| Objective:In this study,graphene oxide(go)/ polycaprolactone(PCL)nanofibers with different concentrations of graphene oxide were prepared by electrospinning,and then the composite matrix nanofibers were formed by combining with rat tail collagen containing gold nanoparticles(Au NPs).The optimal concentration for the growth and differentiation of rat brown adipose stem cells(Bascs)was evaluated in vitro,and the effect of the composite nano materials on the differentiation of Bascs into cardiomyocytes was observed.Methods:1)GO/ PCL composite nanofibers were prepared by electrospinning.The microstructure and particle size of the materials were characterized by TEM and SEM.2)Rat brown adipose stem cells(Bascs)were isolated and cultured,and differentiated cells were identified for cardiomyocyte specificity.3)The biocompatibility of the nanocomposites was evaluated by CCK-8 / live dead staining,and the optimal concentration of graphene oxide for cell growth was selected.4)Rat brown adipose stem cells(Bascs)were inoculated on collagen and go / PCL / Au NPs composite membrane respectively.The expression of intercalated disc associated protein(CX-43,?-actin)was detected by a series of cytology,immunocytochemistry and Western blotting.Results:1)The diameter and three-dimensional porosity of the nanofibers formed by electrospinning with different concentrations of go were not significantly different,which indicated that the materials had certain stability.2.2)The isolated and cultured brown adipose stem cells can spontaneously differentiate into cardiomyocytes and pulsate.3)According to the analysis of CCK-8 and live dead results,go / PCL composite with 0.10mg/ml go is more suitable for cell growth,showing good biocompatibility,and cell growth status is better than other groups.Therefore,go concentration 0.10mg/ml is taken as the experimental group in the follow-up experiment.4)Compared with the collagen group,the go / Au NPs composite nanospinning material can significantly promote the proliferation and differentiation ability of brown adipose stem cells(Bascs),and the differentiated cardiomyocytes have strong spontaneous contraction ability and good spreading ability.Moreover,the composite material can enhance the expression of intercellular intercalated disc related proteins connexin-43 and ?-actinin Compared with collagen group,the expression of CX-43 and ?-actinin in go / Au NPs group was significantly increased,and the difference was statistically significant(P < 0.05).Conclusion:the combination of go / PCL composite nanofibers with gold nanoparticles(Au NPs)and rat brown adipose stem cells(Bascs),the combination of graphene oxide and gold nanoparticles increased the conductivity of the materials for the first time,and confirmed that go / Au NPs composite nanomaterials can promote the differentiation of brown adipose stem cells into cardiomyocytes and enhance cardiomyocytes The expression of specific protein provides a new idea for the application of Bascs and go/ Au NPs composite nanomaterials in the field of myocardial tissue engineering. |
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