CN111607514A - Biological 3D print platform is mixed to compound biomaterial developments - Google Patents
Biological 3D print platform is mixed to compound biomaterial developments Download PDFInfo
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- CN111607514A CN111607514A CN202010467436.7A CN202010467436A CN111607514A CN 111607514 A CN111607514 A CN 111607514A CN 202010467436 A CN202010467436 A CN 202010467436A CN 111607514 A CN111607514 A CN 111607514A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
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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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0655—Chondrocytes; Cartilage
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2513/00—3D culture
Abstract
The invention discloses a composite biological material dynamic mixed biological 3D printing platform. The device comprises a dynamic mixing extrusion device and a 3D printing motion platform, wherein the 3D printing platform is used for controlling the dynamic mixing extrusion device to perform three-dimensional motion so as to realize the preparation of the gel microspheres. The dynamic mixing extrusion device consists of a dynamic mixing printing spray head and micro injection pumps on two sides. The dynamic mixing printing nozzle is mainly used for uniformly mixing the sodium alginate-collagen solution containing the chondrocytes, the micro injection pump is used for controlling the liquid inlet speed and the liquid inlet quantity of the biological material solution, the micro injection pump on one side supplies the sodium alginate solution containing the chondrocytes, and the micro injection pump on the other side supplies the collagen. The invention can print the biological gel material structural body with three structures of point, line and body; the structural material adopted by the mixing cavity part has certain biocompatibility, and biological cells are introduced into the mixing cavity part, so that the initial experiment of the biological 3D printing of the active cells can be carried out.
Description
Technical Field
The invention belongs to the technical field of biological manufacturing-biological 3D printing equipment, and is applied to the field of cell 3D printing. The device specifically comprises a dynamic mixing extrusion device and a three-dimensional motion platform, and realizes the printing and manufacturing of the gel microspheres of the sodium alginate-collagen biological composite material through the assembly of two structures, thereby realizing the 3D culture of chondrocytes.
Background
The traditional biological 3D printing technology mainly uses biological materials without cells to manufacture biological products such as medical models, metal bones, biological ceramics, tissue engineering scaffolds, and the like. The new branch cell 3D printing technology extended from biological 3D printing is to manufacture biocompatible materials, cells and nutrient materials required by the cells into complex three-dimensional functional living tissues in an additive manufacturing mode, and realize the processes from point, line and surface manufacturing dimensionality to three-dimensional printing. Has wide application prospect in the fields of regeneration and reconstruction of tissues such as bones, skin, blood vessels, heart, kidney, liver and the like, drug screening and the like.
The traditional two-dimensional cell culture is carried out on a two-dimensional plane of a culture dish or a culture bottle, and the three-dimensional culture is to use biological materials to construct a corresponding structure to simulate the growth condition and environment of cells in vivo so that the cells can grow in three dimensions under the in vitro condition. The gel microsphere is a simple and symmetrical structure and is widely applied to the aspects of cell wrapping, drug release, biosensors and the like. Unlike medicinal microspheres, microspheres for tissue engineering are generally used as cell scaffolds to provide carriers for adhesion and growth of cells, and the microspheres for tissue engineering should have good biocompatibility and degradability; the preparation method of the microsphere in the prior art mainly comprises an emulsification dispersion method, a coagulation method and a polymerization method.
The raw materials for 3D bioprinting can be broadly divided into two categories: scaffold material and seed cells. The skeleton materials currently used for 3D printing mainly include hydrogel, poly (lactic-co-glycolic acid), acrylate, cellulose, gelatin, alginate, and the like. The seed cells have various types, such as umbilical cord blood endothelial cells, vascular endothelial cells, cardiac muscle cells, fibroblasts, chondrocytes and the like. Biomaterials applied to 3D printing technologies include: 1) biodegradable polymers, one class of which is natural materials, including polysaccharides (starch, alginates, chitin/chitosan, hyaluronic acid derivatives) or proteins (soy, collagen, fibrin glue, etc.), can be used as reinforcing materials. Another class is synthetic biodegradable polymers. Synthetic polymers can be made under controlled conditions; 2) bioactive ceramics, including calcium phosphate ceramics, hydroxyapatite ceramics; 3) the hydrogel is prepared from natural hydrophilic polymer such as polysaccharide (hydrogel of starch, cellulose, alginic acid, hyaluronic acid, chitosan, etc.) and polypeptide (collagen, poly-L-lysine, poly-L-glutamic acid, etc.). The synthetic hydrophilic polymer includes polyethylene, alcohol, acrylic acid and its derivatives (polyacrylic acid, polymethacrylic acid, polyacrylamide, poly-N-polyacrylamide, etc.).
Existing micromixers can be largely divided into dynamic mixers and static mixers, which are distinguished primarily by the presence or absence of active elements. While dynamic micromixers require the mixing power to be obtained by the motion of the active element, static micromixers do not rely on the motion of the element, but rather the mixing effect of the fluid is achieved by the geometry of the mixer itself.
Common static micromixers include T-type and Y-type micromixers, and mixers with complex functions can be obtained by improving simple T-type and Y-type micromixers. Liu et al improve a design micromixer through going on common blender, and its three-dimensional mechanism is similar to snakelike, and the flow of characteristics fluid in bend department has the flow perpendicular with main part flow direction, can make the fluid atress take place the distortion to increase the area of contact between the fluid that waits to mix and made the fluid mixing process strengthened. In the aspect of dynamic micromixers, including ultrasonic mixing, electrokinetic mixing, centrifugal mixing, alternate pulse flow mixing and other modes of micromixers, a Lewis group designs a mechanical mixing type micromixer for printing gradient materials, and structurally comprises a mixing cavity formed by conical dispensing needles, a cylindrical metal stirrer, a liquid inlet joint and a stirring motor.
Disclosure of Invention
The invention designs a composite biomaterial dynamic mixing biological 3D printing platform, and aims to manufacture gel microspheres based on uniform concentration distribution of a sodium alginate-collagen composite biomaterial and realize 3D culture of active chondrocytes. Aiming at a special biological printing system for mixing cells in a sodium alginate-collagen composite solution, a dynamic mixing basic device based on the principle of a dynamic mixer is designed, and the device is directly arranged on a 3D motion platform to move and work, so that the printing and manufacturing of gel microspheres are realized.
The device comprises a dynamic mixing extrusion device and a 3D printing motion platform, wherein the 3D printing platform is used for controlling the dynamic mixing extrusion device to perform three-dimensional motion so as to realize the preparation of the gel microspheres.
The dynamic mixing extrusion device consists of a dynamic mixing printing spray head and micro injection pumps on two sides.
The dynamic mixing printing nozzle is mainly used for uniformly mixing sodium alginate-collagen solution containing chondrocytes, the micro injection pump is used for controlling the liquid inlet speed and the liquid inlet amount of the biological material solution, the micro injection pump on one side supplies the sodium alginate solution containing the chondrocytes, and the micro injection pump on the other side supplies the collagen.
The dynamic mixing printing nozzle comprises a mixing cavity with a discharge hole at the bottom, two feeding ports are symmetrically distributed on two side walls of the cavity, and the feeding ports are butted with Ruhr joints and are used for connecting an outlet of a micro injection pump; a stirrer is arranged in the mixing cavity, and a plurality of stirring blades are arranged on the periphery of the stirrer; the outer side of the top of the cavity is provided with a speed-adjustable stepping motor which is assembled and linked with the stirrer.
Furthermore, the stirrer is replaceable, is made of photosensitive resin and is rapidly manufactured by utilizing a photocuring 3D printing technology; the damage of stirring to cells is reduced and the uniform stirring and mixing of the biological materials are realized by adjusting the rotating speed of the stepping motor; the stirrer is provided with a rotating shaft, namely the rotating shaft and the stirrer are integrated, and the rotating shaft and the speed-adjustable stepping motor are arranged on the stirrer.
Furthermore, the mixing cavity is composed of an upper part and a lower part, and the material of the mixing cavity is photosensitive resin; the upper cavity is provided with a mixing space, a certain gap is reserved between the stirrer and the inner wall of the upper cavity to ensure that the stirrer is stirred and mixed in the mixing space, and a sealing ring is arranged between the rotating shaft and the mixer cavity at one side close to the stirrer to ensure the sealing property of the mixing cavity; the inner wall of the bottom of the lower cavity is in an inverted cone shape and is matched with the bottom of the stirrer, so that the outlet pressure is improved, and the outlet with internal threads is arranged on the outer wall of the lower cavity, so that the sealed installation of the dispensing needle head is realized.
Furthermore, the micro injection pump comprises a ball screw and a lead screw for transmission, a medical injector for loading materials and a thrust clamp, wherein a main shaft of the stepping motor is fixedly connected with the ball screw through a coupler, the rotary motion of the main shaft of the motor is converted into the linear motion of a sliding block, and the linear motion of the sliding block drives the thrust clamp to move, so that the liquid inlet of the materials is controlled.
Furthermore, the micro injection pump adopts a planetary gear reducer between a main shaft of the stepping motor and the ball screw, and is used for effectively controlling the feeding speed.
The invention has the beneficial effects that: the invention realizes the mixed printing of different biological materials and can print biological gel material structural bodies with three structures of points, lines and bodies; the structural material adopted by the mixing cavity part has certain biocompatibility, and biological cells are introduced into the mixing cavity part, so that the initial experiment of active cell biological 3D printing can be carried out, and multiple requirements are met.
Drawings
FIG. 1 is a schematic diagram of the general structure of a biological 3D printing platform;
FIG. 2 is a schematic structural view of a dynamic mixing and extruding apparatus;
FIG. 3 is a schematic view of a micro-syringe pump;
FIG. 4-1 is a schematic diagram of a dynamic hybrid print head configuration;
FIG. 4-2 is a view taken along line A-A of FIG. 4-1;
FIG. 5 is a schematic view of a stirring bar;
FIG. 6 is a schematic diagram of the x-axis motion configuration;
FIG. 7 is another directional view of FIG. 6;
FIG. 8 is a schematic view of the structure of the y-axis and z-axis motion;
FIG. 9 is a schematic diagram of a platform structure.
Detailed Description
The technical solution of the present invention is further described in detail by the accompanying drawings.
The biological 3D printing platform mainly comprises the following parts: dynamic mixing extrusion device, 3D print motion platform. Wherein the extrusion device consists of a dynamic mixing printing nozzle and a micro injection pump; the dynamic mixing printing nozzle is mainly used for uniformly mixing a sodium alginate-collagen solution containing chondrocytes, the micro injection pump is used for controlling the liquid inlet speed and the liquid inlet amount of a biological material solution, and the 3D printing platform is used for controlling the printing nozzle and the micro injection pump to perform three-dimensional movement so as to realize the preparation of gel substances with other structural shapes, such as gel microspheres and the like.
The dynamic mixing printing nozzle comprises a mixing cavity with a discharge hole at the bottom, two feeding ports are symmetrically distributed on two side walls of the cavity, and luer connectors are butted on the feeding ports; a stirrer is arranged in the mixing cavity, a plurality of stirring blades are arranged on the periphery of the stirrer, and a guide chute is arranged between the blades; the outer side of the top of the cavity is provided with a speed-adjustable stepping motor which is assembled and linked with the stirrer, and the working voltage and the current of the speed-adjustable stepping motor are provided by a controller.
The stirrer can be designed into various shapes and can be replaced, installed and used, and the shape is designed to be similar to a gear shape, the material is photosensitive resin, and the stirrer can be quickly manufactured by utilizing a photocuring 3D printing technology. By adjusting the rotating speed of the stepping motor, the damage of stirring to cells is reduced, and the uniform stirring and mixing of the biological materials are realized. The stirring rod is provided with a rotating shaft, namely the rotating shaft and the stirring rod are integrated, and the rotating shaft is directly connected with a driving shaft of the motor to realize rotation.
The mixing cavity is composed of an upper part and a lower part, the upper cavity is provided with a mixing space, a certain gap is reserved between the stirrer and the inner wall of the upper cavity to ensure that the stirrer is stirred and mixed in the mixing space, and a sealing ring is arranged between the rotating shaft and the mixer cavity and close to one side of the stirrer to ensure the sealing property of the mixing cavity; the inner wall of the bottom of the lower cavity is in an inverted cone shape and is matched with the bottom of the stirrer, so that the outlet pressure is improved, and the outlet with internal threads is arranged on the outer wall of the lower cavity, so that the sealed installation of the dispensing needle head is realized.
The speed regulating motor is fixed on the motor fixing frame through bolt connection, and then the printing spray head is integrally assembled on the 3D motion platform. All different parts are connected by bolts to ensure the connection strength. The dual-channel dynamic mixing printing nozzle needs to be cleaned and pre-extruded before the operation of the dual-channel dynamic mixing printing nozzle starts. Luer connectors and sealing rings are selected standard parts. The auxiliary sealing adhesive tape at the threaded matching position is sealed, and the mixing cavity and the stirrer are printed by adopting photocuring 3D.
The mechanical active stirring mixer is adopted, the mixing effect is better than that of a static mixer, the result is simple, and the mixing efficiency is high, so that the uniform mixing of the biological material solutions with different concentrations is better realized.
The micro injection pump mainly comprises a ball screw and a lead screw for transmission, a medical injector for loading materials, a corresponding clamp and a stepping motor for controlling transmission. The injector is a medical injector, so that materials can be conveniently taken and replaced, and the clamps for positioning have corresponding sizes. The stepping motor spindle is fixedly connected with the ball screw through the coupler, the rotary motion of the motor spindle can be converted into the linear motion of the sliding block, and the linear motion of the sliding block drives the thrust clamp to move, so that the liquid inlet of a control material is realized.
The micro injection pump adopts a planet wheel speed reducer between a motor spindle and a ball screw to effectively control the feeding speed. The micro injection pump is simple and compact in structure, high in movement precision and capable of effectively and timely controlling the liquid inlet of the gel solution. Meanwhile, the micro injection pump is installed with the support base on the X axis through screws. Biological 3D printing system adopts two miniature syringe pumps to supply liquid to mixing the cavity, and two syringe pumps are installed in the both sides of mixing the cavity, through with the bottom plate with support the screw connection of base install on 3D motion platform, at the mixed shower nozzle removal in-process, the little syringe pump is followed the motion and is carried out real-time liquid supply.
In the motion design of the 3D printing motion platform, the X, Y shaft motion adopts a synchronous belt to transmit work, and the Z, Y shaft adopts a ball screw to transmit motion.
Dynamic hybrid printing process: firstly, respectively introducing a sodium alginate solution containing cells and collagen into channels on two sides of an upper cavity of a mixer for mixing, controlling the liquid inlet proportion of materials by a micro-injection pump, then driving a stirrer to uniformly mix and stir the materials in a mixing cavity under the drive of a motor, wherein the mixing cavity is tightly sealed, the stirrer can generate enough pressure by rotating, so that the gel solution drops into spherical liquid drops through a conical spray head, and the gel microsphere liquid drops are dropped into a biological culture dish on a platform by matching with a three-dimensional motion platform, wherein the culture dish is provided with a calcium chloride solution, and the gel microsphere liquid drops meet the calcium chloride solution to realize solidification and crosslinking, thereby finally completing the preparation process of the gel microspheres.
Example (b):
as shown in fig. 1, the overall mechanical structure mainly includes: a dynamic mixing extrusion device 1 and a 3D motion platform structure 2.
The dynamic mixing and extruding device realizes the extrusion molding experiment of the biological material, the structure of which is shown in figure 2 and mainly comprises the following components:
the micro injection pump 1-1 comprises a micro injection pump stepping motor 1-1-1; 1-1-2 of a coupler; a rear panel 1-1-3; 1-1-4 of a linear guide rail; 1-1-5 of a screw nut; 1-1-6 of a mobile platform; 1-1-7 parts of a screw; 1-1-8 of a middle plate; 1-1-9 of a thrust clamp; a guide clamp 1-1-10 and an injector 1-1-11.
The dynamic mixing printing nozzle 1-2 comprises a mixing nozzle stepping motor 1-2-1; a step motor fixing seat 1-2-2; luer 1-2-3; 1-2-4 of an upper cavity of the mixing nozzle; 1-2-5 parts of a stirrer; 1-2-6 parts of a rolling bearing; 1-2-7 parts of a sealing ring; 1-2-8 parts of a lower cavity of the mixing nozzle; dispensing needle tube 1-2-9 and dispensing needle head 1-2-10.
The specific structural functions are realized as follows: the micro injection pump is mainly responsible for injecting the biological materials of sodium alginate and collagen into the dynamic mixing printing nozzle, and the mixing proportion of the materials can be controlled by the micro injection pump to realize the mixing of the materials with different concentrations; the dynamic mixing printing spray head stirs and mixes the biogel materials introduced from the two sides of the cavity uniformly, the structural tightness of the spray head is very good, and the pressure generated by the stirring in the spray head is enough to extrude and mold the uniformly mixed materials through the dispensing needle head.
As shown in fig. 3: the stepping motor 1-1-1 drives the coupling 1-1-2 to drive the screw rod 1-1-7 to rotate, the other side of the screw rod is fixed on the back plate 1-1-3 through the middle plate 1-1-8, the screw rod nut 1-1-5 is fixedly installed with the moving platform 1-1-6, the upper side of the moving platform is connected with the thrust clamp 1-1-9 through a bolt, the lower side of the moving platform slides on the linear guide rail 1-1-4, and the injector 1-1-11 extrudes the biological material in the injector under the interaction of the thrust clamp 1-1-9 and the guide clamp 1-1-10 and enters the dynamic mixing printing nozzle 1-2 through a hose. And similarly, the functional principle of the micro-injection pump structure on the other side is also the same.
As shown in fig. 4-1 and 4-2: the mixing nozzle stepping motor 1-2-1 is fixedly arranged on the stepping motor fixing base 1-2-2, the fixing base 1-2-2 is arranged on the extruding device fixing base 3-9 through bolt connection, the mixing nozzle upper cavity 1-2-4 is arranged on the motor fixing base 1-2-2 through bolt connection, the stirrer 1-2-5 is made into a structure as shown in a schematic diagram 5, a blind hole is arranged at the central shaft position and is directly connected and arranged with a driving shaft of the stepping motor 1-2-1, the motor 1-2-1 directly drives the stirrer 1-2-5 to carry out rotary stirring in the mixing cavity, a rolling bearing 1-2-6 is arranged at the position close to the motor driving position, so that the stirring can be carried out more smoothly, a sealing ring 1-2-7 is arranged at the lower side to ensure the sealing property of the mixing nozzle, a part of space is reserved at the lower side of a cavity 1-2-4 on the mixing nozzle, namely the mixing space, to ensure the mixing uniformity of the biological material in the mixing nozzle, two liquid inlet channels are symmetrically distributed at two sides of the cavity 1-2-4 on the mixing nozzle, luer connectors 1-2-3 are respectively arranged and connected to outlets of injectors 1-1-11 through hoses, a through hole with gradient is arranged at a cavity 1-2-8 on the lower side of the mixing nozzle, a dispensing needle tube 1-2-9 is connected with the cavity through screw threads, and a dispensing needle 1-2-10 and the dispensing needle tube 1-2-9 are connected and arranged into a whole through screw threads to complete the printing and molding of the biological.
The three direction's of xyz removal can be realized to 3D motion platform structure, and the primary structure includes x axle motion structure 3, z axle motion structure 4 and y axle motion structure, and concrete structure function realizes:
as shown in fig. 6 and 7: the three-dimensional moving platform supporting structure is built by an aluminum profile frame. An x-axis stepping motor 3-1 is installed on an x-axis bottom plate 3-5 through a stepping motor fixing seat 3-2, the motor 3-1 drives a synchronous belt pulley 3-3 to drive a synchronous belt 3-4, the other side of the synchronous belt 3-4 is connected with a synchronous belt pulley 3-14, and two sides of an x-axis guide shaft 3-7 are fixedly installed between synchronous belt pulley fixing seats 3-6 and 3-10; synchronous pulley fixing seats 3-6 and 3-10 are installed on x-axis bottom plates 3-5 and 3-11 on two sides through bolt connection, the x-axis bottom plates 3-5 and 3-11 on the two sides move in the z-axis direction of a z-axis screw 4-3 and a z-axis guide rod 4-4 through screw nuts 3-12 and linear bearings 3-13 to drive an x-axis movement structure to vertically move up and down, an extrusion device fixing seat 3-9 is fixedly installed on a linear bearing seat 3-8 to linearly move in an x-axis guide shaft 3-7, the extrusion device fixing seat 3-9 is fixedly installed through a synchronous belt connecting plate 3-15, and the synchronous belt 3-5 drives a dynamic mixing extrusion device 1 to move in the x direction.
As shown in fig. 8: the z-axis moving structure mainly comprises a y-axis stepping motor 4-1, a coupler 4-2, a z-axis lead screw 4-3, a z-axis guide rod 4-4, an optical axis base 4-5 and a vertical bearing seat 4-6. The stepping motor 4-1 is installed at the upper top of the aluminum frame structure through a stepping motor fixing seat, the z-axis guide rod 4-4 is installed on the aluminum frame through a z-axis optical axis base 4-5, the stepping motor 4-1 is connected with a z-axis lead screw 4-3 through a coupler 4-2 to realize conversion from rotation to linear motion, and the lower side of the z-axis lead screw 4-3 is fixedly installed on the aluminum frame through a vertical bearing seat 4-6.
As shown in fig. 9: the platform 5-12 is composed of a glass plate 5-12-1, a hot bed plate 5-12-2 and an aluminum substrate 5-12-3, and the horizontal standard adjustment of the experimental platform is adjusted through leveling screws 5-12-4; the movement of the y-axis moving structure is realized by driving a synchronous belt pulley 5-6 to drive a synchronous belt 5-7 to move by a stepping motor 5-8 arranged on a sliding block connecting plate 5-5, and the other side of the synchronous belt is arranged on a base connecting plate 5-11 through a belt pulley fixing nail 5-10; the platform 5-12 is connected with the linear bearing seat 5-3 to realize the movement of the y-axis guide shaft 5-2, and the platform is fixedly connected with a y-axis synchronous belt connecting plate 5-9 to realize the synchronous belt transmission to drive the platform to move in the y-axis direction; two sides of the y-axis guide shaft 5-2 are fixedly connected with an optical axis base 5-1 and a sliding block 5-4 respectively; the petri dish was used to receive the extruded experimental material.
In summary, the present invention is directed to the practical application requirements of gel microspheres in the field of in vitro cell culture and tissue printing, and studies the production of the concentration gradient of gel microspheres, and discusses the influence of the density distribution of cells in the in vitro three-dimensional culture environment on the survival of cells and the expression of tissue functions. A dynamic mixing printing nozzle is designed and manufactured, the two materials are quickly and uniformly mixed by mechanical active stirring, and 3D printing and quick manufacturing of gel microspheres with different material concentrations are realized.
Claims (5)
1. The composite biological material dynamic mixing biological 3D printing platform comprises a dynamic mixing extrusion device and a 3D printing motion platform, wherein the 3D printing platform is used for controlling the dynamic mixing extrusion device to perform three-dimensional motion so as to realize preparation of gel microspheres;
the method is characterized in that:
the dynamic mixing extrusion device consists of a dynamic mixing printing spray head and micro injection pumps on two sides;
the dynamic mixing printing nozzle is mainly used for uniformly mixing a sodium alginate-collagen solution containing chondrocytes, the micro injection pump is used for controlling the liquid inlet speed and the liquid inlet amount of the biological material solution, the micro injection pump on one side supplies the sodium alginate solution containing the chondrocytes, and the micro injection pump on the other side supplies the collagen;
the dynamic mixing printing nozzle comprises a mixing cavity with a discharge hole at the bottom, two feeding ports are symmetrically distributed on two side walls of the cavity, and the feeding ports are butted with Ruhr joints and are used for connecting an outlet of a micro injection pump; a stirrer is arranged in the mixing cavity, and a plurality of stirring blades are arranged on the periphery of the stirrer; the outer side of the top of the cavity is provided with a speed-adjustable stepping motor which is assembled and linked with the stirrer.
2. The composite biomaterial dynamic hybrid biological 3D printing platform as claimed in claim 1, wherein: the stirrer is replaceable, is made of photosensitive resin and is rapidly manufactured by utilizing a photocuring 3D printing technology; the damage of stirring to cells is reduced and the uniform stirring and mixing of the biological materials are realized by adjusting the rotating speed of the stepping motor; the stirrer is provided with a rotating shaft, namely the rotating shaft and the stirrer are integrated, and the rotating shaft and the speed-adjustable stepping motor are arranged on the stirrer.
3. The composite biomaterial dynamic hybrid biological 3D printing platform as claimed in claim 2, wherein: the mixing cavity consists of an upper part and a lower part, and is made of photosensitive resin; the upper cavity is provided with a mixing space, a certain gap is reserved between the stirrer and the inner wall of the upper cavity to ensure that the stirrer is stirred and mixed in the mixing space, and a sealing ring is arranged between the rotating shaft and the mixer cavity at one side close to the stirrer to ensure the sealing property of the mixing cavity; the inner wall of the bottom of the lower cavity is in an inverted cone shape and is matched with the bottom of the stirrer, so that the outlet pressure is improved, and the outlet with internal threads is arranged on the outer wall of the lower cavity, so that the sealed installation of the dispensing needle head is realized.
4. The composite biomaterial dynamic hybrid biological 3D printing platform as claimed in claim 1, wherein: the miniature injection pump comprises a ball screw and a screw rod which are used for transmission, a medical injector for loading materials and a thrust clamp, wherein a main shaft of a stepping motor is fixedly connected with the ball screw through a coupler, the rotary motion of the main shaft of the motor is converted into the linear motion of a sliding block, and the linear motion of the sliding block drives the thrust clamp to move, so that the liquid inlet of the materials is controlled.
5. The composite biomaterial dynamic hybrid biological 3D printing platform as claimed in claim 1, wherein: the miniature injection pump adopts a planetary gear reducer between a main shaft of a stepping motor and a ball screw, and is used for effectively controlling the feeding speed.
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Cited By (5)
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| CN113019262A (en) * | 2021-02-23 | 2021-06-25 | 中国科学院上海硅酸盐研究所 | High-flux material synthesis equipment and method based on micro-fluidic chip |
| CN113459512A (en) * | 2021-06-28 | 2021-10-01 | 浙江工业大学 | 3D printer based on CPVC/PVC gel material |
| CN113499696A (en) * | 2021-07-05 | 2021-10-15 | 南方科技大学 | Liquid mixing apparatus |
| CN114683543A (en) * | 2022-02-25 | 2022-07-01 | 上普博源（北京）生物科技有限公司 | High-throughput biological three-dimensional printing equipment, printing method and application thereof |
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| CN113499696A (en) * | 2021-07-05 | 2021-10-15 | 南方科技大学 | Liquid mixing apparatus |
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| CN114683543A (en) * | 2022-02-25 | 2022-07-01 | 上普博源（北京）生物科技有限公司 | High-throughput biological three-dimensional printing equipment, printing method and application thereof |
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