Introduction: This study is an experimental setup to examinethe effects of splenectomy, applied to multi-trauma patientswho are commonly seen in emergency traumatologypractices and on fracture healing processes with respect tohistopathological, biomechanical, and radiological aspects.Methods: Further, 32 male Sprague Dawley rats (10 monthsold; average weight 394.5±28.3 g) are included in the study.In a dark environment, rats are fed at 22 °C for 12 hourswith standard rodent food ad libitum. They are divided intotwo groups: splenectomy (total splenectomy and long bonefracture healing) and control (only long bone fracture healingn=16). Four months after surgery, rats are killed, and callustissues in fractured femurs were examined histopathologically,radiologically, and biomechanically.Results: In the radiological analysis of the femur materialsin relation to the case and control groups, there was nosubstantial difference in the Goldberg classification score(p˃0.05). Similarly, no substantial difference was observed(p˃0.05) in histopathological examinations and biomechanicalanalysis conducted in femur samples.Conclusion: When all of the results obtained in our studyare evaluated together, it was concluded in the rats to whichsplenectomy is applied that this did not affect fracture healinghistopathologically, biomechanically, and radiologically,despite the modification of immune modulators.