| Spinal cord injury（spinalcordinjury,SCI）is caused by spinal cord compression caused by trauma and disease（such as tumor,hematoma and infection）,resulting in neuronal death and axonal degeneration,resulting in permanent motor dysfunction and loss of sensation below the injury site.It is estimated that 3 million people worldwide suffer from traumatic spinal cord injury（SCI）.In the past 30 years,its global epidemic survey found that the quantity of patients with spinal cord injury has increased from 236 to 1298 per million population.The total cost of life per patient with spinal cord injury is more than ＄3 million,and the calculated annual economic burden in Canada is almost $2.67 billion.Due to the limited spontaneous rehabilitation,there is still no treatment for the injured spinal cord itself,which can only provide supportive relief for patients with permanent disabilities.At present,the difficulty in the treatment of spinal cord injury lies in the spinal cord injury,which directly causes the death of the surrounding nerve cells,which will lead to the cavity in the injured area.Secondly,inflammation appeared in the injured area.Finally,the axons of injured peripheral nerve cells will be degenerated and the proliferation of glial cells will produce scars,which affect the growth of axons.In order to solve these difficulties,we study a new vector to promote the growth and differentiation of neural stem cells in vitro.It expands the visual angle of repairing spinal cord injury.Objective Investigate the effect of modified hyaluronic acid（HA）hydrogel controlled release brain-derived neurotrophic factor（BDNF）on the growth,differentiation,and apoptosis of rat embryonic neural stem cells（rFNSCs）.Methods The experiment took rat embryonic stem cells as the research object.Theexperiments were divided into three groups,blank control group[rFNSCs co-cultured with poly（lactic-co-glycolic acid）（PLGA）and HA],conditional control group（rFNSCs co-cultured with BDNF-PLGA and HA）,and experimental group（rFNSCs cocultured with BDNF-PLGA and modified hyaluronic acid hydrogel that is Dex-HA）.ELISA was used for detecting the release curve of BDNF in the controlled-release scaffold.The Live/Dead assay detected the apoptosis of rFNSCs in each group,CCK-8 experiment was used to test the cell proliferation activity of rFNSCs after coculture with hydrogel.The differentiation ratios of rFNSCs in different groups were detected by immunocytochemical staining.Results ELISA result showed that the controlled-release scaffold could release BDNF for 14 days.Live/Dead result showed the number of apoptosis in BDNF-PLGA/HA group and BDNF-PLGA/Dex-HA group was significantly lower than that in blank control group,and the number of death in BDNF-PLGA/Dex-HA group was the least（P<0.05）.CCK-8 assay showed that the cell proliferation activity of BDNFPLGA/HA and BDNF-PLGA/Dex-HA groups was higher than that of the blank control group,and that of BDNF-PLGA/Dex-HA was also higher than that of BDNFPLGA/HA（P<0.05）.immunocytochemical staining showed that,the BDNFPLGA/Dex-HA hydrogel could promote the neuronal differentiation of rFNSCs and reduce their differentiation into astrocytes in vitro compared with the blank control group and the conditional control group（P<0.05）.Conclusion The porous structure of BDNF-PLGA/Dex-HA hydrogel provides a good three-dimensional microenvironment for the growth,proliferation and cell adhesion of rFNSCs,and the hydrogel can promote the differentiation of rFNSCs into neurons and reduce the differentiation into astrocytes,which solves the problems of lack of neurons and glial scar formation in spinal cord injury area,and lays a good foundation for further animal experiments in the later stage. |
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