Yttrium oxide nanoparticles ameliorates calcium hydroxide and calcium titanate nanoparticles induced genomic DNA and mitochondrial damage, ROS generation and inflammation.

Calcium hydroxide (Ca(OH)2NPs), calcium titanate (CaTiO3NPs) and yttrium oxide (Y2O3NPs) nanoparticles are prevalent in many industries, including food and medicine, but their small size raises concerns about potential cellular damage and genotoxic effects. However, there are very limited studies available on their genotoxic effects. Hence, this was done to investigate the effects of multiple administration of Ca(OH)2NPs, CaTiO3NPs or/and Y2O3NPs on genomic DNA stability, mitochondrial membrane potential integrity and inflammation induction in mouse brain tissues. Mice were orally administered Ca(OH)2NPs, CaTiO3NPs or/and Y2O3NPs at a dose level of 50 mg/kg b.w three times a week for 2 weeks. Genomic DNA integrity was studied using Comet assay and the level of reactive oxygen species (ROS) within brain cells was analyzed using 2,7 dichlorofluorescein diacetate dye. The expression level of Presenilin-1, tumor necrosis factor-alpha (TNF-α) and Interleukin-6 (IL-6) genes and the integrity of the mitochondrial membrane potential were also detected. Oral administration of Ca(OH)2NPs caused the highest damage to genomic DNA and mitochondrial membrane potential, less genomic DNA and mitochondrial damage was induced by CaTiO3NPs administration while administration of Y2O3NPs did not cause any remarkable change in the integrity of genomic DNA and mitochondrial membrane potential. Highest ROS generation and upregulation of presenilin-1, TNF-α and IL-6 genes were also observed within the brain cells of mice administrated Ca(OH)2NPs but Y2O3NPs administration almost caused no changes in ROS generation and genes expression compared to the negative control. Administration of CaTiO3NPs alone slightly increased ROS generation and the expression level of TNF-α and IL-6 genes. Moreover, no remarkable changes in the integrity of genomic DNA and mitochondrial DNA potential, ROS level and the expression level of presenilin-1, TNF-α and IL-6 genes were noticed after simultaneous coadministration of Y2O3NPs with Ca(OH)2NPs and CaTiO3NPs. Coadministration of Y2O3NPs with Ca(OH)2NPs and CaTiO3NPs mitigated Ca(OH)2NPs and CaTiO3NPs induced ROS generation, genomic DNA damage and inflammation along with restoring the integrity of mitochondrial membrane potential through Y2O3NPs scavenging free radicals ability. Therefore, further studies are recommended to study the possibility of using Y2O3NPs to alleviate Ca(OH)2NPs and CaTiO3NPs induced genotoxic effects.

Examining the Effects of Asiaticoside on Dental Pulp Stem Cell Viability and Proliferation: A Promising Approach to Root Canal Treatment.

AIM: This study aims to evaluate the impact of asiaticoside (AC) on the viability and proliferation of dental pulp stem cells (DPSCs), considering the known negative effects of routinely used intracanal medicaments. This evaluation will be compared with the outcomes from using traditional intracanal medicaments, specifically triple antibiotic paste (TAP) and calcium hydroxide [Ca(OH)2]. MATERIALS AND METHODS: The DPSCs were obtained from the third molars of an adult donor. The application of flow cytometry was employed to do a phenotypic analysis on DPSCs using CD90, CD73, CD105, CD34, CD14, and CD45 antibodies. The methylthiazol tetrazolium (MTT) assay was employed to assess cellular viability. The cells were treated with different concentrations of TAP and Ca(OH)2 (5, 2.5, 1, 0.5, and 0.25 mg/mL), along with AC (100, 50, 25, 12.5, and 6.25 µM). A cell proliferation rate was performed at 3, 5, and 7 days. RESULTS: The characterization of DPSCs was conducted by flow cytometry analysis, which verified the presence of mesenchymal cell surface antigen molecules (CD105, CD73, and CD90) and demonstrated the absence of hematopoietic markers (CD34, CD45, and CD14). Cells treated with concentrations over 0.5 mg/mL of TAP and Ca(OH)2 showed a notable reduction in cell viability in comparison to the untreated cells (p < 0.05). Additionally, the cells treated with different concentrations of AC 12.5, 6.25, 25, and 50 µM did not differ significantly from the untreated cells (p > 0.05). Nevertheless, cells treated with concentrations of 100 µM showed a significant reduction in viability compared to the untreated cells (p < 0.05). After a period of 7 days, it was noted that cells exposed to three different concentrations of AC (50, 25, and 12.5 µM) had a notable rise in cell density in comparison to TAP and Ca(OH)2 (p < 0.05). Furthermore, cells that were exposed to a concentration of 12.5 µM exhibited the highest cell density. CONCLUSION: The cellular viability of the AC-treated cells was superior to that of the TAP and Ca(OH)2-treated cells. Moreover, the AC with a concentration of 12.5 µM had the highest degree of proliferation. CLINICAL SIGNIFICANCE: This study underscores the importance of evaluating alternative root canal medicaments and their effects on DPSCs' growth and vitality. The findings on AC, particularly its influence on the survival and proliferation of DPSCs, offer valuable insights for its probable use as an intracanal medication. This research contributes to the ongoing efforts to identify safer and more effective intracanal treatments, which are crucial for enhancing patient outcomes in endodontic procedures. How to cite this article: Alazemi MJ, Badawi MF, Elbeltagy MG, et al. Examining the Effects of Asiaticoside on Dental Pulp Stem Cell Viability and Proliferation: A Promising Approach to Root Canal Treatment. J Contemp Dent Pract 2024;25(2):118-127.

The Effects of Calcium Silicate- and Calcium Hydroxide-based Root Canal Sealers on Postoperative Pain: A Randomized Clinical Trial.

INTRODUCTION: This study aimed to compare the postoperative pain level changes resulting from using calcium silicate- (EndoSeal MTA) and calcium hydroxide-based (Sealapex) root canal sealers in mandibular first and second molar teeth with symptomatic apical periodontitis. METHODS: A total of 60 patients with symptomatic apical periodontitis in their lower molar teeth were randomly allocated into 2 groups according to sealer type (n = 30). Demographic data, including gender, age, and smoking habit, and preoperative pain measures were recorded. Root canal treatments were performed in a single visit. Postoperative pain measurements and analgesic intake were measured at 6, 12, 24, and 48 hours and after 3, 5, and 7 days using the visual analog scale. The data were statistically analyzed using a chi-squared test (to compare gender, age, smoking habit, analgesic intake, and sealer extrusion), the Mann-Whitney U test (to compare pain levels), Friedman tests (for the evaluations of the reduction in pain levels over time), and Spearman's correlation test (to analyze the relationships of age, gender, smoking habit factors with postoperative pain) (P = .05). RESULTS: The statistical analysis showed no significant differences between the groups in postoperative pain and analgesic intake at any of the time intervals evaluated (P > .05). CONCLUSIONS: Patients treated with calcium silicate- and calcium hydroxide-based root canal sealers experienced similar postoperative pain and no statistically significant differences were observed in analgesic intake.

Impact of N-acetylcysteine (NAC) and calcium hydroxide intracanal medications in primary endodontic infection: a randomized clinical trial.

OBJECTIVES: This RCT investigated the impact of N-acetylcysteine (NAC) and calcium hydroxide [Ca(OH)2] intracanal medications (ICMs) in primary endodontic infection with apical periodontitis (PEIAP). MATERIALS AND METHODS: Thirty-six teeth with PEIAP were randomly divided into groups according to the ICM: NAC, Ca(OH)2 + saline solution (SSL), and Ca(OH)2 + 2% chlorhexidine-gel (2% CHX-gel) (all, n = 12). Root canal samples (RCSs) were collected before (s1) and after instrumentation (s2) and after 14 days of ICM (s3). Chemomechanical preparation (CMP) was performed with a Reciproc file and 2.5% NaOCl. Checkerboard DNA-DNA hybridization was used to assess 40 target bacteria species. RESULTS: At s1, bacterial DNA was detected in 100% of RCSs (36/36). All 40 bacterial species were found in PEIAP. The mean number of species per RCS was 17.92 ± 13.18. The most frequent bacteria were S. mitis (65%), E. nodatum (63%), E. faecalis (63%), F. nucl sp vicentii (58%), T. forsythia (58%), and F. periodonticum (56%). CMP reduced the mean number of species per RCS to 6.8 ± 2.36 (p < 0.05). At s3, the intragroup analysis revealed a broader antimicrobial activity for Ca (OH)2 + 2% CHX-gel and NAC than Ca(OH)2 + SSL (p < 0.05). NAC eliminated 8/12 bacteria species resistant to both Ca (OH)2 ICMs, including P. micra, P. nigrescens, T. denticola, A. israelii, P. endodontalis, P. acnes, C. ochracea, and E. corrodens. CONCLUSIONS: Ca (OH)2 + 2% chlorhexidine gel (2% CHX gel) showed a greater bacterial elimination over the number of bacterial species; however, NAC eliminated 8/12 bacteria species resistant to both Ca (OH)2 ICMs (RBR-3xbnnn). CLINICAL RELEVANCE: The use of intracanal medication with a broad antimicrobial activity can optimize root canal disinfection. Ca(OH)2 + 2% CHX gel and NAC showed a broader antimicrobial activity than Ca(OH)2 + SSL against endodontic pathogens in primary root canal infection. TRIAL REGISTRATION: Brazilian Registry of Clinical Trials (REBEC), No. RBR-3xbnnn.

Detoxification of LTA by intracanal medication: analysis by macrophages proinflammatory cytokines production.

The aim of this study was to evaluate in vitro the effect of calcium hydroxide [Ca(OH)2], 2% chlorhexidine gel (CHX) on macrophages (RAW 264.7) to produce pro-inflammatory cytokines and nitric oxide after pretreatment with lipoteichoic acid (LTA) of Enterococcus faecalis. Forty-eight human single-rooted teeth were instrumented with R25.08 (RECIPROC) and sterilized by gamma irradiation. LTA was inoculated in the root canal of each specimen for 96 hours. Specimens were instrumented with 40.06 and 50.05 (RECIPROC) and medicated with: I) Pyrogen-free saline solution (SS); II) 2% CHX gel; III) Ca(OH)2 + SS; or IV) Ca(OH)2 + CHX for 14 days. Three samples (S) were performed of the root canal of each specimen at: S1) immediately after instrumentation; S2) after Ethylenediaminetetraacetic acid (EDTA); S3) after intracanal medication removal. Subsequent quantification of cytokines (IL-1ß, TNF-α, MIP-1α, IP-10, G-CSF and IL-6) by immunosorbent assay (ELISA) and nitric oxide by the Griess method was carried-out. Data were submitted to a normality test and then analyzed with one-way ANOVA and Tukey test with a significance level of 5% using GraphPad Prism 6. Ca(OH)2 + SS and Ca(OH)2 + CHX presented lower levels of TNF-α, TNF-α, IL-6, G-CSF and nitric oxide. Ca(OH)2 + SS was the most effective in reducing MIP-1α. CHX was effective in reducing IL-6 and G-CSF. Therefore, the combined intracanal medication of calcium hydroxide and chlorhexidine is effective in reducing the cytokines TNF-α, IL-1ß, IL-6, G-CSF and nitric oxide.

Antibacterial and anti-inflammatory efficacy of N-acetyl cysteine in endodontic treatment: a scoping review.

BACKGROUND: This scoping review systematically summarized the available evidence about the efficacy of N-acetyl cysteine (NAC) as an intracanal antibacterial and/or anti-inflammatory. METHODS: PubMed, Scopus, Web of Science, and Google scholar search engines/databases were searched up to February 2022 to retrieve relevant studies. The studies were evaluated for eligibility criteria, and identifying relevant studies. RESULTS: Out of 193 studies, 15 fulfilled the inclusion criteria and were processed for data extraction. Thirteen in vitro studies assessed antibacterial/antibiofilm efficacy of NAC, and reported good and promising efficacy: NAC was found as efficacious as the comparators (chlorhexidine, sodium hypochlorite, calcium hydroxide), or even showed higher efficacy. Regarding the anti-inflammatory efficacy of NAC, one in vitro study found it equivalent to, while one clinical trial revealed it more efficacious than calcium hydroxide. CONCLUSIONS: There is accumulating evidence on the anti-microbial and anti-inflammatory efficacy of NAC in context of endodontics. However, further clinical trials with robust methodology and objective and reliable clinical, biological and microbial outcomes are warranted to translate its use for clinical practice on humans.

Pulpal expression of erythropoietin and erythropoietin receptor after direct pulp capping in rat.

This study aimed to evaluate the effect of direct pulp capping on the expression of erythropoietin (Epo) and Epo-receptor (Epor) genes in relation to the expression of inflammatory and osteogenic genes in rat pulp. Dental pulps of the first maxillary molars of Wistar Albino rats were exposed and capped with either calcium hydroxide or mineral trioxide aggregate, or were left untreated. After 4 wk, animals were euthanized, and maxillae were prepared for histological and real-time polymerase chain reaction analysis. Histological scores of pulp inflammation and mineralization, and relative expressions of Epo, Epor, inflammatory cytokines, and pulp osteogenic genes were evaluated. The capped pulps showed higher expressions of Epo, while the untreated pulps had the highest expression of Epor. Both calcium hydroxide and mineral trioxide aggregate downregulated the expression of tumor necrosis factor alpha compared to untreated controls, and upregulated transforming growth factor beta compared to healthy controls. Alkaline phosphatase expression was significantly higher in experimental groups. Relative expression of Epo negatively correlated with pulp inflammation, and positively correlated with pulp mineralization. Pulp exposure promoted expression of Epor and pro-inflammatory cytokines, while pulp capping promoted expression of Epo, alkaline phosphatase, and downregulated Epor and pro-inflammatory cytokines.

Cytokine expression and anti-microbial effectiveness of different calcium hydroxide dilutions: An In Vitro study.

Aims: To determine the cytokine expression by human gingival fibroblasts in response to different calcium hydroxide (Ca(OH)2) dilutions and test the effectiveness of these dilutions in root canal dentin infected with Enterococcus faecalis (E. faecalis). Methods: UltraCal XS Ca(OH)2 dilutions were prepared (60, 10, and 1 mg\mL) and co-cultured with gingival fibroblasts for 24 and 48 hours. Untreated cells were used as controls. Expressions of interleukin (IL-1ß), tumour necrosis factor alpha (TNF-α), transforming growth factor beta (TGF-ß), and IL-10 were analysed with real-time polymerase chain reaction (PCR). Root canals of extracted human teeth were inoculated with E. faecalis. After 21 days, canals were medicated with Ca(OH)2 dilutions for 7 days. Samples were taken to determine bacterial reduction using quantitative PCR. Analysis of variance, Tukey post-test, and Wilcoxon matched pair test were used for statistics. Results: IL-1ß and TNF-α expressions of all Ca(OH)2 dilutions were higher at 24 and 48 hours compared to the control. Similarly, all Ca(OH)2 dilutions induced TGF-ß expression at 24 hours compared to the control and continued to be higher in 60 mg/mL groups at 48 hours. In contrast, IL-10 was constitutively expressed by untreated cells in the control group and was down-regulated significantly by all Ca(OH)2 dilutions at 24 and 48 hours. All dilutions demonstrated a significant E. faecalis reduction (P < 0.001) with no significant difference between dilution groups (P > 0.05). Conclusions: All Ca(OH)2 dilutions had a differential inflammatory effect on fibroblasts and had a down-regulation effect to IL-10. All dilutions tested were effective against E. faecalis, with 60 mg/mL having the highest bacterial reduction.

IL-1ß, IL-6 and TNF-α expression levels of macrophage cells induced by benzydamine hydrochloride, benzydamine hydrochloride with chitosan, calcium hydroxide and chlorhexidine medicaments: An ELISA study.

The aim of the present study was to evaluate IL-1ß, IL-6 and TNF-α expression levels of macrophage cells induced by benzydamine hydrochloride (BNZ), BNZ with chitosan, calcium hydroxide (CH) and chlorhexidine (CHX) medicaments. Half maximal inhibitory concentrations (IC50) were assessed on THP-1, Saos-2, and CRL-2014 cells using MTT assay. THP-1 cells were differentiated into macrophages with phorbol12-myristate13-acetate and activated with lipopolysaccharide. IL-1ß, IL-6 and TNF-α levels in supernatants were determined using enzyme-linked immunosorbent assay (ELISA). The data were examined with one-way ANOVA and Tukey's multiple comparison test (p=0.05). At the selected concentrations, the cell viability was higher than 50% for chitosan and CH, whereas CHX presented lower IC50 values than BNZ and BNZ+chitosan. According to ELISA results, the lowest IL-1ß, IL-6 and TNF-α values were observed with BNZ+Chitosan 50 µg/mL and BNZ 50 µg/mL. BNZ+chitosan 50 µg/mL combination has revealed promising anti-inflammatory effects. Nevertheless, these findings need to be examined in clinical conditions.

Comparative Evaluation of Ozonoid Olive Oil and Calcium Hydroxide as an Indirect Pulp Capping Agent in Primary Mandibular Second Molar: A Randomized Controlled Trial.

AIM: The aim of this study was to evaluate clinical and radiographical success of ozonoid olive oil as an indirect pulp capping (IPC) agent in primary mandibular second molar when compared to calcium hydroxide and to evaluate the antimicrobial efficacy of ozonoid olive oil on Streptococcus mutans and Lactobacilli. MATERIALS AND METHODS: A split-mouth randomized controlled trial was conducted on 30 primary mandibular second molars in 15 children of age 5-9 years with deep dentinal carious lesion. Teeth were randomly allocated to two groups of 15 each. After achieving local anesthesia and rubber dam isolation, an IPC procedure was performed using ozonoid olive oil in group I and calcium hydroxide in group II. Teeth were evaluated clinically and radiographically at 6 and 12 months of follow-up for success or failure of IPC. The bacterial counts of S. mutans and Lactobacilli were measured before and after application of ozonoid olive oil for 60 seconds on dentinal tissue in group I and recorded as colony-forming units per mL (CFU/mL). RESULTS: There were no statistically significant differences found between the materials used for IPC (p >0.05). About 93.33% and 100% clinical and radiographical success rates were seen in group I and group II, respectively. Statistically significant differences were observed for bacterial reduction after the application of ozonoid olive oil (p <0.05) for both the microorganisms. CONCLUSION: The results of this study showed that the success of IPC is independent of capping material. Ozonoid olive oil, an antimicrobial agent, can also be used for IPC. The success of the IPC procedure depends on a reduction in the bacterial count and sealing of the tooth with hermetic restoration. More clinical studies with a larger sample size and longer follow-up duration are required for understanding the efficacy of this material. CLINICAL SIGNIFICANCE: Ozonoid olive oil can be used as an IPC agent in primary molars and also for a bacterial reduction in dentinal caries.

Disinfection of dentinal tubules with diclofenac sodium and N-Acetylcysteine compared with calcium hydroxide as intracanal medicaments against Enterococcus faecalis.

The purpose of this study was to evaluate the disinfection of dentinal tubules with diclofenac sodium (DS), N-acetylcysteine (NAC) and calcium hydroxide (CH). Contaminated dentinal blocks were divided into two control and seven experimental groups (n = 15): CH, DS, NAC, CH + 5% DS, 50% CH + 50% DS, CH + 5% NAC and 50% CH + 50% NAC. After seven days, dentine debris was obtained from two depths of 100 and 200 µm. The bacterial load was assessed by counting the number of colony-forming units (CFUs). Pure DS exhibited maximum antibacterial activity at both depths. At 200 µm, it showed statistically significant differences with all the other groups (P < 0.05). Mixing CH with either 5% or 50% of DS and NAC did not increase the antibacterial efficacy (P > 0.05). Pure DS was most effective in disinfecting dentinal tubules, and mixing CH with DS or NAC is not recommended.

Silver Nanoparticles Alone or in Combination with Calcium Hydroxide Modulate the Viability, Attachment, Migration, and Osteogenic Differentiation of Human Mesenchymal Stem Cells.

This study aimed to evaluate the effect of silver nanoparticles (AgNPs) alone or in combination with calcium hydroxide (Ca(OH)2) on the proliferation, viability, attachment, migration, and osteogenic differentiation of human mesenchymal stem cells (hMSCs). Different concentrations of AgNPs alone or mixed with Ca(OH)2 were prepared. Cell proliferation was measured using AlamarBlue, and hMSCs attachment to dentin disks was evaluated using scanning electron microscopy. Live-dead imaging was performed to assess apoptosis. Wound healing ability was determined using the scratch-migration assay. To evaluate osteogenic differentiation, the expression of Runt-related transcription factor (RUNX2), Transforming growth factor beta-1 (TGF-ß1), Alkaline Phosphatase (ALP), and Osteocalcin (OCN) were measured using real-time reverse transcriptase polymerase chain reaction. ALP staining and activity were also performed as indicators of osteogenic differentiation. AgNPs alone seemed to favor cell attachment. Lower concentrations of AgNPs enhanced cell proliferation. AgNP groups showed markedly less apoptosis. None of the medicaments had adverse effects on wound closure. The expression of TGF-ß1 was significantly upregulated in all groups, and OCN was highly expressed in the AgNP groups. AgNPs 0.06% showed the most enhanced ALP gene expression levels, activity, and marked cytochemical staining. In conclusion, AgNPs positively affect hMSCs, making them a potential biomaterial for various clinical applications.

Synergistic effects of quaternary ammonium compounds and food additive grade calcium hydroxide on microbicidal activities at low temperatures.

The microbicidal activities of mixtures of quaternary ammonium compounds (QACs) and food additive grade calcium hydroxide (FdCa(OH)2) were evaluated in a suspension test at -20°C using an anti-freeze agent (AFA) containing methanol, or at 1°C, with varying contact time, toward avian influenza virus (AIV), Newcastle disease virus (NDV), fowl adenovirus (FAdV), avian reovirus (ARV), Salmonella Infantis (SI) and Escherichia coli (EC). At -20°C, the mixtures could inactivate AIV and NDV within 30 min, FAdV and ARV within 5 sec, and SI and EC within 3 min, respectively. AFA did not inactivate viruses and bacteria within 30 min and 10 min, respectively. At 1°C, the mixtures inactivated FAdV and ARV within 30 sec, AIV within 10 min, and NDV within 30 min. A mixture of slaked lime (SL) and QAC could inactivate FAdV and ARV within 30 sec, but could not inactivate AIV and NDV even after 60 min at 1°C. SL could not substitute FdCa(OH)2 in order to exert the synergistic effects with QAC. Thus, QACs microbicidal activities were maintained or enhanced by adding FdCa(OH)2. It is hence recommended to use QACs with FdCa(OH)2, especially in the winter season.

Comparative Evaluation of Antibacterial Efficacy of Silver and Cadmium Nanoparticles and Calcium Hydroxide against Enterococcus faecalis Biofilm.

AIM: The purpose of this study was to evaluate and compare the antibacterial efficacy of calcium hydroxide medicament, silver (AgNPs) and cadmium nanoparticles (CdSNPs) as medicament against the biofilms of Enterococcus faecalis on dentin sections. E. faecalis is commonly detected in asymptomatic and persistent endodontic infections. METHODS: Twenty standard size dentin sections were prepared. E. faecalis was inoculated on these dentin sections for four weeks to form the bacterial biofilm. Twenty dentin sections were segregated into four different groups with five specimens in each group. Group I was kept as control group, and antibacterial efficacy was tested by treating biofilms with Ca(OH) 2 medicament, 0.02% AgNP and CdSNP gels for 7 days. The ultrastructure of biofilms from each group was examined under scanning electron microscope and was visually evaluated and compared for different groups. RESULTS: Ca(OH)2 exhibited a slight disruption of E. faecalis biofilm. Both AgNP and CdSNP medicaments disrupted E. faecalis biofilm effectively after 7 days of inoculation. AgNPs disrupted the biofilm more effectively than CdSNPs. Biofilms in control group, which was irrigated with saline, did not show any disruption of biofilm, which could be seen as homogenous layer over most of dentin sections. CONCLUSIONS: This study suggests that both AgNP and CdNP gels are effective against E. faecalis and can be used as a medicament to eliminate residual bacterial biofilms during root canal disinfection. AgNP medicament is more effective than CdNP, whereas Ca(OH) 2 is not effective against E. faecalis biofilms. CLINICAL SIGNIFICANCE: Incomplete clearance and the development of antibiotic resistance in E. faecalis are the important factors for failure of root canal treatment. When cationic nanoparticles are introduced for the treatment of biofilms, it can interact with both extracellular polymeric substances and bacterial cells. The initial electrostatic attraction between positively charged nanoparticles and negatively charged bacterial surface leads to bacterial killing via the production of reactive oxygen species. Metal nanoparticles that are effective against E. faecalis have a significant potential role in the prevention and treatment of such cases, as bacteria do not develop resistance against metal nanoparticles.

Simple conversion of 3D electrospun nanofibrous cellulose acetate into a mechanically robust nanocomposite cellulose/calcium scaffold.

Cellulose and its derivatives are widely used as nanofibrous biomaterials, but obtaining 3D cellulose nanofibers is difficult and relevant research is scarce. In the present study, we propose a simple method for converting electrospun 3D cellulose acetate/lactic acid nanofibers via calcium hydroxide treatment into a 3D cellulose/calcium lactate nanocomposite matrix. The conversion resulted in producing a stronger nanofibrous matrix (1.382 MPa vs. 0.112 MPa) that is more hydrophilic and cell-friendly compared to the untreated cellulose acetate/lactic acid group. The successful conversion was verified via FTIR, XPS, TGA, DTG, and XRD. The ability of the scaffolds to provide a suitable environment for cell growth and infiltration was verified by CCK assay and confocal microscopy. The porous nature, mechanical strength, and presence of calcium make the 3D cellulose/calcium lactate matrix a promising material for bone tissue engineering.

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 a calcium hydroxide-based intracanal medicament containing N-2-methyl pyrrolidone as a vehicle against Enterococcus faecalis biofilm

Abstract This study investigated the effect of a calcium hydroxide (CH) paste (CleaniCal®) containing N-2-methyl pyrrolidone (NMP) as a vehicle on Enterococcus faecalis (E. faecalis) biofilms compared with other products containing saline (Calasept Plus™) or propylene glycol (PG) (Calcipex II®). Methodology Standardized bovine root canal specimens were used. The antibacterial effects were measured by colony-forming unit counting. The thickness of bacterial microcolonies and exopolysaccharides was assessed using confocal laser scanning microscopy. Morphological features of the biofilms were observed using field-emission scanning electron microscopy (FE-SEM). Bovine tooth blocks covered with nail polish were immersed into the vehicles and dispelling was observed. The data were analyzed using one-way analysis of variance and Tukey tests (p<0.05). Results CleaniCal® showed the highest antibacterial activity, followed by Calcipex II® (p<0.05). Moreover, NMP showed a higher antibacterial effect compared with PG (p<0.05). The thickness of bacteria and EPS in the CleaniCal® group was significantly lower than that of other materials tested (p<0.05). FE-SEM images showed the specimens treated with Calasept Plus™ were covered with biofilms, whereas the specimens treated with other medicaments were not. Notably, the specimen treated with CleaniCal® was cleaner than the one treated with Calcipex II®. Furthermore, the nail polish on the bovine tooth block immersed in NMP was completely dispelled. Conclusions CleaniCal® performed better than Calasept Plus™ and Calcipex II® in the removal efficacy of E. faecalis biofilms. The results suggest the effect might be due to the potent dissolving effect of NMP on organic substances.

In vitro treatment of Enterococcus faecalis with calcium hydroxide impairs phagocytosis by human macrophages.

OBJECTIVE: Monocyte-derived macrophages (MDMs) ability to phagocytize and produce nitric oxide (NO) was tested against root-canal strains of Enterococcus faecalis submitted to alkaline stress. Root-canal strains were also compared with urine Enterococci. MATERIALS AND METHODS: Enterococcus faecalis were stressed with alkaline-BHI broth and incubated in vitro at a cell/bacteria ratio of 1:5. Phagocytosis was analyzed by fluorescence microscopy using acridine orange stain, and NO concentration was measured in supernatants. RESULTS AND CONCLUSIONS: Alkaline-stress significantly impaired MDMs phagocytosis of E. faecalis strains analyzed, except in ATCC4083 isolated from a pulpless tooth, but NO production was unchanged. Comparison of different strains showed the urine isolate had higher NO levels than root canal strains. Alterations in the bacterial cell wall structures after alkaline-stress possibly made bacteria less recognizable and phagocytized by MDMs but did not affect their ability to activate NO production. Furthermore, root canal strains elicited different responses by immune cells compared with strains from urine. Clinically, impaired phagocytosis of E. faecalis could contribute to their persistence in root canal systems previously treated with calcium hydroxide.

Effect of intracanal medicaments on matrix metalloproteinase-9 and vasoactive intestinal peptide secretion in periapical lesions of re-treated canals: a randomized controlled clinical study.

OBJECTIVE: The aim of this study was to evaluate the effect of calcium hydroxide (Ca[OH]2) and chlorhexidine (CHX) gel on matrix metalloproteinase-9 (MMP-9) and vasoactive intestinal peptide (VIP) secretion in periapical lesions. MATERIALS AND METHODS: A total of 60 patients were randomly divided into two groups that were to receive different medications. Pre-and post-treatment samples were collected from the interstitial fluid of periapical lesions using sterile paper points. VIP and MMP-9 levels were measured by enzyme-linked immunosorbent assay kits, and the data were statistically analyzed. RESULTS: Gender and smoking habits had no effect on the pre- and post-treatment VIP and MMPs levels. Intragroup analyses revealed that in the Ca(OH)2 group, the post-treatment VIP level was found to be significantly higher than the pre-treatment VIP level. In the CHX group, the post-treatment MMP-9 level was significantly higher than the pre-treatment MMP-9 level. CONCLUSION: According to the results of the present study, the type of the medication affected the amount of periapical VIP and MMP-9 secretion. CLINICAL RELEVANCE: VIP is a neuropeptide that promotes new bone formation. Thus, intracanal Ca(OH)2 medication may accelerate the repair process of bone tissue.