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Background: Although traumatic brain injury (TBI) is an important problem, there has been no widespread utilization of neuro-biomarkers to aid the diagnosis of TBI. This study was conducted to evaluate serum S100B and prion protein (PrPC) levels in rats with TBI., Methods: In this study, 15 albino rats were categorized into three groups as follows: sham-operated (1), control (6) and trauma (8) groups. The TBI model was based on the modified free falling model. S100B, PrPC levels were measured using ELISA. Brain specimens were obtained for the pathological examination., Results: Serum S100B and PrPC levels were found to increase in T group at both 2h and 24h after trauma (p<0.002, p<0.002, respectively). We also found higher histopathological injury scores of brain tissues in the T group. Only a positive correlation was found between serum PrPC levels and the extent of brain injury (p=0.039, r=0.731). Using ROC analysis, among the two serum markers investigated, both of them revealed the same sensitivity and specificity for diagnosing TBI., Conclusion: The changes in serum S100B and PrPC levels showed good sensitivity in our experimental model. Therefore, PrPC could be helpful in the early prognostic prediction in patients with TBI. Further studies are needed to test our findings in humans following TBI (penetrating bodies, blunt trauma) to definitively acknowledge it as a reliable biomarker and its subsequent diagnostic utility.