Study Of Carbon Nanomaterials-HUC-MSCs In The Treatment Of Ischemic Stroke In Mice

Objectives:Stroke is one of the three diseases which causes seriously threaten to the physical and mental health of patients.Ischemic stroke,with high morbidity,prevalence and mortality,accounts for 80%of the disease.Currently,the only effective therapy is the drug rt-PA,however,for its narrow time window and bleeding risk,the application of it is still limited.Stem cell has many roles in replacement,induction,anti-inflammatory and angiogenesis,which might be a potential treatment for stroke,whereas the rare resource has dramatically limited clinical use.With the development of carbon nanomaterials and other biological cell engineering,it is provided new idea of stem cell treatment by combing those.In our study,co-cultivated hUC-MSCs on carbon nanomaterials in vitro model and transplanted hUC-MSCs mice stroke model were performed to evaluate its efficacy and potential mechanisms.Methods:Cell viability was determined among hUC-MSCs cultured in vitro,co-cultured with graphene and carbon nano-columns by MTT assay.Meanwhile,Q-PCR and enzyme-linked immunosorbent assay(ELISA)were used to detect neurotrophic factors secreted by hUC-MSCs.The mice were randomly assigned into control group,middle cerebral artery occlusion(MCAO)model group,hUC-MSCs group and carbon nanocage(CNC)group.Mice in hUC-MSCs and CNC group were treated with ischemia and reperfusion respectively.hUC-MSCs or hUC-MSCs co-cultured with CNC was injected into the tail vein of MCAO mice after 60 minutes reperfusion.mNSS scores,cerebral infarct volume,and brain tissue water content were measured to evaluate the therapeutic effect.To further explore the potential mechanism,the expression of neurotrophic in the brain of MCAO mice were also detected.Results:hUC-MSCs after co-cultured with graphene and CNC have no effect on the cell viability.In vitro,the neurotrophins secreted by hUC-MSCs were increased after co-cultured with CNC.After being transplanted CNC in a mouse model of MCAO,the volume of cerebral infarct was reduced,and the expression of NGF(Nerve Growth Factors)were up-regulated.Conclusions:CNC could enhance the cell viability of hUC-MSCs in vitro and increase the contents of neurotrophic factor secreted by hUC-MSCs.The volume of cerebral infarction was reduced after hUC-MSCs transplanting in the MCAO mice,and the neuroprotective effect was more effective after co-cultured with CNC.The potential mechanism be related to the promotion of the secretion of neurotrophic factors.