الفهرس 
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Abstract
The rational of the current research was to realize the therapeutic outcome of BM-MSCs and AD-MSCs against acute and chronic epileptogenic alterations implicated in pilocarpine-induced TLE.
BM-MSCs were obtained from bone marrow of both femur and tibia of male Wistar rats. While, AD-MSCs were isolated from omentum of male Wistar rats. Then mesenchymal stem cells were propagated and characterized by microscopic examination and surface markers expression (CD29, CD73, CD34 and CD45). Additionally, accommodating of the injected MSCs to the injured brain tissues was established by labeling them with PKH26 dye.
One hundred and thirty-five adult male Wistar rats were randomly distributed into nine groups (each group contained 15 rats): group 1: placebo group, in which the rats were injected with all drugs taken by acute epileptic group except pilocarpine that was replaced with equivalent volume of sodium chloride saline (NaCl) 0.9% by intraperitoneal injection, group 2: Acute epileptic group, in which the rats were injected intraperitoneally once with methyl scopolamine (1mg/kg b.wt.) 30 mins prior to single intraperitoneal pilocarpine injection (380 mg/kg b.wt.). After that, all animals received two doses of diazepam separated by one hour (each 4 mg/kg b.wt., orally) 30 mins after seizure onset. These rats were sacrificed after one week from final administration, group 3: Acute epileptic group treated with BM-MSCs, in which the rats were treated as in Gp2, after that they were infused with undifferentiated BM-MSCs (2x106 cells/rat) intravenously and left for two month before being sacrificed, group 4: Acute epileptic group treated with AD-MSCs, in which the rats were treated as in Gp2, after that they were infused with undifferentiated AD-
MSCs (2x106 cells/rat) intravenously and left for two month before being sacrificed, group 5: Acute epileptic group treated with CBZ, in which the rats were treated as in Gp2, after that they were received CBZ in a daily dose of 120 mg/kg b.wt. (40 mg/kg b.wt., orally, three times/day) for two month before being sacrificed, group 6: chronic epileptic group, in which the rats were treated as in Gp2, after that they were sacrificed after two month from final administration, group 7: chronic epileptic group treated with BM-MSCs, in which the rats were treated as in Gp6, after that they were infused with undifferentiated BM-MSCs (4x 106 cells/rat) intravenously and left for four month before being sacrificed, group 8: chronic epileptic group treated with AD-MSCs, in which the rats were treated as in Gp6, after that they were infused with undifferentiated AD-MSCs (4x106 cells/rat) intravenously and left for four month before being sacrificed, group 9: chronic epileptic group treated with CBZ, in which the rats were treated as in Gp6, after that they were received CBZ in a daily dose of 120 mg/kg b.wt. (40 mg/kg b.wt., intragastrically, three times/day) for four month before being sacrificed.
HSP-70, S100β, caspase-8, GABA, Sema4D and galanin concentrations were evaluated in brain via ELISA. Whereas, gene expression levels of hippocampal TLR-4 and synapsin-I were estimated using sqRT-PCR. In addition, histological exploration of the hippocampus and cerebral cortex were carried out.
The results of the current work disclosed that:
1. MSCs were successfully isolated as demonstrated through their morphological spindle shape and the expression of their specific surface signals. MSCs were accommodated in the brain tissues of acute and chronic epileptic rats treated with BM-MSCs or AD-MSCs illustrating that the delivered undifferentiated MSCs were capable of homing to the injured brain tissues.
2. The biochemical findings manifested significant intensification in brain HSP-70, S100β, caspase-8 contents associated with significant decline in brain GABA, galanin and Sema4D concentrations after pilocarpine injection. In contrast, BM-MSCs and AD-MSCs transplantation or CBZ administration constructed significant decline in brain HSP-70, S100β, caspase-8 contents and significant upgrade in GABA, galanin, Sema4D concentrations.
3. Molecular genetic analysis revealed that pilocarpine up-regulates hippocampal TLR-4 mRNA level and down-regulates synapsin-I gene expression level. Moreover, all treatments (BM-MSCs, AD-MSCs and CBZ) decrease hippocampal TLR-4 mRNA level and elevate synapsin-I gene expression level.
4. Optical micrograph of brain tissue sections of rats in acute and chronic epileptic groups revealed nucleus pyknosis and degeneration in most neurons in cerebral cortex and different hippocampal areas. On the opposition, photomicrograph of brain tissue sections of rats in acute and chronic epileptic groups after BM-MSCs transplantation indicated nucleus pyknosis and degeneration of some neurons in cerebral cortex as well as clarified normal histological structure of neurons of subiculum, fascia dentata and hilus in hippocampus. Microscopic investigation of the transverse section through the brain of rats in acute and chronic epileptic groups treated with AD-MSCs identified nucleus pyknosis along with degeneration in some neurons of cerebral cortex, while hippocampal neurons showed normal histological structure of subiculum, fascia dentata and hilus. Photo-documentation of brain tissue sections of rats in acute epileptic group treated with CBZ manifested nucleus pyknosis and degeneration in some neurons of cerebral cortex and different hippocampal regions (subiculum, fascia dentata and hilus). While, microscopic follow up of the transverse section through the brain of rats in the chronic epileptic group treated with CBZ indicated the presence of nucleus pyknosis along with degeneration in most neurons of cerebral cortex, while, the micrograph of subiculum of hippocampus demonstrated normal histological architecture of the neurons. However, photomicrograph of hippocampal fascia dentata and hilus areas clarified nucleus pyknosis and degeneration in some neurons