Engineered artificial structure for vascular network generation and its construction method and application
A construction method and vascularization technology, which are applied in the field of engineered artificial structures and their construction to achieve the effects of excellent biological functions, in vivo regeneration, and wide use.
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Embodiment 1
[0098] Example 1 Preparation of vascularized growth factor sustained-release microspheres
[0099] 1. Sodium alginate (Sigma-Aldrich, A0682) was prepared into a 1.5% (w / w) solution with PBS buffer solution.
[0100] 2. Basic fibroblast growth factor (bFGF, Abcam, AB9596) was uniformly mixed with sodium alginate solution at a concentration of 2 ng / ml and then filled into a disposable needle tube with a volume of 10 ml.
[0101] 3. Connect the high-voltage electrostatic cell microsphere preparation device. The high-voltage electrostatic generating device adopts SA167-Y (Tianjin) high-voltage electric field generator with an output voltage of 15kV; clamp the 10ml syringe in step 2 on the propulsion device: LongerPump Company TS2-60 injection In the pump, the distance between the needle tip and the copper sheet is 20mm, the propulsion speed is 10ml / h, a needle with an inner diameter of 191μm is used, and the collector is a disposable plastic petri dish with a diameter of 60mm (pla...
Embodiment 2
[0104] The preparation of embodiment 2 hydrogel microspheres
[0105] 1. Mix a solution of sodium alginate (Sigma-Aldrich, A0682) and a solution of gelatin (Sigma-Aldrich, G1890) (solvent PBS buffer solution) to a final concentration of 2% (w / w) and 3% (w / w) respectively w) The hydrogel precursor solution.
[0106] 2. Uniformly mix vascular endothelial growth factor (VEGF, Abcam, AB9571) at a concentration of 10 ng / ml with the hydrogel precursor solution obtained in 1, and then fill it into a disposable needle tube with a volume of 10 ml.
[0107] 3. Connect the high-voltage electrostatic spray cell microsphere preparation device. The high-voltage electrostatic generator adopts SA167-Y (Tianjin) high-voltage electric field generator with an output voltage of 10kV; clamp the 10ml syringe in step 2 on the propulsion device: LongerPump Company TS2-60 In the syringe pump, the distance between the needle tip and the copper sheet is 10 mm, the propulsion speed is 20 ml / h, the needl...
Embodiment 3
[0110] Embodiment 3 The preparation of coating material coating layer
[0111]Mix 1ml of collagen solution (Sigma, C7661) with 1ml of hydrogel microspheres and put them into a centrifuge tube. At this time, the cell microspheres sink to the bottom due to their high density. Centrifuge at a speed of 1000r / min for 4min, so that the collagen solution is fully in contact with the unit structure microspheres at the bottom. Static culture in the centrifuge tube for 48-72 hours, so that the collagen solution is evenly distributed on the surface of the microspheres.
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