In Vitro Study Of Electrospun Chitosan/Polycaprolactone Nanofiber Membranes Induced Bone Mesenchymal Cells

Objective Based on the theory of tissue engineering, this study prepared CHS /PCL nanofiber membrane as a scaffold with electrospinning, chitosan and polycaprolactone, the mice bone marrow mesenchymal cells were inoculated in the CHS/PCL nanofiber membrane,evaluating whether the nanofiber membrane promoted rat bone marrow mesenchymal cells into osteoblasts and promoted the proliferation of bone marrow mesenchymal cells, adhesion, growth effects. From the protein level, it explored furtherly osteogenic differentiation capacity of the CHS/PCL nanofiber membranes to mesenchymal cells in vitro. It provided an experimental basis for clinical bone tissue regeneration. Methods Through electrospinning, chitosan and polycaprolactone as the experimental materials,according to the different proportions,to prepare of CHS/PCL nanofiber membranes(0:100,50:50,30:70),by ultraviolet light after sterilization.It tested the line of hydrophilic material and internal morphology.Then in accordance with CHS/PCL nanofiber membrane as a carrier,Bone marrow mesenchymal cells of mice, subculturing from the third to the fifth, the cells were seeded in CHS/PCL nanofiber membranes until after the cells were covered, by SEM(scanning electron microscopy, SEM), DAPI(4,6-diamino-2-phenyl indole, 4,6- diamino- phenylindole) staining was observed in bone marrow mesenchymal cells CHS/PCL nanofibers film adhesion and growth. By MTT(methyl thiazoly terazolium, methyl thiazolyl four salt) experiment to measure proliferation and differentiation of mesenchymal cells in CHS/PCL nanofiber membrane in the first, three, five, seven days. Based on ALP(alkaline phosphatase) experiment,WB(protein immunoblot experiments) and alizarin red staining experiments, evaluation of CHS/PCL nanofiber membranes of mesenchymal cells in vitro osteogenic differentiation levels. By analyzing the results of the above experiment, to explore the promoting capacity of CHS/PCL nanofiber membrane to bone marrow-derived mesenchymal cells in vitro. Results CHS/PCL nanofiber membrane was white film-like material, SEM electron microscope showed CHS/PCL nano fiber membrane, showing filament fiber membrane staggered evenly, sizes porosity inside the membrane, pores having a certain penetration resistance.On the CHS/PCL nanofiber membrane(50: 50,30: 70), it could see particles adhering to nodular spun chitosan the wire and evenly distributing in the spin network architecture.the inter-cultured bone marrow mesenchymal cells were adhered to the different ratios of CHS/PCL nanofiber membranes,it could find nanofiber membranes with good biocompatibility, promoting the adhesion, proliferation and differentiation into bone ability of the cells in nanofiber membranes. Conclusion In tissue engineering,the CHS/PCL nanofiber membrane as a carrier,it had good biocompatibility, plasticity and mechanical properties,and could be degraded and absorbed in vivo.Therefore in tissue engineering,as a scaffold material,cell in vitro in primary and passage culture, and the cells were seeded on the membrane.To observe the growth,adhesion and proliferation of MBSCs,and it also induced bone marrow mesenchymal stem cells to osteoblast differentiation,which were induced into osteoblasts,bone tissue regeneration and repair were obtained.Therefore,chitosan/ polycaprolactone nano fiber membrane was a kind of great potential and prospects for clinical regeneration scaffold materials.This paper discussed the superiority of electrospun CHS/PCL nanofiber membrane,and had laid the experimental foundation for clinical study treatment of oral bone regeneration.