The present study aimed to evaluate the antimicrobial efficacy of silver nanoparticles infused with Azadirachta indica extract and chlorhexidine against red-complex periopathogens. Neem leaf extraction was done followed by standardization to the synthesis of neem-infused silver nanoparticles and fractionation of compounds done by using thin layer chromatography to separate the mixture of neem leaf extract. Characterization of neem-infused silver nanoparticles was done by scanning electron microscopy and UV-Visible spectroscopy. The compound identified in neem-infused silver nanoparticles was gedunin which was confirmed by Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. Determination of antibacterial activity done by disc diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) methods. Group I-99% ethanolic extract, group II-neem-infused silver nanoparticles (NAgNPs), group III-chlorhexidine. The relative inhibitory zone value for Tannerella forsythia (180) in neem-infused silver nanoparticles (group II) was greater when compared with other periopathogens Porphyromonas gingivalis (133) and Treponema denticola (160) than 99% ethanolic extract (group I), chlorhexidine (group III). Neem-infused silver nanoparticles (group III) showed superior antimicrobial activity against T. forsythia (19.3 ± 31.1547) and T. denticola (18±0) when compared with P. gingivalis (17.6 ± 0.5774). On evaluating MIC and minimum bacterial concentrations, P. gingivalis is more resistant than other pathogens in neem-infused silver nanoparticles (group III). Neem-infused silver nanoparticles exhibited superior antibacterial activity as compared with gold-standard chlorhexidine against red-complex periodontal pathogens. For MIC and MBC all the three periopathogens were effective but P. gingivalis was more resistant. Antibiotics are effective against many drug-resistant bacteria. As a ready-made medicine, they can be used to treat many infections. Silver nanoparticles in drug delivery systems generally increase solubility, stability, and biodistribution, thereby increasing their effectiveness. Green synthesis using plant extracts as precursors to synthesize nanoparticles has proven to be environmentally non-hazardous combined with remarkably improved efficacy against bacterial and viral diseases. So neem-infused silver nanoparticles can be utilized as a drug delivery system. Hence, it can be used as a potential antibacterial ingredient in formulations for periodontal use like mouthwashes and gels for local drug delivery. How to cite this article: Krishnappan S, Ravindran S, Balu P, et al. Comparative Evaluation of Antimicrobial Efficacy of Silver Nanoparticles Infused with Azadirachta indica extract and Chlorhexidine Against Red-Complex Pathogens. J Contemp Dent Pract 2024;25(6):547-553.