Construction Of Vascularized Tissue Engineering Bone Based On 3D Printing And Electrospinning Technology

Objective: Using 3D printing technology and high-voltage electrostatic textile technology to construct a tissue engineering scaffold-cell complex,and to verify its advantages in the construction of vascularized tissue engineering bone.Methods: The three-dimensional model with strong personalized appearance was developed from the handling,editing and making of the Computed Tomography(CT)by using medical image three-dimensional reconstruction software(Mimics)and 3D printer.While printing on the model,using coaxial high-voltage electrostatic to make the /P3HB4 HB fibers with P3HB4 HB as its core layer,PVA and HMSC(PVA-cells)as its shell.The primary stent-cell complex was induced to the osteogenic direction for 14 days and implant into the composite endothelial cells.In vitro experiment,observed the mechanical properties of P3HB4 HB external stent made by 3D printing through mechanical test;observed the microstructures and biocompatibility of PVA / P3HB4 HB scaffolds made by electrostatic textile technology through light microscope,transmission electron microscope,hydrophobicity test and scanning electron microscope;observed the adhesion and proliferation of the cells in scaffolds through scanning electron microscopy,DAPI fluorescence staining,acridine orange fluorescence staining and CCK-8 test.In vivo experiment,the scaffold-cell complex was used as experimental group and the cell-free scaffolds as controls,specimens were removed after 12 weeks and 24 weeks,made staining of HE,VonKossa,Sirius red,equine triplicate,CD31 immunohistochemistry and type?collagen immunohistochemistry,and observed the capacity of scaffold-cell complex and vascular.Results: The model with strong personalized appearance can be made by 3D printing technology.Models can provide some mechanical support for tissue engineering scaffolds.The PVA / P3HB4 HB stent with electrostatic High porosity,good biocompatibility and three-dimensional structure of biomimetic extracellular matrix made by electrostatic textile technology can be observed through the scanning electron microscopy and hydrophilicity experiments.The cells still have adhesion and proliferation capability after their composition with scaffolds through the reveal of acridine orange fluorescence staining,DAPI fluorescence staining,scanning electron microscopy and CCK-8 test,achieved a molding and precise planting of material and the cell.In vivo experiment,HE staining,Von Kossa staining,Sirius red staining,Masson trichrome staining,CD31 immunohistochemical staining,type?collagen immunohistochemical staining were performed at 12 and 24 weeks respectively.The results were positive and the number and quality of bone tissue and vascular structure increased significantly over time,the control group was negative.Conclusion: The scaffold-cell complexes constructed by 3D printing technology and high-voltage electrostatic textile technology have certain mechanical properties and structure and function of bionic extracellular matrix.After the composition of endothelial cells,the tissue engineering bone containing blood vessels can be constructed in vivo.