based on antimicrobial agents used (Group 1: Ozonated olive oil gel; Group 2: Chlorhexidine gel; Group 3: Fluoridated mouthwash). Elastomeric modules from brackets were collected at T0 (Fresh samples) and T1 (2nd week) and T2 (4th week) for assessment of the microbial growth. These collected modules were cultured and evaluated for the presence of Streptococcus Mutans and Lactobacilli and numbers of colonies were counted at each interval. Data obtained was subjected to statistical analysis using SPSS software (Version 20.0). Level of significance was kept at 5%. Intra-group and inter-group comparison between pretreatment, 2nd week and 4th week was done for each group using Wilcoxon signed rank test and Mann-Whitney U test. RESULTS: There was presence of Streptococcus Mutans and Lactobacilli during orthodontic treatment which progressively increased from To to T1 and then declined from T1 to T2. The colony counts were maximum for Fluoridated mouthwash and least for Chlorhexidine and the results were statistically significant (P < 0.05). CONCLUSION: All three antimicrobial agents used were effective against Streptococcus mutans and Lactobacillus. Chlorhexidine proved to be more efficacious whereas Fluoridated mouthwash proved to be least effective against both Streptococcus mutans and Lactobacillus bacteria.

Inter-comparison of antimicrobial photodynamic therapy, LASER, and an antifungal agent as adjunct intracanal irrigation techniques to standard disinfection protocols in reducing Candida albicans counts in the root canals of primary teeth: A pilot study.

BACKGROUND: Despite endodontic infections being a common problem in the primary dentition, some of the infected primary teeth can remain functional until the exfoliation through endodontic treatment. The primary endodontic treatment goal must be to optimize root canal disinfection and to prevent reinfection. AIM: The aim of this study is to compare the antifungal efficacy of antimicrobial photodynamic therapy, LASER, and 1% clotrimazole as irrigants by reducing the colony-forming units (CFU) of Candida albicans in the infected root canals of primary teeth. MATERIALS AND METHODS: Pulp therapy was performed on deciduous mandibular second molars of 20 healthy children aged 5-8 years, which were divided into four groups of intracanal disinfection protocols: Group 1 (n = 5) - Sodium Hypochlorite, Group 2 (n = 5) - Photodynamic, Group 3 (n = 5) - Normal saline with LASER, and Group 4 - (n = 5) Antifungal. Disinfection potential was compared by collecting samples one just after access opening and the other after the experimental groups using paper points. The samples were cultured and incubated for 48 h to check change in CFU of the fungi. RESULTS: Data were analyzed statistically using the Shapiro-Wilk's test, Mann-Whitney U-test; Kruskal-Wallis test, and Wilcoxon signed-ranks test. No significant difference (P > 0.05) was found between the different groups. However, complete inhibition of C. albicans CFU was seen with Group 4. CONCLUSION: One percent Clotrimazole (antifungal) can be utilized as an adjunct in the endodontic irrigation protocol for better success of pulpectomy in children.

Biochemical analysis of cytokeratin fragment 21-1 concentration and expression of cytokeratin 19 in oral potentially malignant disorders.

INTRODUCTION: Cytokeratin fragment 21-1 (CYFRA21-1), a constituent of the intermediate filament protein is known to be elevated in cancer. In vitro cleavage of cytokeratin 19 (CK19) protein results in the release of it's fragments into the supernatants of premalignant cell lines. This study was designed with the aim to investigate the concentrations of CYFRA21-1 in serum and saliva of oral potentially malignant disorders (OPMD), to evaluate CK19 expression in tissues of the same patients and to correlate the levels of CYFRA21-1 concentration in serum and saliva with CK19 expression in OPMDs, and to compare it with oral squamous cell carcinoma (OSCC), which was taken as positive control. MATERIALS AND METHODS: Concentration of CYFRA21-1 was measured in saliva and serum of 30 OPMD cases with five patients having OSCC using ELISA technique and analysis of CK19 protein expression in the tissue of same patients using immunohistochemical technique was done. RESULTS: Concentration of CYFRA21-1 in saliva and serum with regard to CK19 protein expression in tissues was significantly higher in control group than in study groups. CONCLUSION: CYFRA21-1 can be used as a promising diagnostic molecule and as an adjunctive marker for early detection, disease staging, and monitoring.

Molecular Analysis as a Guide to Determining the Extent and Pathophysiology of Perilesional Tissues in Oral Epithelial Dysplasias.

INTRODUCTION: Clinically and histologically normal appearing perilesional mucosa of epithelial dysplasia may harbor early genetic changes. Hence, the present study is designed to determine the early molecular changes in the form of p16 and EGFR immunoexpressions in perilesional tissues. OBJECTIVES: To analyze immunohistochemical expressions of p16 and EGFR individually and percentage change of immunoexpressions in oral dysplastic lesions and their perilesional tissues. MATERIALS AND METHODS: Forty formalin-fixed paraffin-embedded tissues of oral epithelial dysplasia with perilesional tissue marked by India ink were included in this study. Immunohistochemical staining was performed using anti-p16 and anti-EGFR monoclonal antibodies (BioGenex) using squamous cell carcinoma of uterine cervix and breast carcinoma as the positive controls, respectively. RESULTS: p16 and EGFR expressions were assessed based on the presence, intensity, extent and immunolocalization of positive cells. Out of 40 cases, p16 immunoexpression was positive in 82.5% cases of lesional tissues and in 62.5% cases of perilesional tissues (p ≤ 0.05); however, EGFR immunoexpression was positive in 90% cases of both lesional and perilesional tissues (p > 0.05). CONCLUSION: The disease status and progression based on p16 and EGFR expressions and co-expressions can be used as an effective guide to evaluating the progression of normal epithelium to dysplastic epithelium in otherwise clinically normal mucosa.