Abstract：ObjectiveTo observe the effect of Hyperthermia preconditioning olfactory ensheathing cells (OECs) combined with neural stem cells (NSCs) on spinal cord injury, and to provide reference for clinical treatment of spinal cord injury.MethodsNSCs were cultured with normal or 40℃ pretreated OECs supernatant respectively, and the number of NSCs differentiation into neurons were compared between the two groups. The spinal cord semi-transection injury model was established, then the rata with spinal cord injury were treated by combined transplantation of NSCs with OECs or hyperthermia pretreated OECs . Control group was injected with PBS. The motor recovery and pathological changes of the rats were observed, and the effect of hyperthermia preconditioning OECs on the repair of spinal cord injury was analyzed.ResultsIn the cell experiments, the proportion of NSCs differentiation into neurons in the hyperthermia preconditioning group was significantly higher than that in the control group. In animal experiments, the BBB scores of motor function in the hyperthermia preconditioning group increased rapidly 2 weeks after transplantation, and entered the slow rising stage 6 weeks after transplantation. The increase amplitude and speed were significantly higher than those in the other two groups, and the motor function recovered obviously. The results of rod climbing score and rotation time were similar to the BBB score. The pathological section of spinal cord showed that the hyperthermia preconditioning group had the best recovery effect of spinal cord tissue structure. BrdU tracer staining showed that the transplanted cells were widely distributed in the damaged area after repair.ConclusionHyperthermia preconditioning of OECs can promote the differentiation of NSCs into neurons. Combined transplantation of Hyperthermia preconditioning OECs and NSCs can effectively treat spinal cord injury.
[1] Kadoya K, Lu P, Nguyen K, et al.Spinal cord reconstitution with homologous neural grafts enables robust corticospinal regeneration[J]. Natu Med, 2016, 22(5): 479-487. [2] Zhou P, Guan J, Xu P, et al.Cell Therapeutic Strategies for Spinal Cord Injury[J]. Adv Wound Care (New Rochelle), 2019, 8(11): 585-605. [3] Lu P, Ceto S, Wang Y, et al.Prolonged human neural stem cell maturation supports recovery in injured rodent CNS[J]. J Clin Invest, 2017, 127(9): 3287-3299. [4] Wang G, Ao Q, Gong K, et al.Synergistic effect of neural stem cells and olfactory ensheathing cells on repair of adult rat spinal cord injury[J]. Cell Transplant, 2010, 19(10): 1325-1337. [5] Shpargel KB, Jalabi W, Jin Y, et al. Preconditioning paradigms and pathways in the brain [J]. Cleve Clin J Med, 2008, 75 Suppl 2: S77-82. [6] Chen C R, Li Y C, Young T H.Gallium nitride induces neuronal differentiation markers in neural stem/precursor cells derived from rat cerebral cortex[J]. Acta biomater, 2009, 5(7): 2610-2617. [7] Nash H H, Borke R C, Anders J J.New method of purification for establishing primary cultures of ensheathing cells from the adult olfactory bulb[J]. Glia, 2001, 34(2): 81-87. [8] Basso D M, Beattie M S, Bresnahan J C.Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection[J]. Exp Neurol, 1996, 139(2): 244-256. [9] Brailowsky S, Knight RT, Blood K, et al.γ-Aminobutyric acid-induced potentiation of cortical hemiplegia[J]. Brain Res, 1986, 362(2): 322-330. [10] Kleiser B, Van Reempts J, Van Deuren B, et al.Favourable effect of flunarizine on the recovery from hemiparesis in rats with intracerebral hematomas[J]. Neurosci Lett, 1989, 103(2): 225-228. [11] Curtis E, Martin J R, Gabel B, et al. A First-in-Human, Phase I Study of Neural Stem Cell Transplantation for Chronic Spinal Cord Injury [J]. Cell Stem Cell, 2018, 22(6): 941-950. e6. [12] Xie J, Li Y, Dai J, et al.Olfactory Ensheathing Cells Grafted Into the Retina of RCS Rats Suppress Inflammation by Down-Regulating the JAK/STAT Pathway[J]. Front Cell Neurosci, 2019, 13: 341 [13] Li Y, Huo S, Fang Y, et al.ROCK Inhibitor Y27632 Induced Morphological Shift and Enhanced Neurite Outgrowth-Promoting Property of Olfactory Ensheathing Cells via YAP-Dependent Up-Regulation of L1-CAM[J]. Front Cell Neurosci, 2018, 12: 489. [14] Wright AA, Todorovic M, Tello-Velasquez J, et al.Enhancing the Therapeutic Potential of Olfactory Ensheathing Cells in Spinal Cord Repair Using Neurotrophins[J]. Cell Transplant, 2018, 27(6): 867-878. [15] Beckingham LJ, Todorovic M, Tello Velasquez J, et al.Three-dimensional cell culture can be regulated by vibration: low-frequency vibration increases the size of olfactory ensheathing cell spheroids[J]. J Biol Eng, 2019, 13: 41. [16] Pespeni M, Hodnett M, Pittet JF.In vivo stress preconditioning[J]. Methods, 2005, 35(2): 158-164. [17] Du F, Zhu L, Qian ZM, et al.Hyperthermic preconditioning protects astrocytes from ischemia/reperfusion injury by up-regulation of HIF-1 alpha expression and binding activity[J]. Biochim Biophys Acta, 2010, 1802(11): 1048-1053. [18] Redaelli CA, Wagner M, Kulli C, et al.Hyperthermia-induced HSP expression correlates with improved rat renal isograft viability and survival in kidneys harvested from non-heart-beating donors[J]. Transpl Int, 2001, 14(6): 351-360. [19] Xu H, Aibiki M, Nagoya J.Neuroprotective effects of hyperthermic preconditioning on infarcted volume after middle cerebral artery occlusion in rats: role of adenosine receptors[J]. Crit Care Med, 2002, 30(5): 1126-1130. [20] Koffler J, Zhu W, Qu X, et al.Biomimetic 3D-printed scaffolds for spinal cord injury repair[J]. Nat Med, 2019, 25(2): 263-269.
2023,Vol. 20
