Antimicrobial effect of Pentaclethra Macroloba plant extract against Enterococcus Faecalis.

This study evaluated the antimicrobial efficacy of a new intracanal drug based on Pentaclethra macroloba extract, a plant of Amazonian origin, against Enterococcus faecalis using macrodilution test and intratubular evaluation with Confocal Laser Scanning Microscopy (CLSM). The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the pure extract of Pentaclethra macroloba andits association with calcium hydroxide and ultracall were determined. Then, thirty-three dentin cylinders were prepared and inoculated with E. faecalis, to evaluate the antimicrobial effect of the medications on the dentinal tubules with CLSM. The data was analyzed using the Kruskal-Wallis and Dunn tests. The extract in association with calcium hydroxide showed a lower CBM, and in the intratubular test all tested medications were effective against E. faecalis (P >0.05). The new intracanal drug based on P. macroloba extract has an antimicrobial effect against E. faecalis and further studies are needed for its clinical use.

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 disinfection procedures performed prior to regenerative endodontic therapy: An integrative review.

The aim of this integrative review was to identify whether the disinfection procedures performed prior to regenerative endodontic treatment were effective on biofilm removal from the root canals. The research was based on PubMed, Latin American and Caribbean Health Sciences Literature (Lilacs) and Scientific Electronic Library Online (SciELO) databases. Four articles were selected; one of the studies was in vivo and the others ex vivo. Different disinfection procedures were studied, characterised mainly by the use of intracanal medication, highlighting the double antibiotic paste, triple antibiotic paste and calcium hydroxide paste. Disinfection ability was evaluated against Enterococcus faecalis and multispecies biofilms by using the fluorescence technique and colony forming unit counting, for 7 to 21 days. Double antibiotic paste and triple antibiotic paste demonstrated excellent antibiofilm activity, unlike CH paste that showed limited disinfection, even when associated with different antimicrobial agents. Triple antibiotic paste was the most effective medication against biofilm.

Potential of Bouea macrophylla kernel extract as an intracanal medicament against mixed-species bacterial-fungal biofilm. An in vitro and ex vivo study.

OBJECTIVE: To investigate the antimicrobial activity of B. macrophylla kernel extract against mixed-species biofilms of E. faecalis, S. gordonii and C. albicans in vitro. To evaluate the efficacy of the extract as an intracanal medicament compared with Ca(OH)2 and chlorhexidine in ex vivo tooth model. METHODS: The antibiofilm effect of B. macrophylla kernel extract was determined by AlamarBlue™ assay and the effect on biofilms was visualized by LIVE/DEAD® BacLight™ viability test. Mixed-species biofilms were incubated into the tooth model (N = 42) for 21 days. The teeth were randomly divided into 4 medicament groups for 7 days: (i) normal saline, (ii) calcium hydroxide (Ca(OH)2), (iii) chlorhexidine gel, (iv) B. macrophylla kernel extract. Dentine samples were collected, qPCR with PMA was used to quantify the viability and species composition of each sample. SEM was used to visualize the effect of medicament on biofilm structure. RESULTS: The MBIC was 6.25 mg/mL and the MBEC was 50 mg/mL. The integrity of microbial cells was progressively compromised as concentration increased, resulting in greater cell death. Ex vivo tooth model revealed that biofilm treated with 50 mg/mL of the B. macrophylla extract demonstrated a significantly higher proportions of dead cells than in Ca(OH)2, chlorhexidine and normal saline groups (p < 0.01). Disruption of biofilm structure and enlargement of dentinal tubules was observed in B. macrophylla group on SEM. CONCLUSION: The extract of B. macrophylla kernel exhibited significant antibiofilm effect against the mixed-species biofilms of E. faecalis, S. gordonii and C. albicans.

Comparative evaluation of the antibacterial effect of Allium sativum, calcium hydroxide and their combination as intracanal medicaments in infected mature anterior teeth: A randomized clinical trial.

AIM: The purpose of this study was to compare the antibacterial effects of Allium sativum (garlic extract), calcium hydroxide (Ca [OH]2 ) and their combination as intracanal medicaments in infected mature anterior teeth using real-time PCR. METHODOLOGY: This prospective double-blind, controlled, parallel, superiority, randomized clinical trial was carried out on 66 permanent, necrotic incisors associated with asymptomatic apical periodontitis in 66 male patients. Patients were randomly divided into three groups (n = 22) according to the intracanal medications used. After access preparation, four microbiological samples (S) were taken using sterile absorbent paper points as follows: S1: before canal instrumentation and S2: after cleaning and shaping. The third sample (S3) and fourth sample (S4) were taken after the placement of the tested intracanal medications into their corresponding canals for 7 and 14 days, respectively. Total DNA was extracted from microbiological samples and relative quantitative real-time PCRs were done to quantify the relative gene expression fold change (FC) for Enterococcus faecalis and Streptococcus species. At significance level p ≤ .05, the data were statistically analysed in SPSS software using Kruskal-Wallis and Freidman's tests, followed by Dunn-Bonferroni post hoc test for pairwise comparisons. RESULTS: Both bacterial mean FC decreased significantly after mechanical instrumentation (S1 to S2) in all groups. However, no statistically significant differences were found after intracanal medicament placement (from S2 to S3 and from S3 to S4) except in the garlic group. Garlic significantly reduced Enterococcus faecalis FC in S3 and S4 when compared to Ca (OH)2 and Ca (OH)2 + garlic combination. However, garlic and Ca (OH)2 reduced Streptococcus bacteria in S3 similarly. Whilst in S4, garlic showed significantly more reduction than Ca (OH)2 . The combination of Ca (OH)2 with garlic extract showed the least significant bacterial reduction. CONCLUSION: Within the study limitations, garlic intracanal medicament has a comparable anti-Streptococcus efficiency to Ca (OH)2 , whilst it is more effective against Enterococcus faecalis species. When Ca (OH)2 and garlic are combined, their antibacterial effectiveness is reduced. Increasing the time of application for tested intracanal medicaments by more than one week has no additional antibacterial effectiveness.

Hepatic enzymes and immunoinflammatory response to Bio-C Temp bioceramic intracanal medication implanted into the subcutaneous tissue of rats.

Our purpose was to evaluate the biocompatibility and hepatotoxicity of a new bioceramic intracanal medicament, Bio-C Temp (BIO). The biological properties of BIO were compared with calcium hydroxide-based intracanal medicament (Calen; CAL), used as gold pattern. Polyethylene tubes filled with BIO or CAL, and empty tubes (control group, CG) were implanted into subcutaneous tissue of rats. After 7, 15, 30 and 60 days, the samples were embedded in paraffin for morphological, quantitative and immunohistochemistry analyses. At 7 and 60 days, blood samples were collected for analysis of serum glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) levels. The data were submitted to two-way ANOVA and Tukey's test (p ≤ 0.05). No significant difference was detected in serum GOT and GPT levels among BIO, CAL and CG specimens. In all periods, BIO specimens exhibited lower number of inflammatory cells and immunoexpression of IL-6, a pro-inflammatory cytokine, than CAL specimens. The reduction of these parameters was accompanied by significant increase in the collagen content and in the immunoexpression of IL-10, a cytokine involved in the tissue repair, over time. Our findings indicate that Bio-C Temp is biocompatible and had no hepatotoxicity effect.

Antimicrobial action, cytotoxicity, calcium ion release, and pH variation of a calcium hydroxide-based paste associated with Myracrodruon urundeuva Allemão extract.

The extract of Myracrodruon urundeuva Allemão (aroeira), as a vehicle, associated with calcium hydroxide (CH) paste was evaluated based on cell viability, antimicrobial action, calcium ion release, and pH variation. Calcium hydroxide with propylene glycol was used as control. The pH variation was measured at 3, 24, 72, 168, 140, 360, and 720 h and calcium ion release was measured on days 7, 15, and 30. Cell viability was assessed with NIH/3T3 cells using MTT and crystal violet assays, after 24, 48, and 72 h. Antibacterial activity was determined by the disc diffusion method, while microbial reduction (Enterococcus faecalis) was evaluated using the time-kill test. The CH paste formulated with aroeira showed antibacterial activity against Enterococcus faecalis and further did not interfere with pH, calcium ion release, or cell viability; moreover, the formulation had antimicrobial activity and could serve as a vehicle for CH paste.

Comparative Evaluation of Efficacy of Calcium Hydroxide, Propolis, and Glycyrrhiza glabra as Intracanal Medicaments in Root Canal Treatment.

AIM: To evaluate and compare the antimicrobial efficacy of Ca(OH)2, 25% propolis, and 25% Glycyrrhiza glabra as intracanal medicaments in root canal treatment. MATERIALS AND METHODS: Total 60 freshly extracted permanent incisors were decoronated and chemomechanical preparation of root canal was performed. Samples were inoculated with a pure culture of Enterococcus faecalis and incubated for 21 days. Colony-forming units (CFUs) were recorded before medication. Incubated samples were randomly categorized into three groups, namely, Ca(OH)2, propolis, and G. glabra, with 20 samples in each group. Antibacterial activity was assessed by evaluating the variance in the CFUs on Day 7. Paired "t" test and Post-hoc Tukey's test were applied to analyze the data. RESULTS: Reduction of CFUs was noticed in all the groups (p <0.001), however the reduction was more predominant in the propolis group. CONCLUSION: Propolis is more effective against E. faecalis, when compared to G. glabra and Ca(OH)2. CLINICAL SIGNIFICANCE: Propolis could be used as an effective medicament in root canal treatment.

Antibacterial Efficacy of Triple Antibiotic Medication With Macrogol (3Mix-MP), Traditional Triple Antibiotic Paste, Calcium Hydroxide, and Ethanol Extract of Propolis: An Intratubular Dentin Ex Vivo Confocal Laser Scanning Microscopic Study.

INTRODUCTION: This study aimed to assess the effectiveness of antibacterial activity of medications used in regenerative endodontic treatment. METHODS: Sixty-seven dentin cylinders of single-rooted teeth were contaminated with a culture of Enterococcus faecalis (ATCC 29212; American Type Culture Collection, Manassas, VA) for 5 days. Samples were divided into 1 control group and the following experimental groups according to the medication applied: traditional triple antibiotic paste (TAP), clindamycin-modified TAP (mTAP), triple antibiotic medication with macrogol (3Mix-MP), clindamycin-modified 3Mix-MP (m3Mix-MP), calcium hydroxide (CH), and ethanol extract of propolis (EEP). After 14 days, the medications were removed, and the samples were submitted to confocal laser scanning microscopic analysis to quantify the percentage of viable bacteria. The distribution of data was confirmed by the Shapiro-Wilk test. The Kruskal-Wallis and Dunn tests were used for intergroup comparisons, and the Wilcoxon test was used for comparison between superficial and deep antibacterial efficacy for the same medication. The level of significance was set at P < .05. RESULTS: 3Mix-MP and m3Mix-MP presented significantly higher antibacterial efficacy compared with the other tested medications (P < .05), except for mTAP. mTAP was more effective than TAP (P < .05). The antibacterial efficacy of EEP and CH did not differ significantly from TAP and mTAP (P > .05). All medications showed effective antibacterial action compared with the control group (P < .05). CONCLUSIONS: 3Mix-MP and m3Mix-MP, which present extremely high concentrations of antibiotics (1500 mg/mL), were not more effective than mTAP at the concentration recommended by the American Association of Endodontists (5 mg/mL). Moreover, CH and EEP were as effective as TAP and mTAP.

Selenium intracanal dressing: effects on the periapical immune response.

OBJECTIVES: To evaluate the selenium (Se) behavior when used as an endodontic dressing in teeth with pulp necrosis. Additionally, its effects was also compared with the calcium hydroxide (C.H.), which is used globally as a root canal dressing, and the combination of the C.H. with Se (C.H. + Se). MATERIALS AND METHODS: The sample consisted of 60 patients requiring endodontic treatment who were divided into groups, i.e., without intracanal medication (empty) and with medications as follows: selenium (Se), calcium hydroxide (C.H.), and calcium hydroxide + selenium (C.H. + Se) (n = 15). After the coronary opening, three absorbent paper points were placed in the RCS and maintained for 2 min for microbial evaluation. Following the cleaning and shaping procedures, new paper points were introduced into the root canal system, passing passively through the root apex (2 mm) into the periapical tissues for 2 min, for immune evaluation. The collections were performed again 15 days later. Real-time PCR quantified the expression of the prokaryotic 16S ribosomal RNA. The 16S mRNA was evaluated before the cleaning and shaping procedures and 15 days later in the groups treated with or without medication. RESULTS: A significant reduction in the microbial load was observed only in the groups that received endodontic dressing (p < 0.05). The cytokines IFN-γ, TNF-α, IL-1α, IL-17A, IL-10, IL-6 and MCP-1, were also quantified by real-time PCR. There was an increase in the gene expression level of the cytokines (T15) TNF-α and IL-10 in the C.H. group compared to the other groups (p < 0.05). The IFN-γ mRNA expression was reduced in the groups treated with the medications (Se, C.H., and C.H. + Se). CONCLUSIONS: The findings of the present study indicate that in the case of treatment over multiple sessions, the use of root canal dressing is essential to avoid the root canal system (RCS) microbial recolonization. Selenium potentiated the effects of calcium hydroxide inducing an anti-inflammatory response in periapical tissues. CLINICAL RELEVANCE: Se is a mineral essential for the formation of the amino acid selenocysteine, which is directly involved in the maintenance of the immune response. Selenium has been widely used in the medical field in the treatment of cancer, as an activator of bone metabolism, and as a stimulator of the immune system. In this study, it was shown that the incorporation of Se, whether as intracanal medication alone or in conjunction with other medications, may potentiate periapical tissue repair after RCS cleaning and shaping procedures.

Evaluation of Resin-Based Material Containing Copaiba Oleoresin (Copaifera Reticulata Ducke): Biological Effects on the Human Dental Pulp Stem Cells.

The purpose of this study was to analyze in vitro the biological effects on human dental pulp stem cells triggered in response to substances leached or dissolved from two experimental cements for dental pulp capping. The experimental materials, based on extracts from Copaifera reticulata Ducke (COP), were compared to calcium hydroxide [Ca(OH)2] and mineral trioxide aggregate (MTA), materials commonly used for direct dental pulp capping in restorative dentistry. For this, human dental pulp stem cells were exposed to COP associated or not with Ca(OH)2 or MTA. Cell cytocompatibility, migration, and differentiation (mineralized nodule formation (Alizarin red assay) and gene expression (RT-qPCR) of OCN, DSPP, and HSP-27 (genes regulated in biomineralization events)) were evaluated. The results showed that the association of COP reduced the cytotoxicity of Ca(OH)2. Upregulations of the OCN, DSPP, and HSP-27 genes were observed in response to the association of COP to MTA, and the DSPP and HSP-27 genes were upregulated in the Ca(OH)2 + COP group. In up to 24 h, cell migration was significantly enhanced in the MTA + COP and Ca(OH)2 + COP groups. In conclusion, the combination of COP with the currently used materials for dental pulp capping [Ca(OH)2 and MTA] improved the cell activities related to pulp repair (i.e., cytocompatibility, differentiation, mineralization, and migration) including a protective effect against the cytotoxicity of Ca(OH)2.

Effect of Calcium Hydroxide Combinations with Green Tea Extract and Cocoa Pod Husk Extract on p38 MAPK and Reparative Dentine.

AIM AND OBJECTIVE: The aim of this research is to analyze the effect of calcium hydroxide combinations with green tea extract and the combination of calcium hydroxide with cocoa pod husk extract on the activation of p38 MAPK and wide area of reparative dentin in mice dental. MATERIALS AND METHODS: This study used 36 rats that were randomly divided into three treatment groups: positive control group was applied calcium hydroxide and aquades (group I), the test group was applied calcium hydroxide combined with cocoa pod husk extract (group II), and the next test group was applied using calcium hydroxide combined with green tea extract (group III); all the cavities were restored with RMGIC. On day 7 and 28, experimental animals from each treatment group were killed by peritoneal injection to see the activation of p38 MAPK, while reparative dentin was only seen on day 28. RESULTS: The result of data analysis using Multiple Comparison Tukey HSD test showed significant difference between the positive control group and the test groups for the average p38 MAPK activation value on day 7 and 28. But there was no significant difference between two test groups. The same thing was obtained in the calculation of the average area of reparative dentin, where group I had the lowest value compared to groups II and III on day 28 with a significant difference. There was no significant difference between groups II and III. CONCLUSION: The use of combination calcium hydroxide with green tea extract and combination calcium hydroxide with cocoa pod husk extract have significant effect on p38 MAPK activation and wide area of reparative dentin in mice dental. CLINICAL SIGNIFICANCE: The use of combination calcium hydroxide with green tea extract and combination calcium hydroxide with cocoa pod husk extract have been proven to activate more p38 and form a wider reparative dentin.

The Susceptibility to Calcium Hydroxide Modulated by the Essential walR Gene Reveals the Role for Enterococcus faecalis Biofilm Aggregation.

INTRODUCTION: Enterococcus faecalis is considered a predominant pathogen for persistent periapical infections and in addition is reportedly resistant to calcium hydroxide medication. The WalRK 2-component system of E. faecalis is essential for environmental adaptation, survival, and virulence. The goal of this study was to investigate the potential roles of walR in the regulation of biofilm aggregation, alkaline stress, and susceptibility to calcium hydroxide (CH) medication. METHODS: Antisense walR RNA (aswalR) overexpression strains were constructed. Exopolysaccharide (EPS) production and bacterial viability of E. faecalis biofilms were evaluated by confocal laser scanning microscopy. Quantitative real-time polymerase chain reaction was used to investigate the expressions of virulent factor genes. The proportion of viable bacteria and EPS production in dentin were assessed after CH medication. RESULTS: We showed that walR interference by aswalR RNA leads to a reduction in the dextran-dependent aggregation in E. faecalis biofilm. The overexpression of aswalR reduced the transcripts of the virulence genes and alkaline stress tolerance ability. Furthermore, the down-regulation of walR sensitized E. faecalis in infected canals to CH medication associated with inhibiting EPS synthesis. CONCLUSIONS: The data suggest a role for the walR regulator in the susceptibility to CH associated with dispelling the EPS matrix, which could be explored as a potential supplementary therapy for the management of root canal infection.

Effects of a Chimeric Lysin against Planktonic and Sessile Enterococcus faecalis Hint at Potential Application in Endodontic Therapy.

Enterococcus faecalis is a commensal opportunistic pathogen found in the intestine, mouth, and vaginal tract of humans. As an invasive pathogen in the oral cavity, E. faecalis is one of the leading causes of periapical endodontic lesions. However, due to the strong biofilm-forming capacity and tolerance of E. faecalis to conventional antibiotics and treatments, limited therapeutic options are available. In the present study, we investigated the activity of ClyR, a chimeric lysin with extended streptococcal lytic spectrum, against planktonic and sessile E. faecalis cells in vitro and in an ex vivo dental model. Our results showed that ClyR has robust and rapid lytic activity against multiple E. faecalis strains, killing >90% planktonic cells within 1 min at a concentration of 50 µg/mL. The biochemical experiments combined with microscopy analysis revealed that ClyR degrades E. faecalis biofilm with high efficacy in a dose-dependent manner, reducing the survival rate to 90% viable bacteria within biofilms at a low dose of 50 µg/mL, which is much better than ampicillin and similar to calcium hydroxide, the extensively used routine intracanal medicament in the treatment of endodontics and dental traumatology. The robust activity of ClyR against both planktonic and sessile E. faecalis suggests the potential of ClyR in treating endodontic infections caused by E. faecalis.

Detection and function of lipopolysaccharide and its purified lipid A after treatment with auxiliary chemical substances and calcium hydroxide dressings used in root canal treatment.

AIM: To investigate the influence of auxiliary chemical substances (ACSs) and calcium hydroxide [Ca(OH)2 ] dressings on lipopolysaccharides (LPS)/lipid A detection and its functional ability in activating Toll-like receptor 4 (TLR4). METHODOLOGY: Fusobacterium nucleatum pellets were exposed to antimicrobial agents as following: (i) ACS: 5.25%, 2.5% and 1% sodium hypochlorite solutions (NaOCl), 2% chlorhexidine (CHX) (gel and solution) and 17% ethylenediaminetetraacetic acid (EDTA); (ii) intracanal medicament: Ca(OH)2 paste for various periods (1 h, 24 h, 7 days, 14 days and 30 days); (iii) combination of substances: (a) 2.5% NaOCl (1 h), followed by 17% EDTA (3 min) and Ca(OH)2 (7 days); (b) 2% CHX (1 h), afterwards, 17% EDTA (3 min) followed by Ca(OH)2 (7 days). Saline solution was the control. Samples were submitted to LPS isolation and lipid A purification. Lipid A peaks were assessed by matrix-assisted laser desorption ionization time-of-flight mass spectrom (MALDI-TOF MS) whilst LPS bands by SDS-PAGE separation and silver staining. TLR4 activation determined LPS function activities. Statistical comparisons were carried out using one-way anova with Tukey-Kramer post-hoc tests at the 5% significance level. RESULTS: Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of control lipid A demonstrated the ion cluster at mass/charge (m/z) 1882 and an intense band in SDS-PAGE followed by silver staining of control LPS. In parallel, LPS control induced a robust TLR4 activation when compared to ACS (P ≤ .001). 5.25% NaOCl treatment led to the absence of lipid A peaks and LPS bands, whilst no changes occurred to lipid A/LPS after treatment with others ACS. Concomitantly, 5.25% NaOCl-treated LPS did not activate TLR4 (P < .0001). As for Ca(OH)2 , lipid A was not detected by MALDI-TOF nor by gel electrophoresis within 24 h. LPS treated with Ca(OH)2 was a weak TLR4 activator (P < .0001). From 24 h onwards, no significant differences were found amongst the time periods tested (P > 0.05). The addition of Ca(OH)2 for 7 days to cells treated either with 2.5% NaOCl or 2% CHX led to the absence of lipid A peaks and LPS bands, leading to a lower activation of TLR4. CONCLUSION: 5.25% NaOCl and Ca(OH)2 dressings from 24 h onwards were able to induce both, loss of lipid A peaks and no detection of LPS bands, rendering a diminished immunostimulatory activity through TLR4.

Effects of mineral trioxide aggregate, BiodentineTM and calcium hydroxide on viability, proliferation, migration and differentiation of stem cells from human exfoliated deciduous teeth.

The aim of the study was to evaluate the effects of the capping materials mineral trioxide aggregate (MTA), calcium hydroxide (CH) and BiodentineTM (BD) on stem cells from human exfoliated deciduous teeth (SHED) in vitro. SHED were cultured for 1 - 7 days in medium conditioned by incubation with MTA, BD or CH (1 mg/mL), and tested for viability (MTT assay) and proliferation (SRB assay). Also, the migration of serum-starved SHED towards conditioned media was assayed in companion plates, with 8 µm-pore-sized membranes, for 24 h. Gene expression of dentin matrix protein-1 (DMP-1) was evaluated by reverse-transcription polymerase chain reaction. Regular culture medium with 10% FBS (without conditioning) and culture medium supplemented with 20% FBS were used as controls. MTA, CH and BD conditioned media maintained cell viability and allowed continuous SHED proliferation, with CH conditioned medium causing the highest positive effect on proliferation at the end of the treatment period (compared with BD and MTA) (p<0.05). In contrast, we observed increased SHED migration towards BD and MTA conditioned media (compared with CH) (p<0.05). A greater amount of DMP-1 gene was expressed in MTA group compared with the other groups from day 7 up to day 21. Our results show that the three capping materials are biocompatible, maintain viability and stimulate proliferation, migration and differentiation in a key dental stem cell population.

Antimicrobial activity of Psidium cattleianum associated with calcium hydroxide against Enterococcus faecalis and Candida albicans: an in vitro study.

OBJECTIVE: Evaluate, in vitro, the antimicrobial activity of Psidium cattleianum leaf extracts combined with calcium hydroxide against Enterococcus faecalis and Candida albicans biofilm. MATERIALS AND METHODS: Dentin specimens obtained from extracted bovine incisors were infected during 14 days with E. faecalis ATCC 29212 and C. albicans ATCC 10231. The specimens were filled with calcium hydroxide pastes prepared with the following vehicles: Psidium cattleianum ethanolic, Psidium cattleianum propylene glycolic, distilled water, and saline as control. After 24 h, 3, 7, and 14 days, the canals were irrigated with sterile saline and dried. Dentin samples were collected from the canals with burs of increasing diameters. To determine the number of colony-forming units (CFU), samples were inoculated onto BHI agar supplemented with yeast extract (0.5%), at 37 °C, for 48 h, in CO2 enriched atmosphere. Comparisons among the groups for the variation factors were performed by ANOVA and Tukey's test. RESULTS: Ethanolic and propylene glycolic extracts showed significantly higher antimicrobial activity against E. faecalis (p < 0.01) when compared with distilled water. The ethanolic extract exhibited in 24 h the same antibacterial activity that propylene glycolic extract and distilled water after 7 and 14 days. For C. albicans, all were effective in reducing the number of CFU at all periods. CONCLUSION: The P. cattleianum ethanolic extract presented the fastest and highest antimicrobial activity against E. faecalis, significantly reducing the microbial load in 24 h. All medications were effective against C. albicans. CLINICAL RELEVANCE: The antibacterial potential of P. cattleianum and its biological compatibility associated with calcium hydroxide indicate promising applications in the field of dentistry.

Use of sodium trimetaphosphate in the inhibition of dentin matrix metalloproteinases and as a remineralizing agent.

OBJECTIVES: Because of its ability to act as an antiproteolytic agent, the effect of sodium trimetaphosphate (STMP) against specific enzymes extracted from sound dentin and its performance under acidic challenge on demineralized dentin were investigated. METHODS: The antiproteolytic potential of STMP (0.5%, 1.0%, and 1.5%) was assessed in triplicate by zymography. For the evaluation of remineralization activity, 50 bovine-root dentin specimens were selected and randomly divided into 5 groups (n=10). Three areas were determined for each specimen: 1) control (no treatment); 2) demineralized (artificial caries-like challenge); 3) treated (demineralized and subjected to pH-cycling for 7days, and treated for 10min with 1.5% STMP, 1.5% STMP+calcium hydroxide (Ca[OH]2), 1.5% STMP+sodium fluoride (NaF), NaF, or deionized H2O). The dentin specimens were analyzed for superficial hardness (SH) and cross-sectional hardness (CSH) at different depths (10, 30, 50, 70, 90, 110, and 220µm) using a Knoop penetrator (10g/10s). Statistical analyses were performed with analysis of variance (ANOVA) and Tukey tests (p<0.05). RESULTS: The zymographic analysis showed that 1.5% STMP promoted complete inhibition of gelatinolytic activity. Therefore, 1.5% STMP was investigated in association with supplemented calcium or fluoride; a combination of 1.5% STMP and Ca(OH)2 significantly increased the mechanical properties of the treated dentin. CONCLUSION: 1.5% STMP serves as an antiproteolytic agent against matrix metalloproteinases extracted from human dentin. Furthermore, when supplemented with Ca(OH)2, 1.5% STMP may potentially induce remineralization. CLINICAL SIGNIFICANCE: STMP can be introduced as a novel strategy that combines enzymatic inhibition and remineralizing potential, which can serve to strengthen dentin and improve stability. STMP may have potential in the treatment of demineralized dentin lesions, especially when supplemented with calcium.

Effects of mineral trioxide aggregate, BiodentineTM and calcium hydroxide on viability, proliferation, migration and differentiation of stem cells from human exfoliated deciduous teeth

Abstract Objective: The aim of the study was to evaluate the effects of the capping materials mineral trioxide aggregate (MTA), calcium hydroxide (CH) and BiodentineTM (BD) on stem cells from human exfoliated deciduous teeth (SHED) in vitro. Material and Methods: SHED were cultured for 1 - 7 days in medium conditioned by incubation with MTA, BD or CH (1 mg/mL), and tested for viability (MTT assay) and proliferation (SRB assay). Also, the migration of serum-starved SHED towards conditioned media was assayed in companion plates, with 8 μm-pore-sized membranes, for 24 h. Gene expression of dentin matrix protein-1 (DMP-1) was evaluated by reverse-transcription polymerase chain reaction. Regular culture medium with 10% FBS (without conditioning) and culture medium supplemented with 20% FBS were used as controls. Results: MTA, CH and BD conditioned media maintained cell viability and allowed continuous SHED proliferation, with CH conditioned medium causing the highest positive effect on proliferation at the end of the treatment period (compared with BD and MTA) (p<0.05). In contrast, we observed increased SHED migration towards BD and MTA conditioned media (compared with CH) (p<0.05). A greater amount of DMP-1 gene was expressed in MTA group compared with the other groups from day 7 up to day 21. Conclusion: Our results show that the three capping materials are biocompatible, maintain viability and stimulate proliferation, migration and differentiation in a key dental stem cell population.

Calcium hydroxide associated with a new vehicle: Psidium cattleianum leaf extracts. Tissue response evaluation.

The aim of this study was to evaluate edemogenic activity and subcutaneous inflammatory reaction induced by Psidium cattleianum leaf extracts associated with Ca(OH)2. Thirty male Wistar rats, split equally into three groups [aqueous extract + Ca(OH)2; ethanolic extract + Ca(OH)2; and propylene glycol + Ca(OH)2], were assessed every 3 h or 6 h (five animals in each period). Under general anesthesia, 0.2 mL of 1% Evans blue per 100 g of body weight was injected into the penile vein and each combination to be evaluated was subcutaneously injected into the dorsal region 30 min thereafter. Edemogenic activity was analyzed by spectrophotometry (λ=630 nm). For inflammatory reaction analysis, 50 rats received four polyethylene tubes (three experimental groups) and an empty tube (control group). The assessments were made at 7, 15, 30, 60, and 90 days, followed by hematoxylin-eosin staining and by the assignment of scores for evaluation of tissue response intensity. Ethanolic extract + Ca(OH)2 yielded the largest edemogenic activity at 3 h. Intergroup differences at 6 h were not significant. The histological analysis showed progressive repair over time (p<0.05) and aqueous and ethanolic extracts produced similar responses to those of the control and Ca(OH)2 + propylene glycol groups. Psidium cattleianum leaf extracts used as Ca(OH)2 vehicles evoked similar tissue response when compared to Ca(OH)2 associated with propylene glycol.