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Resin-based root canal sealer can bond to dentin and establish a hermetic seal. The aim of this study was to evaluate the physicochemical properties, sealing ability, cytotoxicity and bond strength of a new resin-based root canal sealer (NRCS). Methods: The physicochemical properties were assessed by the flow, setting time, solubility, film thickness, radiopacity and dimensional changes. Sixty premolar root samples were filled with either Resilon/NRCS or Resilon/Epiphany and sectioned perpendicularly at the long axis at 2 mm below the cemento-enamel junction into 1-mm serial slices. The bond strength was tested by a universal testing machine. The glucose microleakage model was used to test the sealing ability. Elutes of NRCS and Epiphany were co-cultured with human periodontal ligament cells to test the cytotoxicity. Results: All the physicochemical properties of NRCS conformed to ISO 6876:2001(E). The root samples filled with Resilon/NRCS had significantly less leakage (p < 0.01) and greater bond strength (p < 0.001) than the Resilon/Epiphany group had. Environmental scanning electron microscopy showed that Resilon/NRCS filling material was intimately bonded to the root dentin. Although NRCS was slightly toxic to human periodontal ligament cells, its cytotoxicity was significantly less than that of Epiphany (p < 0.01) Conclusion: NRCS has better physicochemical and sealing properties, as well as lower cytotoxicity and microleakage than Epiphany has