Comparative Analysis of the Cytotoxic Effects of Modified Triple Antibiotic Hydrogel: Insights From Experimental Models.

INTRODUCTION:  The effectiveness of intracanal medicaments (ICMs) in root canal therapy is critical for successful dental treatments, yet their cytotoxic effects pose significant challenges. This research uses zebrafish embryos and dental pulp stem cells (DPSCs) to identify the optimal concentration that balances antibacterial efficacy with minimal toxicity. AIM:  This study aims to address the need for effective ICMs in dentistry by formulating and assessing the embryotoxicity and cytocompatibility of a novel carrageenan-based modified triple antibiotic paste (MTAP) hydrogel at different concentrations (1, 5, and 10 mg/mL) using a zebrafish model and cell culture assay. MATERIALS AND METHODS:  The hydrogel was formulated by combining antibiotic solutions (ciprofloxacin, metronidazole, and amoxicillin) with carrageenan and xanthan gum. Zebrafish embryos were exposed to varying concentrations of MTAP hydrogel, chlorhexidine (CHX), calcium hydroxide (CaOH2), and plain carrageenan to assess developmental toxicity, survival rate, heart rate, hatching rate, and macrophage migration. The cytotoxicity against DPSCs was examined within a timeframe of 6, 24, and 72 hours with the use of the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay. RESULTS:  The analysis revealed developmental toxicity with malformations observed at higher concentrations of MTAP hydrogel, CaOH2, and CHX medicaments, indicating potential toxicity. Significant impacts on survival, heart rate, and hatching rate were noted in the CaOH2 and CHX groups, as well as at higher MTAP hydrogel concentrations, emphasizing the importance of dosage considerations. The neutral red assay confirmed toxicity, with macrophage migration observed in CaOH2, CHX, and higher MTAP hydrogel concentrations. Lower concentrations, particularly at 1 mg/mL, showed no adverse effects on zebrafish embryos and larvae. These findings align with cell viability investigations, which demonstrated that higher antibiotic concentrations resulted in decreased cell proliferation and viability over time. Conversely, at a lower concentration of 1 mg/mL, cell proliferation notably increased after 72 hours. Plain MTAP and CHX exhibited the highest toxicity levels in the MTT assay. CONCLUSION:  The study concludes that while higher concentrations of MTAP hydrogel exhibit toxic effects, the hydrogel at 1 mg/mL demonstrates no adverse impact on zebrafish embryos, larvae, and DPSCs. These findings underscore the necessity of optimizing ICM concentrations to balance antibacterial efficacy and minimal cytotoxicity.

Exploring Periostracum as an Alternative Root Canal Irrigant: Insights From Zebrafish Embryo Experiments.

Objectives Root canal treatments aim to eliminate biofilms effectively. Considering the limitations of chemical irrigants, there is growing interest in natural alternatives like periostracum due to their antibacterial and fouling-resistant properties. This study aimed to assess periostracum's toxicity as a root canal irrigant by investigating its effects on zebrafish embryos' heart rate, survival rate, and hatching rate, as well as inflammation studies using neutral red assays comparing it to standard irrigants like ethylenediaminetetraacetic acid (EDTA), chlorhexidine (CHX), and sodium hypochlorite (NaOCl). Materials and methods Zebrafish embryos were exposed to varying concentrations of periostracum irrigant and standard irrigants. Heart rate, survival rate, and hatching rate were evaluated as indicators of developmental toxicity using microscopy. Statistical analysis, utilizing GraphPad Prism software (version 5.03, GraphPad Software, LLC, San Diego, California, United States), involved one-way ANOVA and Tukey's post-hoc test to determine significance levels (p < 0.05) across control and other groups based on triplicate means and standard deviation. Results The periostracum irrigant demonstrated superior survival rates, heart rates, and hatching rates at specific concentrations compared to standard irrigants (p < 0.01), maintaining favorable heart rates and hatching rates at those concentrations. However, higher concentrations resulted in diminished hatching rates (p < 0.05). Additionally, this study revealed increased inflammation when larvae were treated with NaOCl, EDTA, and CHX. Conversely, no inflammation was observed when subjected to periostracum irrigants. These findings suggest potential advantages of periostracum as a root canal irrigant due to its increased biocompatibility. Conclusion Periostracum displayed promising attributes in zebrafish embryo experiments, such as stable heart rate, hatching rate, and survival rate, along with reduced developmental toxicity and inflammation, indicating potential advantages as a root canal irrigant, including reduced toxicity compared to conventional agents. Further research involving diverse demographics and long-term effects is crucial to validate periostracum's clinical applicability and safety in endodontic therapies.

Comparison of Antibacterial Efficacy of Triple Antibiotic-Loaded Hydrogel Versus Modified Triple Antibiotic-Loaded Hydrogel as Intracanal Medicament Against Enterococcus faecalis: An In vitro Study.

OBJECTIVE: Triple antibiotic paste (TAP) is known to have an essential role in the success of endodontic treatment by eliminating pathogens from the root canal system. Unfortunately, it causes discolouration and cytotoxicity at high concentrations. The objective of this research was to assess and compare the antimicrobial effectiveness of various concentrations (1 mg, 5 mg, 10 mg) of TAP, TAP hydrogel (TAPH), M-TAP, and M-TAP hydrogel (MTAPH) against Enterococcus faecalis. METHODS: The agar well diffusion method was used to assess the antibiotic sensitivity of the following intracanal medicaments: TAP (ciprofloxacin, metronidazole, and minocycline) mixed in a ratio of 1: 1: 1; TAPH, M-TAP (ciprofloxacin, metronidazole, and amoxicillin), M-TAPH and plain hydrogel. Each tested medicament was individually evaluated for its antimicrobial activity against Enterococcus faecalis. Structural and topographical characterisation were analysed using a Scanning Electron Microscope (SEM) and interpreted using ImageJ software. A microdilution broth test was performed to examine the minimum inhibitory concentration and minimum bactericidal concentration (MBC) of M-TAP and TAP. RESULTS: Except for the plain hydrogel, M-TAP and hydrogel and TAP and hydrogel showed significantly varied inhibitory zones at different concentrations. M-TAPH showed the highest mean zone of inhibition of 21.6, 33.33 and 38.0 mm at a concentration of 1, 5, and 10 mg/mL when compared to TAPH, which showed a mean zone of inhibition of 3.3 mm,12.3 mm, 21.3 mm at the respective concentrations. The MIC study shows that more than 75% of Enterococcus faecalis growth was inhibited by M-TAP at a concentration of 5 µg/mL, whereas TAP showed inhibition at a concentration of 35 µg/mL. MBC results indicate that almost 99.9% of the bacterial population was killed at a concentration of 100 µg/mL (10-1) for TAP and 10 µg/mL (10-2) for M-TAP. CONCLUSION: The antibacterial efficacy of M-TAP was significantly higher than TAP. Application of M-TAP at lower doses is advised to overcome the disadvantages seen with TAP.

Biomimetic Approach to Counter Streptococcus mutans Biofilm: An In Vitro Study on Seashells.

Aim This study aimed to investigate the anti-adherent property of the seashell surface and periostracum to prevent the formation of Streptococcus mutans biofilm. Materials and methods The seashells were initially collected from the natural urban beach, and an antibiofilm assay of the shells with and without periostracum was performed against Streptococcus mutans. Furthermore, the seashells were analyzed with a stylus profilometer (Mitutoyo Surftest SJ-301, Mitutoyo America Corporation, Illinois, USA), atomic force microscope (AFM; Nanosurf Easyscan 2, Nanosurf Inc., USA), contact angle assessment, Fourier-transform infrared (FTIR) spectroscopy analysis, and scanning electron microscopy (SEM; JEOL USA, Inc., FE-SEM IT800, Massachusetts, USA) analysis. The ability of seashells to prevent the attachment of Streptococcus mutans and form a biofilm with and without periostracum was studied by crystal violet assay. Results The results revealed that shells without periostracum promoted higher biofilm formation when compared to those having intact periostracum (by 15%, p<0.001). Shell 1 showed the highest biofilm formation, whereas shell 3 showed the least biofilm formation due to the differences in their surface morphologies. The remaining shells (4, 2, 6, and 5) showed interspersed biofilm formation. Conclusion In summary, our study was able to correlate the topologies of the shell surface with the biofilm formed by Streptococcus mutans with the wetting behavior of those shell surfaces and their roughness. More hydrophobic surfaces (with intact periostracum) were observed to lead to less attachment (correlation coefficient=-0.67). This study can pave the way for designing such biomimetic surfaces to prevent bacterial attachment.

Influence of Sucrose and Arenga pinnata Solutions on Enamel Surface Demineralization: A Profilometric Study.

Background Dental caries is a multifactorial disease that has the potential to impact individuals across various life stages. The influential role of sugar as a contributing risk element in the inception and advancement of dental caries is significantly pronounced.  Aim The research aim was to analyze and compare the enamel surface roughness in teeth exposed to sucrose and Arenga pinnata (palm sugar) solutions by using a stylus profilometer Materials and methods In this investigation, 34 freshly extracted anterior teeth were obtained and they were split into two groups depending on the solution in which they were immersed. Group A consists of 17 teeth immersed in 1% sucrose solution supplemented in brain heart infusion (BHI) broth solution and Group B consists of 17 teeth immersed in 1% Arenga pinnata BHI broth. Each sample served as its own control. Streptococcus mutans was inoculated into these groups and they were immersed in their respective solution for five days. A stylus profilometer was utilized to measure the surface roughness of the teeth in this study. Data analysis involved paired t-tests for intragroup comparisons and independent t-tests for intergroup comparisons using SPSS software version 23. Results After five days of exposure to palm sugar or sucrose, it was observed that there was demineralization of the enamel surface on both samples. Although there was no statistical significance (p<0.05) when an independent t-test was conducted among these samples, there was a visible increase in the numerical values of Ra, Rq, Rz of teeth exposed to sucrose compared to palm sugar with a p-value of 0.529, 0.122 and 0.357, respectively. Conclusion From this study, it was concluded that although both sucrose and Arenga pinnata cause demineralization of enamel, it was shown that the latter caused lesser demineralization when compared to refined sugars to a certain extent. This study establishes a foundation for forthcoming investigations that could potentially explore the utilization of natural sugars as a substitute for sucrose, while also evaluating the mechanistic aspects underlying the impact of these sugars on enamel demineralization.