1. A three-dimensional printing system of ECM hydrogel mixed with a cross-linking agent, comprising: including ECM aquogel syringe needle (1) and cross-linking agent syringe needle (2), ECM aquogel syringe needle (1) one end is provided with first syringe pump (3), first syringe pump (3) external connection has first flow rate controller (4), cross-linking agent syringe needle (2) one end is provided with second syringe pump (5), second syringe pump (5) external connection has second flow rate controller (6), ECM aquogel syringe needle (1) and cross-linking agent syringe needle (2) one end are connected with T type syringe needle (7), T type syringe needle (7) one end is connected with adjustable length and extrudes head (8), adjustable length is extruded head (8) inside and is provided with spiral blending ware (9), adjustable length is extruded head (8) bottom and is provided with print platform (10).

2. The three-dimensional printing system of ECM hydrogel mixed with cross-linking agent of claim 1, wherein: the ECM hydrogel sample injection needle (1) is filled with ECM hydrogel, one end of the ECM hydrogel sample injection needle (1) is provided with a first sample injection pump (3) for extruding the ECM hydrogel from the inside of the ECM hydrogel sample injection needle (1) at a constant speed, and the first sample injection pump (3) is electrically connected with an external power supply through a lead.

3. The three-dimensional printing system of ECM hydrogel mixed with cross-linking agent of claim 1, wherein: the cross-linking agent sample injection needle (2) is filled with a cross-linking agent, one end of the cross-linking agent sample injection needle (2) is provided with a second sample injection pump (5) for extruding the cross-linking agent from the inside of the cross-linking agent sample injection needle (2) at a constant speed, and the second sample injection pump (5) is electrically connected with an external power supply through a lead.

4. The three-dimensional printing system of ECM hydrogel mixed with cross-linking agent of claim 1, wherein: the first flow rate controller (4) is electrically connected with the first sample feeding pump (3) through a lead.

5. The three-dimensional printing system of ECM hydrogel mixed with cross-linking agent of claim 1, wherein: the second flow rate controller (6) is electrically connected with the second sample pump (5) through a lead.

6. The three-dimensional printing system of ECM hydrogel mixed with cross-linking agent of claim 1, wherein: t type syringe needle (7) top and ECM aquogel sampling needle (1) through connection, T type syringe needle (7) side and cross-linking agent sampling needle (2) through connection, T type syringe needle (7) bottom is connected with adjustable length and extrudes head (8).

7. The three-dimensional printing system of ECM hydrogel mixed with cross-linking agent of claim 1, wherein: the adjustable length extrusion head (8) is fixed in a nut mode, and the adjustable length extrusion head (8) is sleeved at the bottom end of the T-shaped needle head (7) and is connected with the T-shaped needle head (7) through threads.

8. A method of three-dimensional printing of ECM hydrogel mixed with a cross-linking agent, comprising: a three-dimensional printing system based on mixing of ECM hydrogel with cross-linking agent according to claims 1-7, the three-dimensional printing method comprising the following steps,

step one, placing an ECM hydrogel material to be printed into an ECM hydrogel sample injection needle (1), and using a first sample injection pump (3) to advance so that the ECM hydrogel in the ECM hydrogel sample injection needle (1) enters from the upper part of a T-shaped needle head (7);

placing the ECM cross-linking agent into the cross-linking agent sample injection needle (2), and pushing the ECM cross-linking agent into the cross-linking agent sample injection needle (2) from a side channel of the T-shaped needle head (7) by using a second sample injection pump (5);

step three, the first sample injection pump (3) and the second sample injection pump (5) of the two injection pumps respectively use independent flow rate controllers, namely a first flow rate controller (4) and a second flow rate controller (6) to control the flow rate of the corresponding sample injection needle, so that two components of the ECM printing ink can be mixed in a certain proportion;

step four, a spiral blending device (9) arranged in the adjustable length extrusion head (8) can enable printing ink to be correspondingly extruded and uniformly mixed when the mixed ECM printing ink enters the adjustable length extrusion head (8);

and step five, extruding the printing ink from the lower part of the length-adjustable extrusion head (8) and forming on the printing platform (10).

Background

ECM hydrogel is a natural material derived from living organisms, and contains various types of collagen as a main component, but when a general collagen solution is thermally crosslinked, crosslinking speed is too slow and crosslinking strength is not high, and thus ECM is crosslinked using a crosslinking agent externally added.

The prior art has the following defects and shortcomings:

however, in the current processing method, the cross-linking agent is premixed with ECM and then added into the storage cylinder, but some cross-linking agents react with ECM materials in the process of uniform mixing, so that the ECM printing process becomes uncontrollable over time, therefore, the cross-linking agent and ECM raw materials need to be stored separately and mixed in the printing process to accurately control the printing process, and meanwhile, the printing ink with a single formula cannot simulate the mechanical properties of anisotropy or gradient properties of human tissues and organs, so that a device capable of immediately adjusting the formula of the printing ink is needed to accurately control the formula of the printed ECM ink.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a three-dimensional printing method for mixing an ECM hydrogel with a cross-linking agent, which can solve the problem that the existing three-dimensional printing method cannot simulate the mechanical performance of anisotropy or gradient property of human tissues and organs by using printing ink with a single formula, and the problem that the cross-linking agent is premixed with ECM and then added into a storage cylinder in the existing processing method, but some cross-linking agents react with ECM materials in the uniform mixing process, so that the ECM printing process can become uncontrollable along with the time; the invention effectively solves the problems in the prior art by arranging the three-dimensional printing method for mixing the ECM hydrogel and the cross-linking agent.

In order to realize the purpose of the three-dimensional printing method for mixing the ECM hydrogel and the cross-linking agent, the invention provides the following technical scheme: the utility model provides a three-dimensional printing system that ECM aquogel and cross-linking agent mix, includes ECM aquogel syringe and cross-linking agent syringe, ECM aquogel syringe one end is provided with first syringe pump, first syringe pump external connection has first flow rate controller, cross-linking agent syringe one end is provided with the second syringe pump, second syringe pump external connection has second flow rate controller, ECM aquogel syringe and cross-linking agent syringe one end are connected with T type syringe needle, T type syringe needle one end is connected with adjustable length and extrudes the head, adjustable length is extruded the inside spiral blending machine that is provided with of head, adjustable length is extruded the head bottom and is provided with print platform.

Preferably, the ECM hydrogel sample injection needle is filled with ECM hydrogel, one end of the ECM hydrogel sample injection needle is provided with a first sample pump for extruding the ECM hydrogel from the inside of the ECM hydrogel sample injection needle at a constant speed, and the first sample pump is electrically connected with an external power supply through a lead.

Preferably, the cross-linking agent sample injection needle is filled with the cross-linking agent, one end of the cross-linking agent sample injection needle is provided with a second sample injection pump for extruding the cross-linking agent from the inside of the cross-linking agent sample injection needle at a constant speed, and the second sample injection pump is electrically connected with an external power supply through a lead.

Preferably, the first flow rate controller is electrically connected with the first sample pump through a lead.

Preferably, the second flow rate controller is electrically connected with the second sample pump through a lead.

Preferably, the top end of the T-shaped needle head is in through connection with an ECM hydrogel sample injection needle, the side end of the T-shaped needle head is in through connection with a cross-linking agent sample injection needle, and the bottom end of the T-shaped needle head is connected with an extrusion head with adjustable length.

Preferably, the adjustable length extrusion head is fixed in a nut mode, and the adjustable length extrusion head is sleeved at the bottom end of the T-shaped needle head and is in threaded connection with the T-shaped needle head.

Further, a three-dimensional printing method of mixing ECM hydrogel with a cross-linking agent comprises the following steps,

step one, placing an ECM hydrogel material to be printed into an ECM hydrogel sample injection needle, and pushing by using a first sample injection pump to enable the ECM hydrogel in the ECM hydrogel sample injection needle to enter from the upper part of a T-shaped needle head;

step two, placing the ECM cross-linking agent into a cross-linking agent sample injection needle, and pushing the ECM cross-linking agent into the cross-linking agent sample injection needle from a side channel of the T-shaped needle head by using a second sample injection pump;

step three, controlling the flow rate of the corresponding sampling needle by using a first flow rate controller and a second flow rate controller of the two injection pumps respectively so that two components of the ECM printing ink can be mixed in a certain proportion;

step four, a spiral mixer is arranged in the adjustable-length extrusion head, and when mixed ECM printing ink enters the adjustable-length extrusion head, the printing ink can be correspondingly extruded and uniformly mixed;

and step five, extruding the printing ink from the lower part of the length-adjustable extrusion head, and forming on a printing platform.

Compared with the prior art, the invention provides a three-dimensional printing method for mixing ECM hydrogel and a cross-linking agent, which has the following beneficial effects:

the ECM hydrogel and the needed cross-linking agent are separately arranged in two storage devices, independent extrusion control devices are respectively used for controlling the sampling speeds of the two raw materials, and a T-shaped extrusion head is used for uniformly mixing the two raw materials;

the printing method can add the crosslinking agent with the required content in real time in the printing process of the ECM hydrogel biological ink, avoid failure in the printing process caused by early solidification and molding of the ECM biological ink after the crosslinking agent is directly mixed, and simultaneously can control the content of the crosslinking agent added in the printing process of the ECM hydrogel biological ink in real time, accurately control the components of the printing ink in real time and realize the preparation of the anisotropic biological scaffold.

Drawings

FIG. 1 is a schematic diagram of a three-dimensional printing system according to the present invention.

In the figure: 1. an ECM hydrogel sample injection needle; 2. a crosslinking agent sample injection needle; 3. a first sample injection pump; 4. a first flow rate controller; 5. a second sample injection pump; 6. a second flow controller; 7. a T-shaped needle head; 8. an adjustable length extrusion head; 9. a spiral blender; 10. a printing platform.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Referring to fig. 1, a three-dimensional printing system for mixing ECM hydrogel and a crosslinking agent includes an ECM hydrogel sample injection needle 1 and a crosslinking agent sample injection needle 2, a first sample injection pump 3 is disposed at one end of the ECM hydrogel sample injection needle 1, a first flow rate controller 4 is connected to an external portion of the first sample injection pump 3, a second sample injection pump 5 is disposed at one end of the crosslinking agent sample injection needle 2, a second flow rate controller 6 is connected to an external portion of the second sample injection pump 5, a T-shaped needle 7 is connected to one ends of the ECM hydrogel sample injection needle 1 and the crosslinking agent sample injection needle 2, an adjustable length extrusion head 8 is connected to one end of the T-shaped needle 7, a spiral mixer 9 is disposed inside the adjustable length extrusion head 8, and a printing platform 10 is disposed at a bottom of the adjustable length extrusion head 8.

In summary, specific examples of the three-dimensional printing method in which ECM hydrogel is mixed with a crosslinking agent are given below;

the ECM hydrogel is prepared from skin by the following steps: dissolving 500 mu l of ECM stock solution in 1XPBS aqueous solution to prepare ECM printing hydrogel printing ink, placing the ECM printing hydrogel printing ink into an ECM hydrogel sampling needle 1, and filling the cinchepin solution into a cross-linking agent sampling needle 2 to serve as a cross-linking agent of the printing ink. Extruding the ECM printing ink in the ECM hydrogel sampling needle 1 at the flow rate of 140 mul/min, adjusting the Jinniping solution to be 5-10 mul/min according to the required requirement, simultaneously starting two injection pumps, simultaneously extruding the ECM ink and a cross-linking agent into a T-shaped needle 7, uniformly mixing the two solutions by a spiral uniform mixer 9 in the T-shaped needle 7, adjusting the length of the lower section of an extrusion head 8 with adjustable length, adjusting the proper pre-mixing time, and extruding the printing ink to realize final forming on a printing platform 10.

The working use process and the installation method of the invention are that the crosslinking agent addition in the ECM is controlled in a double-cylinder mixing extrusion mode when the three-dimensional printing method for mixing the ECM hydrogel and the crosslinking agent is used.

The printing system for realizing the method mainly comprises an ECM hydrogel sample injection needle 1, a cross-linking agent sample injection needle 2, a first sample injection pump 3, a first flow rate controller 4 and a second flow rate controller 6 which correspond to the first flow rate controller, a T-shaped needle head 7 and a spiral mixer 9 inside the T-shaped needle head, wherein the ECM hydrogel material to be printed is firstly placed into the ECM hydrogel sample injection needle 1, and the ECM hydrogel sample injection needle 1 is pushed by using the first sample injection pump 3 to enable the ECM hydrogel in the ECM hydrogel sample injection needle 1 to enter from the upper part of the T-shaped needle head 7; the ECM cross-linking agent is placed into a cross-linking agent sample injection needle 2, the ECM cross-linking agent in the cross-linking agent sample injection needle 2 enters from a side channel of a T-shaped needle head 7 by pushing of a second sample injection pump 5, a first sample injection pump 3 and the second sample injection pump 5 respectively control the flow rate of corresponding sample injection needles by using independent flow rate controllers, two components of ECM printing ink can be mixed according to a certain proportion, a spiral blending device 9 is arranged inside the T-shaped needle head 7, when the mixed ECM printing ink enters the T-shaped needle head 7, the printing ink can be correspondingly extruded and uniformly mixed, an adjustable length extrusion head 8 is fixed by a screw cap, the printing ink can be extruded from the lower part of the adjustable length extrusion head 8 and is formed on a printing platform 10.

The printing method can add the crosslinking agent with the required content in real time in the printing process of the ECM hydrogel biological ink, avoid failure in the printing process caused by early solidification and molding of the ECM biological ink after the crosslinking agent is directly mixed, and simultaneously can control the content of the crosslinking agent added in the printing process of the ECM hydrogel biological ink in real time, accurately control the components of the printing ink in real time and realize the preparation of the anisotropic biological scaffold.

It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.