CN107214957B - Composite biological material 3D printing system and printing method - Google Patents
Composite biological material 3D printing system and printing method Download PDFInfo
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- CN107214957B CN107214957B CN201710486777.7A CN201710486777A CN107214957B CN 107214957 B CN107214957 B CN 107214957B CN 201710486777 A CN201710486777 A CN 201710486777A CN 107214957 B CN107214957 B CN 107214957B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y30/00—Apparatus for additive manufacturing; Details thereof or accessories therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y10/00—Processes of additive manufacturing
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- Manufacturing & Machinery (AREA)
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- Spinning Methods And Devices For Manufacturing Artificial Fibers (AREA)
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Abstract
The invention provides a composite biological material 3D printing system and a printing method. The 3D printing system for the composite biological material comprises a first charging barrel and a second charging barrel, wherein an electrostatic spinning device for generating continuous reinforcing fibers is arranged below the first charging barrel, the electrostatic spinning device comprises a high-voltage nozzle arranged at the bottom of the first charging barrel and a grounding ring arranged below the high-voltage nozzle, and an electric field is formed between the high-voltage nozzle and the grounding ring; the utility model discloses a mixing nozzle, including ground ring, second feed cylinder, mixing nozzle, adapter tube, second feed cylinder, the ground ring below is equipped with mixing nozzle, the second feed cylinder with be equipped with between the mixing nozzle, adapter tube one end is connected the bottom of second feed cylinder, the other end is connected mixing nozzle's upper end. The invention has simple structure and convenient use, can realize the accurate molding of various materials and cells, improves the mechanical property and the cell activity of the printing structure body, and forcefully promotes the attachment, migration and proliferation of the cells.
Description
Technical Field
The invention relates to the technical field of biological tissue engineering, in particular to a composite biological material 3D printing system and a printing method.
Background
Tissue engineering is a discipline of fusion engineering, life sciences, and materials sciences that constructs and cultures biologically active structures in vitro by mimicking the process of human tissue organ formation. Among these, 3D printing technology is the most powerful research means in the field of tissue engineering because it can use various materials to form any complex three-dimensional structure. The principle of the 3D printing technology is layered manufacturing and layer-by-layer accumulation. The traditional cell printing spray head is to mix cells with biological materials and extrude the mixture to form a strip of silk threads, and then repeatedly form surfaces, and the surfaces are piled up to form a corresponding three-dimensional structure. The typical spray head is a motor-assisted spray head.
FIG. 1 is a schematic drawing showing the three-dimensional structure of a traditional motor-assisted spray head, a linear stepping motor is fixed on a moving device bracket, a screw rod can apply certain pressure to forming materials in the spray head under the drive of the linear stepping motor, the forming materials are extruded from a spray nozzle, and a heating rod and a heat-insulating jacket are arranged at the lower section of the spray head to keep the forming materials in the spray head at set temperatures.
This design, while simple and reliable, has the following disadvantages: 1) The mechanical property of the printed structure body is poor, and the biological natural polymer material which is mixed with cells and has good biocompatibility, such as gelatin, collagen, fibrinogen and the like, has weak mechanical property, and can not be even taken out for detection after being soaked in a culture solution for a long time after being printed; 2) The cell activity is not high, the traditional printing spray head is pushed and extruded by a motor, and the volumetric control method generates great shearing force to cells when the cells pass through a narrow nozzle, so that cell membranes are broken and dead; 3) In order to provide a certain molding property to the relevant material, the concentration of the biological material of the mixed cells such as gelatin is high, and in this case, migration and proliferation of the cells therein are difficult.
Disclosure of Invention
The invention provides a 3D printing system for composite biological materials, which is simple in structure and convenient to use, and can realize accurate molding of various materials and cells.
Another object of the present invention is to provide a printing method of the composite biomaterial.
In order to solve the technical problems, the invention adopts the following technical scheme: the 3D printing system for the composite biological material comprises a first charging barrel and a second charging barrel, wherein an electrostatic spinning device for generating continuous-state reinforced fibers is arranged below the first charging barrel, the electrostatic spinning device comprises a high-voltage nozzle arranged at the bottom of the first charging barrel and a grounding ring arranged below the high-voltage nozzle, an electric field is formed between the high-voltage nozzle and the grounding ring, a mixing nozzle is arranged below the grounding ring, a transfer pipe is arranged between the second charging barrel and the mixing nozzle, one end of the transfer pipe is connected with the bottom of the second charging barrel, the other end of the transfer pipe is connected with the upper end of the mixing nozzle, the first charging barrel is filled with a biological synthetic polymer material solution for generating continuous-state electrostatic spinning fibers, and because an electric field is formed between the high-voltage nozzle and the grounding ring, the biological synthetic polymer material solution forms micron-level jet flow through the high-voltage nozzle based on an electrostatic spinning principle, namely the final continuous-state electrostatic spinning fibers enter the mixing nozzle through the grounding ring, and the biological synthetic polymer material has good mechanical property, and the generated continuous-state electrostatic spinning fibers play a role in supporting role in a composite biological fiber skeleton; the second charging barrel is filled with a mixed solution of cells and biological natural polymer materials for generating bioactive materials, the biological natural polymer materials comprising the cells have good biocompatibility, a proper living microenvironment can be provided for the cells, the mixed solution enters the mixing nozzle through the switching pipe and wraps the periphery of the electrostatic spinning fiber from the first charging barrel, and finally the mixed solution and the mixed solution are sprayed out of the mixing nozzle together.
Further, the first temperature control device and the second temperature control device are arranged on the periphery of the first charging barrel and the periphery of the second charging barrel respectively, and the first temperature control device and the second temperature control device can provide the most suitable temperature environment for the inside of the first charging barrel and the second charging barrel respectively so as to ensure the optimal activity of materials and cells contained in the first charging barrel and the second charging barrel.
Further, the first temperature control device comprises a first heating ring arranged on the outer wall of the first charging barrel and a first heat preservation layer coated on the peripheries of the first heating ring, the first charging barrel and the mixing nozzle. The second temperature control device comprises a second heating ring arranged on the outer wall of the second charging barrel and a second heat preservation layer coated on the second heating ring and the periphery of the second charging barrel. In this way, the first and second cartridges are each provided with an independent heating ring and heat insulating layer, and therefore the temperatures inside the first and second cartridges are independently controlled, that is, the first and second temperature control devices are independently controlled from each other.
Further, the distance between the high-voltage nozzle and the grounding ring is 3-30mm.
Further, the pushing mode of the feed liquid contained in the first feed cylinder and the second feed cylinder is high-pressure gas pushing.
The invention also provides a 3D printing method of the composite biological material, which comprises the following steps:
s1, selecting proper biological synthetic polymer materials to prepare a solution, loading the solution into a first charging barrel for standby, selecting proper cells and biological natural polymer materials, preparing a cell mixed solution according to proper proportion, and loading the solution into a second charging barrel for standby;
s2, starting a first temperature control device and a second temperature control device, and respectively setting the temperatures of the first charging barrel and the second charging barrel at proper gears;
s3, starting a control program to enable the 3D printing system to work, forming micron-sized jet flow by the biosynthetic high polymer material solution in the first charging barrel through a high-voltage nozzle based on the electrostatic spinning principle, then enabling the jet flow to enter a mixing nozzle through a grounding ring, and enabling the cell mixed solution in the second charging barrel to enter the mixing nozzle through a transfer pipe and surround the micron-sized jet flow formed by the biosynthetic high polymer material solution;
s4, spraying the solution of the biosynthesis polymer material and the cell mixed solution through a mixing nozzle to form the biological composite material containing the electrostatic spinning fiber.
In step S1, the bio-synthetic polymer material is one or more of polyurethane, polylactic acid, polyglycolide, and polylactic acid-glycolic acid copolymer. The cells are any one of adipose stem cells, bone marrow mesenchymal stem cells, cardiac muscle cells, schwann cells, liver cells, cancer cells and the like; the biological natural polymer material is any one or more than two of fibrinogen, collagen, gelatin, silk fibroin, alginate, hyaluronic acid, chitosan and the like.
Compared with the prior art, the invention has the beneficial effects that:
according to the 3D printing system, the first charging barrel and the second charging barrel are arranged, the high-voltage nozzle and the grounding ring are arranged at the bottom of the first charging barrel, the first charging barrel is filled with the biological synthetic polymer material solution with good mechanical properties, the second charging barrel is filled with the mixed solution of cells and biological natural polymer materials, and based on the combination of the electrostatic spinning technology and the high-pressure pneumatic technology, the accurate formation of various materials and cells can be realized, the mechanical properties of a forming structure are further improved, the good mechanical properties of the biological synthetic polymer materials and the good biocompatibility of the biological natural polymer materials are combined, the mechanical properties and the cell activity of a printing structure are improved, and the attachment, migration and proliferation of cells can be forcefully promoted.
The 3D printing system provided by the invention uses high-pressure gas to push the cell mixed liquid to extrude, is pressure type control, can weaken the shearing force affecting the activity of cells to the minimum, greatly improves the activity of the formed cells, and has high pneumatic spraying precision and high response speed.
Drawings
Fig. 1 is a schematic diagram showing the three-dimensional structure subdivision of a conventional motor-assisted spray head.
FIG. 2 is a graph showing the effect of the electrospun fiber-containing biocomposite material of the present invention.
FIG. 3 is a schematic cross-sectional view of a biocomposite material containing electrospun fibers of the present invention.
Fig. 4 is a schematic diagram of the overall structure of the 3D printing head according to the present invention.
In the figure: 201-a screw; 202-a linear stepper motor; 203-heating rod; 204-heat insulation jacket; 205-nozzle; 601-biological natural polymer material; 602-cells; 603—biosynthesis of polymeric materials; 1-a first barrel; 2-a second barrel; 3-high voltage nozzle; 4-a ground ring; 5-a mixing nozzle; 6, connecting a pipe; 7-a first heating ring; 8-a first heat preservation layer; 9-a second heating ring; 10-a second heat preservation layer.
Detailed Description
The drawings are for illustrative purposes only and are not to be construed as limiting the present patent; for the purpose of better illustrating the embodiments, certain elements of the drawings may be omitted, enlarged or reduced and do not represent the actual product dimensions; it will be appreciated by those skilled in the art that certain well-known structures in the drawings and descriptions thereof may be omitted. The positional relationship depicted in the drawings is for illustrative purposes only and is not to be construed as limiting the present patent.
Example 1
As shown in fig. 4, a 3D printing system for composite biological materials, which comprises a first charging barrel 1 and a second charging barrel 2, wherein an electrostatic spinning device for generating continuous reinforced fibers is arranged below the first charging barrel 1, the electrostatic spinning device comprises a high-voltage nozzle 3 arranged at the bottom of the first charging barrel 1 and a grounding ring 4 arranged below the high-voltage nozzle 3, thus an electric field is formed between the high-voltage nozzle 3 and the grounding ring 4, a mixing nozzle 5 is arranged below the grounding ring 4, a transfer tube 6 is arranged between the second charging barrel 2 and the mixing nozzle 5, one end of the transfer tube 6 is connected with the bottom of the second charging barrel 2, the other end of the transfer tube is connected with the upper end of the mixing nozzle 5, the first charging barrel 1 is filled with a biological synthetic polymer material solution 603 for generating continuous electrostatic spinning fibers, and because the electric field is formed between the high-voltage nozzle 3 and the grounding ring 4, based on the electrostatic spinning principle, the biological synthetic polymer material solution forms a jet of micron-sized through the high-voltage nozzle 3, namely, the final continuous electrostatic spinning fibers enter the biological spinning ring 5 to have good mechanical properties, and the biological polymer material solution enters the composite spinning frame 603 through the high-mechanical properties of the high-voltage nozzle 5, and the biological spinning frame 4 has good mechanical properties; the second material cylinder 2 contains a mixed solution of cells 602 and a biological natural polymer material 601 for generating bioactive materials, the biological natural polymer material 601 comprising the cells 602 has good biocompatibility, a proper living microenvironment can be provided for the cells 602, the mixed solution enters the mixing nozzle 5 through the switching tube 6, wraps the periphery of the electrostatic spinning fiber coming out of the first material cylinder 1, and finally the mixed solution and the biological natural polymer material 601 are sprayed out of the mixing nozzle 5 together.
As shown in fig. 4, the first temperature control device and the second temperature control device are respectively disposed at the periphery of the first barrel 1 and the second barrel 2, and the first temperature control device and the second temperature control device can respectively provide the most suitable temperature environment for the inside of the first barrel 1 and the second barrel 2, so as to ensure the optimal activity of the materials and cells contained in the first barrel 1 and the second barrel 2.
As shown in fig. 4, the first temperature control device includes a first heating ring 7 disposed on the outer wall of the first barrel 1, and a first heat insulation layer 8 covering the first heating ring 7, the first barrel 1, and the periphery of the mixing nozzle 5. The second temperature control device comprises a second heating ring 9 arranged on the outer wall of the second charging barrel 2 and a second heat preservation layer 10 coated on the second heating ring 9 and the periphery of the second charging barrel 2. In this way, the first and second cartridges 1 and 2 are each provided with an independent heating ring and insulation layer, and thus the temperatures inside the first and second cartridges 1 and 2 are independently controlled, that is, the first and second temperature control devices are independently controlled from each other.
In this embodiment, the distance between the high voltage nozzle 3 and the ground ring 4 is 3-30mm.
In this embodiment, the pushing manner of the feed liquid contained in the first feed cylinder 1 and the second feed cylinder 2 is high-pressure gas pushing.
Example 2
A method of 3D printing of composite biological materials, comprising the steps of:
s1, selecting polyurethane as a biological synthetic polymer material 603, dissolving polyurethane in tetraethylene glycol to prepare a solution with the concentration of 5% (w/v), loading the solution into a first charging barrel 1 for standby, selecting adipose-derived stem cells as cells 602, selecting fibrinogen as a biological natural polymer material 601, wherein the fibrinogen solution is prepared by dissolving fibrinogen powder in a DMEM solution with the mass concentration of 0.1% (w/v), and mixing the adipose-derived stem cells into the fibrinogen solution with the density of 1 multiplied by 10 6 Adding endothelial cell growth factor (50 ng/mL) into the mixture to prepare a cell mixture, and filling the cell mixture into a second charging barrel 2 for later use;
s2, starting a first temperature control device and a second temperature control device, and respectively setting the temperatures of the first charging barrel 1 and the second charging barrel 2 at proper gears;
s3, starting a control program to enable the 3D printing system to start to work, forming micron-sized jet flow by the solution of the biological synthetic polymer material 603 in the first charging barrel (1) through the high-voltage nozzle 3 based on the electrostatic spinning principle, then enabling the solution to enter the mixing nozzle 5 through the grounding ring 4, and enabling the cell mixed solution in the second charging barrel 2 to enter the mixing nozzle 5 through the transfer pipe 6 and surround the micron-sized jet flow formed by the solution of the biological synthetic polymer material 603;
s4, spraying the solution of the biosynthesis polymer material 603 and the cell mixed solution through a mixing nozzle 5 to form the composite biomaterial containing the electrostatic spinning fiber shown in fig. 2 and 3.
In this embodiment, the bio-synthetic polymer material 603 may be any one or more of polyurethane, polylactic acid, polyglycolide, and polylactic acid-glycolic acid copolymer. The cell 602 may be any one of adipose stem cells, bone marrow mesenchymal stem cells, cardiac muscle cells, schwann cells, liver cells, cancer cells, and the like; the biological natural polymer material 601 may be any one or more of fibrinogen, collagen, gelatin, silk fibroin, alginate, hyaluronic acid, chitosan, and the like.
It is to be understood that the above examples of the present invention are provided for clarity of illustration only and are not limiting of the embodiments of the present invention. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. Any modification, equivalent replacement, improvement, etc. which come within the spirit and principles of the invention are desired to be protected by the following claims.
Claims (9)
1. The composite biological material 3D printing system is characterized by comprising a first charging barrel (1) and a second charging barrel (2), wherein an electrostatic spinning device for generating continuous reinforcing fibers is arranged below the first charging barrel (1), the electrostatic spinning device comprises a high-voltage nozzle (3) arranged at the bottom of the first charging barrel (1) and a grounding ring (4) arranged below the high-voltage nozzle (3), and an electric field is formed between the high-voltage nozzle (3) and the grounding ring (4); the utility model discloses a mixing nozzle, including ground ring (4), second feed cylinder (2), mixing nozzle (5) are equipped with switching pipe (6) between second feed cylinder (2), switching pipe (6) one end is connected the bottom of second feed cylinder (2), the other end is connected the upper end of mixing nozzle (5).
2. The composite biomaterial 3D printing system according to claim 1, wherein the first and second barrels (1, 2) are provided with a first and a second temperature control device, respectively, at the periphery thereof, and the first and second temperature control devices are controlled independently of each other.
3. The composite biomaterial 3D printing system according to claim 2, wherein the first temperature control device comprises a first heating ring (7) arranged on the outer wall of the first charging barrel (1) and a first heat preservation layer (8) coated on the peripheries of the first heating ring (7), the first charging barrel (1) and the mixing nozzle (5).
4. The composite biomaterial 3D printing system according to claim 2, wherein the second temperature control device comprises a second heating ring (9) arranged on the outer wall of the second charging barrel (2) and a second heat insulation layer (10) coated on the periphery of the second heating ring (9) and the second charging barrel (2).
5. A composite biomaterial 3D printing system according to claim 1, characterized in that the distance between the high voltage nozzle (3) and the ground ring (4) is 3-30mm.
6. A composite biomaterial 3D printing system according to any of claims 1-5, wherein the pushing means of the feed liquid contained in the first and second cartridges (1, 2) is high pressure gas pushing.
7. A method of 3D printing of a biocomposite material comprising electrospun fibers, comprising the steps of:
s1, selecting a proper biological synthetic polymer material (603) to prepare a solution, loading the solution into a first charging barrel (1) for standby, selecting proper cells (602) and a biological natural polymer material (601), preparing a cell mixed solution according to a proper proportion, and loading the solution into a second charging barrel (2) for standby;
s2, starting a first temperature control device and a second temperature control device, and respectively setting the temperatures of the first charging barrel (1) and the second charging barrel (2) at proper gears;
s3, starting a control program to enable the 3D printing system to start to work, forming micron-sized jet flow by the solution of the biological synthetic polymer material (603) in the first charging barrel (1) through the high-voltage nozzle (3) based on the electrostatic spinning principle, then enabling the solution to enter the mixing nozzle (5) through the grounding ring (4), and meanwhile enabling the cell mixed solution in the second charging barrel (2) to enter the mixing nozzle (5) through the switching tube (6) and surrounding the micron-sized jet flow formed by the solution of the biological synthetic polymer material (603);
s4, spraying the solution of the biosynthesis polymer material (603) and the cell mixed solution through a mixing nozzle (5) to form the biological composite material containing the electrostatic spinning fiber.
8. The printing method according to claim 7, wherein in step 1, the bio-synthetic polymer material (603) is any one or more of polyurethane, polylactic acid and polylactic acid-glycolic acid copolymer.
9. The printing method according to claim 7, wherein in step 1, the cells (602) are any one of adipose stem cells, bone marrow mesenchymal stem cells, cardiomyocytes, schwann cells, hepatocytes and cancer cells; the biological natural polymer material (601) is any one or more than two of fibrinogen, collagen, gelatin, silk fibroin, alginate, hyaluronic acid and chitosan.
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| CN108042854A (en) * | 2017-12-16 | 2018-05-18 | 诺迈尔（苏州）医学科技有限公司 | For the production technology of the gelatin fiber guide tissue regeneration film of Dental implant surgery |
| CN108527841A (en) * | 2018-02-26 | 2018-09-14 | 南昌大学 | A kind of method that multi-modal biological printing system and biological printing system prepare biomimetic scaffolds |
| CN109732903A (en) * | 2019-01-25 | 2019-05-10 | 上海交通大学医学院附属第九人民医院 | A kind of high-precision biology 3 D-printing method and device based on electrostatic spinning technique |
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| CN207310537U (en) * | 2017-06-23 | 2018-05-04 | 广州迈普再生医学科技有限公司 | A kind of composite biological material 3D printing system |
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Address after: 510663 Guangdong Guangzhou high tech Industrial Development Zone, Science City, 80 E third district. Applicant after: Medprin Regenerative Medical Technologies Co.,Ltd. Address before: 510670 Guangzhou District, Guangdong, new high tech Industrial Development Zone, Science City, 80 E third district. Applicant before: Medprin Regenerative Medical Technologies Co.,Ltd. |
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