Effect of Curcumin on the Head and Neck Squamous Cell Carcinoma Cell Line HN5.

BACKGROUND: Curcumin has been isolated from the rhizomes of Curcuma longa. Over the years, it has shown outstanding therapeutic potential in various human disorders, including cancers. OBJECTIVE: The aim is to study curcumin's effects on the apoptosis signaling pathway in the head and neck squamous cell carcinoma (HNSCC) cell line HN5. METHODS: The cytotoxicity of curcumin on HN5 cells were assessed. In addition, HN5 cells were also treated with curcumin to evaluate its effect on the caspase-8, -9, Bcl-2, Bax, and Stat3 gene expressions. RESULTS: The results exhibited that cell viability reduced following curcumin treatment in a concentration- dependent manner. Curcumin treatment caused decreased expression of Bcl2, with simultaneous upregulation of the Bax/Bcl2 ratio. Curcumin increased caspase-9 expression, did not affect caspase-8, and decreased Stat3 expression. The induction of the mitochondria-dependent apoptosis pathway of curcumin happened by modulating the expression of Bcl2 and Bax genes, resulting in the caspase-9 activation. Furthermore, curcumin decreased the expression of the Stat3 in HN-5 cells. CONCLUSIONS: In conclusion, curcumin showed marked anticancer effects in the HN-5 cell line by modulating Stat-3; Bax/Bcl-2 expression in vitro.

Effect of Ferula persica plant methanol extract on the level of Cox-2 in induced squamous cell carcinoma (SCC) in rat tongue.

Background: More than 90% of oral cancers are cases of squamous cell carcinoma. Standard treatment of cancer includes a combination of surgery, chemotherapy and radiotherapy. Each of these treatments, however, brings about certain problems and side effects. Today herbal medicine, has become a more preferable option in dealing with health problems or preventing them because this type of medicine has better compatibility with the body and does not cause undesirable side effects. In this study , the effect of Ferula persica plant methanol extraction on Cox-2 levels in SCC induced rat tongue is conducted in vivo. Methods: In this lab research, 75 rats from SD race in the age - range of 2/5 - 3 months were selected and put in five groups. In order to induce tongue carcinoma, 4- Nitroquinoline 1 (4 NQO) powder was used 3 times a week for each rat. Furthermore, Ferula persica extract was given to each of the groups in order to examine Cox-2 changes in the blood. Results: Comparison of Cox-2 average in various groups resulted in the observation that there was significant difference between the Cox-2 levels in the groups which had only received carcinogen and the other groups. In this group, Cox-2 level was less and in the group that had received Ferula extract (500 mg) along with carcinogen , Cox-2 level was found to be more than other groups. Conclusion: Ferula persica extract does not have reducing effect on serum Cox-2.

Effect of Beta-Blockers on Number of Osteoblasts and Osteoclasts in Alveolar Socket Following Tooth Extraction in Wistar Rats.

STATEMENT OF THE PROBLEM: Various researchers have suggested the use of ß2-adrenergic receptor antagonists in prevention or treatment of bone resorption. PURPOSE: This study aimed to evaluate the effect of ß2-adrenergic receptor antagonists on number of osteoclasts and osteoblasts involved in the healing of extraction socket of maxillary first molar in rats. MATERIALS AND METHOD: Maxillary first molars of 40 rats were extracted and divided into two groups. The test group received 0.1 mg/kg propranolol intraperitoneally daily. The controls received normal saline. At days 7, 14, 21 and 28 post-extraction, 5 rats were euthanized from each group. Maxillary bone was resected and the mean number of osteoblasts and osteoclasts in tooth socket was measured. RESULTS: After 1 week, the number of osteoclasts in the controls was significantly higher than the test group. A significant increase in the number of osteoclasts in both groups at week 1 was observed compared to the following weeks (p< 0.05). The number of osteoblasts in the controls at second week reached its maximum rate but stayed constant in the 3rd and 4th weeks. Osteoblasts in the test group increased similar to the controls but reached its maximum at 3rd week and showed a significant increase compared to the controls (p< 0.05). CONCLUSION: ß2 adrenergic receptor antagonists decrease the number of osteoclasts and increase the number of osteoblasts during extraction socket healing.

IL-4 induces the formation of multinucleated giant cells and expression of Beta5 integrin in central giant cell lesion

BACKGROUND: It is now well established that IL-4 has a central role in the development of monocytes to multinucleated giant cells (MGCs) by inducing the expression of integrins on the surface of monocytes. The aim of this study was to investigate the potential role of IL-4 in induction of β5 integrin expression in the peripheral blood samples of patients with giant cell granuloma. MATERIAL AND METHODS: Monocytes were isolated from peripheral blood samples of patients with central giant cell granuloma (CGCG) and healthy controls using human Monocyte Isolation Kit II. Isolated monocytes were then cultured in the absence or presence of IL-4 (10 and 20 ng/mL), and following RNA extraction and cDNA synthesis, Real-time PCR was performed to determine the level of β5 integrin expression. The formation of CGCGs and morphological analyses were done under light microscopy. For confirmation of CGCGs, immunocytochemistry technique was also carried out by anti-RANK (receptor-activator of NF-κB ligand) antibody. RESULTS: In both patient and control groups, β5 levels were significantly enhanced by increasing the IL-4 dose from 10 to 20 ng/mL. In addition, these differences were significant between patient and control groups without IL-4 treatment. On the other hand, the number of cells which expressed RANK and therefore the number of giant cells were significantly higher in the patient group in comparison to controls, as assessed by immunohistochemistry evaluations. CONCLUSIONS: In this study, we showed an elevation in the expression levels of β5 integrin when stimulated by IL-4. It is strongly indicated that this integrin acts as an important mediator during macrophage to macrophage fusion and development of giant cells

Tissue Reaction and Biocompatibility of Implanted Mineral Trioxide Aggregate with Silver Nanoparticles in a Rat Model.

INTRODUCTION: Biocompatibility and antimicrobial activity of endodontic materials are of utmost importance. Considering the extensive applications of mineral trioxide aggregate (MTA) in dentistry and antimicrobial properties of silver nanoparticles, this study aimed to evaluate the subcutaneous inflammatory reaction of rat connective tissues to white MTA with and without nanosilver (NS) particles. METHODS AND MATERIALS: Polyethylene tubes (1.1×8 mm) containing experimental materials (MTA and MTA+NS and empty control tubes) were implanted in subcutaneous tissues of seventy-five male rats. Animals were divided into five groups (n=15) according to the time of evaluation: group 1; after 7 days, group 2; after 15 days, group 3; after 30 days, group 4; after 60 days and group 5; after 90 days. The inflammatory reaction was graded and data was analyzed using the Kruskal-Wallis and Mann-Whitney U tests. Statistical significance was defined at 0.05. RESULTS: Comparison of cumulative inflammatory reaction at all intervals revealed that the mean grade of inflammatory reaction to MTA, MTA+NS and control samples were 3, 2 and 2, respectively. According to the Mann-Whitney analysis there were no significant differences between MTA+NS and MTA (P=0.42). CONCLUSION: Incorporation of 1% nanosilver to MTA does not affect the inflammatory reaction of subcutaneous tissue in rat models.

Therapeutic Efficacy of Orally Delivered Doxorubicin Nanoparticles in Rat Tongue Cancer Induced by 4-Nitroquinoline 1-Oxide.

PURPOSE: Oral cancer is one of the most significant cancers in the world, and squamous cell carcinoma makes up about 94% of oral malignancies. The aim of the present study was to compare the efficacy of doxorubicin plus methotrexate - loaded nanoparticles on tongue squamous cell carcinoma induced by 4NQO and compare it with the commercial doxorubicin and methotrexate delivered orally on seventy SD male rats. METHODS: 70 rats were divided into five groups. During the study, the animals were weighed by a digital scale once a week. Number of mortalities was recorded in the data collection forms. At the end of the treatment, biopsy samples were taken from rat tongues in order to evaluate the severity of dysplasia and the extent of cell proliferation. The results were analyzed using ANOVA, descriptive statistics and chi-square test. RESULTS: No statistically significant difference was found in the mean weight of five groups (p>0.05). No significant relationship was found between groups and mortality rate (P = 0. 39). In addition, there was a significant relationship between groups and the degree of dysplasia (P <0.001). The statistical analysis showed a significant relationship between groups and the rate of cell proliferation (p <0.001). CONCLUSION: The results of the present study showed that the use of doxorubicin plus methotrexate - loaded nanoparticles orally had more therapeutic effects than commercial doxorubicin plus methotrexate.

Garlic ((Allium sativum)) Fresh Juice Induces Apoptosis in Human Oral Squamous Cell Carcinoma: The Involvement of Caspase-3, Bax and Bcl-2.

Background and aims. There is no report on the apoptotic impact of Allium sativum L.(Garlic) on the oral squamous cell carcinoma (KB); hence, this study was designed to survey the apoptotic effects of garlic fresh juice (GFJ) on the KB cells. Materials and methods. MTTassay (MicrocultureTetrazolium Assay) was carried out to evaluate the cytotoxicity of GFJ on KB cells. Furthermore, TUNEL(Terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling)and DNA fragmentation tests were performed to determine if GFJ is able to induce apoptosis in KB cells. Also a standard kit was used to assess caspase-3 activity in KB cells. Also western blotting was employed to evaluate the effect of GFJ on Bax:Bcl-2 ratio. Results. Significant cytotoxic effects were observed for the minimum used concentration (1µg/mL) as calculated to be 77.97±2.3% for 24 h and 818±3.1% for 36h of incubation (P < 0.001). Furthermore, TUNEL and DNA fragmentation tests corroborated the apoptosis inducing activity of GFJ. Consistently, after treating KB cells with GFJ(1µg/mL), caspase-3 activity and Bax:Bcl-2 ratio were raised by 7.3±0.6 and (P <0.001) folds, respectively. Conclusion. The results of this study advanced that GFJ induces apoptosis in the KB cells through increasing caspase-3 activity and Bax:Bcl2 ratio which could be attributed to its organo-sulfurcomponents.

Molecular Targeting of Her-2/neu Protein Is Not Recommended as an Adjuvant Therapy in Oral Squamous Cell Carcinoma and Oral Lichen Planus.

PURPOSE: Target therapy against molecular markers of EGFR family has been recently used in the treatment protocol of several diseases. However, the role of Her-2/neu in OSCC is a matter of controversy and more studies are required to clarify the role of Her-2/neu in OSCC. We aimed to investigate the role of Her-2/neu in the process of carcinogenesis in oral epithelium as ELP is a premalignant and OSCC is a malignant lesion, but RLP shows no evidence of premalignancy and malignancy.To our Knowledge, this is the first study comparing Her-2/neu expression in erosive lichen planus (ELP), reticular lichen planus (RLP), and oral squamous cell carcinoma (OSCC). METHODS: 60 samples, including 20 cases of RLP, 20 cases of ELP, and 20 cases of OSCC were evaluated immunohistochemically in this study. RESULTS: Our findings showed that there was not a significant increase in Her-2/neu expression from RLP to ELP and from ELP to OSCC. Therefore, Her-2/neu had no role in differentiating between RLP, ELP and OSCC and this protein does not contribute to the process of carcinogenesis in the oral epithelium. CONCLUSION: The lack of Her-2/neu overexpression indicates that molecular targeting of Her-2/neu protein is not recommended as an adjuvant therapy in OSCC and OLP.

P53 gene codon 72 polymorphism in patients with oral squamous cell carcinoma in the population of northern Iran

OBJECTIVES: Squamous cell carcinoma is the most common cancer of the oral cavity, and several etiologic factors are involved in its development. Single nucleotide polymorphism (SNP) of the P53 gene codon 72 (P53c72) changes the structure of the protein and affects its activity. The prevalence of P53c72 different genotypes, which seems to vary with race and geographic location, has shown a strong correlation with many types of human cancers. The aim of this study was to investigate the correlation between P53c72 polymorphism and risk of oral squamous cell carcinoma (OSCC) in the heavily populated Gilan Province in northern Iran. Design of Study: This case-control study was done on 55 paraffin-embedded samples from OSCC patients and 100 samples of non-dysplastic oral cavity lesions. The P53c72 genotypes were determined using the ARMS-PCR method. SPSS-15 software was used for statistical analysis. RESULTS: There were no significant statistical differences found between the prevalence of different P53c72 genotypes in the OSCC group vs. the control. However, the Pro/Pro genotype in OSCC samples showed a strong correlation with age, as 70% of such patients were below 50 years old. Interestingly, a large portion (40%) of the patients with the Pro/Pro genotype had the tumor in the lip area. CONCLUSIONS: Although P53c72 polymorphism does not appear to be a predisposing factor for OSCC in the population of Northern Iran, the Pro/Pro genotype could be considered as a risk factor for OSCC in adults below 50 years old and the anatomical location of the tumor

A review of antibacterial agents in endodontic treatment.

Microorganisms play a major role in initiation and perpetuation of pulpal and periapical diseases. Therefore, elimination of the microorganisms present in the root canal system is the fundamental objective of endodontic treatment. The use of mechanical debridement, chemical irrigation or other antimicrobial protocols and intra-canal medicaments are critical to attain this goal. The aim of this article was to review the antimicrobial agents and their properties in endodontics.

P53 gene codon 72 polymorphism in patients with oral squamous cell carcinoma in the population of northern Iran.

OBJECTIVES: Squamous cell carcinoma is the most common cancer of the oral cavity, and several etiologic factors are involved in its developing. Single nucleotide polymorphism (SNP) of the P53 gene codon 72 (P53c72) changes the structure of the protein and affects its activity. The prevalence of P53c72 different genotypes, which seems to vary with race and geographic location, has shown a strong correlation with many types of human cancers. The aim of this study was to investigate the correlation between P53c72polymorphism and risk of oral squamous cell carcinoma (OSCC) in the heavily populated Gilan Province in northern Iran. DESIGN OF STUDY: This case-control study was done on 55 paraffin-embedded samples from OSCC patients and 100 samples of non-dysplastic oral cavity lesions. The P53c72 genotypes were determined using the ARMS-PCR method. SPSS-15 software was used for statistical analysis. RESULTS: There were no significant statistical differences found between the prevalence of different P53c72 genotypes in the OSCC group vs. the control. However, the Pro/Pro genotype in OSCC samples showed a strong correlation with age, as 70% of such patients were below 50 years old. Interestingly, a large portion (40%) of the patients with the Pro/Pro genotype had the tumor in the lip area. CONCLUSIONS: Although P53c72 polymorphism does not appear to be a predisposing factor for OSCC in the population of Northern Iran, the Pro/Pro genotype could be considered as a risk factor for OSCC in adults below 50 years old and the anatomical location of the tumor.

Induction of Apoptosis and Cytotoxic Activities of Iranian Orthodox Black Tea Extract (BTE) Using in vitro Models.

PURPOSE: Plant-derivate therapeutic agents can perform cancer chemotherapeutic activity through triggering apoptotic cell death. Our aim was to investigate the cytotoxic effects, induction of apoptosis, and the mechanism of cell death of Iranian orthodox black tea extracts (BTEs) and hydro methanolic purified fractions (40, 60, 80 and 100%) in KB cells (oral squamous cell carcinoma). METHODS: In order to analyze the cytotoxic activity of the BTEs, MTT (3-(4, 5- dimetylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide) and Trypan-blue assays were performed in oral squamous cell carcinoma (KB). Furthermore, the apoptosis inducing action of the extracts was determined by TUNEL, DNA fragmentation and cell death detection analysis. RESULTS: Dichloromethane BTE and hydro methanol fractions (40 and 60%) extract showed no cytotoxic effects; however, hydro methanol crude and hydro methanol fractions of BTE (80 and 100%) significantly inhibited cell growth and viability in a dose and time dependent manner. In addition, Cell death assay, TUNEL, and DNA fragmentation indicated induction of apoptosis by hydro methanol 80 and 100% fractions of BTE in KB cells. Statistical significance was determined by analysis of variance (ANOVA), followed by Duncan test and p value ≤0.05 was considered significant. CONCLUSION: The results from the present study suggests that the hydro methanol crude and hydro methanol fractions of BTE (80 and 100%) are significant source of compounds with the anti proliferative and cytotoxic activities, and this may be useful for developing potential chemo preventive substances.

Induction of apoptosis by grape seed extract (Vitis vinifera) in oral squamous cell carcinoma.

Development of novel therapeutic modalities is crucial for the treatment of oral squamous cell carcinoma (OSCC). Recent scientific studies have been focused on herbal medicines as potent anti-cancer drug candidates. This study is the first to investigate the cytotoxic effects and the mechanism of cell death induced by grape seed extract (GSE) in oral squamous cell carcinoma (KB cells). MTT (3-(4,5-dimetylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) and trypan blue assays were performed in KB cells as well as human umbilical vein endothelial cells (HUVEC) were used to analyze the cytotoxic activity of GSE. Furthermore, the apoptosis-inducing action of the extract was determined by TUNEL, DNA fragmentation and cell death analysis. Statistical significance was determined by analysis of variance (ANOVA), followed by Duncan's test at a significance level of P≤0.05. The results showed apoptotic potential of GSE, confirmed by significant inhibition of cell growth and viability in a dose- and time- dependent manner without inducing damage to non-cancerous cell line HUVEC. The results of this study suggest that this plant contains potential bioactive compound(s) for the treatment of oral squamous cell carcinoma.

Correlation of histopathologic features with demographic, gross and radiographic findings in giant cell granulomas of the jaws.

Background and aims. The correlation between morphology of giant cells in peripheral granulomas of the jaws and the aggressive behavior of the lesion is unknown. This study investigated the correlation between the histopathologic features with demographic, gross and radiographic findings in giant cell granulomas. Materials and methods. In this analytical study, data from 23 cases of central giant cell granuloma (CGCG) and 42 cases of peripheral giant cell granuloma (PGCG) were analyzed, focusing on age, gender, location, and gross and radiographic features. For each patient, microscopic slides were assessed in terms of histologic features of giant cells and stroma. Results. No significant differences were found in the mean number of nuclei or the size of nuclei and giant cell distribution patterns between the jaws and genders in both lesions (P >0.05). Correlation between the mean number of nuclei and age was positively significant and correlation between the size of nuclei and age was negatively significant (P < 0.05). In addition, correlation between the mean number and size of nuclei and the size of the lesion was significant (P < 0.05). Correlation between stroma and aggressiveness of CGCGs was not statistically significant. Correlation between histopathologic features and radiographic findings was not statistically significant (P > 0.05). Conclusion. There were correlations between the mean number of nuclei per giant cell and the size of the lesion and age, and between the size of nuclei and size of the lesion. No relation was observed between histopathologic and radiographic features.

Inverted papilloma of the palate with malignant transformation.

The inverted papilloma is a unilateral sinonasal benign tumor which is characterized by aggressive local invasion, high recurrence rate, and transformation into malignancy. The etiology of inverted papilloma is still unknown. Possible causes include allergy, chronic sinusitis, occupational exposure to dusts and aerosols, tobacco, and viral infections. Treatment is complete surgical excision and close postoperative follow-up is necessary. Here we report a case of inverted papilloma arising from the hard palate with malignant transformation in a 41-year-old female. Clinical and histological features and treatment are discussed with the review of literature.

Effects of various mixing techniques on push-out bond strengths of white mineral trioxide aggregate.

INTRODUCTION: The aim of this study was to evaluate the effect of 3 different mixing methods on push-out bond strength of white tooth-colored mineral trioxide aggregate (MTA). METHODS: Sixty 2-mm-thick root sections were prepared from 60 single-rooted human teeth and filled with MTA. The samples were randomly divided into 3 groups (n = 20) of MTA mixed by ultrasonic, trituration, or conventional methods and then allowed to set for 72 hours in a synthetic tissue fluid environment. Push-out bond strength values of the specimens were measured by a universal testing machine and examined under a stereomicroscope at × 4 to determine the nature of the bond failure. RESULTS: The means of push-out bond strength values of white MTA in the ultrasonic, conventional, and trituration groups were 105.67 ± 12.79, 118.95 ± 12.76, and 99.60 ± 14.27 MPa, respectively. The differences between the 3 groups were not statistically significant. CONCLUSIONS: On the basis of the results of this study, it can be concluded that different mixing methods have no significant effect on push-out bond strength of white MTA.

Comparison of two histopathologic methods for evaluating subcutaneous reaction to mineral trioxide aggregate

Objectives: One of the most important factors for suitable materials for pulp therapy is biocompatibility. Two histopathologic methods of Cox and Federation Dentaire International (FDI) were used to evaluate inflammation. In Cox method, density of inflammatory cells, tissue reactions like fibrosis, vascular responses like congestion and fibrin extravasation have been used to evaluate inflammatory reactions. The aim of this study was to compare the accuracy of pathologists’ interpretations using two different methods. Study design: Three pathologists observed the degree of inflammation in 225 histopathologic sections. These sections showed inflammation in subcutaneous connective tissue of rats adjacent to polyethylene tubes, filled with white or gray mineral trioxide aggregate. Empty tubes served as controls. Samples were harvested after 7-, 15-,30-, 60-, and 90-days. All pathologists examined the sections under a light microscope (Carl Zeiss, Oberkochen,Germany) at ×400 magnifications. Chi-Square test was used to evaluate the difference between inflammation grades when one pathologist used two methods. Cohen’s Kappa value was used to measure agreement of three pathologists to recognize the degrees of inflammations when using one of the methods(..) (AU)

Comparison of two histopathologic methods for evaluating subcutaneous reaction to mineral trioxide aggregate.

OBJECTIVES: One of the most important factors for suitable materials for pulp therapy is biocompatibility. Two histopathologic methods of Cox and Federation Dentaire International (FDI) were used to evaluate inflammation. In Cox method, density of inflammatory cells, tissue reactions like fibrosis, vascular responses like congestion and fibrin extravasation have been used to evaluate inflammatory reactions. The aim of this study was to compare the accuracy of pathologists' interpretations using two different methods. STUDY DESIGN: Three pathologists observed the degree of inflammation in 225 histopathologic sections. These sections showed inflammation in subcutaneous connective tissue of rats adjacent to polyethylene tubes, filled with white or gray mineral trioxide aggregate. Empty tubes served as controls. Samples were harvested after 7-, 15-, 30-, 60-, and 90-days. All pathologists examined the sections under a light microscope (Carl Zeiss, Oberkochen, Germany) at ×400 magnifications. Chi-Square test was used to evaluate the difference between inflammation grades when one pathologist used two methods. Cohen's Kappa value was used to measure agreement of three pathologists to recognize the degrees of inflammations when using one of the methods. RESULTS: There were no significant differences between the two methods when one of the pathologist used these methods to report the degree of inflammation (p=0.054). However, two other pathologists reported significant differences between two methods (p=0.005, p=0.001). In the FDI method, there was an acceptable agreement between first and second, and first and third pathologist in terms of the degree of inflammation, and intermediate agreement existed between the second and third pathologist. With the Cox method, no agreement among the pathologists could be found. CONCLUSION: The results of three pathologists in terms of rating inflammation with the FDI method showed better agreement than with the Cox method. Therefore, FDI method is more reliable than the Cox method to evaluate inflammation.

Endodontic treatment of a hypertaurodont mandibular second molar: a case report.

A 28-year old patient was referred for endodontic treatment of his left mandibular second molar. The patient had no history of systemic diseases and his chief complaint was spontaneous pain in left posterior region of mandible. Clinically, there was a deep caries in the left mandibular second molar. Radiographic examination of this tooth revealed a long crown containing large pulp chamber and two short roots with a apically located furcation, indicating hypertaurodontism. After coronal access preparation, five orifices were found including mesiolingual, mesiobuccal, distobuccal, mid-distal and distolingual. Subsequent to root canal preparation, a modified filling technique was used for canal obturation. After one year the treated tooth was symptom free.

White sponge nevus: a case report.

White sponge nevus (WSN) is a rare hereditary dyskeratotic hyperplasia of mucous membranes. It is an autosomal dominant disorder with variable penetrance. We report a case of WSN in a healthy 21-year-old male with no history of familial involvement. A white smooth plaque with no erythema or other structural abnormalities was observed, which confirmed the diagnosis of WSN histopathologically.