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The success of endodontic treatment depends on the thorough removal of microorganisms from the root canal system. The search for new ways to eliminate the microorganisms is therefore justified. Nd:YAP is a laser that uses yttrium aluminum perovskite, doped with neodymium crystal, as active laser medium. We used the Nd:YAP laser in an in vitro experiment to evaluate the bactericidal effect of three parameters of Nd:YAP laser-activated irrigation on biofilms of Enterococcus faecalis in root canals. The canals of 45 extracted human single-root teeth were prepared on a #35 Mtwo instrument and contaminated with E. faecalis for 14 days. Forty infected single-root teeth were then randomly divided into four groups according to the irrigation agitation protocols as follows: 5.25% sodium hypochlorite (NaOCl), Nd:YAP laser (180 mJ) + NaOCl, Nd:YAP laser (280 mJ) + NaOCl, and Nd:YAP laser (360 mJ) + NaOCl. The remaining bacteria were counted immediately using the cell count method. Teeth were firstly spilt and one half examined by scanning electron microscopy (SEM). The other half involved examination of bacterial colonization in dentinal tubules using confocal laser scanning microscopy (CLSM). Nd:YAP laser (280 mJ) + NaOCl and Nd:YAP laser (360 mJ) + NaOCl completely removed the E. faecalis biofilms from the root canal walls and made it the cleanest among the treatment groups. Bacterial reductions in the treatment groups for dentinal tubules are presented in a descending order as follows: Nd:YAP laser (360 mJ) (53.7%), Nd:YAP laser (280 mJ) (51.5%) > Nd:YAP laser (180 mJ) (45.3%) > 5.25% NaOCl (31.9%) > control (19.3%) (p < 0.05). Nd:YAP laser of 280 mJ and 360 mJ showed effective bactericidal effect in removing E. faecalis biofilm from the root canal walls and dentinal tubules.

Catheter-Associated Hospital-Acquired Infections (HAI's) are caused by biofilm-forming bacteria. Using a novel approach, we generated anti-infective barrier on catheters by charging them with Nitric Oxide (NO), a naturally-produced gas molecule. NO is slowly released from the catheter upon contact with physiological fluids, and prevents bacterial colonization and biofilm formation onto catheter surfaces. The aim of the study was to assess the anti-infective properties of NO-charged catheters exposed to low concentration (up to 103 CFU/ml) of microbial cells in-vitro. We assessed NO-charged tracheal tubes using Pseudomonas aeruginosa, dialysis and biliary catheters using Escherichia coli, and urinary catheters using E. coli, Candida albicans or Enterococcus faecalis. Safety and tolerability of NO-charged urinary catheters were evaluated in a phase 1 clinical study in 12 patients. Six patients were catheterized with NO-charged catheters (NO-group), followed by 6 patients catheterized with regular control catheters (CT-group). Comparison of safety parameters between the study groups was performed. NO-charged tracheal, dialysis biliary and urinary catheters prevented P. aeruginosa, E. coli and C. albicans attachment and colonization onto their surfaces and eradicated corresponding planktonic microbial cells in the surrounding media after 24-48 hours, while E. faecalis colonization onto urinary catheters was reduced by 1 log compared to controls. All patients catheterized with an NO-charged urinary catheter successfully completed the study without experiencing NO-related AE's or serious AE's (SAE's). These data highlight the potential of NO-based technology as potential platform for preventing catheter-associated HAI's.

Objective This study investigated the associations among endodontic instruments, ultrasonic tips and various final irrigation protocols for removing intracanal and intratubular biofilms in long oval canals. Methodology One hundred mandibular premolars inoculated with Enterococcus faecalis were divided into two groups: the control group (CG: n  = 10), which received no treatment; and the test groups ( n  = 30), which included saline (SS), sodium hypochlorite (2.5% NaOCl) and chlorhexidine (2% CHX). The samples were prepared with Reciproc ® Blue 25/0.08, Flatsonic, Clearsonic and Reciproc Blue 40/0.06. Each test group was divided into three subgroups ( n  = 10): conventional irrigation (CI) with syringes and needles, passive ultrasonic irrigation (PUI) and continuous ultrasonic irrigation (CUI) with an Irrisonic tip. The root canals were examined using confocal laser scanning microscopy. Mann‒Whitney and Kruskal‒Wallis tests followed by Dunn post hoc tests ( p  < 0.05) were used for statistical analysis. Results NaOCl and CHX were effective for intracanal and intratubular decontamination. Saline showed significant decontamination in the intratubular areas when CUI was used. The CI method was the least effective for intracanal decontamination. Conclusion The use of Flatsonic and Clearsonic ultrasonic tips is promising for root canal disinfection. Clinical relevance The disinfection techniques in endodontic treatment focus on the removal of biofilms from long oval canals, which is crucial for effective cleaning during root canal procedures.

Background In recent years, the diode laser (810 nm) has been used for root canal disinfection, which plays an important role in endodontic therapy. This study was undertaken to evaluate the disinfecting ability of a diode laser in experimentally infected root canals of primary teeth. Methods Human retained mandibular primary anterior teeth without apical foramen resorption were selected and contaminated with Enterococcus faecalis for 21 days. The specimens were randomly divided into four groups: the negative group (no treatment), positive group (5.25% NaOCl), diode laser group (diode laser), and diode-NaOCl group (diode laser combined with NaOCl). The disinfecting abilities of the treatments were measured by the numbers of bacteria, scanning electron microscopy and confocal laser microscopy (live-dead staining). Results Eighty teeth were selected. After irradiation and irrigation, the elimination of bacteria and the smear layer in the laser groups and positive group were significantly superior, compared with the negative group ( p  < 0.01). In the diode-NaOCl group, bacterial reduction reached nearly 100% on the surfaces of root canals; live bacteria were rarely observed, even in deeper dentinal tubules. Conclusion Use of a diode laser, especially in combination with NaOCl, was effective for disinfecting infected root canals of primary teeth.

Objective The success of root canal treatment and regenerative endodontics relies on thorough disinfection and dentin integrity preservation to ensure long-term tooth survival. This study evaluates the pH stability, material characteristics, microhardness and antimicrobial effects of an allicin-incorporated GO-AgNP hydrogel compared to conventional intracanal medicaments. Methods An allicin-incorporated GO-AgNP hydrogel was synthesized using allicin extract, GO-AgNPs, and sodium alginate. Characterization was performed via FTIR, SEM, and EDX. pH stability of AllGOAgNP, CaOH, CHX, and TAP was assessed at 5 min, 24 h, and 7 days using a digital pH meter. A total of 120 extracted human premolars were randomly assigned to four groups: (1) Control, (2) CaOH, (3) TAP, and (4) AllGOAgNP. Medicaments were applied and incubated at 37 °C with 100% humidity for 1 week, 1 month, and 3 months. Dentin microhardness was evaluated using a Vickers microhardness tester before and after treatment across the coronal, middle, and apical thirds. Additionally, antimicrobial efficacy against E. faecalis and C. albicans was assessed using the disc diffusion method, with inhibition zones measured for each medicament. Statistical analysis was performed using one-way ANOVA and Tukey’s post-hoc test ( p  < 0.05). Results FTIR analysis confirmed the successful incorporation of allicin, GO, AgNPs, and sodium alginate. SEM images showed a uniform nanoparticle distribution in the hydrogel, and EDX confirmed the presence of key elements, including silver and sulfur. The Allicin-GO-AgNP hydrogel maintained a near-neutral pH (mean 7.083), while CaOH (mean 12.297) and TAP (mean 12.683) exhibited highly alkaline pH levels. ANOVA results demonstrated significant differences in microhardness across groups and regions ( p  < 0.05). The Allicin-GO-AgNP hydrogel exhibited significantly higher microhardness than CaOH and TAP across all regions ( p  < 0.001), with no significant difference from the control in the coronal and middle thirds ( p  > 0.05). SEM-EDX analysis of treated dentin confirmed minimal structural alterations in the Allicin-GO-AgNP hydrogel group compared to the control. In antimicrobial testing, the hydrogel demonstrated moderate efficacy with inhibition zones of 20 mm against E. faecalis and 13 mm against C. albicans , outperforming calcium hydroxide after 24 h. Conclusion The Allicin-GO-AgNP hydrogel demonstrated superior dentin preservation compared to conventional intracanal medicaments. Its near-neutral pH, structural stability, and microhardness retention make it a promising alternative for endodontic applications, particularly in regenerative endodontics. Future studies should focus on its long-term biocompatibility and antimicrobial effectiveness in clinical settings. Clinical significance The Allicin-GO-AgNP hydrogel preserves dentin integrity better than conventional medicaments, reducing the risk of structural weakening. Its near-neutral pH minimizes collagen degradation, making it a promising option for regenerative endodontics. This novel hydrogel offers a biocompatible alternative with potential long-term clinical benefits.

Objective. To evaluate the effect of the combination of calcium hydroxide (Ca(OH)2) and a novel electrolyzed superoxidized solution at neutral pH, known as OxOral® on Enterococcus faecalis growth in root canals. Methods. Sixty human teeth were used, from which root canals were infected and randomly divided into the following treatment groups: saline solution, saline solution plus Ca(OH)2, OxOral®, and OxOral® plus Ca(OH)2. Results. A permanent reduction in bacterial growth was observed at days 1, 6, 12, and 18 after OxOral® plus Ca(OH)2 treatment from 4.4±0.074 log10 CFU/mL to 0.0±0.001 log10 CFU/mL. In addition, alkaline conditions maintenance was observed from application time (pH=12.2±0.033) to 18 d posttreatment (pH=12.6±0.083). Conclusion. The combination of OxOral® and Ca(OH)2 provides an alkaline pH and inhibits E. faecalis growth into the root canals. Our study opens the possibility for further research on the use of OxOral® in endodontic therapy.

Objectives Successful endodontic therapy is based on the reduction of infecting bacteria by cleaning, shaping, and disinfecting of the root canal system, thus the use of intracanal dressing is necessary for optimal success of root canal treatment. This study was designed to evaluate the effect of chitosan and propolis as intracanal medicaments against Enterococcus faecalis compared to calcium hydroxide in primary root canals. Material and Methods Ninety‐six extracted primary second molars were collected. Teeth preparation was completed to size 30 K‐file. They were randomly divided into four groups; (A): chitosan, (B): propolis, (C): calcium hydroxide, and (D): control group (saline). The tooth specimens were inoculated with E. faecalis. Then, tested materials were applied for all groups in accordance to the groups each tooth belonged to. Following this, the bacterial colonies were counted after 24 h, 72 h, and 1 week of applying dressing materials and incubation. Finally, one–way analysis of variance and Fisher's least significant difference tests were used for statistical comparisons between the groups at a significance level of .05. Results No statistically significant difference was found between groups A, B, and C for both 24 h and a week (p ≥ .05). Yet, a statistical difference between groups A, B, C, and D after 72 h and 1 week were seen (p ≤ .05). Conclusions Chitosan and propolis medicaments were as effective as calcium hydroxide against E. faecalis in primary root canal treatment and might be considered as an alternative dressing material between treatment sessions.

Background: Sodium hypochlorite (NaOCl) is the most commonly used irrigant in endodontics . The purpose of this study was to evaluate the effect of NaOCl solution (2.5%) and gel (3%) with/without passive ultrasonic irrigation (PUI) on Enterococcus faecalis, Escherichia coli, and their endotoxins, lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Methods: 40 human lower premolars were contaminated with E. coli (ATCC 25922) for 28 days and E. faecalis (ATCC 29212) for 21 days. Specimens were randomly divided into four groups: (1) 2.5% NaOCl irrigating the canals without PUI activation; (2) 2.5% NaOCl with PUI; (3) 3% NaOCl gel irrigating the canals without PUI; and (4) 3% NaOCl gel with PUI. 40 mL of irrigant was used for each group. PUI activation was carried out using E1-Irrisonic stainless-steel tip at 10% frequency. After treatment, all specimens were filled with 3mL of 17% ethylenediaminetetraacetic acid (EDTA) for 3min and then washed with nonpyrogenic saline solution. Three samples were collected from the canals: S1, at baseline to confirm biofilm formation; S2 after treatment; and S3 after EDTA. Samples were assessed for E. coli and E. faecalis colony forming units, and LPS and LTA were assessed using chromogenic kinetic LAL assay and ELISA, respectively. Data were analyzed by Kruskal-Wallis, Friedmann and Dunn tests with α≤0.05. Results: All groups were effective in reducing the microbial load of E. coli and E. faecalis after treatment without a significant difference among the groups. NaOCl and NaOCl gel groups had no significant difference in reducing LPS and LTA. Statistically increased reduction was seen for NaOCL + PUI and NaOCl gel + PUI compared for groups without PUI. Conclusions: NaOCl gel has the same antimicrobial action of NaOCl solution and can partially detoxify endotoxins. PUI improves NaOCl (gel or solution) action over E. faecalis and E. coli and their endotoxins.

For a One-Health investigation of antimicrobial resistance (AMR) in Enterococcus spp., isolates from humans and beef cattle along with abattoirs, manured fields, natural streams, and wastewater from both urban and cattle feedlot sources were collected over two years. Species identification of Enterococcus revealed distinct associations across the continuum. Of the 8430 isolates collected, Enterococcus faecium and Enterococcus faecalis were the main species in urban wastewater (90%) and clinical human isolates (99%); Enterococcus hirae predominated in cattle (92%) and feedlot catch-basins (60%), whereas natural streams harbored environmental Enterococcus spp. Whole-genome sequencing of E. faecalis (n = 366 isolates) and E. faecium (n = 342 isolates), revealed source clustering of isolates, indicative of distinct adaptation to their respective environments. Phenotypic resistance to tetracyclines and macrolides encoded by tet(M) and erm(B) respectively, was prevalent among Enterococcus spp. regardless of source. For E. faecium from cattle, resistance to β-lactams and quinolones was observed among 3% and 8% of isolates respectively, compared to 76% and 70% of human clinical isolates. Clinical vancomycin-resistant E. faecium exhibited high rates of multi-drug resistance, with resistance to all β-lactam, macrolides, and quinolones tested. Differences in the AMR profiles among isolates reflected antimicrobial use practices in each sector of the One-Health continuum.

Purpose Plants extracts are used in urology to manage urinary tract infections. We aimed to evaluate the efficacy of a preparation with solidago, orthosiphon, birch and cranberry extracts (CISTIMEV PLUS ® ) in reducing microbial colonization and biofilm development in patients with indwelling urinary catheters. Methods All consecutive outpatients attending our department between January and June 2010 for the substitution of indwelling catheters were considered for this single-blinded, randomized and controlled pilot study to test superiority of the preventative management (CISTIMEV PLUS ® , 1 tablet daily for 30 days) in respect to no treatment. A sample size of 10–40 participants per group was considered adequate. All patients underwent urine culture the same day of the catheter substitution and were then randomized into test group ( n  = 48) and control group ( n  = 35). Ultrastructural analysis was also performed. After 30 days, the catheter was replaced and the analysis repeated. The primary outcome was the rate of positive urinary culture at the end of the entire study period. Results Ten patients abandoned the study. At 30 days, according to per-protocol analysis, the groups statistically differed regarding the rate of positive urine cultures: test group 10/43 and control group 16/30 ( p  = 0.013) (−30.1 % [95 % CI −51.94 to −8.21]). The most common isolated bacteria were Escherichia coli and Enterococcus faecalis. Conclusions The use of solidago, orthosiphon, birch and cranberry extracts resulted in a significant reduction of microbial colonization in patients with indwelling urinary catheters. Larger clinical trials are needed to demonstrate that the effects here reported are sufficient to reduce symptomatic catheter-associated urinary tract infections.