The Study Of Novel Functional Three-dimensional Nanofibrous Scaffold For Cell Culture Application

Compared with traditional two-dimensional cell culture,the three-dimensional culture technology that develops with tissue engineering,can provide cells with an environment which is similar to the natural extracellular matrix and maintain the state of cells in the living body,so it has a wide range of applications in drug screening,tissue engineering scaffolds,cell biology,etc.The scaffolds based on electrospinnng technology which are more similar to the fibrous structure of matrix ECM are widely applied in study of three-dimensional cell culture.However,most current nanofiber scaffolds only provide a macroscopic three-dimensional growth environment.In the micro-state,the cells still combine with scaffolds in nearly two-dimensional growth mode which fail to maintain the true three-dimensional cell culture state.In order to establish the ideal three-dimensional culture scaffolds,a three-dimensional nanofiber scaffold with high micropore size was prepared and functionalized.Then Hep G2 cells and RGC5 cells were applied to construct the cell-scaffold three-dimensional culture system with the exploration of cell morphology and proliferation in the scaffolds.The research processes were as follows:(1)We introduced magnetic nanoparticles into polyacrylonitrile spinning liquid and the water phase was used as receiving device during the process of electrospinning.The nanofibers could break through the surface tension of water with the attraction of neodymium iron boron magnet which was at the bottom of water to magnetic nanoparticles that are in nanofibers.As a result,the nanofibers accumulated in water to form highly discrete three-dimensional nanofiber scaffolds;(2)A layer of poly 3,4-ethylenedioxythiophere conductive polymer was loaded with the method of in-situ polymerization.Subsequently,the graphene oxide sheets were adsorbed on the outermost layer of nanofibers by the contribution of electrostatic adsorption force.Besides,the mechanical,electrical properties and thermodynamic stability of scaffolds were explored;(3)Finally,Hep G2 cells were co-cultured with non-functionalized nanofiber scaffolds to observe the morphology of the cells.The RGC5 cells were co-cultured with the functionalized nanofiber scaffolds.The cell morphology and proliferation were observed and the apoptosis status was detected and analyzed.Based on the research,the conclusions are drawn as follows:(1)With the introduction of Fe3O4 magnetic nanopraticles,three-dimensional nanofiber scaffolds with micropore which is suitable for cell migration can be prepared by electrospinning and received by water with assistance of magnetic field,besides Fe3O4 can be relatively uniformly dispersed in the namofibers;(2)The functional modification of nanofibers was made by in-suit polymerization of poly 3,4-ethylenedioxythiophere on the surface of nanofibers and electrostatic adsorption of graphene oxide on the outermost layer.As a result,the functionalized nanofiber scaffolds exhibited excellent electrochemical properties;(3)As a cell culture scaffold,the functional modified three-dimensional nanofiber scaffolds could significantly provide three-dimensional growth environment for cells,and cells were tightly combined with nanofibers,which was conductive to information transmission between cells and promoting cell proliferation in the form of agglomeration communities,and ultimately,constitute the ideal scaffold-cell three-dimensional culture system.This study provides a universal method for the preparation of three-dimensional cell culture scaffolds and has significant effect on the construction of three-dimensional cell culture system.It is of great significance for the construction of cell model in vitro which is similar to organisms and the development of tissue engineering scaffolds.