Evaluation of neuroprotective effects of different treatments in TBI induced Epileptogenesis in rats:
Model standardizing studies confirmed the altered neurobehavioral and brain complications results in futuristic PTE. Fasudil Hydrochloride (FH, 10mg/kg i.p.), Flufenamic Acid (FFA, 20mg/kg oral), valproic acid (VPA, 350mg/kg i.p.), and Valproic Acid + Flufenamic Acid (VFA, 175mg/kg i.p. + 10mg/kg oral) treatment groups were compared against disease and normal healthy controls to check the efficacy in TBI progression. TBI was induced into 6 treatment groups in which 2 were TBI control, EPLT group, and 4 were selected for treatment studies. One control and one surgery group i.e. Sham were also considered for comparisons to conclude the surgery effect. All the groups were analyzed for bodyweight difference, neurological severity score, and motor coordination on Day0, Day1, Day3, Day7, and Day14 before proceeding towards molecular estimation.
Animal Body Weight Difference: The weight severity profile was observed followed by the head injury. The line graph of weight difference on Day3, Day7, and Day14 showed a significant difference in weight difference (grams). All treatment groups i.e. FH, FFA, VPA, and VFA were found to be significantly effective to maintain the rat weight compared to disease controls i.e. TBI/EPLT groups on Day7. There was no significant difference observed between intra-treatment groups on Day7 (Figure-2).
Neurological Severity Scale Estimation: The NSS observed in the VFA group was more significant to restoring the neurological score on Day3, Day7, and Day14 when compared to the TBI and EPLT group. The other treatment groups showed a significant increase in NSS scores on Day3, Day7, and Day14 when compared to disease controls (Figure-2).
Rotarod Test for Motor Co-ordination: The latency to fall (in seconds) was found to be significant for all treatment groups when compared to disease controls on Day7 and Day14 (Figure-2).
Seizure Score assessment after Brain Trauma: A sub-convulsive dose of PTZ(35mg/kg) was injected on day14 and injured rats were observed for seizure scoring for 2hours. Increased susceptibility for seizures was confirmed in the EPLT group with a low seizure threshold. EPLT group showed behavioral arrests, complex partial, dileptic, and tonic-colonic seizures. But no seizures were observed in any treatment groups (Table in Figure-2).
Brain Infarction by TTC Staining:The infarction was visible in the striatum and cortex leaving the hippocampus. Control and Sham group observed deep red in staining with normal histology with no infarction in the brain. TBI and EPLT groups were showed hypoperfusion significantly in the area of infarct (white pale color) when compared to normal controls. FH, FFA, VFA treatment groups showed a significant difference to the disease control in red staining i.e. less hypoperfusion and rich red stain when compared to TBI and EPLT groups. But VPA significantly shows nearly red staining of injured brain tissue with very less area of infarct (Figure-3).
Evan’s Blue concentration: Evan’s blue (2%, 4mL/kg i.p.) was administered in rats and an abrupt increase of Evan’s blue was seen in TBI and EPLT groups. But the Evans concentration in FH, FFA, VPA, and VFA groups was found significantly less compared to injury controls. VPA alone and in combination with FFA was found very effective as compared to FH and FFA alone (Figure-3).
Brain Edema difference:Surgically removed rat brains were weighed just after the sacrifice and after 24 and 48 hours when placed in an incubator at 70oC. The brain edema in all the treatment groups showed no significant differences when compared to disease controls after 24 hours but there is a significant difference after 48 hours i.e. the value of water content in the treatment groups was found less as compared to the TBI/EPLT groups (Figure-3).