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Making method of three-dimensional vascularized musculocutaneous flap based on biological 3D printing

A 3D printing and manufacturing method technology, applied in biochemical equipment and methods, vascular endothelial cells, microorganisms, etc., can solve problems such as the difficulty in realizing the independent culture and differentiation of different cell components, and the difficulty in realizing the integration of blood flow and perfusion of different cell components. , to achieve the effect of high biocompatibility and improved survival rate

Active Publication Date: 2021-06-04
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Existing technologies include 3D printing, mold forming, etc., but it is difficult to realize the independent culture and differentiation of different cell components, and it is difficult to realize the integrated perfusion of blood flow and the assembly of different cell components

Method used

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  • Making method of three-dimensional vascularized musculocutaneous flap based on biological 3D printing
  • Making method of three-dimensional vascularized musculocutaneous flap based on biological 3D printing
  • Making method of three-dimensional vascularized musculocutaneous flap based on biological 3D printing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A method for making a three-dimensional vascularized myocutaneous flap based on biological 3D printing, the specific steps are as follows: (1) according to the mass volume ratio of 5% GelMA (methacryloyl gelatin), 2% sodium alginate, 10% PEGDA ( Polyethylene glycol diacrylate) and 0.05% LAP were dissolved in 25 mM 4-hydroxyethylpiperazineethanesulfonic acid buffer solution (HEPES). Add vascular smooth muscle cells to make the cell concentration 10^7 cells / mL. Ink was stored at 37°C until use. Prepare 50mM CaCl 2 solution as an ionic crosslinker. Three-dimensional tubular structures are printed using a two-channel coaxial system. Inner layer is 50mM CaCl 2 Solution, using a 20mL syringe, the extrusion speed is 0.1mm / min. The outer layer is bio-ink, using a 5mL syringe with an extrusion speed of 0.56mm / min. The bioprinted constructs were first treated with CaCl 2 solution cross-linked and then exposed to blue light (395nm, 8mW / cm 2 Power) 5min for further cross-li...

Embodiment 2

[0034] A method for making a three-dimensional vascularized myocutaneous flap based on biological 3D printing, the specific steps are as follows: (1) 7% GelMA (methacrylated gelatin), 1% sodium alginate, 15% PEGDA (polyethylene glycol Diacrylate) and 0.1% LAP were dissolved in HEPES (25mM). Add vascular smooth muscle cells to make the cell concentration 10^7 cells / mL. Ink was stored at 37°C until use. Prepare 30mM CaCl 2 solution as an ionic crosslinker. Three-dimensional tubular structures are printed using a two-channel coaxial system. The inner layer is 30mM CaCl 2 Solution, using a 20mL syringe, the extrusion speed is 0.15mm / min. The outer layer is bio-ink, using a 5mL syringe with an extrusion speed of 0.84mm / min. The bioprinted constructs were first treated with CaCl 2 solution cross-linked and then exposed to blue light (395nm, 8mW / cm 2Power) 7min for further cross-linking. Connect the stent through a 26G indwelling needle, and inject 0.5 ml of vascular endothe...

Embodiment 3

[0038] A method for making a three-dimensional vascularized myocutaneous flap based on biological 3D printing, the specific steps are as follows: (1) 5% GelMA (methacrylated gelatin), 3% sodium alginate, 10% PEGDA (polyethylene glycol Diacrylate) and 0.05% LAP were dissolved in HEPES (25 mM). Add vascular smooth muscle cells to make the cell concentration 10^7 cells / mL. Ink was stored at 37°C until use. Prepare 75mM CaCl 2 solution as an ionic crosslinker. Three-dimensional tubular structures are printed using a two-channel coaxial system. The inner layer is 75mM CaCl 2 Solution, using a 20mL syringe, the extrusion speed is 0.1mm / min. The outer layer is bio-ink, using a 5mL syringe with an extrusion speed of 0.56mm / min. The bioprinted constructs were first treated with CaCl 2 solution cross-linked and then exposed to blue light (395nm, 8mW / cm 2 Power) 7min for further cross-linking. Connect the stent through a 26G indwelling needle, and inject 0.8 ml of vascular endot...

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Abstract

The invention provides a making method of a three-dimensional vascularized musculocutaneous flap based on biological 3D printing. Coaxial biological 3D printing is utilized; a calcium chloride solution is taken as an inner layer; methacrylic anhydride gelatin, sodium alginate, polyethylene glycol diacrylate and vascular smooth muscle cells are mixed to serve as outer-layer biological ink; a printed scaffold is thoroughly cross-linked with blue light through calcium chloride; a three-in-one perfusate scaffold is finally formed, so that blood supply is established; vascular endothelial cells are poured into the scaffold; vascular cell components are simulated; fibroblasts, adipose tissue-derived stromal cells and skeletal muscle myoblasts are respectively inoculated on a three-layer scaffold; a fibroblast culture medium, an adipogenesis induction culture medium and a myoblast culture medium are respectively added and put into a reactor for culture; and the three-layer scaffold is folded and bond through rat tail collagen to form the stacked scaffold. According to the invention, integrated perfusion of blood flow and independent culture differentiation and assembly of different cell components are realized.

Description

technical field [0001] The invention relates to the technical field of biological materials, in particular to a method for manufacturing a three-dimensional vascularized myocutaneous flap based on biological 3D printing. Background technique [0002] Myocutaneous flap is a kind of composite tissue flap, which is taken from a certain muscle (or part of muscle) of the body together with its superficial subcutaneous tissue and skin, and transferred with the blood vessel entering the muscle as the pedicle. It is used for larger wounds. Repair of defects or reconstruction of muscle function. The musculocutaneous flap has abundant blood supply, strong anti-infection ability, easy survival, and rich tissue volume. It is one of the commonly used tissue flaps in plastic and reconstructive surgery. [0003] Existing technologies include 3D printing, mold forming, etc., but it is difficult to realize the independent culture and differentiation of different cell components, and it is d...

Claims

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Application Information

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IPC IPC(8): A61L27/40A61L27/02A61L27/22A61L27/20A61L27/18A61L27/38A61L27/50B33Y70/10B33Y80/00C12N5/071C12N5/077C12N5/0775
CPCA61L27/025A61L27/222A61L27/20A61L27/18A61L27/3826A61L27/3808A61L27/3804A61L27/3834A61L27/3895A61L27/3873A61L27/3886A61L27/50B33Y70/10B33Y80/00C12N5/0697A61L2300/412A61L2430/30C12N2502/1347C12N2502/28C12N2502/1323C12N2502/1382C12N2502/1335C12N2533/54C12N2533/74C12N2533/40C08L5/04C08L71/02
Inventor 牟珊汪振星孙家明周莹芊郭亚琪锁丽涛陈雳风周牧冉刘绍恺罗超曾宇阳方慧敏慈海侯金飞谢昕芳孙谛张郭刘剑王冰倩倪若飘王斌陈佳龙李志鹏姜文彬揭君津赵阳
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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