Polymerase chain reaction-based identification of various serotypes of Streptococcus mutans in adults with and without dental caries.

Background: Dental caries is a multistep process which initiates the development of plaque' defined as a structured biofilm containing microbial communities. Teeth provide unique surfaces for bacterial colonization. Serotypes of Streptococcus mutans implicate the development of dental caries. Aim: The aim of the study was to determine the prevalence and association of serotypes of S. mutans in groups with and without dental caries. Materials and Methods: One hundred and fifty adults aged between 18 and 35 years were included in the study. Supragingival plaque samples were collected, followed by deoxyribonucleic acid extraction. Polymerase chain reaction was performed to identify S. mutans and its serotypes. Proportions of S. mutans and its serotypes were correlated with caries-active (CA) and caries-free (CF) groups. Results: CA group showed 66.7% positivity for S. mutans and CF group showed only 42.7% of positivity. Serotype C showed a higher proportion followed by E' F, and K in the CA group, whereas in the CF group, higher proportion was observed with K followed by C' E, and F. 70.8% cases showed single serotype in the CA group and 83.3% in CF group. Multiple serotypes were seen in 29.2% in the CA group and 16.7% in the CF group. Conclusions: The study clearly established variation in proportions of S. mutans and its serotypes between CA and CF groups. Positive correlation was observed in the CA group for S. mutans and its serotypes.

Association of F. alocis and D. pneumosintes with Periodontitis Disease Severity and Red Complex Bacteria.

Oral biofilms are considered the principal etiological agent in the development of periodontitis. Novel species that may contribute to periodontitis and dysbiosis have been identified recently. The study aims to evaluate the presence of F. alocis and D. pneumosintes in healthy and diseased patients and their association with clinical parameters and with red complex bacteria. The study included 60 subjects, with 30 patients each in the healthy and periodontitis groups. The clinical parameters were noted, and samples were subjected to DNA extraction followed by a polymerase chain reaction. Statistical analysis was performed using the Graph Pad Prism software. Results: F. alocis and D. pneumosintes were detected at a significantly higher percentage in the periodontitis group compared to the healthy group (p < 0.05). D. pneumosintes was significantly associated with T. forsythia in the periodontitis group (p < 0.05). Both of these organisms were present in sites with higher clinical attachment loss (p < 0.05). This study demonstrated that both F. alocis and D. pneumosintes were detected at a significantly higher percentage in periodontitis subjects and were detected more frequently in sites with a greater clinical attachment loss. It was also evident that both F. alocis and D. pneumosintes can be present independently of other putative periodontal pathogens.

Evaluation of antimicrobial efficacy of ethanolic extract of Cuminium Cyminium as intracanal medicament on common bacteria of endodontic infections: An in vitro study.

Context: Antimicrobial intracanal medicaments play a vital role in successful outcome of any endodontic procedure. One such plant extract Cuminium cyminium, as intracanal medicaments needs to be researched. Aims: The purpose of this study was in vitro assessment of the antibacterial activity of ethanol extract of C. Cyminium in comparison to Calcium hydroxide (Ca[OH]2) as intracanal medicament against the pathogens of endodontic infection, at an interval 1 h, 24 h, 48 h, and 72 h. Settings and Design: The study was conducted in the central research laboratory of our institute. Freshly prepared C. cyminium extract was procured from AYUSH approved laboratory and direct contact test (DCT) was utilized, which is based on turbidometric determination of microbial growth in a 96-well microplate, carrying 6 times for each bacteria. Methodology: Three groups were assigned for each material in a 96 microwell plate for DCT. Bacterial growth kinetics was monitored at intervals of 1 h, 24 h, 48 h, and 72 h using spectrophotometer at 595 nm. The optical density of T2 (Test group), P2 (Positive control), and N2 (Negative control) was considered. Statistical Analysis Used: After compiling the data, based on the normality of data, further statistical analysis was conducted using Kolmogorov-Smirnov test, Paired t-test, and pairwise comparisons by Turkey's multiple post hoc procedures. The level of statistical significance was set at P = 0.05. Results: The comparison of mean optical density values of C. cyminium in comparison with Ca(OH)2 against the microorganisms of endodontic origin showed a statically significant decrease in bacterial viability at the end of 24 h, 48 h, and 72 h. Conclusion: Based on the results of the study, it can be concluded that C. cyminium has significant antibacterial action against endodontic origin, at interval of 24 h, 48 h, and 72 h.

"Efficacy of cytotoxic effect of green tea catechins on the human periodontal fibroblasts and human dental pulp fibroblasts -An in vitro study".

Background: Inflammation of tooth-supporting tissue and the pulp tissue is followed by wound healing and regeneration process that involves the specific type of connective tissue cells, the fibroblasts. During periodontitis and pulpitis, the inflammation of the tissue causes damage to the fibroblasts. These fibroblasts secrete collagen proteins and maintain the structural framework; along with this the inflammatory process moves toward healing where in the specific cells such as the fibroblast cells play important roles. Green tea catechins epigallocatechin-3-gallate (EGCG) being one of the major catechins is known to have multiple beneficial effects on human fibroblasts. Objective: To assess the in vitro cytotoxicity of green tea catechins on the human periodontal ligament (PDL) fibroblasts and human dental pulp fibroblasts. Materials and Methods: Human PDL fibroblasts (hPDLFs) and human dental pulp fibroblasts were isolated from the two extracted premolar teeth that were indicated for orthodontic treatment. The fibroblasts were then seeded in 96 well tissue culture plate for cell viability study. EGCG was used at different concentration to treat the cells. After 48 h; (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) (MTT) assay was performed to determine the cell viability. Results: The vitality of hPDLFs and human dental pulp fibroblasts was found to be inversely proportional to EGCG concentrations. Conclusions: hPDLFs have shown 37% proliferation at lowest concentration of EGCG used and human dental pulp fibroblasts have shown 99% viability at lowest concentration of EGCG used.

Assessment of the effects of hydrogen water on human gingival fibroblast cell culture in patients with chronic periodontitis.

Background: Activated inflammatory cells produce reactive oxygen species (ROS) to eliminate pathogens. Under normal conditions, the pathogens are taken care of, and tissues are repaired. However, in periodontal disease, persistent inflammation causes increased ROS release and impaired healing. Therefore, removal of overproduced ROS using antioxidants is necessary. Hydrogen water has an antioxidative effect on cells and impedes oxidative stress-related disorders. Aim: To study the effect of hydrogen water on cell viability, migration, and its antioxidative potential in fibroblasts obtained from chronic periodontitis patients. Materials and Methods: The gingival tissue samples were obtained from 26 subjects (13 periodontally healthy individuals and 13 chronic periodontitis patients) and processed. The human gingival fibroblasts were cultured and the assays were commenced once adequate growth was detected. The effect of hydrogen water on cell viability was checked by neutral red assay, while the migration potential was assessed by transwell migration assay. The antioxidative potential of hydrogen water was evaluated by CUPRAC assay. Statistical Analysis: Intergroup comparison was done using Mann-Whitney U-test. Intragroup comparison was done using Wilcoxon signed-rank test. Results: Hydrogen water was nontoxic to the fibroblasts at 24 h and 48 h. The intergroup comparison of the cell viability between hydrogen water-treated periodontally healthy gingival fibroblasts (HF) and fibroblasts from patients with chronic periodontitis (CF) showed a statistically significant (P = 0.00) difference at 24 h and 48 h. Hydrogen water also positively influenced the migratory capacity. Hydrogen water-treated fibroblasts obtained from chronic periodontitis patients showed more migration in comparison to the healthy group (P = 0.00). Hydrogen water showed an antioxidative potential. The maximum potential was seen in relation to the fibroblasts obtained from chronic periodontitis patients at 48 h. Conclusion: Hydrogen water was nontoxic, increased the migratory capacity, and showed an antioxidative potential on human fibroblasts obtained from periodontally healthy individuals and patients with chronic periodontitis.

Simultaneous detection and evaluation of Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, and Prevotella melaninogenica in subgingival plaque samples of chronic periodontitis and healthy individuals through multiplex polymerase chain reaction.

Background: Prevotella is a Gram-negative anaerobic bacilli. The phenotypic characteristics of the various species of Prevotella are similar, which often makes it difficult in routine differentiation and identification of all the species. Aim: The purpose of the study was to detect and compare presence of Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica, and Prevotella loescheii in subgingival plaque samples of chronic periodontitis and healthy individuals. Materials and Methods: Two hundred and thirty-six subjects were considered consisting of chronic periodontitis (128) and healthy (108) individuals. Subgingival plaque sample was collected in reduced transport fluid and analyzed. DNA extraction and polymerase chain reaction (PCR) were performed for genus Prevotella followed by positive samples were considered for the detection of selected species through multiplex PCR using specific primers. Results: Out of 236 samples, 94.1% were positive for genus Prevotella. Out of 222 cases P. nigrescens showed the highest number of cases positive (59.5%) followed by P. melaninogenica (57.2%), P. intermedia (55.4%), and P. loescheii (40.1%). Species were analyzed individually between chronic periodontitis and healthy, P. intermedia, P. nigrescens, and P. loescheii showed greater positivity in healthy compared to chronic periodontitis. Positivity for P. melaninogenica was high in chronic periodontitis compared to healthy. Conclusion: The number of positive cases for species, when correlated with clinical parameters showed an increase in mean score for all clinical parameters assessed, suggesting the presence of variation in the prevalence of Prevotella species and geographic variation do exist in oral microflora. Findings suggest that they can be normal commensals and opportunistic.

Regeneration of periodontal ligament fibers around mini dental implants and their attachment to the bone in an animal model: A radiographic and histological study.

Background: Tissue-engineered periodontal ligament (PDL) around a dental implant by using PDL stem cells (PDLSCs) may be useful in periodontal regeneration and can reduce or eliminate certain shortcomings of dental implants. Materials and Methods: PDLSCs were isolated from extracted human PDL cells and cultured in a bioreactor. They were identified using markers CD45, CD73, CD90, CD105, and CD146. After the formation of multiple cellular layers, they were then attached on titanium mini dental implants and placed in rabbit tibia. The rabbits were sacrificed after 9 months, and the implants were analyzed histologically and radiographically by Cone beam computed tomography (CBCT). Results: Isolated PDLSCs obtained from human premolars showed a colony-forming ability on the 7th day and 14th day. Immunocytochemistry revealed that cells had taken up the adequate positive stains for primary antibodies CD73, CD90, CD105, and CD146 and negative staining for CD45. The histological sections obtained from sacrificed rabbits, when viewed under the light microscope, clearly showed the presence of PDL around dental implants. CBCT examination showed that the implant was well within the bone and did not migrate. The site appeared to be normal without any lytic changes in the bone. Conclusion: It can safely be postulated from the present study that tissue engineering of PDL can be achieved around dental implants using PDLSCs. Important inter-tissue interactions like the formation of a functional PDL around the implantation site, and induction of bone formation in the vicinity of the implants may be possible. Future research in humans is required for further research.

Evaluation of Presence of Enterococcus faecalis in Root Canals of Deciduous Molars with Necrotic Pulp by Agar Culture and Polymerase Chain Reaction.

Objective: To assess the presence of Enterococcus faecalis in root canals of deciduous molars with necrotic pulp by agar culture and polymerase chain reaction (PCR) assay. Materials and methods: This is an experimental study, where a total of 120 endodontic samples were taken from deciduous molars with necrotic pulps. The presence of Enterococcus faecalis was assessed by culture, using Enterococcus confirmatory agar, and by PCR assay. Statistical analysis of the results was performed using McNemar's test. Results: The presence of Enterococcus faecalis was detected in 20 samples (16.67% of total) by microbial culture and in 45 samples (37.5% of total) by PCR assay, with a statistically significant difference between the two methods (p < 0.001). Microbial culture and PCR both detect Enterococcus faecalis, with the latter detecting an additional 25 positive samples. Conclusion: In this study, PCR assay was significantly more sensitive than agar culture method in detecting the presence of Enterococcus faecalis in root canals of deciduous molars with necrotic pulp, that is, 37.5% of all samples. Clinical significance: Importance of presence of Enterococcus faecalis in necrotic pulps of deciduous teeth, as it is primarily responsible for failure of endodontic treatment, thus helping clinicians to advocate the use of local drug delivery in primary teeth endodontics and also aids clinicians in choosing the most effective intracanal medication. How to cite this article: Nalawade TM, Bhat KG, Kale AD, et al. Evaluation of Presence of Enterococcus faecalis in Root Canals of Deciduous Molars with Necrotic Pulp by Agar Culture and Polymerase Chain Reaction. Int J Clin Pediatr Dent 2023;16(6):816-819.

Synthesis, crystal structure and biological properties of a fluorophore-labelled mixed-ligand copper(II) complex incorporating N-hydroxynaphthalene-1,8-dicarboximide.

The mixed-ligand fluorophore-labelled copper(II) complex aqua[2,4-dioxo-3-azatricyclo[7.3.1.05,13]trideca-1(12),5,7,9(13),10-pentaen-3-olato-κ2O2,O3](1,10-phenanthroline-κ2N,N')copper(II) nitrate, [Cu(C12H6NO3)(C12H8N2)(H2O)]NO3·CH3OH or [Cu(L)(phen)(H2O)]NO3·CH3OH (where phen is 1,10-phenanthroline and HL is N-hydroxynaphthalene-1,8-dicarboximide), (1), was synthesized and structurally characterized. The structure of (1) was confirmed by single-crystal X-ray structure determination. The complex crystallized in the triclinic space group P-1. The geometry around the copper centre is distorted square pyramidal, with the apical position occupied by a water molecule. The complex is highly fluorescent in organic and aqueous solutions. It has good anticancer activity, with an IC50 value of 17 µM, which is almost five times greater than cisplatin (IC50 = 82 µM) under identical experimental conditions.

Curcumin-Encapsulated Nanomicelles Improve Cellular Uptake and Cytotoxicity in Cisplatin-Resistant Human Oral Cancer Cells.

Oral cancer has a high mortality rate, which is mostly determined by the stage of the disease at the time of admission. Around half of all patients with oral cancer report with advanced illness. Hitherto, chemotherapy is preferred to treat oral cancer, but the emergence of resistance to anti-cancer drugs is likely to occur after a sequence of treatments. Curcumin is renowned for its anticancer potential but its marred water solubility and poor bioavailability limit its use in treating multidrug-resistant cancers. As part of this investigation, we prepared and characterized Curcumin nanomicelles (CUR-NMs) using DSPE-PEG-2000 and evaluated the anticancer properties of cisplatin-resistant cancer cell lines. The prepared CUR-NMs were sphere-shaped and unilamellar in structure, with a size of 32.60 ± 4.2 nm. CUR-NMs exhibited high entrapment efficiency (82.2%), entrapment content (147.96 µg/mL), and a mean zeta potential of -17.5ζ which is considered moderately stable. The cellular uptake and cytotoxicity studies revealed that CUR-NMs had significantly higher cytotoxicity and cellular uptake in cisplatin drug-resistant oral cancer cell lines and parental oral cancer cells compared to plain curcumin (CUR). The DAPI and FACS analysis corroborated a high percentage of apoptotic cells with CUR-NMs (31.14%) compared to neat CUR (19.72%) treatment. Conclusively, CUR-NMs can potentially be used as an alternative carrier system to improve the therapeutic effects of curcumin in the treatment of cisplatin-resistant human oral cancer.

Detection and comparison of prevalence of Porphyromonas gingivalis through culture and Real Time-polymerase chain reaction in subgingival plaque samples of chronic periodontitis and healthy individuals.

Introduction: The micro-flora of oral cavity is a myriad of micro-organism. Any infection of oral cavity leads to diseased condition which is a transitional transformation of the micro-organism in a specific paradigm depending upon the diseased condition. Periodontitis is one of the predominant chronic diseases which is a multifactorial infection. Porphyromonas gingivalis is a key etiological agent in causing periodontitis. To study the predominance of these bacteria in the diseased condition is important to detect, quantify and to find its efficacy by comparing different methods for identification. Aim and Objectives: The aim of the study is to determine the prevalence of P. gingivalis by anerobic culture and by real-time polymerase chain reaction (PCR) from subgingival plaque samples of chronic periodontitis and healthy individual and to compare efficacy of two methods. Materials and Methods: A total of 400 subjects were considered, and subgingival plaque was collected using paper points. Individual were equally divided into two groups: chronic periodontitis (200) and healthy individuals (200). Each plaque sample collected was divided into two aliquots of which the first aliquot was subjected for anerobic culture to isolate P. gingivalis. Phenotypical identification was done morphologically and biochemically further quantification of P. gingivalis was done by colony-forming unit. The second aliquot was subjected for DNA extraction and real-time PCR was conducted to detect and quantify P. gingivalis using specific primer. Results: Out of 400 samples, 73% showed detection of P. gingivalis by culture method and through reverse transcription-PCR (RT-PCR), the detection was 75%. Individual detection of P. gingivalis by culture in chronic periodontitis was 89.5% and 54.4% in healthy individuals, while detection by RT-PCR was found to be 91.5% in chronic periodontitis and 58% in healthy individuals. However, comparison between two techniques in detection of P. gingivalis was statistically insignificant. Conclusion: When we compared RT-PCR with culture RT-PCR showed higher positivity. RT-PCR is more sensitive and requires less time to detect. However, in the present study, culture also showed good positivity, suggesting proper dilution and with extended incubation, the specificity of culture can be improved to a great extent.

Comparison of culture and polymerase chain reaction-restriction fragment length polymorphism for identification of various Capnocytophaga species from subgingival plaque samples of healthy and periodontally diseased individuals.

Introduction: Capnocytophaga are facultative anaerobic Gram-negative bacilli and recognized as opportunistic pathogens of various extraoral infections. Only a few studies attempted to identify all the seven species of Capnocytophaga phenotypically and genotypically in healthy individuals and patients with chronic periodontitis. Studies to determine the prevalence of Capnocytophaga in subgingival plaque samples from healthy individuals, chronic gingivitis and periodontitis among Indian population are lacking. Aim: The aim of this study was to identify and compare the presence of Capnocytophaga species phenotypically through microbial culture and biochemical tests and genotypically through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in subgingival plaque of healthy individuals and patients with chronic gingivitis and chronic periodontitis. Materials and Methods: A total of 300 subjects, 100 each with gingivitis, periodontitis and periodontally healthy gingiva subjected, were included. Subgingival plaque was collected and was cultured for phenotypic identification (microbial culture and biochemical test), and for genotypic identification, DNA extraction was done and PCR-RFLP analysis was performed to identify the genus Capnocytophaga and also to identify different species of Capnocytophaga. Results: Of 300 individuals, Capnocytophaga species were identified from 237 (79%) individuals by PCR and 82 (27.33%) by culture. The prevalence of Capnocytophaga ochracea was found to be higher with both the methods followed by Capnocytophaga gingivalis and Capnocytophaga granulosa. Capnocytophaga genospecies, Capnocytophaga leadbetteri and Capnocytophaga Sputigena were isolated only by culture with very low prevalence that is 1.33%, 1.33% and 0.66%, respectively. We could not get any isolate of Capnocytophaga haemolytica by any of the two methods. Conclusion: Capnocytophaga species could be found in gingival sulci as well as periodontal pockets and can be detected by culture and PCR-RFLP. However, higher prevalence of these species in healthy compared to disease requires further analysis to determine their role in healthy and diseased periodontium.

Cancer Stem Cell Traits in Tumor Spheres Derived from Primary Laryngeal Carcinoma Cell Lines.

OBJECTIVE: Cancer stem cells (CSCs) belong to a subpopulation of undifferentiated cells present within tumors that have the potential to regenerate, differentiate, maintenance of pluripotency, drug resistance, and tumorigenicity when transplanted into an innate host. These can influence the growth and behavior of these tumors and are used to investigate the initiation, progression, and treatment strategies of laryngeal cancer. Research on CSC science and targeted therapies were hinge on their isolation and/or enrichment procedures. The object of the study is to isolate cancer stem cells from primary laryngeal carcinoma (CSCPLC) by tumor spheres enrichment. We checked the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance. MATERIALS AND METHODS: We performed tumor sphere formation assay (primary, secondary, and tertiary) chemotherapy resistance by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay were performed to evaluate the CSC cells. Immunofluorescence for stem cell markers (CD133+, CD44+) and gene expression of stem cell markers for CD133+, CD44+, OCT4, SOX2, and NANOG was done using the real-time polymerase chain reaction technique. RESULTS: We were able to isolated CSC subpopulations from PLC cell lines by the tumor sphere method. These cells exhibited good primary, secondary, and tertiary tumor sphere formation efficiency and also disclosed a resistant index of more than 2. Immunofluorescence for stem cell markers (CD133+ and CD44+) confirms the presence of CSC. There was significantly higher mRNA expression of stem cell markers in CSC enriched subpopulations compared to the parental cell lines. CONCLUSION: We conclude that tumor spheres enrichment is an efficient, economical, and reliable approach for the isolation and characterization of CSC from PLC cell lines. These cells demonstrated the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance.

Solitary Median Maxillary Central Incisor: A Case Report with 3-Year Follow-Up and Literature Review.

A solitary median maxillary central incisor (SMMCI) is a rare anomaly that can occur alone or be associated with other systemic abnormalities. Early diagnosis of SMMCI is crucial as it might indicate the presence of an associated congenital or developmental abnormality. The prevalence of live-born children with SMMCI is determined to be 1:50,000 and is more common among females. The purpose of this paper was to report an unusual case of a 9-year-old girl with SMMCI who had no growth deficiency or any other systemic involvement. Since pediatricians and dentists are the first professionals to evaluate an SMMCI's patient in most cases, it is important that they be aware of the possibility of other related systemic problems that require systemic care. Appropriate treatment, diagnosis, and referral should also include neuropediatric evaluation, genetic testing, and craniofacial profile analysis along with multidisciplinary approach.

Green tea catechins showed antibacterial activity on streptococcus mutans -An in vitro study.

BACKGROUND: The main bacterial aetiological agents in caries formation are the α-haemolytic Streptococcal species Streptococcus mutans, which has been found to be the initiator of most dental caries. The leaves of Camellia sinensis known as green tea, has properties, such as antibacterial and anti-cariogenic. Epigallocatechin-3-gallate (EGCG) one of the most abundant catechins found in green tea is known to contribute to these effects. AIM: To evaluate the antibacterial effect of green tea catechins namely EGCG on S. mutans with two different methods at different concentrations. OBJECTIVES: 1) To assess the antimicrobial efficacy of EGCG by disc diffusion test at concentrations of 100, 75, and 50 µg/mL. 2) To assess the antimicrobial efficacy of EGCG by Minimum inhibitory concentration (MIC) test at concentrations ranging from 0.2 to 100 µg/mL. METHODOLOGY: Commercially available purest form of green tea polyphenol EGCG was used in the study. Disc diffusion test on agar medium and MIC test was used to determine the susceptibility of the S. mutans to green tea catechins EGCG. RESULTS: The results of the agar well diffusion method showed that the EGCG extract has shown zones of inhibition against S. mutans at concentrations of 100 µg/mL (28.67 mm), 75 µg/mL (15.33 mm), 50 µg/mL (10.33 mm) while that of MIC test of EGCG extract of concentrations ranging from 0.2 to 100 µg/mL against S. mutans shows that the mean MIC value was 1.07. CONCLUSION: Catechins in the tea are potentially anti-cariogenic agents which can reduce bacterial presence in the oral cavity and have the potential to be further used for the preparation of dentifrice and mouthwash.

Coculture method for in vitro cultivation of uncultured oral bacteria.

PURPOSE: The purpose of the study is to culture uncultured oral bacteria with helper strains using the coculture method from the subgingival plaque samples of chronic periodontitis patients. MATERIALS AND METHODS: The samples were processed and inoculated on a blood agar medium enriched with hemin and Vitamin K. A helper strain Propionibacterium acnes (ATCC 6919) was cross-streaked across the inoculums to facilitate coculture. The plates were then incubated for 7 days with subsequent subculturing and further incubation. RESULTS: Satellite colonies around helper strain showed one colony type of Porphyromonas gingivalis, one was of nonpigmented Prevotella, three were of Fusobacterium nucleatum and five isolates remained unidentified. CONCLUSIONS: Coculture could be used effectively as one of the methods in the isolation and in vitro cultivation of oral bacteria. Incubation using the anaerobic jar technique was found to be economical and efficient for the growth of anaerobic oral bacteria.

Isolation and immunohistochemical characterization of periodontal ligament stem cells: A preliminary study.

CONTEXT: It is a known fact that periodontal tissue regeneration can be achieved by the use of periodontal ligament stem cells (PDLSCs). Current mainstay of periodontal treatment is focusing on stem cell tissue engineering as an effective therapy, making it important to isolate PDLSCs from periodontal tissues. AIMS: The present research endeavor was undertaken to elucidate a technique for isolating PDLSCs for in vivo reconstructing the natural PDL tissue. SETTINGS AND DESIGN: The study design involves In vitro prospective study. MATERIALS AND METHODS: Premolar teeth were extracted from 12 patients who were under orthodontic treatment. PDL cells were scraped from their roots. Using 10 ml of Dulbecco's modified Eagle's medium with pH 7.2, the specimens of the periodontal tissue were transferred to laboratory where cell culture was done. Isolated stem cells were grown on 24-well microtiter plates-containing cover slips. They were incubated overnight at approximately 37°C in 95% air and 5% humidification. Anti-CD 45, CD73, CD90, CD105, and CD146 antibodies were used. After staining, cells were observed under phase-contrast microscopy and in inverted microscope. RESULTS: The cells showed a marked growth and 90% confluence at day 6. Cells presented thin and long fibroblastic spindle morphology. Isolated PDLSCs showed colony-forming ability at the 14th day after seeding. Immunohistochemical staining of PDLSCs showed positive uptake for CD146, CD90, CD73, CD105, and negative uptake for CD45. CONCLUSIONS: The human PDLSCs can be clearly isolated and characterized by using CD90, CD73, CD146, and CD105 markers of stem cells.

Site specificity and expression profile of miR-21 in oral squamous cell carcinoma.

BACKGROUND: Oral Squamous Cell Carcinoma (OSCC) is the most common epithelial malignancy of the oral cavity which has evolved globally as a grave and growing health problem. It shares a wide geographic variation with respect to the incidence rate and exhibits anatomic adaptation to oral environment with varied clinical presentation along with a spectrum of histological mélange. Besides, in recent cancer research, both genetics and epigenetics add on at the molecular level and accounts for this diversification and tumor heterogeneity of OSCC and thereby substantiates to the miRNA expression profiling in OSCC. AIMS AND OBJECTIVES: In the present study, subsite specificity of miR-21 expression in tissue specimens of OSCC of Tongue, Buccal mucosa, and Gingivo buccal (GB) sulcus were analyzed. MATERIALS AND METHODS: Quantification of miR-21 was done on 30 tissue samples of OSCC using real-time polymerase chain reaction (RT-PCR). RESULTS: Results indicated that miR-21 expression was significantly expressed at the subsites. Out of 30 samples, 22 showed upregulation, and 8 showed down-regulation with reference to endogenous control. The comparative Ct method was used to analyze the differences in subsite specific expression of miR-21 in OSCC cases. It was significantly upregulated in the buccal mucosa (p=0.002), followed by GB sulcus (p=0.01) and Tongue (p=0.25). CONCLUSION: In conclusion, the study could identify the differential miR-21 expression at sub-sites, indicating that it may serve as a diagnostic marker with further elaboration on a larger sample size..

Assessment of antibacterial effect of hydrogen water on plaque from patients with chronic periodontitis.

BACKGROUND: Periodontitis is an inflammatory disease causing destruction of tissues surrounding the teeth. The primary etiological factor for periodontitis is plaque. An inference could be drawn that an overall reduction in microorganisms halts disease progression. It is desirable to have natural agents with minimal side effects to reduce the microbial load. AIM: The aim of the study is to assess the effect of hydrogen water on microbial count in plaque obtained from chronic periodontitis patients and to determine the antibacterial activity of hydrogen water at various time intervals. MATERIALS AND METHODS: A total of twenty chronic periodontitis patients were included after obtaining approval from the institutional ethical committee. Written informed consent was obtained from all the twenty participants. Plaque samples were collected and exposed to hydrogen water at baseline, 1 min, 2 min 30 s, and 5 min. Samples were then cultured on blood agar and incubated in aerobic and anaerobic conditions. The colony forming units and total bacterial count were recorded after 24-48 h. STATISTICAL ANALYSIS: Intragroup pair-wise comparison was done using Wilcoxon sign-ranked test. RESULTS: Hydrogen water showed antibacterial activity against aerobic and anaerobic organisms associated with chronic periodontitis. There was a statistically significant difference in the number of colony forming units from baseline to 1 and 2.5 min for the aerobic culture and also for baseline to 1, 2.5, and 5 min for the anaerobic culture. CONCLUSION: The data of the present study indicate that hydrogen water has an antibacterial effect on microorganisms associated with chronic periodontitis.

8-Hydroxyquinoline derived p-halo N4-phenyl substituted thiosemicarbazones: Crystal structures, spectral characterization and in vitro cytotoxic studies of their Co(III), Ni(II) and Cu(II) complexes.

The current paper deals with 8-hydroxyquinoline derived p-halo N4-phenyl substituted thiosemicarbazones, their crystal structures, spectral characterization and in vitro cytotoxic studies of Co(III), Ni(II) and Cu(II) complexes. The molecular structures of the ligands, (E)-4-(4-halophenyl)-1-((8-hydroxyquinoline-2-yl)methylene)thiosemicarbazones (halo = fluoro/chloro/bromo) are determined by single crystal X-ray diffraction method. The crystal structures reveal that the ligands are non-planar and exist in their thioamide tautomeric forms. The various physicochemical investigations of the synthesized complexes reveal metal to ligand stoichiometry to be 1:2 in Co(III) complexes whereas 1:1 in Ni(II) and Cu(II) complexes. The ligands coordinate in a tridentate NNS fashion around Co(III) centers to form an octahedral geometry and square planar geometry around Ni(II) and Cu(II) metal centers. The oxidation of Co(II) to Co(III) is observed on complexation. The synthesized compounds are subjected to in vitro cytotoxicity studies. When compared to bare ligands, the complexes show enhancement of the antiproliferative activity against MCF-7, breast cancer cell lines. The Co(III) complexes of fluoro and bromo derivatives of ligands have displayed remarkable results with roughly two fold increase in their activity in correlation to the standard drug, Paclitaxel. Moreover, the fluorescence microscopy images of cells stained with acridine orange-ethidium bromide suggest an apoptotic mode of cell death.