Manufacturing method of three-dimensional spiral directional active ventricular structure

Abstract

The invention relates to a manufacturing method of a three-dimensional spiral directional active ventricular structure, which comprises the following steps of: inducing cells and hydrogel fibers to generate directional arrangement by utilizing internal stress generated by a three-dimensional driving bracket encapsulated in hydrogel; a spiral directional ventricle structure consistent with the natural ventricle in arrangement direction is designed and constructed through the macroscopic shape and the structural unit of the three-dimensional driving stent, active ventricle tissue manufacturing with systolic and diastolic, blood pumping and electrical conduction functions is achieved through in-vitro dynamic culture, and the requirements of tissue repair and in-vitro drug testing are met.

Description

Technical Field

[0001] The invention relates to the field of advanced manufacturing and biomedical technology, and in particular to a method for manufacturing a three-dimensional spiral oriented active ventricular structure. Background Art

[0002] The muscle tissue of the natural heart is composed of a highly ordered spiral oriented structure, which is essential for the natural heart to achieve specific functions and electrophysiological performance. The oriented myocardial cells and collagen fibers in the left ventricle provide a structural basis for the regular contraction and relaxation of the heart, making the heart beat with strong ductility. Therefore, reconstructing the spiral oriented structure in the natural ventricle is a necessary condition for constructing functional in vitro heart tissue.

[0003] At present, a variety of methods have been proposed for the construction of directional structures, including magnetic field induction, electric field induced mechanical stretching, directional freezing and self-assembly. Although these methods have successfully constructed hydrogel directional structures, they can only construct simple unidirectional directional structures. It is difficult to prepare multi-layer directional structures at different angles in the hydrogel, and it is impossible to simulate the multi-directional layered directional structure of the natural ventricle, and it is even more impossible to reconstruct the macroscopic shape of the ventricle. In addition, mechanical stretching can only be done by unidirectional stretching through the mold. After stretching, the hydrogel and the mold are difficult to separate, and the separation process will affect the orientation. Only myocardial cell bundle structures can be constructed; electric field induction, directional freezing, etc. have a great impact on cell activity, and it is difficult to construct active tissues, such as the patent application named "A directional arrangement of cryogel fiber scaffolds and its preparation method and application" (publication number: CN115382024A). Submicron-scale filament structures arranged in different directions are prepared by electrospinning and air-fluid jet spinning technology, which can guide cells to arrange along the filaments to form a multi-directional oriented structure, such as the patent application named "A polylactic acid / polycaprolactone oriented porous fiber membrane and its application" (publication number: CN118512655A); however, due to the limited controllability of the fiber deposition position and direction, it is difficult to accurately control the fiber orientation direction. Even if the shape of the ventricle can be constructed by spinning on a three-dimensional profiling mold, the mechanical properties of this structure composed of filaments are not enough to maintain a three-dimensional shape during cell culture, resulting in the inability to effectively simulate the shape of the natural ventricle. In addition, the micron-scale fiber scaffold prepared by this method has the problem of being too dense in structure. Cells tend to adhere to the surface of the scaffold and it is difficult to penetrate into it. Cells can only be guided to form a directional structure on the surface.

[0004] Electrostatic printing technology has achieved controllable manufacturing of micron-scale fiber scaffolds, and can construct fiber scaffolds with multiple layers of different orientations to guide cells to form layered directional structures, such as the patent application entitled "A high-density oriented micron fiber conductive nerve guide and controllable electrostatic printing method" (publication number: CN118340947A); however, due to the repulsion effect between fibers caused by charge accumulation during electrostatic printing, it is impossible to construct a highly oriented structure, and it is difficult to construct a large-volume tissue structure; in addition, these methods of guiding cell orientation through filaments can only induce cells to proliferate and migrate along the filaments to achieve directional arrangement, and cannot achieve the orientation of hydrogel fibers.

[0005] Hydrogel 3D printing technology provides great freedom for the manufacture of complex multi-directional active tissues. Cell-containing hydrogels are directly printed through extrusion-based 3D printing technology, and the shear stress applied to the extruded material by the printing nozzle can make the printed cells and hydrogel fibers produce orientation along the printing direction. By adjusting the printing direction in the horizontal plane (XY plane), oriented structures in different directions can be produced, such as the patent application named "A 3D printing method for forming oriented filament structures in situ" (publication number: CN117532871A); however, since the extruded filaments are usually around 100 microns, the directional trend is not obvious, and the material will experience stress relaxation after extrusion, resulting in further weakening of the directional trend. This method can only construct a three-dimensional structure along the direction of the printed filament and based on the principle of layer-by-layer stacking. The adjustability of the directional direction is limited, and it is difficult to achieve continuous oriented structures in different directions in the XZ and YZ planes, and it is impossible to construct the layered spiral oriented structure of the natural ventricle.

[0006] Hydrogel electrostatic printing technology combines the advantages of 3D extrusion printing and electric field induction, and can produce more refined cell-hydrogel oriented structures with more obvious directional trends, such as the patent application entitled "A high-precision biocomposite 3D printing device and printing method" (publication number: CN106222085A); however, electrostatic printing technology takes a lot of time to construct large-scale oriented structures, and cells in the printing barrel have sedimentation problems, resulting in uneven distribution of cells in the printed structure; long-term printing will also affect the activity of cells in the printed structure and the barrel, which is not conducive to the construction of cell-carrying active oriented structures; in addition, since 3D printing technology is mainly based on the layer-by-layer stacking principle to construct three-dimensional oriented structures, the orientation of the constructed oriented structure can only change along the printing filament direction within a single layer plane and between each layer plane, and it is difficult to construct three-dimensional oriented active tissues with different spatial orientation angles. Summary of the invention

[0007] In order to overcome the shortcomings of the above-mentioned prior art, the purpose of the present invention is to provide a method for manufacturing a three-dimensional spirally oriented active ventricular structure, which utilizes a three-dimensional driving scaffold encapsulated in a hydrogel to generate internal stress to induce cells and hydrogel fibers to produce directional arrangement, and constructs a spirally oriented ventricular structure consistent with the natural ventricular arrangement direction through the macroscopic appearance and structural unit design of the three-dimensional driving scaffold, and realizes the manufacturing of active ventricular tissue with contraction and relaxation, blood pumping and electrical conduction functions through in vitro dynamic culture, thereby meeting the needs of tissue repair and in vitro drug testing.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is:

[0009] A method for manufacturing a three-dimensional spiral oriented active ventricular structure comprises the following steps:

[0010] 1) Aiming at the layered spirally arranged myocardial bundle structure in natural ventricular tissue, a three-dimensional driven scaffold structural unit is designed based on finite element simulation analysis, so that the internal stress distribution in the structural unit is consistent with the orientation of the natural ventricle; in addition, the three-dimensional driven scaffold has elastic deformation ability through the design of the structural unit, and can deform synchronously with the cells under the drive of myocardial cells to meet the requirements of ventricular diastole and contraction as well as blood pumping; the overall shape of the three-dimensional driven scaffold is designed by further combining the structural unit with the macroscopic appearance of the natural ventricle, so that the cells and hydrogels planted on the three-dimensional driven scaffold have the shape of the ventricle during the gelation process and the subsequent cell culture and ventricular functionalization process;

[0011] 2) selecting the material of the 3D driving scaffold to be used according to the mechanical and functional properties of natural ventricular tissue, and preparing the 3D driving scaffold by 3D printing, casting, machining or laser cutting;

[0012] 3) designing and manufacturing a casting shell and an inner core of the cell-hydrogel according to the natural tissue morphology and the size of the three-dimensional driving scaffold manufactured in step 2);

[0013] 4) sterilizing and hydrophilizing the three-dimensional driving bracket prepared in step 2) and the casting shell and inner core manufactured in step 3);

[0014] 5) preparing sterile hydrogel and mixing it evenly with the digested cardiomyocytes;

[0015] 6) applying external force to controllably deform the three-dimensional driving scaffold processed in step 4), and then placing it into the casting shell of the cell-hydrogel processed in step 4);

[0016] 7) adding the cell-loaded hydrogel prepared in step 5) into the cell-hydrogel casting shell containing the three-dimensional driving scaffold obtained in step 6) until it covers the entire three-dimensional driving scaffold, and then placing the inner core treated in step 4) and letting it stand to allow the hydrogel to reach a semi-gelled state;

[0017] 8) removing the inner core to allow the three-dimensional driving scaffold to gradually recover its deformation. During this process, the three-dimensional driving scaffold drives the cell-hydrogel to deform accordingly;

[0018] 9) After the hydrogel is completely gelled, the constructed three-dimensional directional active tissue is taken out from the cell-hydrogel casting shell and placed in a culture medium for culture.

[0019] The structural unit of the three-dimensional driving support in the step 1) is one or more of polygonal structures such as oblique lines, curves, parallelograms, regular hexagons, four-pointed stars, five-pointed stars, etc., or one or more of structural units such as negative Poisson's ratio, zero Poisson's ratio, positive Poisson's ratio, etc. The structural unit can meet 20%-100% elastic deformation and provide mechanical support during tissue culture.

[0020] The step 1) optimizes the structural unit through finite element simulation analysis and design, so that the stress distribution in the structural unit is consistent with the spiral orientation direction of the ventricle, and through layered directional structure design, the layered spiral directional structure identical to the natural ventricle is reconstructed, so that the constructed active ventricle has spiral contraction ability and can achieve regular blood pumping and electrical conduction functions.

[0021] The 3D printing of the three-dimensional driving bracket in the step 2) is one or more of fuse forming technology, selective laser sintering technology, photocuring forming technology, surface exposure forming technology, suspension printing technology, and coaxial printing technology.

[0022] The material of the three-dimensional driving bracket in the step 2) includes thermoplastic polymers such as polycaprolactone (PCL), polylactic acid (PLA) and polylactic acid-glycolic acid copolymer (PLGA), thermosetting polymers such as polyglycerol sebacate (PGS), PDMS, Ecoflex, polyurethane, one or more of hydrogel materials such as polyvinyl alcohol (PVA), polyacrylamide (PAAM), polyacrylic acid (PAA), photo-crosslinked gelatin (GelMA), and also includes one or more of materials composited with conductive materials such as carbon nanotubes, graphite, graphene, polythiophene, polypyrrole, polyaniline, etc.

[0023] In the step 2), the three-dimensional driving scaffold material includes degradable materials and non-degradable materials according to the ultimate use target of the ventricle. When degradable materials are used, the three-dimensional driving scaffold is completely degraded after cell functionalization, thereby realizing the manufacture of a bionic ventricle composed only of cells and hydrogels. When non-degradable materials such as conductive materials are selected as the three-dimensional driving scaffold material, the cells in the ventricle are electrically stimulated by the conductive scaffold, so that the electrophysiological properties of the ventricle and the electrical conduction mechanism can be studied.

[0024] The preparation method of the cast shell in step 3) is one or more of 3D printing, casting, and machining, and the shell material is one or more of PDMS, Ecoflex, agarose, photocurable resin and the like.

[0025] The hydrogel in step 5) includes one or more composite materials selected from materials such as gelatin, GelMA, sodium alginate, collagen, bovine fibrin, Matrigel, decellularized matrix, hyaluronic acid, silk protein, chitosan, and ionic hydrogel.

[0026] The cells in step 5) include one or more cells selected from the group consisting of cardiomyocytes, Schwann cells, glial cells, neural stem cells, mesenchymal stem cells, induced pluripotent stem cells, extended pluripotent stem cells, skeletal muscle cells, endothelial cells, and epithelial cells.

[0027] The deformation modes of the three-dimensional driven scaffold in step 6) include compression, stretching, torsion and the like, and the deformation timing includes applying deformation before the cell-hydrogel is added or applying deformation after the cell-hydrogel is added.

[0028] The step 8) can also use an inner core or external force to further deform the three-dimensional driving stent to meet the orientation requirements of the target tissue.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] (1) The present invention adopts a method for stress-induced hydrogel fiber and cell orientation within the structural unit. The orientation process is simple and rapid, and can achieve efficient construction of large-volume active oriented structures. It breaks through the limitations of existing oriented structure construction methods in terms of orientation direction, scale and activity, and provides a new way to construct biomimetic active tissues.

[0031] (2) The present invention adopts a three-dimensional driving support structure unit design, which can not only realize regionalized directional control within the same plane, but also realize layered spiral orientation across different heights and directions. This design makes the directional direction widely adjustable. Combined with the overall bionic shape of the three-dimensional driving support, the integrated bionic manufacturing of the ventricular macroscopic shape and the layered spiral directional microstructure is realized.

[0032] (3) The three-dimensional driving scaffold of the present invention is encapsulated inside the cell-hydrogel, which can not only support the hydrogel to maintain the structural shape, but also does not affect the constructed ventricular structure to achieve functions such as beating and pumping blood; when degradable materials are used to manufacture the three-dimensional driving scaffold, a ventricular structure completely composed of hydrogel and cells can be achieved after the three-dimensional driving scaffold is degraded.

[0033] (4) The present invention proposes a concept for manufacturing three-dimensional spiral oriented active ventricular tissue. Based on the influence of internal stress on the arrangement direction of cells and hydrogel fibers, a structural unit capable of achieving three-dimensional spiral orientation is designed on the basis of simulation analysis. Through the design of a layered oriented structure, a layered spiral oriented structure identical to that of a natural ventricle is reconstructed, so that the constructed active ventricle has the ability of spiral contraction, can achieve regular blood pumping and electrical conduction functions, and makes the in vitro cultured ventricular tissue closer to natural tissue. This can provide an effective in vitro model for drug testing and pathological research, and help to deeply understand the pathogenesis of heart disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is a schematic diagram of a method for manufacturing a three-dimensional spiral oriented active ventricular structure according to an embodiment of the present invention.

[0035] Figure 2 This is a flow chart of a method for manufacturing a three-dimensional spiral oriented active ventricular structure according to an embodiment of the present invention.

[0036] Figure 3 Graph showing the internal stress distribution in different structural units according to an embodiment of the present invention.

[0037] Figure 4 It is an internal stress distribution diagram of the partitioned and oriented structure and a diagram of the partitioned and oriented effect of hydrogel fibers and cells in an embodiment of the present invention.

[0038] Figure 5 It is a simulation model of the three-dimensional layered spiral oriented ventricular structure, a three-dimensional driving bracket and a hydrogel fiber orientation effect diagram of an embodiment of the present invention.

[0039] Figure 6 This is a characterization diagram of the blood pumping and electrical conduction functions of the three-dimensional spirally oriented active ventricular structure according to an embodiment of the present invention. DETAILED DESCRIPTION

[0040] The present invention is further described below in conjunction with examples and accompanying drawings. The examples provided are only for illustrating the present invention, but not for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0041] Reference Figure 1A method for manufacturing a three-dimensional spirally oriented active ventricular structure, the principle of which is to design the shape of a three-dimensional driven bracket and the structural unit of a three-dimensional driven bracket that can achieve controllable deformation according to the macroscopic shape and microscopic fiber and cell arrangement direction of natural ventricular tissue, and to manufacture the three-dimensional driven bracket by 3D printing and other methods. The three-dimensional driven bracket is caused to produce spiral contraction deformation by an external driving device, and after cell-hydrogel is implanted therein, the hydrogel fiber is caused to produce spiral deformation by internal stress, and the three-dimensional driven bracket is restored to shape, and spiral orientation in different directions can be achieved by adjusting the unit structure and structural parameters of the three-dimensional driven bracket.

[0042] Reference Figure 2 , a method for manufacturing a three-dimensional spiral oriented active ventricular structure, comprising the following steps:

[0043] 1) Aiming at the layered spirally arranged myocardial bundle structure in natural ventricular tissue, a three-dimensional driven scaffold structural unit is designed based on finite element simulation analysis, so that the internal stress distribution in the structural unit is consistent with the orientation of the natural ventricle; in addition, the three-dimensional driven scaffold has elastic deformation ability through the design of the structural unit, and can deform synchronously with the cells under the drive of myocardial cells to meet the requirements of ventricular diastole and contraction as well as blood pumping; the overall shape of the three-dimensional driven scaffold is designed by further combining the structural unit with the macroscopic appearance of the natural ventricle, so that the cells and hydrogels planted on the three-dimensional driven scaffold have the shape of the ventricle during the gelation process and the subsequent cell culture and ventricular functionalization process;

[0044] In this embodiment, the shape of a three-dimensional driving stent and a structural unit of a three-dimensional driving stent capable of achieving controllable deformation are designed according to the macroscopic shape of natural ventricular tissue and the myocardial bundle structure arranged in gradients in different directions. The structural unit is optimized through finite element simulation analysis so that the internal stress distribution in the structural unit is consistent with that of natural tissue.

[0045] This embodiment optimizes the structural unit through finite element simulation analysis, can achieve three-dimensional spiral orientation, and through the design of layered spiral orientation structure, reconstructs the same layered spiral orientation structure as the natural ventricle, so that the constructed active ventricle has spiral contraction ability and can achieve regular blood pumping and electrical conduction functions; the structural unit adopts a parallelogram structure, the structural unit meets 60% elastic deformation, and provides mechanical support during tissue culture;

[0046] 2) According to the mechanical and functional properties of natural ventricular tissue, biodegradable PGS was selected as the 3D driving scaffold material, and the 3D driving scaffold structure was prepared by 3D printing;

[0047] 3) designing and manufacturing a casting shell and an inner core of the cell-hydrogel according to the natural tissue morphology and the size of the three-dimensional driving scaffold manufactured in step 2);

[0048] In this embodiment, a resin mold is prepared using photocuring printing technology, Ecoflex is poured into the mold for remolding, and the mold is cured in an oven at 60°C for 8 hours, and the final cast shell and inner core are taken out;

[0049] 4) After sterilizing the PGS three-dimensional drive bracket, the casting shell and the inner core, they are placed in a plasma cleaning machine for hydrophilic treatment;

[0050] 5) Prepare a collagen hydrogel with a concentration of 4%-20%, adjust the pH to neutral, resuspend the digested and centrifuged cardiomyocytes, and mix well;

[0051] 6) After applying 60% deformation to the PGS three-dimensional driving scaffold, the cell-hydrogel treated in step 4) is poured into the casting shell;

[0052] 7) adding the cell-loaded hydrogel prepared in step 5) into the cell-hydrogel casting shell containing the three-dimensional driving scaffold obtained in step 6) until the entire three-dimensional driving scaffold is covered, and then placing the inner core treated in step 4) and standing at 37° C. for 5-10 minutes to allow the collagen hydrogel to reach a semi-crosslinked state;

[0053] 8) removing the inner core to gradually restore the deformation of the three-dimensional driving scaffold. During this process, the three-dimensional driving scaffold drives the cells and the hydrogel to deform accordingly;

[0054] 9) After the hydrogel is completely gelled, the constructed three-dimensional directional active tissue is removed from the cell-hydrogel casting shell, and the three-dimensional driving scaffold coated with the hydrogel is slowly released. The tissue is allowed to stand for 10-20 minutes, and the culture medium is added and placed in a 37°C incubator for culture. After the PGS is degraded and the cardiomyocytes mature, ventricular function characterization and subsequent drug testing are performed.

[0055] like Figure 3 As shown, in this embodiment, by designing different structural units, the internal stress in the structural unit can be regulated to be distributed in different directions; the parallelogram structural unit in the figure can make the internal stress distributed along the inclined angle, and the internal stress angle can be regulated by controlling the parallelogram angle; the multi-angle oblique line structural unit and the negative Poisson's ratio structure in the figure can make the internal stress distributed in a zoned and directional manner; the four-pointed star structural unit in the figure can not only make the internal stress distributed in a zoned manner, but also make the internal stress distributed in a ring shape.

[0056] like Figure 4 As shown, the present embodiment designs a symmetrical parallelogram structural unit. From the simulation structure, it can be seen that the internal stress directions in region 1 and region 2 are also symmetrically distributed. After planting cells and hydrogels in the scaffold, a partitioned directional structure can be constructed. As shown in the figure, the hydrogel fibers and cells are symmetrically distributed along the internal stress direction.

[0057] like Figure 5As shown, in this embodiment, a three-dimensional layered driving stent with a ventricular shape and different inclination angles of the inner and outer layers is designed based on a parallelogram structural unit. The internal stress simulation model verifies that the internal stress of the three-dimensional layered driving stent is distributed in a layered spiral, and the internal stresses of the inner and outer layers are arranged in opposite directions. PGS is selected as the three-dimensional driving stent material, and the driving stent is manufactured by melt printing technology. The confocal microscope proves that the direction of the hydrogel fibers in the inner and outer layers of the prepared ventricular structure is the same as the simulation result, and has the effect of layered spiral orientation;

[0058] like Figure 6 As shown, in this embodiment, the blood pumping function of the constructed three-dimensional spiral oriented active ventricle structure is characterized. The movement trajectory of the fluorescent particles at the ventricular outlet is analyzed by the PIV equipment, and it can be seen that the particles are pumped out of the ventricle when the ventricle contracts, and the particles are pumped into the ventricle when the ventricle relaxes; the electrical conduction on the ventricular surface is characterized by the cursor measurement equipment, and it can be seen that the electrical signal propagates in a spiral, which proves that the three-dimensional spiral oriented active ventricle manufactured in this embodiment has blood pumping and electrical conduction functions.

[0059] The above description of the embodiments is to facilitate the understanding and use of the invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments, and improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the present invention should be within the scope of protection of the present invention.

Claims

1. A method for manufacturing a three-dimensional spiral oriented active ventricular structure, characterized in that: The following steps are involved: 1) Aiming at the layered spirally arranged myocardial bundle structure in natural ventricular tissue, a three-dimensional driving stent structure unit is designed based on finite element simulation analysis, so that the internal stress distribution in the structure unit is consistent with the natural ventricular orientation; In addition, the design of the structural unit enables the three-dimensional driving scaffold to have elastic deformation ability, and deform synchronously with the cells under the drive of the myocardial cells to meet the requirements of ventricular relaxation and contraction and blood pumping. The overall shape of the three-dimensional driving scaffold is further designed by combining the shape of the structural unit with the macroscopic appearance of the natural ventricle, so that the cells and hydrogels planted on the three-dimensional driving scaffold have the shape of the ventricle during the gelation process and the subsequent cell culture and ventricular functionalization process. 2) According to the mechanical and functional characteristics of natural ventricular tissue, the material of the 3D drive scaffold to be used is selected, and the 3D drive scaffold is prepared by 3D printing, casting, machining or laser cutting. 3) designing and manufacturing a casting shell and an inner core of the cell-hydrogel according to the natural tissue morphology and the size of the three-dimensional driving scaffold manufactured in step 2); 4) sterilizing and hydrophilizing the three-dimensional driving bracket prepared in step 2) and the casting shell and inner core manufactured in step 3); 5) preparing sterile hydrogel and mixing it evenly with the digested cardiomyocytes; 6) applying external force to controllably deform the three-dimensional driving scaffold processed in step 4), and then placing it into the casting shell of the cell-hydrogel processed in step 4); 7) adding the cell-loaded hydrogel prepared in step 5) into the cell-hydrogel casting shell containing the three-dimensional driving scaffold obtained in step 6) until it covers the entire three-dimensional driving scaffold, and then placing the inner core treated in step 4) and letting it stand to allow the hydrogel to reach a semi-gelled state; 8) removing the inner core to allow the three-dimensional driving scaffold to gradually recover its deformation. During this process, the three-dimensional driving scaffold drives the cell-hydrogel to deform accordingly; 9) After the hydrogel is completely gelled, the constructed three-dimensional directional active tissue is taken out from the cell-hydrogel casting shell and placed in a culture medium for culture.

2. The manufacturing method according to claim 1, characterized in that: The structural unit of the three-dimensional driving support in the step 1) is one or more of the polygonal structures of oblique lines, curves, parallelograms, regular hexagons, four-pointed stars, and five-pointed stars, or one or more of the structural units with negative Poisson's ratio, zero Poisson's ratio, and positive Poisson's ratio. The structural unit can meet 20%-100% elastic deformation and provide mechanical support during tissue culture.

3. The manufacturing method according to claim 1, characterized in that: The step 1) optimizes the structural unit through finite element simulation analysis and design, so that the stress distribution in the structural unit is consistent with the spiral orientation direction of the ventricle, and through layered directional structure design, the layered spiral directional structure identical to the natural ventricle is reconstructed, so that the constructed active ventricle has spiral contraction ability and can achieve regular blood pumping and electrical conduction functions.

4. The manufacturing method according to claim 1, characterized in that: The 3D printing of the three-dimensional driving bracket in the step 2) is one or more of fuse forming technology, selective laser sintering technology, photocuring forming technology, surface exposure forming technology, suspension printing technology, and coaxial printing technology.

5. The manufacturing method according to claim 1, characterized in that: The material of the three-dimensional driving support in the step 2) is a thermoplastic polymer of polycaprolactone (PCL), polylactic acid (PLA) and polylactic acid-glycolic acid copolymer (PLGA), a thermosetting polymer of polyglyceryl sebacate (PGS), PDMS, Ecoflex, polyurethane, a hydrogel material of polyvinyl alcohol (PVA), polyacrylamide (PAAM), polyacrylic acid (PAA), and photo-crosslinked gelatin (GelMA), or one or more of the materials composited with a conductive material of carbon nanotubes, graphite, graphene, polythiophene, polypyrrole, and polyaniline.

6. The manufacturing method according to claim 1, characterized in that: The materials of the three-dimensional driving scaffold in the step 2) include degradable materials and non-degradable materials according to the ultimate use target of the ventricle; when degradable materials are used, the three-dimensional driving scaffold is completely degraded after cell functionalization to achieve a bionic ventricle composed only of cells and hydrogels; when non-degradable materials of conductive materials are selected as the three-dimensional driving scaffold materials, the cells in the ventricle are electrically stimulated through the conductive scaffold to study the electrophysiological properties of the ventricle and the electrical conduction mechanism.

7. The manufacturing method according to claim 1, characterized in that: The preparation method of the cast shell in step 3) is one or more of 3D printing, casting, and machining, and the shell material is one or more of PDMS, Ecoflex, agarose, photocurable resin and the like.

8. The manufacturing method according to claim 1, characterized in that: The hydrogel in step 5) is one or more composite materials selected from the group consisting of gelatin, GelMA, sodium alginate, collagen, bovine fibrin, Matrigel, decellularized matrix, hyaluronic acid, silk protein, chitosan, and ionic hydrogel; The cells in step 5) include one or more cells selected from the group consisting of cardiomyocytes, Schwann cells, glial cells, neural stem cells, mesenchymal stem cells, induced pluripotent stem cells, extended pluripotent stem cells, skeletal muscle cells, endothelial cells, and epithelial cells.

9. The manufacturing method according to claim 1, characterized in that: The deformation modes of the three-dimensional driven scaffold in step 6) include compression, stretching, and torsion, and the deformation timing includes applying deformation before the cell-hydrogel is added or applying deformation after the cell-hydrogel is added.

10. The manufacturing method according to claim 1, characterized in that: The step 8) can also use an inner core or external force to further deform the three-dimensional driving stent to meet the orientation requirements of the target tissue.

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