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Introduction: The successful treatment of infected or inflamed endodontic tissues requires chemomechanical debridement of the canal spaces and proper sealing of the coronal and apical canal openings. Only a few methods are available to further disinfect areas or initiate regeneration of local tissues. In this study, we assessed the ability of 255-nm and 405-nm light-emitting diode (LED) treatment to kill planktonic cultures of Enterococcus faecalis and induce the production of cellular biomarkers related to endodontic tissue regeneration., Methods: We determined the antimicrobial effects of 255-nm and 405-nm LED treatment on E. faecalis and the effects of 255-nm and 405-nm LED treatment on the production of osteoinductive, angiogenic, proliferative, and proinflammatory biomarkers from human embryonic palatal mesenchyme (HEPM) cells and gingival fibroblasts., Results: We showed that 255-nm LED but not 405-nm LED treatment killed E. faecalis; the 255-nm LED and sodium hypochlorite more efficiently killed E. faecalis; neither 255-nm nor 405-nm LED treatment affected the viability of HEPM cells and gingival fibroblasts; and 255-nm LED treatment, alone or in combination with 405-nm LED treatment, of HEPM cells and gingival fibroblasts induced the production of biomarkers related to endodontic tissue regeneration., Conclusions: The results of this study suggest a new treatment modality using short periods of 255-nm LED treatment as an adjunct to chemomechanical debridement for the disinfection of inflamed sites and the production of biomarkers related to endodontic tissue regeneration., (Copyright © 2019 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)