A proper bond between root canal filling materials and dentin surface is essential to resist dislodgement and guarantee long-term success. Blood exposure is likely to occur in various clinical situations in which calcium silicate-based materials are used; therefore, it is fundamental to render data concerning the influence of blood on bond strength. The present study aims to evaluate the effect of blood contamination on the push-out bond strength obtained with three different biomaterials to root canal dentin; Ninety extracted human mono-radicular permanent teeth were selected. The root canals were prepared with Gates Glidden burs until a diameter of 1.10 mm was achieved. Teeth were then randomly divided into six experimental groups (n = 15) according to the presence/absence of blood contamination and biomaterial used for root canal filling (ProRoot® MTA, BiodentineTM, and TotalFill® BC Putty). After one week, each root was sectioned in three segments (coronal, middle, and apical regions). Specimens were then submitted to push-out bond strength tests. Fracture pattern evaluation was performed. The significance level was set at 5%.; Blood contamination did not affect the push-out bond strength of any of the three tested calcium silicate-based cements (p > 0.05). Regardless of blood contamination, TotalFill showed statistically higher push-out bond strength when compared with Biodentine (p = 0.040) and MTA (p = 0.004). Biodentine exhibited higher bond strength than MTA (p = 0.043). Biomaterials’ comparison within each radicular segment revealed statistically superior bond strength of both Biodentine and TotalFill over MTA (p < 0.05) in the coronal segment. TotalFill presented higher push-out bond strength regarding the apical segment compared to Biodentine (p = 0.003). Fractures were mostly adhesive.; Overall results indicate TotalFill presents the highest push-out bond strength values, followed by Biodentine and, lastly, MTA. Blood contamination did not affect the dislodgement resistance. Biomaterials’ comparison within each radicular segment revealed both TotalFill and Biodentine as the preferable alternatives for application in the coronal region. TotalFill might be the biomaterial of choice for placement in the apical region.