Photo-activated disinfection using photosensitizers has been proposed as an adjunct to conventional endodontic therapy for additional disinfection and improved treatment outcomes. This systematic review aims to investigate the effectiveness of photosensitizers used in laser endodontics disinfection. A comprehensive literature search is conducted to identify in vitro and in vivo studies involving photo-activated disinfection in endodontic treatment. Publications are selected based on predetermined eligibility criteria. From 121 articles, 34 met the inclusion criteria, comprising eight polymicrobial and 26 bacteria-specific studies. A total of 10 photosensitizers were investigated, including methylene blue, tolonium chloride, polyethylenimine and chlorin (e6), phenothia zinium, chitosan, rose bengal, malachite green, indocyanine green, chlorine p6 and curcumin. Photo-activated disinfection using tolonium chloride or methylene blue can be an effective adjunct to conventional root canal therapy. More studies are needed to assess the performance of newer photosensitizers to optimize photoactivated disinfection of the complex root canal space.