Olfactory Ensheathing Cells Group Research Articles

Effect of multifactorial therapeutic approach on axonal regeneration and cell viability in an in-vitro model of spinal-derived neural injury.

The highly debilitated nature of spinal cord injuries (SCI) creates an inhibitory repair environment that limits the recovery rate and therefore single interventional treatment has been resulted in incomplete recovery. A multifactorial approach that combines several therapeutic approaches may address diverse aspects of SCI pathology and enhance the recovery rate over single therapy. Accordingly, in this study, we aimed to investigate the effect of combined olfactory ensheathing cells (OECs) (to transport trophic factor, mediate immunomodulation, provide a suitable environment for cell survival), G-CSF (to establish a favorable environment for cell survival) and lipopolysaccharide (LPS) (to boost the protective activity of OEC) therapy on the cell viability after a scratch injury caused by a cataract knife on cells in an in-vitro model of spinal-derived neural injury. In this study, we used mixed neuronal-glial cultures, which are widely used for an in vitro study of neuronal damage. Scratch insult was made on cells using a cataract knife. The cells were divided into 8 groups (two control groups with and without olfactory ensheathing cells (OECs) treatment, injury group, three injury groups with single therapy by using super low dose of LPS (SLD-LPS) (100pg/ml), OEC group, and G-CSF (100ng/ml) group, and two injury groups with combined therapy (OEC with SLD-LPS and with all three treatments)). We found a significant decrease in the survival rate of injured cells (p < 0.001) 24h after scratching insult. Our results indicated morphological alterations in cells in the acute phase (1, 2 and 6h) after injury, with significant increased gap size at 6h after induction of injury. Our combined therapy, significantly prevented cell death and decreased the size of the gap over time. We found that combined therapy promoted cell survival following spinal injury by providing a neuroprotective environment for cells. Therefore, our findings provide new insight into the combined therapy, which can be considered for promising preclinical therapeutic strategy for SCI toward clinical trials.

Minocycline Enhance the Restorative Ability of Olfactory Ensheathing Cells by the Upregulation of BDNF and GDNF Expression After Spinal Cord Injury

Introduction:Spinal Cord Injury (SCI) is a global public health issue that results in extensive neuronal degeneration, axonal and myelin loss, and severe functional deficits. Neurotrophic factors are a potential treatment for reducing secondary damage, promoting axon growth; they are responsible for inducing myelination after injury. Olfactory Ensheathing Cells (OECs) and minocycline have promoted locomotor function after SCI. The present study investigated the neuroprotective effects of combined treatment with minocycline and OECs on spinal cord injury related to Brain-Derived Neurotrophic Factor (BDNF) and Glial Derived Neurotrophic Factor (GDNF) expressions after SCI.Methods:Adult female rats were used to experimental SCI by weight compression method. Rats received an intraperitoneal minocycline injection (90 mg/kg) immediately after SCI and 24 h after injury. OECs were transplanted one week after the injury. The hindlimb function was assessed using Basso Beattie Bresnahan (BBB) locomotor rating scale and Electromyography (EMG). After 5 weeks, the spinal cord segment centered at the injury site was removed for histopathological analysis. Immunohistological and western blot assays were performed to observe the expression of NeuN, BDNF, GDNF, and Myelin Basic Protein (MBP).Results:SCI induced the loss of locomotor function with decreased BDNF and GDNF expressions in the injury site. Minocycline+OECs increased the score of the BBB locomotor scale and increased spared tissue in the injury site. Immunohistochemical results suggested that NeuN expression significantly increased in the minocycline+OECs group than other groups. Moreover, electromyography amplitude in treated rats was increased compared to the control group. BDNF, GDNF, and MBP expressions and the number of ventral motor neurons increased further by minocycline+OECs in SCI rats.Conclusion:The present study provides evidence that minocycline may facilitate recovery of locomotor function by OECs by increasing BDNF and GDNF expressions following SCI.HighlightsCombined treatment with Minocycline and OECs increased the locomotor function.The results showed that BDNF and GDNF expression increased by combined treatment with minocycline and OECs.Plain Language SummaryThis study examined the effect of combined treatment with minocycline and olfactory ensheathing cell on the BDNF and GDNF expression after spinal cord injury model in rat. The results showed that injection of minocycline before transplantation of OECs enhances expression of neurotrophic factors that lead to an appropriate environment for transplanted OECs and increases neuronal survival that promotes tissue sparing and functional recovery.

Olfactory ensheathing cells in chronic ischemic stroke: A phase 2, double-blind, randomized, controlled trial

Olfactory ensheathing cells (OECs) have shown promising results for patients with neurologic diseases in non-double-blind, placebo control studies. Thirty patients with a unilateral ischemic stroke of more than a year were enrolled in a phase 2, multicenter, randomized, double-blind, and placebo-controlled cell therapy trial with a subsequent 12-month follow-up. The primary therapeutic objective has shown that after 12 months, there were significant differences in National Institutes of Health Stroke Scale (NIHSS), modified Rankin Scale (mRS) and Barthel Index (BI) assessment scores among the OEC group, Schwann cell group and placebo medium group at one-year follow-up. The second therapeutic objective found that there were significant differences in NIHSS, mRS, and BI assessment scores when comparing the endpoint data with the baseline data in the OEC group. There was neither hypersensitivity reaction nor adverse event. The results of this multicenter, randomized, double-blind, and placebo-controlled study indicate that injecting OECs into the olfactory sub-mucosa have neurorestorative effects, which can improve the quality of life for patients with chronic ischemic strokes without serious side effects.

Olfactory ensheathing cells improve the survival of porcine neural xenografts in a Parkinsonian rat model.

Parkinson's disease (PD) features the motor control deficits resulting from irreversible, progressive degeneration of dopaminergic (DA) neurons of the nigrostriatal pathway. Although intracerebral transplantation of human fetal ventral mesencephalon (hfVM) has been proven effective at reviving DA function in the PD patients, this treatment isclinically limited by availability of hfVMand the relatedethical issues. Homologous tissuesto hfVM, such as porcine fetal ventral mesencephalon (pfVM) thus present a strong clinical potential if immune response following xenotransplantation could be tamed. Olfactory ensheathing cells (OECs) are glial cells showing immunomodulatory properties. It is unclear but intriuging whether these properties can be applied to reducing immune response following neural xenotransplantation of PD. To determine whether OECs may benefit neural xenografts for PD, different compositions of grafting cellswere transplanted into striatum of the PD model rats. We used apomorphine-induced rotational behavior to evaluate effectiveness of the neural grafts on reviving DA function. Immunohistochemistry was applied to investigate the effect of OECs on the survival of neuroxenografts and underlyingmechanisms ofthis effect. Four weeks following the xenotransplantation, we found that the PD rats receiving pfVM+OECs co-graft exhibited a better improvement in apomorphine-induced rotational behavior compared with those receiving only pfVM cells. This result can be explained by higher survival of DA neurons (tyrosine hydroxylase immunoreactivity) in grafted striatum of pfVM+OECs group. Furthermore, pfVM+OECs group has less immune response (CD3+ T cells and OX-6+ microglia) around the grafted area compared with pfVM only group. These results suggest that OECs may enhance the survival of the striatal xenograftsvia dampeningtheimmune responseat the grafted sites. Using allogeneic OECs as a co-graft material for xenogeneic neural grafts could be a feasible therapeutic strategy to enhance results and applicability of the cell replacement therapy for PD.

Microencapsulated olfactory ensheathing cell transplantation reduces P2X4 receptor overexpression and inhibits neuropathic pain in rats

ObjectivesThe aim of this study was to determine the role of microencapsulated olfactory ensheathing cells (MC-OECs) transplantation in rats with sciatic nerve injury-induced neuropathic pain, and its relationship with P2X4 receptor expression in the L4-5 spinal cord segment. MethodsOlfactory bulb tissues of healthy Sprague Dawley (SD) rats were collected to culture olfactory ensheathing cells (OECs) using differential attachment methods. Seventy-two healthy SD rats were randomly assigned to four groups: the sham, chronic constriction injury (CCI), OECs, and MC-OECs groups. Mechanical paw withdrawal thresholds were measured on days 7, 14 and 21 after surgery. The expression of P2X4 receptor genes in the L4-5 spinal cord segment was detected by in situ hybridization, western blotting and RT-PCR. ResultsOn post-surgical days 7, 14 and 21. The mechanical paw withdrawal thresholds of rats from low to high were the CCI, OECs, MC-OECs and sham groups. The expression level of P2X4 receptor mRNA and protein from low to high were the sham, MC-OECs, OECs and CCI groups. Compared with the OECs group, the mechanical paw withdrawal thresholds of rats were increased, and the expression level of P2X4 receptor mRNA and protein were decreased in the MC-OECs group. In addition, the mechanical paw withdrawal thresholds of rats were higher and the expression level of P2X4 receptor were lower in the MC-OECs group on post-surgical days 21 than that on days 7 and 14. All differences between groups were statistically significant (P value < 0.05). ConclusionsOECs and MC-OECs transplantation can reduce the expression level of P2X4 receptor and relieve pain. The therapeutic efficacy was better in the MC-OECs group than in the OECs group. MC-OECs transplantation have the better effects of treatment with the increase of MC-OECs transplantation time.

Olfactory Ensheathing Cells Grafted Into the Retina of RCS Rats Suppress Inflammation by Down-Regulating the JAK/STAT Pathway

The inflammatory microenvironment in the retina plays a vital role in the pathogenesis and progression of retinitis pigmentosa (RP). Microglial inflammatory cytokines production leads to gliosis and apoptosis of retinal neurons, and ultimately, visual loss. Cell-based therapies using grafted olfactory ensheathing cells (OECs) have demonstrated modulation of degenerative microenvironments in the central nervous system (CNS), in a number of animal models. However, mechanisms by which grafted OECs can reduce degeneration in the retina are not well understood. In the present study, we set up an in vitro OEC/BV2 microglia co-culture system, and an in vivo royal college of surgeons (RCS) rat model, used cell transplantation, immunohistochemistry, RT-PCR, western blot to explore the mechanisms by which OECs affect expression of pro- or anti-inflammatory cytokines and polarization of M(IL-6) and M(Arg1) type microglial activation in the retina. We found that compared with the LPS (Lipopolysaccharide) and olfactory nerve fibroblast (ONF), the OEC and BV2 co-culture group modulate microglial cytokines releasing toward the anti-inflammation, and away from the pro-inflammation, which was followed by higher IL-4 and IL-10 and lower TNF-a and IL-6 in their expression levels. In vivo, the transplantation group significantly reduced activated resident microglia/infiltrated macrophage, and expression of pro-inflammatory cytokines in RCS rats retina, increased anti-inflammatory cytokines in transplantation area. Additionally, we found that OECs expressed SOCS3 and down-regulated the JAK2/STAT3 (Janus Kinase 2/Signal Transducer and Activator of Transcription 3) pathway. Thirdly, OEC transplantation reduced Caspase-3 expression, protected inner retinal neurons and photoreceptors and therefore, delayed the visual function degeneration. In conclusion, our data suggest that OECs delay retinal degeneration in RP, at least in part through immunomodulation of microglia via the JAK/STAT pathway.

Ginsenoside Rg1 Promotes the Migration of Olfactory Ensheathing Cells via the PI3K/Akt Pathway to Repair Rat Spinal Cord Injury.

The aim of this study was to determine the effects of ginsenoside Rg1 on the migration of olfactory ensheathing cells (OECs) in vitro, and its influence on the therapeutic efficacy of OECs transplanted in vivo for the treatment of spinal cord injury (SCI). Primary cultured and purified OECs (prepared from rats) were treated with ginsenoside Rg1. The wound healing test indicated that ginsenoside Rg1 promoted the migration of OECs. Real-time RT-PCR demonstrated that ginsenoside Rg1 upregulated the expression of migration-related factors of OECs, including matrix metalloproteinases-2 (MMP-2), MMP-9, and neural cell adhesion molecule 1 (NCAM1). Moreover, Western blot analysis indicated that ginsenoside Rg1 significantly promoted the migration of OECs via the phosphatidylinositol 3-kinase (PI3K)/Akt pathway. An SCI rat model was induced in vivo using a revised Allen's method. The Basso, Beattie, and Bresnahan (BBB) scores and histological analysis demonstrated that OECs, which were treated with ginsenoside Rg1, exhibited significant improvement in SCI compared with both the control group and the OEC group. Thus, ginsenoside Rg1 may represent a novel treatment target for SCI.

Olfactory ensheathing glia cell therapy and tubular conduit enhance nerve regeneration after mouse sciatic nerve transection

The regenerative potential of the peripheral nervous system (PNS) is widely known, but functional recovery, particularly in humans, is seldom complete. Therefore, it is necessary to resort to strategies that induce or potentiate the PNS regeneration. Our main objective was to test the effectiveness of Olfactory Ensheathing Cells (OEC) transplantation into a biodegradable conduit as a therapeutic strategy to improve the repair outcome after nerve injury. Sciatic nerve transection was performed in C57BL/6 mice; proximal and distal stumps of the nerve were sutured into the collagen conduit. Two groups were analyzed: DMEM (acellular grafts) and OEC (1×105/2μL). Locomotor function was assessed weekly by Sciatic Function Index (SFI) and Global Mobility Test (GMT). After eight weeks the sciatic nerve was dissected for morphological analysis. Our results showed that the OEC group exhibited many clusters of regenerated nerve fibers, a higher number of myelinated fibers and myelin area compared to DMEM group. The G-ratio analysis of the OEC group showed significantly more fibers on the most suitable sciatic nerve G-ratio index. Motor recovery was accelerated in the OEC group. These data provide evidence that the OEC therapy can improve sciatic nerve functional and morphological recovery and can be potentially translated to the clinical setting.

Ginsenosides Rg1 enhance the ability of olfactory ensheathing cells to repair spinal cord injury: An experimental study

Objective To study the effects of ginsenoside Rg1 on effects of olfactory ensheathing cells (OECs) in spinal cord injury as well as their expression of neurotrophic factors like ciliary neurotrophic factor (CNTF), vascular endothelial growth factor (VEGF). Methods OECs were primary cultured and purified from olfactory bulb of newborn SD rats. Purified cells were divided into 2 groups, the experimental group was treated with Rg1 in concentration of 40 μg/ml for 72 h (the optimal concentration and action time which have been proven by our previous study), the control group received an equal volume of Dulbecco's Modified Eagle Media: Nutrient Mixture F-12 (DMEM/F12) medium. The models of spinal cord injury was established by using Allen's combat, and were randomly divided into 3 groups, OECs group (inject the control group cells), Rg1+ OECs group (inject the experimental group cells) and control group (inject an equal volume of DMEM/F12 medium). Using reverse transcriptase-polymerase chain reaction (RT-PCR) to determine mRNA expression of CNTF and VEGF in OECs group and Rg1+ OECs group. Results The Basso Beattie Bresnahan (BBB) score of Rg1+ OECs group was higher than the other two groups (15.0±1.0 vs.12.0±2.0, 7.4±0.9, P<0.05). Histology revealed that Rg1+ OECs group had larger number of neurons (64±3, 38±2, 26±2, P<0.01), lower level of GFAP expression and less damage of spinal cord tissue than OECs group and control group (8.65±0.32, 4.93±0.25, 2.21±0.17, P<0.01). RT-PCR analysis demonstrated that Rg1 significantly up-regulated mRNA expression of CNTF and VEGF (0.78±0.03, 0.39±0.03; 0.86±0.03, 0.61±0.02, P<0.05). Conclusion Ginsinoside Rg1 obviously improve the effects of olfactory ensheathing cell transplantation for spinal cord injury by the up-regulation of CNTF and VEGF. Key words: Ginsenoside Rg1; Olfactory ensheathing cells; Spinal cord injury; Cell transplantation; Gene expression

The Effects of Co-transplantation of Olfactory Ensheathing Cells and Schwann Cells on Local Inflammation Environment in the Contused Spinal Cord of Rats.

Inflammatory response following spinal cord injury (SCI) is important in regulation of the repair process. Olfactory ensheathing cells (OECs) and Schwann cells (SCs) are important donor cells for repairing SCI in different animal models. However, synergistic or complementary effects of co-transplantation of both cells for this purpose have not been extensively investigated. In the present study, we investigated the effects of co-transplantation of OECs and SCs on expression of pro- or anti-inflammatory factor and polarization of macrophages in the injured spinal cord of rats. Mixed cell suspensions containing OECs and SCs were transplanted into the injured site at 7days after contusion at the vertebral T10 level. Compared with the DMEM, SC, or OEC group, the co-transplantation group had a more extensive distribution of the grafted cells and significantly reduced number of astrocytes, microglia/macrophage infiltration, and expression of chemokines (CCL2 and CCL3) at the injured site. The co-transplantation group also significantly increased arginase+/CD206+ macrophages (IL-4) and decreased iNOS+/CD16/32+ macrophages (IFN-γ), which was followed by higher IL-10 and IL-13 and lower IL-6 and TNF-α in their expression levels, a smaller cystic cavity area, and improved motor functions. These results indicate that OEC and SC co-transplantation could promote the shift of the macrophage phenotype from M(IFN-γ) to M(IL-4), reduce inflammatory cell infiltration in the injured site, and regulate inflammatory factors and chemokine expression, which provide a better immune environment for SCI repair.

Synergetic effects of ciliary neurotrophic factor and olfactory ensheathing cells on optic nerve reparation

This study is to investigate the effect of the combination of the olfactory ensheathing cells (OEC) transplantation and intravitreous injection of ciliary neurotrophic factor (CNTF) on the retinal ganglia cells' (RGC) apoptosis and axonals' reparation and regeneration. In this study, the supraorbital margin exposure of the optic nerves was used to establish adult SD rats' optic nerve inhausted injury model as control group. Then the purified OECs were injected into the optic nerve sheaths, and CNTF was injected into the vitreous body simultaneously. The rats were divided into control group, CNTF group, OECs group, and OEC+CNTF combined group. At 4 weeks postoperatively, a cholera toxin B subunit (CTB) anterograde tracing technique and fluorescence (FG) biotinylated dextran amine (BDA) retrograde tracing technique were adopted to evaluate the survival of RGC and the regeneration of optic nerve axons. The number of survival neurons of the same vision field and the density of neurons were evaluated by analysis of variance. At the one and three quadrant distance between optic disc 2 mm spot, the number of the RGC in the control group was significantly (F = 633.38, P < 0.01) decreased compared with the normal group (46.00 ± 1.42, 34.80 ± 1.34, 25.00 ± 1.61, 15.40 ± 2.30). The survived RGC in OEC group was significantly more than that in CNTF group, while the combined treatment with CNTF and OEC had strongest repair effect. The neuron axon density showed a statistically significant difference in the average optical density value at distance between foramen opticum 2 mm spot (OEC+CNTF: 3.18 ± 0.26, OEC: 2.96 ± 0.28, CNTF: 2.83 ± 0.37, and control: 2.75 ± 0.12, respectively, F = 17.66, P < 0.01) . Optic nerves damage can be repaired and regenerated by the combined treatment with OEC and CNTF suggesting that CNTF and OEC have synergistic effect on the treatment of optic nerve injury and repair. Transplantation of OECs may genetically modify the secretion of human CNTF and promote optic nerve injury repair.

Comparison of Schwann Cells and Olfactory Ensheathing Cells for Peripheral Nerve Gap Bridging

Introduction: Previously, we introduced the biogenic conduit (BC) as a novel autologous nerve conduit for bridging peripheral nerve defects and tested its regenerative capacity in a short- and long-term setting. The aim of the present study was to clarify whether intraluminal application of regeneration-promoting glial cells, including Schwann cells (SC) and olfactory ensheathing cells (OEC), displayed differential effects after sciatic nerve gap bridging. Material and Methods: BCs were generated as previously described. The conduits filled with fibrin/SC (n = 8) and fibrin/OEC (n = 8) were compared to autologous nerve transplants (NT; n = 8) in the 15-mm sciatic nerve gap lesion model of the rat. The sciatic functional index was evaluated every 4 weeks. After 16 weeks, histological evaluation followed regarding nerve area, axon number, myelination index and N ratio. Results: Common to all groups was a continual improvement in motor function during the observation period. Recovery was significantly better after SC transplantation compared to OEC (p < 0.01). Both cell transplantation groups showed significantly worse function than the NT group (p < 0.01). Whereas nerve area and axon number were correlated to function, being significantly lowest in the OEC group (p < 0.001), both cell groups showed lowered myelination (p < 0.001) and lower N ratio compared to the NT group. Discussion: SC-filled BCs led to improved regeneration compared to OEC-filled BCs in a 15-mm-long nerve gap model of the rat.

Combined Transplantation of Neural Stem Cells and Olfactory Ensheathing Cells Improves the Motor Function of Rats with Intracerebral Hemorrhage

To investigate the effects of combined transplantation of neural stem cells (NSC) and olfactory ensheathing cells (OEC) on the motor function of rats with intracerebral hemorrhage. In three days after a rat model of caudate nucleus hemorrhage was established, NSCs and OEC, NSC, OEC (from embryos of Wistar rats) or normal saline were injected into hematomas of rats in combined transplantation group, NSC group, OEC group, and control group, respectively. Damage of neural function was scored before and in 3, 7, 14, 30 days after operation. Tissue after transplantation was observed by immunocytochemistry staining. The scores for the NSC, OEC and co-transplantation groups were significantly lower in 14 and 30 days after operation than in 3 days after operation (P < 0.05). The scores for the NSC and OEC groups were significantly lower than those for the control group only in 30 days after operation (P < 0.05), while the difference for the NSC-OEC group was significant in 14 days after operation (P < 0.05). Immunocytochemistry staining revealed that the transplanted OEC and NSC could survive, migrate and differentiate into neurons, astrocytes, and oligodendrocytes. The number of neural precursor cells was greater in the NSC and combined transplantation groups than in the control group. The number of neurons differentiated from NSC was significantly greater in the co-transplantation group than in the NSC group. Co-transplantation of NSC and OEC can promote the repair of injured tissue and improve the motor function of rats with intracerebral hemorrhage.

The repair of acute spinal cord injury in rats by olfactory ensheathing cells graft modified by glia cell line-derived neurotrophic factor gene in combination with the injection of monoclonal antibody IN-1

To research the repair effect of transplantation of glial cell line-derived neurotrophic factor (GDNF) modified olfactory ensheathing cells (OECs) combination with injecting axonal growth inhibiting protein antibody (IN-1) in vivo. To construct lentivirus vector with GDNF gene and infect OECs in vitro, use the immunoblotting (Western Blot) to observe the expression of GDNF was detected through Western Blot. Fifty adult female SD rats which to establish thoracic spinal cord transection injury model were randomly divided into A (control group), B (IN-1 antibody group), C (OECs group), D (GDNF-OECs group), and E (GDNF-OECs+IN-1 group) 5 groups of each 10 rats. To observe regeneration of the impaired nerve axon by NF200 immunohistochemistry, Biotinylated dextran amine (BDA) anterograde tracing corticospinal tract. Basso, Beattie and Bresnahan (BBB) score was used to evaluating hindlimb motor function recovery. Add up to 13 rats died post operation. OECs labeled by hoechst still survived and migrated in spinal cord 8 weeks post operation. Lots of confused and disorderly regenerated axons which crossing the injured region of spinal cord were displayed between spinal cord stumps in GDNF-OECs+IN-1 group and GDNF-OECs group; some of axons existed in OECs group, but there is no obviously continue nerve fibers crossing the injured region of spinal cord;in contrast to IN-1 and control groups, few of regenerated axons and atrophy of spinal cord stumps were observed. The results of BBB hindlimb motor rating scale were 7.70+/-0.24, 7.89+/-0.15, 10.50+/-0.25, 11.43+/-0.23 and 12.81+/-0.40 for the control group, IN-1 group, OECs group, GDNF-OECs group and the allied treatment group respectively. The transplantation of GDNF-OECs combination with IN-1 antibody may benefit the survival and regeneration of the injured axons, and accelerate the repair of the injured spinal cord and functional recover of hindlimb locomotor in rats in a more efficient way than that with OECs or IN-1 alone.

Transplantation of olfactory ensheathing cells irradiated by low power laser irradiation to repair spinal cord injury in rats

Objective To investigate the effects of transplantation of olfactory ensheathing cells (OECs) irradiated by low power laser irradiation(LPLI) on the repair of transversal spinal cord injury in rats. Methods Twenty-four SD rats were randomized into 3 even groups after the animal models of spinal cord completely transected at T12 had been established for 4 weeks. OECs(group A), OECs ir-radiated by LPLI(group B) and DMEM fluid(empty control group) were transplanted into the caudal zone of the spinal cord in the 3 groups(n=8) respectively. The functional repair was evaluated by Bundle branch block (BBB) score, pathology and Flumgold label, respectively. The BBB scores before and after transplantation were statistically analyzed by one-factor analysis of variance or repeated measurement anal-ysis of variance, respectively (α=0.05). Results The methods of transplantation and the evaluation time had significant effects on the BBB scores in the 3 groups (P=0.000), and the BBB scores were significantly different among the 3 groups (P=0.000). Anti-NGRFp75 and anti-GFAP staining positive OECs were observed in the cephalic and caudal areas in group B, only anti-GFAP staining positive OECs were observed in group A, but nothing was found in the empty control group. The neural fibers labeled by Flurogold passed through the lesion area and extended into the cephalic and caudal areas in groups A and B, but were not present in the empty control group. Conclusion Although transplantation with both OECs and OECs irradiated by LPLI can promote the repair of spinal cord injury in rats, OECs irradiated by LPLI may be more effective. Key words: Olfactory ensheathing cells; Spinal cord injury; Cell transplantation; Laser therapy, low-level

The treatment of olfactory ensheathing cells-neurotrophin-3 gene engineering cell transplantation on experimental allergic encephalomyelitis

Objective To explore the repair mechanism of olfactory ensheathing cells (OECs)-neurotrophin-3 (NT-3) gene engineering cell on neuron myeline and axon of experimental allergic encephalomyelitis (EAE).Methods OECs-NT-3 gene engineering cell, constructed by ueurotrophin-3 transinfecting OECs inducted by retrovirus, was transplanted into lateral ventricle.The migration and distribution were observed and compared with control group and OECs transplantation group.Then myeline repair and axon regeneration were evaluated in the aspects of function score, morphological structure, SYN grey level Results (1) OECs-NT-3 could survive, diffuse, migrate with axons, spread in the focus diffusely on the 28th day after transplantation.(2) OECs-NT-3 survived and migrated to the transcription level of NT-3 mRNA in transgene group, being (212.3±16.1)×10-2, significantly higher than OECs group ((1.98±0.19)×10-2) and the contrast group ((1.23±0.13)×10-2, t = - 31.161, -31.928, P < 0.01).(3) The myeline of transgene group was kept complete and the number of inflamatory focus was lower than those of other groups (t = 11.388-22.728, P <0.01).(4) The SYN grey level of transgene group was obviously higher (P < 0.01).Conclusion OECs-NT-3 cell expresses NT-3 in EAE stably and effectively, which contributes to the repair of myeline and the regeneration of axon. Key words: Encephalomyelitis, autoimmune, experimental; Gene therapy; Neurotrophic 3; Gene transfer techniques

Evaluation of Olfactory Ensheathing and Schwann Cells after Implantation into a Dorsal Injury of Adult Rat Spinal Cord

Olfactory ensheathing cells (OECs) and Schwann cells (SCs) obtained from adult transgenic rats expressing alkaline phosphatase (AP) were studied following implantation into intact spinal cord and after dorsal column crush (DCC) injury, either within the lesion or near the lesion borders. We observed no evidence of migration of AP OECs or AP SCs after lesion site injections, with most cells remaining in or nearby the injection/lesion site. Acute injection of either cell type outside of the lesion site resulted in the presence of cells in the lesion even two hours after injection. However, after a 2-week delay between DCC injury and cell injection, only OECs injected 2.5-mm outside of a DCC lesion entered the lesion, while SCs did not pass a region of increased astroglial immunoreactivity. GFAP-immunoreactivity also revealed differences in the astroglial scar at the lesion border with openings apparent in this region only in the OEC group. SCs induced greater ingrowth of CGRP-positive axons within the lesion, two weeks post-injury. Equivalent numbers of GAP-43-positive axons grew within the lesion after SC or OEC implantation. These findings show that, although there is no active migration for either cell type, both OECs and SCs are able to support axonal regrowth and/or sprouting into the lesion. The openings in the astroglial boundary at the lesion site may give OECs a potential advantage over SCs in promoting axonal growth through the astroglial scar.