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Aim:To assess the effect of chemo-mechanical root canal preparation using ProTaper Next and Videya N rotary files with and without root canal filling on the fracture resistance of roots.Methodology: A total of 40 freshly extracted mandibular premolars were decoronated and randomly divided into 5 equal groups (n=8) according to the files used as follows; group 1 prepared using ProTaper Next files without filling; group 2 prepared using ProTaper Next files and filled with gutta percha and Totalfilbioceramic sealer; group 3 prepared using Videya N files without filling; group 4 prepared using Videya N files and filled with gutta percha and Totalfilbioceramic sealer; and group 5 without preparation or filling as a control. All roots were subjected to load using universal testing machine till fracture and load needed was recorded in Newtons. The data were statistically analyzed using one-way ANOVA test, and post hoc analysis. Results:The 4 experimental groups significantly lowered the roots' fracture resistance compared to the control group without a statistically significant difference between the experimental groups.The filled groups showed a non-statistically significant increased fracture resistance compared to the non-filled ones. Conclusion:Chemo-mechanical root canal preparation decreases the roots' fracture resistance.Root canal filling using bioceramic sealers might enhance the roots' fracture resistance.