| Objective White matter damage is one of the common forms of brain damage in premature infants.Currently,there is no effective treatment.Surviving children often have residual neurological sequelae,including cerebral palsy,cognitive or behavioral defects.This article intends to study the effect of transplanting human oligodendrocyte precursor cells via lateral ventricular and nasal pathways on rats of white matter damage in preterm infants and to provide clinical basis for stem cell treatment of white matter damage in preterm infants.Methods1.The establishment and identification of PWMI model :PWMI model was prepared by occluding the right common carotid artery of 3-day-old Sprague Dawley(SD)rats with hypoxia for 90 min.Behavioral and morphology methods were used to evaluate the extent of damage to the model.2.The culture and identification of hOPCs: hOPCs were induced using neural stem cell lines from human fetal brain established in this laboratory and we identify the purity of cells by immunofluorescence staining of specific markers.3.Verification of transplantation path: four days after modeling,normal unlabeled hOPCs were transplanted through the lateral ventricle and the brain was taken to track the cells 7 days after transplantation by stem121 immunofluorescence staining to confirm the accuracy of the transplant site;Three days after modeling,CM-Dil and normal unlabeled hOPCs were first grafted by nasal transplantation and the brain was taken to track the cells 3 days after transplantation to confirm the feasibility of the method.4.Forty 3-day-old SD rats were divided into 4 groups: Sham operation group,PWMI group,intracerebroventricular transplantation group,and nasal transplantation group,each with 10rats;the cells were observed by stem121 and human nuclear antigen(h NA)immunofluorescence staining at 12 w after transplantation.Migration,differentiation,and proliferation,applied behavioral,Luxol fast blue(LFB)myelin staining,myelin basic protein(MBP)fluorescence staining,and transmission electron microscopy to verify neuroreparable effects of intanasal and intracerebroventricular transplantation.Results1.Neurological deficits are apparent after white matter damage in preterm infants.On the 7th day after modeling,compared with the normal side,the positive area of MBP decreased and myelin sheath was lost in the cingulate,corpus callosum,and outer capsule of the affected side of the PWMI model.The results of HE staining showed that the corpus callosum and hippocampal tissues were loose and the inflammatory cells were infiltrated.2.The growth status of hOPCs is good and the specific markers A2B5,NG2,PDGFR-α,SOX10,O4,and Olig2 are all positive for immunofluorescence staining,immunofluorescence staining of the astrocyte marker GFAP and neuron marker tuj-1 was rarely positive.3.Cells were detected in the lateral ventricle four days after intracerebroventricular transplantation.Cells were detected near the cortex,hippocampus and lateral ventricles in the injured areas of the brain three days after the nasal transplantation.4.At 12 w after cell transplantation,cells migrated extensively through the lateral ventricle transplantation group;partly differentiated into mature oligodendrocytes,undifferentiated into astrocytes and neurons;Very few cells are in a proliferative state capable of division.The defect of nerve function was improved obviously,the structure of myelin sheath was tight,and the thickness of myelin sheath was increased.In the nasal transplantation group,the nerve function defect was significantly improved,the myelin structure was tight,and the myelin sheath thickness was increased.Conclusions1.The WMI model of premature SD rats is stable and reliable and the system for judging the injury degree of the model is preliminarily established.2.Transplanted cells through the lateral ventricle can promote myelin repair and regeneration and improve the functional behavior of rats.3.Transplanted cells through the nose can enter the brain tissues of rats and migrate around the injured side,promoting myelin sheath repair and improving the functional behavior of rats,which proved that the route of nasal transplantation was feasible4.Transnasal transplantation of hOPCs can be used as a new method for the treatment of white matter injury in premature infants. |
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