WO2023113354A1 - Bioink composition kit for tubular biotissue, fabrication method therefor, method for constructing tubular biotissue, using same, and tubular biotissue constructed by same method - Google Patents

Abstract

The present invention relates to a technology for constructing a tubular biotissue, such as a blood vessel, a nerve conduit, and the like and, more specifically, to a bioink composition kit for a tubular biotissue, a fabrication method therefor, a method for constructing a tubular biotissue, and a tubular biotissue constructed by the method, wherein a bioink composition kit composed of two solutions having different in physical properties to each other due to their different components, and tubular biotissues having various shapes can be easily constructed by using the bioink composition with different physical properties in the shell and core of the coaxial nozzle.

Description

Bioink composition kit for tubular living tissue, manufacturing method thereof, manufacturing method of tubular living tissue using the same, and tubular living tissue produced by the same

The present invention relates to a tubular biological tissue such as a blood vessel or a nerve conduit and a method for manufacturing the same, and more specifically, to form a tubular biological tissue, two solutions having different physical properties by having different components are formed, i.e., a core portion, which is an inner core. Prepare a bioink composition kit composed of a sacrificial solution to form a tubular structure surrounding the core and a hydrogel forming a tube, and use the bioink composition kit to print two heads provided in a 3D printer under different process conditions. It relates to a bioink composition kit for tubular biological tissues that can be easily produced by simultaneous operation to produce tubular biological tissues having various shapes, a method for manufacturing the same, a method for manufacturing tubular biological tissues using the same, and a tubular biological tissue produced by the method .

Tubular biological tissues have different diameters, lengths, shapes, curvatures, changes in diameters, and constituent cells depending on the organ site, and there are various types such as tubular structures composed of multi-layered tissue structures such as 2 and 3 layers as well as single-layer structures. .

Since it is difficult to make a multi-layer tubular biological tissue in the past, a biocompatible material with high viscosity is supported on a mold, etc., several linear structures are fabricated and embedded using a sacrificial composition, and the internal structure of the cavity remains after removing the sacrificial composition. A method of coating various cells was used.

Recently, a method of manufacturing a blood vessel or the like using a coaxial nozzle has emerged. This method puts a small-diameter nozzle inside a large-diameter nozzle, outputs removable bio-ink to the innermost nozzle (core), and outputs curable bio-ink to the gap (shell) between the nozzles and nozzles. Shapes can be created immediately. In addition, there is an advantage in that single-layer or multi-layer tubular shapes can be manufactured in various ways by overlapping several nozzles having different diameters.

However, the conventional coaxial nozzle-based tubular biological tissue manufacturing method has poor self-standing ability due to weak mechanical properties of the hydrogel after printing and curing. Such a decrease in shape-retaining ability causes difficulty in maintaining a circular tubular cross-sectional shape, yield reduction when manufacturing a curved pipe shape, and difficulty in connecting a perfusate for culturing tubular biological tissue.

Therefore, there is a need to develop a tubular biological tissue manufacturing technology that can solve these problems.

The present inventors have completed the present invention by developing a tubular biological tissue manufacturing technology with improved mechanical properties as a result of numerous studies.

Accordingly, an object of the present invention is to provide bioink in which two solutions having different physical properties due to different components, that is, a sacrificial solution for forming a core portion, which is the inner center, and a tube-forming hydrogel forming a tubular structure surrounding the core portion are separated and packaged. It is to provide a composition kit and a manufacturing method thereof.

Another object of the present invention is to simultaneously drive two heads provided in a 3D printer under different process conditions using a bioink composition kit to easily form tubular biological tissues having various shapes with a 3D printer, thereby enabling various bends. It is to provide a method for manufacturing tubular biological tissue using 3D printing, which can increase the yield when manufacturing the shape.

Another object of the present invention is to improve self-standing ability by increasing mechanical properties, so that it is easy to maintain a circular tubular cross-sectional shape after printing and curing, and it is easy to connect perfusate for culturing tubular biological tissue. It is to provide a tubular biological tissue.

The object of the present invention is not limited to the objects mentioned above, and other objects not mentioned will be clearly understood by those skilled in the art from the description below.

In order to achieve the object of the present invention described above, first, the present invention is LithiumPhenyl (2,4,6-trimethylbenzoyl) phosphinate (LAP) 0.03 to 0.07% by weight, Gelatin methacrylate (GelMA) 5 to 15% by weight and alginate 2 to Conduit-forming hydrogel containing 3% by weight; and a sacrificial hydrogel containing 0.05 to 0.15N calcium chloride and 30 to 50% by weight of pluronic F-127.

In a preferred embodiment, the conduit-forming hydrogel contains phosphate buffered saline (PBS).

In a preferred embodiment, the conduit-forming hydrogel and the sacrificial-forming hydrogel are separately packaged and stored in a refrigerator.

In addition, the present invention is a conduit-forming hydrogel preparation step; And a sacrificial formation hydrogel preparation step; provides a method for manufacturing a bioink composition kit for tubular biological tissues independently performed.

In a preferred embodiment, the conduit-forming hydrogel preparation step is to prepare a 0.05 to 0.15% by weight LAP solution by dissolving LithiumPhenyl (2,4,6-trimethyl benzoyl) phosphinate (LAP) in PBS; Preparing 15 to 25% by weight of GelMA hydrogel by dissolving Gelatin methacrylate (GelMA) in the LAP solution; Preparing an alginate hydrogel of 4 to 5.5% by weight by dissolving alginate in distilled water; and preparing a conduit-forming hydrogel by mixing the GelMA hydrogel and the alginate hydrogel at a volume ratio of 1:1.

In a preferred embodiment, the dissolution is performed by selectively repeating one or more of vortexing, storage in a water bath at 37 ± 0.5 ° C, and water bath at 180 ± 0.5 ° C.

In a preferred embodiment, the mixing of the GelMA hydrogel and the alginate hydrogel is performed by repeatedly performing 37 ± 0.5 ° C water bath storage and vortexing.

In a preferred embodiment, the step of preparing the sacrificial hydrogel may include preparing a 0.05 to 0.15N calcium chloride solution by adding calcium chloride to distilled water; and adding 30 to 50% by weight of pluronic F-127 to the 0.05 to 0.15N calcium chloride solution and mixing.

In a preferred embodiment, the mixing step is performed by repeatedly performing low-temperature storage and vortexing.

In addition, the present invention is a 3D printer of the duct-forming hydrogel and the sacrificial hydrogel of any one of the above-described bioink composition kits for tubular biological tissues or the bioink composition kit for tubular biological tissues prepared by any one of the above-described manufacturing methods. loading into; A pipeline structure formed by simultaneously driving the two heads provided in the 3D printer under different process conditions to form a sacrificial body, which is a core portion, with the sacrificial hydrogel, and forming a tubular structure surrounding the core portion with the conduit-forming hydrogel. outputting; Curing the tubular structure by UV irradiating the surface of the pipeline structure; and cutting both ends of the cured pipeline structure and then removing the sacrificial material to form a hollow core portion.

In a preferred embodiment, the process conditions of the head for outputting the tubular structure with the tube-forming hydrogel in the outputting step are 26±0.5° C. and 220 to 230 kPa, and the head for outputting the sacrificial material with the sacrificial hydrogel. The process conditions are 35±0.5℃ and 120~130kPa.

In a preferred embodiment, the UV irradiation is performed for 150 seconds to 210 seconds so that the amount of light reaching the surface of the pipeline structure is 65 to 75 mW/cm 2 .

In a preferred embodiment, the step of removing the sacrificial material comprises dissolving the sacrificial material by injecting PBS or distilled water into one end of the cut pipeline structure; and inhaling the liquid present in the hollow of the pipeline structure.

In a preferred embodiment, the size of the hollow in the pipeline structure and the thickness of the tubular structure are determined according to the diameter of the nozzle used in the 3D printing.

In addition, the present invention provides a tubular biological tissue produced by the above-described manufacturing method.

In the above-described bioink composition kit of the present invention, two solutions having different physical properties due to different components, that is, a sacrificial solution for forming the core part, which is the inner center, and a tube-forming hydrogel for forming a tubular structure surrounding the core part are separated and packaged. Therefore, it maintains its physical properties well and is convenient to use.

In addition, the method for producing tubular biological tissue using 3D printing of the present invention simultaneously drives two heads provided in a 3D printer under different process conditions using a bioink composition kit to 3D print tubular biological tissues having various shapes. It can be easily formed, so the yield can be increased when manufacturing various bends.

In addition, according to the tubular biological tissue of the present invention, the mechanical properties are increased and the self-standing ability is excellent, so it is easy to maintain the circular tubular cross-sectional shape after printing and curing, and the perfusate for culturing the tubular biological tissue Easy to connect.

Effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description below.

1a to 1d are photographs showing one embodiment of the step of preparing a LAP solution performed during the preparation step of the hydrogel to form a conduit in the method for manufacturing a bioink composition kit according to various embodiments of the present invention.

2a to 2d are photographs showing one embodiment of a step of preparing a GelMA solution.

3a to 3c are photographs showing one embodiment of the step of preparing an alginate solution.

4a to 4c are photographs showing one embodiment of a step of preparing a tubular hydrogel.

5 is a photograph showing one embodiment of the sacrificial material formation hydrogel preparation step in the bioink composition kit manufacturing method according to various embodiments of the present invention.

Figure 6a is a photograph showing that the conduit-forming hydrogel prepared according to Figs. 4a to 4c is cured by UV irradiation, and Fig. 6b is a photograph showing that it is not cured without UV irradiation.

7 is a schematic diagram showing the implementation principle of a method for manufacturing tubular biological tissue using 3D printing according to various embodiments of the present invention.

Figure 8a is a photograph showing the durability of the output produced by the manufacturing method of the present invention can be picked up with tweezers, and Figs. 8b to 8d are photographs showing various types of tubular biological tissues manufactured by the manufacturing method of the present invention.

9a and 9b are photographs of the result of evaluating the perfusion performance of the straight conduit manufactured by the manufacturing method of the present invention.

10a to 10c are photographs of the results of evaluating the perfusion performance of the U-shaped conduit manufactured by the manufacturing method of the present invention.

Terms used in the present invention are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this application, terms such as "comprise" or "having" are intended to indicate that there is a feature, number, step, operation, component, part, or combination thereof described in the description of the invention, but one or more other It should be understood that it does not preclude the possibility of addition or existence of features, numbers, steps, operations, components, parts, or combinations thereof.

Terms such as first and second may be used to describe various components, but the components should not be limited by the terms. These terms are only used for the purpose of distinguishing one component from another. For example, a first element may be termed a second element, and similarly, a second element may be termed a first element, without departing from the scope of the present invention.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and unless explicitly defined in the present invention, they should not be interpreted in an ideal or excessively formal meaning. don't

In interpreting the components, even if there is no separate explicit description, it is interpreted as including the error range.

In the case of a description of a temporal relationship, for example, when a temporal precedence relationship is described as 'after', 'continue to', 'after ~', 'before', etc., 'immediately' or 'directly' Including non-consecutive cases unless ' is used.

Hereinafter, the technical configuration of the present invention will be described in detail with reference to preferred embodiments.

However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Like reference numbers used throughout to describe the invention indicate like elements.

The technical feature of the present invention is a bioink composition kit that maintains physical properties and is convenient to use because the sacrificial solution forming the core part, which is the inner center, and the tube-forming hydrogel forming the tubular structure surrounding the core part are separated and packaged. Tubular biological tissues having various shapes can be easily formed with a 3D printer by simultaneously driving the two heads provided in the 3D printer under different process conditions, thereby increasing the yield when producing various bends using a 3D printer. Tubular biotissue manufacturing method and improved self-standing ability due to increased mechanical properties, so it is easy to maintain a circular tubular cross-sectional shape after printing and curing, and it is easy to connect perfusate for culturing tubular living tissue. It is in tubular tissue.

That is, the present invention is a bioink composition of a new composition that can compensate for the disadvantages of the existing coaxial nozzle-based tubular biological tissue manufacturing method, which has poor self-standing ability due to weak output and mechanical properties of the hydrogel after curing. Because the kit was developed.

Therefore, the bioink composition kit for tubular living tissue of the present invention contains 0.03 to 0.07% by weight of LithiumPhenyl (2,4,6-trimethylbenzoyl)phosphinate (LAP), 5 to 15% by weight of Gelatin methacrylate (GelMA), and 2 to 3% by weight of alginate. Conduit-forming hydrogel containing % and the remaining amount of solvent; and a sacrificial hydrogel containing a 0.05 to 0.15N calcium chloride solution and a 30 to 50 wt % pluronic F-127 aqueous solution.

Here, alginate or the like included in the conduit-forming hydrogel enhances the viscosity before curing. In particular, since alginate can be cured to create a small-sized egg-box structure by adding ions, an interpenetrated network (IPN) in which the two materials cross each other during the subsequent photocuring reaction of GelMA, a photoreactive biomaterial By forming, it is possible to further increase the mechanical strength.

The duct-forming hydrogel may contain phosphate buffered saline (PBS), which may be an aqueous salt solution containing disodium hydrogen phosphate and sodium chloride of known composition, or an aqueous salt solution containing potassium chloride and potassium dihydrogen phosphate. there is.

Since the sacrificial hydrogel can be printed with a 3D printer and must have properties that can be dissolved in an aqueous solvent for a short time in the printed state, it is implemented to include 0.05 to 0.15N calcium chloride solution and 30 to 50% by weight of pluronic F-127. did In particular, calcium chloride (CaCl ₂ ) contained in the sacrificial hydrogel can immediately cause ionic curing of alginate contained in the conduit-forming hydrogel.

The conduit-forming hydrogel and the sacrificial-forming hydrogel may be separately packaged and refrigerated.

Next, the method for manufacturing a bioink composition kit for tubular biological tissues of the present invention includes the preparation step of a hydrogel forming a conduit; and sacrificial solution preparation step; performed independently.

First, the conduit-forming hydrogel preparation step is to prepare a 0.05 to 0.15% by weight LAP solution by dissolving LithiumPhenyl (2,4,6-trimethyl benzoyl) phosphinate (LAP) in PBS; Preparing 15 to 25% by weight of GelMA hydrogel by dissolving Gelatin methacrylate (GelMA) in the LAP solution; Preparing an alginate hydrogel of 4 to 5.5% by weight by dissolving alginate in distilled water; and preparing a conduit-forming hydrogel by mixing the GelMA hydrogel and the alginate hydrogel at a volume ratio of 1:1.

Dissolution performed in each step can be achieved by selectively repeating one or more of vortexing, storage in a water bath at 37 ± 0.5 ° C, and hot water at 180 ± 0.5 ° C. That is, as shown in Figures 1a to 3c, as one embodiment, the step of preparing the LAP solution is performed only vortexing, and the step of preparing the GelMA hydrogel is vortexing and 37 ± 0.5 ℃ water bath storage alternately GelMA It can be repeated until it is completely dissolved, and the step of preparing the alginate hydrogel can be performed with water bath at 180 ± 0.5 ° C. after vortexing.

In addition, in the step of preparing the conduit-forming hydrogel, the mixing of the GelMA hydrogel and the alginate hydrogel can be performed by repeatedly performing 37 ± 0.5 ° C water bath storage and vortexing, as shown in FIGS. 4a to 4c.

The sacrificial material formation hydrogel preparation step is to prepare a 0.05 to 0.15N calcium chloride solution by adding calcium chloride to distilled water; and adding 30 to 50% by weight of pluronic F-127 to the 0.05 to 0.15N calcium chloride solution and mixing. Here, the mixing step may be performed by repeatedly performing low temperature storage and vortexing as shown in FIG. 5 .

Next, the method for producing tubular biological tissues using 3D printing of the present invention is a conduit of any one of the above-described bioink composition kits for tubular biological tissues or a bioink composition kit for tubular biological tissues manufactured by any one of the above-described manufacturing methods. Loading the formed hydrogel and the sacrificial hydrogel into a 3D printer; A pipe formed by simultaneously driving the two heads provided in the 3D printer under different process conditions to form a sacrificial object, which is the inner center portion, with the sacrificial hydrogel and forming a tubular structure surrounding the inner center portion with the conduit-forming hydrogel. outputting the line structure; Curing the tubular structure by UV irradiating the surface of the pipeline structure; and cutting both ends of the cured pipeline structure and then removing the sacrificial material to form a hollow in the inner central portion.

In the step of loading the tube-forming hydrogel and the sacrificial-forming hydrogel into the 3D printer, the tube-forming hydrogel and the sacrificial-forming hydrogel are injected into two syringes, and a coaxial nozzle is inserted into the syringe into which the sacrificial-forming hydrogel is placed. It can be performed by attaching a plastic nozzle to a syringe into which hydrogel is injected and then attaching each to a 3D printer head.

The output step is a step of outputting a pipeline structure composed of a core portion and a tubular structure through a coaxial nozzle connected by simultaneously driving two heads provided in the 3D printer, which is a process of a head that outputs a tubular structure with a conduit-forming hydrogel Conditions are 26 ± 0.5 ℃, 220 ~ 230 kPa, process conditions of the head outputting the sacrificial material to the sacrificial hydrogel may be 35 ± 0.5 ℃, 120 ~ 130 kPa.

In the curing step, UV irradiation may be performed for 150 seconds to 210 seconds so that the amount of light reaching the surface of the pipeline structure is 65 to 75 mW/cm 2 .

The step of removing the sacrificial object may include dissolving the sacrificial object by injecting PBS or distilled water into one end of the cut pipeline structure; and inhaling the liquid present in the hollow of the pipeline structure.

The implementation principle of the method for manufacturing a tubular biological tissue using 3D printing according to various embodiments of the present invention having the above configuration is shown in FIG. 3 . As shown in Figure 7, it is possible to print a long cylindrical tube with different inner and outer materials by simultaneously using the two heads provided in the 3D printer at different air pressures and different temperatures. In particular, when alginate and Ca ions meet Using the property of curing, it can be seen that the inner wall of the conduit is cured by CPF-127 (Pluronic F127/CaCl2) and the outer wall of the conduit is cured by UV.

Finally, since the tubular biological tissue of the present invention is manufactured by the above-described manufacturing method, as can be seen through the experimental examples described later, the mechanical properties are increased and the self-standing ability is excellent, so that after printing and curing, it has a circular shape. It is easy to maintain the tubular cross-sectional shape, and it is easy to connect the perfusate for culturing the tubular biological tissue.

Example 1

1. Conduit-forming hydrogel preparation step

(1) Preparing LAP solution

40mL of 1X PBS and 0.04g of LAP were put in a 50mL conical tube, wrapped in foil, and vortexed until the LAP was completely dissolved to prepare a 0.1% by weight LAP solution.

(2) Preparing GelMA hydrogel

4g of GelMA was added to 20mL of 0.1% by weight LAP solution and vortexed. After putting it in a 37 ℃ water bath for 30 minutes, it was vortexed. A 20 wt% GelMA hydrogel was prepared by repeating the above process until GelMA was completely dissolved.

(3) preparing an alginate hydrogel

After putting 20mL of DW and 0.96g of alginate in a 50mL conical tube and vortexing, a 4.8% by weight alginate hydrogel was prepared by heating in a hot plate at 180 ° C for about 4 hours.

(4) Preparing a conduit-forming hydrogel

20mL of 20% by weight GelMA hydrogel was added to 20mL of prepared 4.8% by weight alginate hydrogel, and the final concentration was 2.4% by weight of alginate, 0.05% by weight of LAP, and 10% by weight of GelMA. After putting it in a 37 ℃ water bath for 30 minutes, the process of vortexing was repeated until the alginate hydrogel and the GelMA hydrogel were completely mixed without showing lumps. A rotater was used during the mixing process.

2. Hydrogel preparation step for sacrificial formation

Calcium chloride was diluted in DW to prepare 0.1N CaCl₂. 40% Pluronic F-127 was added to 0.1N CaCl₂ solution and vortexed. A hydrogel for sacrificial formation was prepared by storing at a low temperature in a refrigerator and repeating vortexing until completely mixed. If necessary, a rotater can be used during the mixing process.

Example 2

1. Preparing for printing

3ㅇ3/16 Silicon tubes were cut by 9 cm and stored in a 50mL conical tube with 70% ethanol, and 17/21G coaxial nozzles and 15G plastic nozzles were prepared. As the 3D printer, a 3D bioprinter from TNR Biofab was used, and a prepared silicon tube was installed by connecting it between the side protrusion of the coaxial nozzle and the plastic nozzle.

2. Step of loading the conduit-forming solution and the sacrificial-forming solution into the 3D printer

The hydrogel for conduit formation and the hydrogel for sacrificial formation prepared in Example 1 were loaded on the two heads provided in the 3D printer, respectively. On the other hand, GelMA included in the hydrogel for forming a conduit is in a liquid state at high temperature and hardens close to solid when stored in a refrigerator, and the hydrogel for forming a sacrificial material exists in a liquid state at a low temperature, so each hydrogel contained in a syringe is a printable jelly. It took about 30 to 1 hour to adapt to the print head temperature until it changed to the same state.

Step 3. Outputting the pipeline structure

The process conditions of the head loaded with the hydrogel for formation of the conduit were 26° C. and 220 to 230 kPa, and the process conditions of the head loaded with the hydrogel for formation of the sacrificial body were adjusted to 35° C. and 120 to 130 kPa. After that, the 3D printer was driven to output the pipeline structure.

4. Curing step

UV irradiation was performed for 180 seconds so that the amount of light reaching the surface of the output pipeline structure was 70 mW/cm 2 .

As shown in FIG. 8A, a conduit having a desired shape can be obtained as a printing output, and it can be seen that it has sufficient durability to be picked up with tweezers after curing.

5. Steps to remove the victim

Cut both ends of the hardened pipeline structure, i.e., the conduit, with scissors. The conduit was erected obliquely, and CPF-127 was removed by flowing 1X PBS or DW several times with a pipet aid in the part filled with the hydrogel (CPF-127) for sacrificial formation. To remove all the liquid inside the conduit, the liquid was sucked into the suction pipet.

Through the above-described method, it is possible to obtain various types of tubular biological tissue, such as the straight conduit shown in FIG. 9A as well as the tubular biological tissue having the structure shown in FIGS. 8B to 8D.

Experimental Example 1

In order to check the UV curing of the hydrogel for tube formation obtained in Example 1, the hydrogel for tube formation was placed in two conical tubes, and UV irradiation was performed for 180 seconds so that the light intensity of one was 70 mW / cm 2, and the other One did not undergo UV irradiation.

As a result, it can be confirmed that curing occurs when UV irradiation is performed, as shown in FIG. 6a, and curing does not occur when UV irradiation is not performed, as shown in FIG. 6b.

Experimental Example 2

The perfusion performance of the tubular biological tissue prepared in Example 2 was evaluated using a syringe pump as follows, and the results are shown in FIGS. 9a to 10c.

A nozzle connected to a syringe pump was connected to the end of the hollow conduit, and the colored liquid was flowed at a constant flow rate.

As shown in FIG. 9A, the possibility of perfusion was confirmed by connecting a nozzle to one end of the straight conduit, and as shown in FIG. 10A, the possibility of perfusion was confirmed by connecting nozzles to both ends of the U-shape conduit.

As shown in FIGS. 9b, 10b, and 10c, it can be seen that the perfusion performance is very good.

Although not shown, as a result of testing the perfusion performance several times with different conduits, it was confirmed that the colored liquid did not leak through the conduit wall but flowed out to the end of the conduit on the other side.

Although the present invention has been shown and described with preferred embodiments as described above, it is not limited to the above embodiments, and to those skilled in the art within the scope of not departing from the spirit of the present invention Various changes and modifications will be possible.

Claims (15)

LithiumPhenyl (2,4,6-trimethylbenzoyl) phosphinate (LAP) 0.03 to 0.07% by weight, gelatin methacrylate (GelMA) 5 to 15% by weight and alginate 2 to 3% by weight containing a hydrogel forming a conduit; and 0.05 to 0.15N calcium chloride and 30 to 50% by weight of a sacrificial hydrogel containing pluronic F-127; tubular bio-tissue bioink composition kit comprising a. According to claim 1, The conduit-forming hydrogel is a bioink composition kit for tubular biological tissue, characterized in that it contains phosphate buffered saline (PBS). According to claim 1, The bioink composition kit for tubular biological tissue, characterized in that the conduit-forming hydrogel and the sacrificial-forming hydrogel are separately packaged and stored in a refrigerator. Conduit-forming hydrogel preparation step; and A method for producing a bioink composition kit for tubular biological tissue independently performing the step of preparing a sacrificial material-forming hydrogel. The method of claim 4, wherein the conduit-forming hydrogel preparation step Preparing a 0.05 to 0.15% by weight LAP solution by dissolving LithiumPhenyl (2,4,6-trimethyl benzoyl)phosphinate (LAP) in PBS; Preparing 15 to 25% by weight of GelMA hydrogel by dissolving Gelatin methacrylate (GelMA) in the LAP solution; Preparing an alginate hydrogel of 4 to 5.5% by weight by dissolving alginate in distilled water; and Preparing a tube-forming hydrogel by mixing the GelMA hydrogel and the alginate hydrogel at a volume ratio of 1: 1; According to claim 5, The dissolution is a method for producing a bioink composition kit for tubular biological tissue, characterized in that by repeatedly performing at least one of vortexing, storage in a water bath at 37 ± 0.5 ° C, and water bath at 180 ± 0.5 ° C. According to claim 5, The mixing of the GelMA hydrogel and the alginate hydrogel is a method for producing a bioink composition kit for tubular biological tissue, characterized in that it is made by repeatedly performing 37 ± 0.5 ° C water bath storage and vortexing. The method of claim 4, wherein the step of preparing the sacrificial hydrogel Preparing a 0.05 to 0.15N calcium chloride solution by adding calcium chloride to distilled water; and 30 to 50% by weight of pluronic F-127 is added to the 0.05 to 0.15N calcium chloride solution and mixed. According to claim 8, The mixing step is a method for producing a bioink composition kit for tubular biological tissue, characterized in that performed by repeatedly performing low temperature storage and vortexing. The tubular bio-ink composition kit for tubular living tissue of any one of claims 1 to 3 or the tubular bio-ink composition kit for tubular living tissue prepared by the manufacturing method of any one of claims 4 to 9, and Loading the sacrificial hydrogel into a 3D printer; A pipeline structure formed by simultaneously driving the two heads provided in the 3D printer under different process conditions to form a sacrificial body, which is a core portion, with the sacrificial hydrogel, and forming a tubular structure surrounding the core portion with the conduit-forming hydrogel. outputting; Curing the tubular structure by UV irradiating the surface of the pipeline structure; and Cutting both ends of the cured pipeline structure and then removing the sacrificial material so that the core part is hollow. According to claim 10, In the outputting step, the process conditions of the head for outputting the tubular structure with the tube-forming hydrogel are 26±0.5° C. and 220 to 230 kPa, and the process conditions for the head for outputting the sacrificial material with the sacrificial hydrogel are 35±0.5° C. Tubular biological tissue manufacturing method using 3D printing, characterized in that 0.5 ℃, 120 ~ 130 kPa. According to claim 10, The UV irradiation is a tubular biological tissue manufacturing method using 3D printing, characterized in that carried out for 150 seconds to 210 seconds so that the amount of light reaching the surface of the pipeline structure is 65 to 75 mW / cm 2 . 11. The method of claim 10, wherein removing the sacrifice comprises Dissolving the sacrificial object by injecting PBS or distilled water into one end of the cut pipeline structure; and Tubular biological tissue manufacturing method using 3D printing, characterized in that carried out including; sucking the liquid present in the hollow of the pipeline structure. According to claim 10, Tubular biological tissue manufacturing method using 3D printing, characterized in that the size of the hollow in the pipeline structure and the thickness of the tubular structure are determined according to the diameter of the nozzle used in the 3D printing. A tubular biological tissue prepared by the method of claim 10.

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