| Background：Intracerebral hemorrhagic stroke (ICH) is a huge burden for health care system and individualfamily for the high mortality and mobility. There was no FDA approved drugs for ICH. Cellbased therapy hold a great potential for central nervous system disorders. As a novel approachto cure hemorrhage stroke stem cell transplantation has been accepted gradually. Among stemcells that are candidates for transplantation, bone marrow mesenchymal stem cells (BMSCs) arepreferred because they are available from autologous donation therefore eliminates ethicaldisputes and other concerns related to graft rejection. Still there have been a lot of challengingissues that hinder the efficacy and efficiency of cell-based therapy need to be explored.Aim：In this study we tested the hypothesis that intranasally delivered hypoxia-preconditionedBMSCs could reach the brain, promote tissue repair and improve functional recovery after ICHin mice.Methods：There were4parts of this study totally.1. Hemorrhagic stroke was induced in adult C57/B6mice by injection of different dose ofCollagenase type IV into the striatum, morphology and behavior tests were used to test thestablity of the ICH model.2. BMSCs were isolated and cultured from SD rats, cells were incubated under normoxicconditions or in a finely controlled ProOx C-chamber system (0.5%oxygen) for24h. CellViability was tested by MTT, BMSCs migration ability in vitro was measured with scratchwound healing assay, Western blot analysis was applied to measure the expression ofneurotrophic factors.3. Assessment of transplanted BMSCs: Brain sections were analyzed by fluorescencemicroscopy. Sections were checked starting from the olfactory bulb to other regions of the brainlike cortex, basal ganglia to test the reliable of this deliver method. 4. To evaluate the therapeutic effect of HP-BMSCs: These animals were randomly divided into3groups, at3days and7days after ICH HP-BMSCs or PBS was administration intranasally.(1)A set of behavior tests including mNss, Rotarod, sticky dot and Open Field Test wereperformed at different time points after ICH to evaluate the neurological function deficits.(2)Western blot analysis was used to test the expression of growth factors VEGF (VasclarEndothelial Growth Factor), BDNF (Brain-Derived Nerotrophic Factor) and GDNF (Glial cellline-Derived Neurotrophic Factor). Immunohistochemistry was applied to identify neurogenesisaround hemotoma and SVZ.(4) Nissle stainig was used to assess the morphology change afterICH.Results：1.0.03U Collagenase type IV were stereotaxically injected into stratum to generate ICH. Thismodel is reliable which has significant behavior deficits and lower mortality.2. HP-BMSCs showed a higher level of expression of trophic/growth factors, including EPO,EPOR, SDF-1. HP-BMSCs exhibited enhanced migratory capacities in vitro.3. HP-BMSCs reached the olfactory bulb, ischemic cortex and mostly deposited outside ofhematoma after administration.4. The HP-BMSCs treated group showed markedly improvement of functional performancecompared with the control group. More BrdU and DCX collable cells were found in SVZregion and more BrdU and NeuN collable cells were found around hematoma. Western blotindicated that growth factor like GDNF, BDNF, VEGF were significantly increased inHP-BMSCs treatment group.Conclusion：This study shows that intranasal delivery stem cell can enter the brain and migrate to peri-injurelocation also secret growth factors. These data also suggest that intranasally deliver HP-BMSCscould improve neurological outcome and enhance neurogenesis after ICH in mice, providing anovel therapeutic approach for hemorrhagic stroke. |
