Biochemical properties of novel Carbon nanodot-stabilized silver nanoparticles enriched calcium hydroxide endodontic sealer

Abstract

Calcium Hydroxide-based endodontic sealer loaded with antimicrobial agents have been commonly employed in conventional root canal treatment. These sealers are not effective against E. faecalis due to the persistent nature of this bacterium and its ability to evade the antibacterial action of calcium hydroxide. Therefore, endodontic sealer containing Carbon nanodots stabilized silver nanoparticles (CD-AgNPs) was proposed to combat E. faecalis. The therapeutic effect of CD-AgNPs was investigated and a new cytocompatible Calcium Hydroxide-based endodontic sealer enriched with CD-AgNPs was synthesized that exhibited a steady release of Ag+ ions and lower water solubility at 24 hours, and enhanced antibacterial potential against E. faecalis. CD-AgNPs was synthesized and characterized morphologically and compositionally by Scanning Electron Microscopy, Fourier Transform Infrared Spectroscopy (FTIR), and UV-Vis Spectroscopy, followed by optimization via minimum inhibitory concentration (MIC) determination against E. faecalis by broth microdilution technique and Cytotoxicity analysis against NIH3T3 cell lines via Alamar Blue assay. Calcium hydroxide in distilled water was taken as control (C), Calcium hydroxide with to CD-AgNPs (5mg/ml and 10mg/ml) yielded novel endodontic sealers (E1 and E2). Morphological and chemical analysis of the novel sealers were done by SEM and FTIR; followed by in vitro assessment for antibacterial potential against E. faecalis via agar disc diffusion method, release of Ag+ ions for 21 days by Atomic Absorption Spectrophotometry and water solubility by weight change for 21 days. CD-AgNPs were 15–20 nm spherical-shaped particles in uniformly distributed clusters and revealed presence of constituent elements in nano-assembly. FTIR spectra revealed absorption peaks that correspond to various functional groups. UV-Vis absorption spectra showed prominent peaks that correspond to Carbon nanodots and Silver nanoparticles. CD-AgNPs exhibited MIC value of 5mg/ml and cytocompatibility of 84.47% with NIH3T3 cell lines. Novel endodontic sealer cut-discs revealed irregular, hexagonal particles (100–120 nm) with aggregation and rough structure with the presence of constituent elements. FTIR spectra of novel endodontic sealers revealed absorption peaks that correspond to various functional groups. Novel endodontic sealers exhibited enhanced antibacterial potential where E-2 showed greatest inhibition zone against E. faecalis (6.3±2 mm), a steady but highest release of Ag+ ions was exhibited by E-1 (0.043±0.0001 mg/mL) and showed water solubility of <3% at 24 hours where E-2 showed minimal weight loss at all time intervals. Novel endodontic sealers were cytocompatible and showed enhanced antibacterial potential against E. faecalis, however, E2 outperformed in this study in all aspects.