Self-powered biological 3D printing bio-ink, 3D printing bioactive scaffold and preparation method of 3D printing bioactive scaffold

Abstract

According to the self-energized biological 3D printing bio-ink, the 3D printing bioactive scaffold and the preparation method of the self-energized biological 3D printing bio-ink, the controllable-release self-energized biological 3D printing bio-ink is prepared by mixing the organic high-molecular polymers with different molecular weights and the energy metabolism promoting substances, and the ink is good in biocompatibility; by changing the content of energy supply substances, the ink can be designed to'provide 'and'accept' different cell metabolism regulation clues in an adjustable time regulation manner, better migration and adhesion of cells are realized, and functions are exerted, and meanwhile, the ink has controllable slow release of energy metabolism promoting substances and generates micro-nano pores when being used for constructing a three-dimensional bracket through 3D printing, so that the energy metabolism promoting effect is improved. The hydrophilicity of the scaffold is improved, the biological energy metabolism activity of the composite material is enhanced, the energy metabolism promoting substance can provide a sufficient nutrition environment for cell growth, the cell energy metabolism process is regulated and controlled, and adhesion, growth, proliferation and functionalization of cells are facilitated.

Description

TECHNICAL FIELD

[0001] The present application relates to biomedical engineering, in particular to a self-powered biological 3D printing biological ink, a 3D printing bioactive scaffold and a preparation method thereof. BACKGROUND

[0002] The regulation of cellular energy metabolism can be a determinant of cell survival, proliferation, differentiation and specific functions, and thus, energy metabolism plays a crucial role in tissue repair and regeneration. Adenosine triphosphate (ATP) is the main source of cellular energy and plays a role in many biological processes. Metabolic factors such as glucose have been shown to regulate the energy requirements of stem cells during osteoblast differentiation, and energy deficiency can largely impair bone formation. Studies have shown that when fibroin hydrogel loaded with hMSCs is implanted subcutaneously in mice, the glycolysis and ATP reserves of the cells decrease by 66% and 40% respectively within 24 hours, and a large number of cells die (-94%) within the first 3 days of implantation. This requires the scaffold to provide the necessary nutrients for MSCs to prolong their survival after implantation. By regulating oxidative metabolism to increase the ATP level of cells, it can help to address the high energy requirements associated with enhanced anabolic, mitotic and migratory activities in tissue repair / regeneration. The latest progress in cellular bioenergetics (CBE) highlights the potential of methods that allow the delivery of bioenergy for therapeutic purposes, and a common approach is to increase the oxidation of alternative energy substrates available in the microenvironment, such as amino acids, fatty acids and metabolites (such as lactic acid, citric acid) to meet their energy requirements. However, these applications only provide benefits in in vitro models or relatively thin superficial tissues (such as skin) when continuously dosed. So far, it has been difficult to achieve three-dimensional scaffolds with long-term bioenergetic effects to stimulate the repair of critical size defects in complex tissues such as bone. This is largely due to the inability to improve the stability and related activity of ATP using existing scaffold manufacturing techniques, as well as the sensitivity of ATP to the physiological environment.

[0003] Cells can perceive and respond to the surface topography of biomaterials, and topographical cues such as geometry, roughness and shape, in turn, profoundly influence cell morphology, migration and differentiation. The development of metabolic regulation for biomaterial design is of great significance not only for cell therapy, but also for advanced biomaterial design, although it is still in its infancy, and a comprehensive understanding of the mechanisms of the material cue-metabolic state-cell behavior axis is still in its infancy. For example, citrate-based biomaterials, whose physical properties are fine-tuned, can be designed to "provide" and "accept" different metabolic regulation cues in a tunable and temporal manner to meet the dynamic needs for optimal regeneration results. These biomaterials consciously guide cell metabolism and subsequent cell behavior to enhance and predictable regeneration results.

[0004] A common approach in existing technologies is to increase the oxidation of available alternative energy substrates in the microenvironment, such as amino acids, fatty acids, and metabolites (such as lactate and citric acid) to meet their energy needs. However, these applications only provide benefits in in vitro models or when continuously administered to relatively thin superficial tissues (such as skin), making it difficult to achieve long-term energy supply from the materials. Secondly, the traditional use of nanocomposites and bioactive fillers (such as bioceramics) as additives to increase scaffold bioactivity often fails to respond quickly to cell behavior and cannot control the regulation of cell metabolism. Summary of the Invention

[0005] In view of this, it is necessary to provide a self-powered biological 3D printing bio-ink, 3D printing bioactive scaffold and its preparation method that can controllably release and provide energy and regulate cell metabolism to address the current defects of uncontrollable cell metabolism and the inability to achieve long-term energy supply.

[0006] To solve the above problems, this application adopts the following technical solutions:

[0007] One of the purposes of this application is to provide a method for preparing a self-powered biological 3D printing bio-ink, comprising the following steps:

[0008] The self-powered biological 3D printing bio-ink is prepared by a solvent dispersion method, specifically comprising:

[0009] An organic high molecular polymer is mixed with an organic solvent to prepare a high molecular solution, an energy metabolism promoting substance is dissolved in the high molecular solution, and the mixture is stirred at room temperature to obtain a viscous high molecular mixed solution, and the viscous high molecular mixed solution is vacuum dried to obtain the self-powered biological 3D printing bio-ink; or

[0010] The self-powered biological 3D printing bio-ink is prepared by a melt dispersion method, specifically comprising:

[0011] The organic high molecular polymer and the energy metabolism promoting substance are mixed, dissolved at 100-120° C., and then stirred evenly to room temperature to obtain the self-powered biological 3D printing biological ink.

[0012] In some embodiments, the mass ratio of the organic high molecular polymer to the energy metabolism promoting substance is 100:0.1-100:50.

[0013] In some embodiments, the organic high molecular polymer includes at least one of polycaprolactone, gelatin, alginate, chitosan, polyethylene glycol, polyurethane, polylactic acid, polylactic acid copolymer and polyhydroxy fatty acid.

[0014] In some embodiments, the energy metabolism promoting substance includes at least one of pyruvate, citrate, fatty acid, glutamine or glucose.

[0015] In some embodiments, the pyruvate comprises at least one of sodium pyruvate, potassium pyruvate, and calcium pyruvate.

[0016] In some embodiments, the organic solvent comprises dichloromethane, chloroform, and acetone.

[0017] The second object of the present application also provides a self-powered biological 3D printing biological ink prepared by any of the preparation methods.

[0018] The second object of the present application also provides a printing method of a 3D printing bioactive scaffold, wherein the self-powered biological 3D printing biological ink is loaded into a 3D extrusion printer cartridge, heated and adjusted printing parameters to obtain the 3D printing bioactive scaffold.

[0019] In some embodiments, the printing method adopts single-channel biological printing or multi-channel alternating biological printing.

[0020] The third object of the present application also provides a 3D printing bioactive scaffold, which is printed by the printing method of the 3D printing bioactive scaffold.

[0021] The technical scheme adopted by the present application has the following beneficial effects:

[0022] The self-powered biological 3D printing biological ink, the 3D printing bioactive scaffold and the preparation method thereof provided by the present application are prepared by mixing organic high molecular polymers with different molecular weights and energy metabolism promoting substances to obtain a self-powered biological 3D printing biological ink with controllable release. The ink has good biocompatibility, and can be designed to "provide" and "accept" different cell metabolism regulation clues in a time-adjustable manner by changing the content of the energy-providing substance, so as to realize better migration and adhesion of cells and play functions. Meanwhile, the ink is used to construct a three-dimensional scaffold by 3D printing, generates micro-nano pores by controllably releasing the energy metabolism promoting substance, improves the hydrophilicity of the scaffold, enhances the biological energy metabolism activity of the composite material, and the energy metabolism promoting substance can provide sufficient nutrient environment for cell growth, regulate the energy metabolism process of cells, and is conducive to the adhesion, growth, proliferation and functionalization of cells. BRIEF DESCRIPTION OF DRAWINGS

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following will briefly introduce the drawings needed to be used in the embodiments of the present application or the prior art description. Obviously, the drawings described below are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor on the basis of these drawings.

[0024] Figure 1Structure schematic diagram of 3D printing bioactive scaffold provided by the embodiment of the present application, wherein (A) is a single-channel 3D printing pyruvate / polycaprolactone scaffold physical diagram; (B) is a double-channel alternating bio 3D printing pyruvate / polycaprolactone / hydrogel (cell loading) scaffold physical diagram.

[0025] Figure 2 Schematic diagram of pyruvate sodium release kinetics in the scaffold with different pyruvate sodium addition amounts provided by the embodiment 1 of the present application.

[0026] Figure 3 Schematic diagram of surface water contact angle of the scaffold with different pyruvate sodium addition amounts provided by the embodiment 1 of the present application.

[0027] Figure 4 Surface morphology (SEM) of the scaffold with different pyruvate sodium addition amounts provided by the embodiment 1 of the present application, wherein (A) is the surface morphology of the 3D printing pyruvate / polycaprolactone scaffold; (B) is the surface morphology of the 3D printing pyruvate / polycaprolactone scaffold after 30 days in PBS buffer.

[0028] Figure 5 Schematic diagram of degradation kinetics of the scaffold with different pyruvate sodium addition amounts in PBS buffer at 37℃ provided by the embodiment 1 of the present application.

[0029] Figure 6 Schematic diagram of cytotoxicity results of the scaffold with different pyruvate sodium addition amounts at 1, 3, 5 days provided by the embodiment 1 of the present application.

[0030] Figure 7 Schematic diagram of real-time heat release monitoring of the scaffold and cell culture provided by the embodiment 1 of the present application. DETAILED DESCRIPTION

[0031] The embodiments of the present application are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar notations represent the same or similar elements or elements with the same or similar functions throughout. The embodiments described below by referring to the accompanying drawings are exemplary and are intended to explain the present application, and cannot be understood as a limitation of the present application.

[0032] In the description of the present application, it is understood that the terms "upper", "lower", "horizontal", "inner", "outer" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present application and simplifying the description, and therefore cannot be understood as indicating or implying that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation of the present application.

[0033] In addition, the terms "first", "second", "third", etc. are used only for descriptive purposes and are not to be construed as indicating or implying relative importance or an ordered ranking of the indicated technical features. Thus, features defined with "first", "second" or "third" can explicitly or implicitly include one or more of the features. In the description of the present application, the meaning of "a plurality of" is two or more, unless otherwise expressly and specifically limited.

[0034] In order to make the purpose, technical solutions and advantages of the present application more clear, the present application is further described in detail below in combination with the drawings and examples.

[0035] The preparation method of the self-powered biological 3D printing biological ink provided by the present embodiment specifically includes the following steps:

[0036] The self-powered biological 3D printing biological ink is prepared by using a solvent dispersion method, specifically including:

[0037] The organic high polymer is mixed with the organic solvent to prepare a high molecular solution, the energy metabolism promoting substance is dissolved in the high molecular solution, the viscous high molecular mixed solution is obtained by stirring at room temperature, and the self-powered biological 3D printing biological ink is obtained by vacuum drying treatment of the viscous high molecular mixed solution.

[0038] Or the self-powered biological 3D printing biological ink is prepared by using a melting dispersion method, specifically including:

[0039] The organic high polymer is mixed with the energy metabolism promoting substance, and then dissolved and stirred uniformly at 100-120℃ to room temperature to obtain the self-powered biological 3D printing biological ink.

[0040] Specifically, the organic high polymer is placed in a polytetrafluoroethylene beaker to form a high molecular solution, the energy metabolism promoting substance is dissolved in the high molecular solution, and then dissolved and stirred uniformly at 100-120℃ to room temperature to obtain the self-powered biological 3D printing biological ink.

[0041] In the present embodiment, the organic high polymer includes at least one of polycaprolactone, gelatin, alginic acid, chitosan, polyethylene glycol, polyurethane, polylactic acid, poly(lactic-co-glycolic acid) and polyhydroxyalkanoate.

[0042] In the present embodiment, the energy metabolism promoting substance includes at least one of pyruvate, citrate, fatty acid, glutamine or glucose. The pyruvate includes at least one of sodium pyruvate, potassium pyruvate and calcium pyruvate.

[0043] In the present embodiment, the organic solvent includes dichloromethane, chloroform and acetone.

[0044] In the embodiment, the mass ratio of the organic polymer and the energy metabolism promoting substance is 100:0.1-100:50.

[0045] It can be understood that the types and molecular weights of the organic polymer and the energy metabolism promoting substance mentioned in the preparation method described above have very high selectivity and can be selected according to the use requirements.

[0046] It can be understood that the release rate is controlled by changing the addition amount and type of the energy metabolism promoting substance in the application, so as to change the surface micro-morphology (micro-nano-pore) of the scaffold, improve the degradation rate, improve the biological activity of the 3D printed scaffold, promote cell proliferation and adhesion.

[0047] It should be noted that the self-powered 3D printing biological ink provided in the embodiment can be used for single-channel extrusion type biological 3D printing, and can also be applied to multi-channel alternating biological 3D printing (one or more channels can be hydrogel loaded with cells).

[0048] The application also provides a printing method of a 3D printed biological active scaffold, wherein the self-powered biological 3D printing biological ink is loaded in a 3D extrusion type printer barrel, heated and adjusted printing parameters to obtain the 3D printed biological active scaffold.

[0049] In the embodiment, the printing method adopts single-channel biological printing or multi-channel alternating biological printing.

[0050] Please refer to Figure 1 , the structure schematic diagram of the active scaffold printed by the printing method of the 3D printed biological active scaffold provided in the above embodiment, wherein (A) is a single-channel 3D printed pyruvate / polycaprolactone scaffold real object diagram; (B) is a double-channel alternating biological 3D printed pyruvate / polycaprolactone / hydrogel (loaded with cells) scaffold real object diagram.

[0051] The 3D printed biological active scaffold and the preparation method provided by the application, by mixing organic polymers with different molecular weights and energy metabolism promoting substances, a controllable release self-powered biological 3D printing biological ink is prepared, the ink has good biocompatibility, by changing the content of the energy substance, it can be designed to "provide" and "accept" different cell metabolism regulation clues in a adjustable and time manner, to realize better migration and adhesion of cells, and to play functions, at the same time, the ink is used to construct a three-dimensional scaffold by 3D printing, controllable slow-release energy metabolism promoting substances are generated, micro-nano-pores are generated, the hydrophilicity of the scaffold is improved, the biological energy metabolism activity of the composite material is enhanced, the energy metabolism promoting substance can provide sufficient nutrient environment for cell growth, regulate cell energy metabolism process, and is conducive to cell adhesion, growth, proliferation and functionalization.

[0052] The above technical solutions of the present application are described in detail below in combination with specific embodiments.

[0053] Example 1

[0054] A. Preparation of 3D printing polycaprolactone self-powered biomaterial ink (Mn = 45000) by melt dispersion method

[0055] First, 10 g of polycaprolactone was placed in a polytetrafluoroethylene beaker, and then 0.1, 0.5, 0.8, 1, 2, and 3 g of sodium pyruvate powder were dissolved in the polymer, respectively, at 100-120°C, and stirred uniformly to room temperature (the mass ratio of polycaprolactone to sodium pyruvate was 100:1, 100:5, 100:8, 100:10, 100:20, and 100:30). The operation was repeated three to five times.

[0056] B. Preparation of self-powered scaffold by 3D printing

[0057] The self-powered biomaterial ink obtained in step A was subjected to 3D printing, and the mixed ink was loaded into the barrel of a 3D extrusion printer, heated to 90°C, and the printing parameters were adjusted to obtain the final 3D structure scaffold.

[0058] The scaffold was subjected to sodium pyruvate drug release determination, and the results are shown in Figure 2 The release rates of different sodium pyruvate addition groups were different, and the sodium pyruvate release time of each group was 1% (6h), 5% (24h), 10% (72h), 20% (7d), and 30% (>7d). The water contact angle of the scaffold was determined Figure 3 ), the addition of sodium pyruvate reduced the water contact angle and improved the hydrophilicity of the scaffold. The surface morphology of the scaffold was characterized by SEM Figure 4 ), the addition of sodium pyruvate increased the surface roughness of the scaffold, and due to the release of sodium pyruvate, different sizes of micro-nano pores were formed on the surface of the scaffold. After immersion in PBS buffer at 37°C for 30 days, the surface roughness of the scaffold with added sodium pyruvate increased, and the micro-nano pores also increased. At the same time, the degradation rate of 3D scaffolds with different amounts of sodium pyruvate in PBS buffer at 37°C was monitored Figure 5

[0059] Finally, the prepared scaffold was co-cultured with human bone marrow mesenchymal stem cells (BMSC), and the cell cytotoxicity and proliferation were determined, Figure 6 The CCK-8 results showed that the 1%, 5%, 8%, and 10% addition groups promoted cell survival within 1 day, and better cell proliferation at 3 days and 5 days; the 20% and 30% addition groups showed cytotoxicity due to high solubility. Figure 7 ​The real-time heat release monitoring results show that the 10% added amount of the scaffold can provide long-term energy supply for the energy required by cell life activities.

[0060] Example 2

[0061] A. Preparation of 3D printing polycaprolactone self-powered biomaterial ink (Mn = 14000) by melt dispersion method

[0062] First, 10 g of polycaprolactone was placed in a polytetrafluoroethylene beaker, and then 0.1, 0.5, 0.8, 1, 2, and 3 g of calcium pyruvate powder were dissolved in the polymer, respectively, and stirred uniformly to room temperature after dissolution at 100-120°C (the mass ratio of polycaprolactone to calcium pyruvate is 100:1, 100:5, 100:8, 100:10, 100:20, and 100:30). The operation was repeated three to five times.

[0063] B. Preparation of self-powered scaffold by 3D printing by melt extrusion

[0064] The self-powered biomaterial ink obtained in step A was subjected to 3D printing, and the mixed ink was loaded into the barrel of a 3D extrusion printer and heated to 65°C. The printing parameters were adjusted to obtain the final 3D structure scaffold. The prepared scaffold was co-cultured with human bone marrow mesenchymal stem cells (BMSCs), and cell cytotoxicity and proliferation assays were performed. The CCK-8 results showed that the 1%, 5%, 8%, and 10% added amount groups promoted cell survival within 1 day and promoted cell proliferation better at 3 days and 5 days; the 20% and 30% added amount groups showed cytotoxicity due to high solubility.

[0065] Example 3

[0066] A. Preparation of 3D printing methacrylated gelatin (GelMA) self-powered biomaterial ink by solvent dispersion method

[0067] First, 10 g of GelMA was placed in a polytetrafluoroethylene beaker, and then 0.5 g of sodium pyruvate and 0.5 g of calcium pyruvate, 0.5 g of sodium pyruvate and 0.8 g of calcium pyruvate, 0.5 g of sodium pyruvate and 1 g of calcium pyruvate, 0.8 g of sodium pyruvate and 0.5 g of calcium pyruvate, 0.8 g of sodium pyruvate and 0.8 g of calcium pyruvate, and 0.8 g of sodium pyruvate and 1 g of calcium pyruvate powder were dissolved in the polymer, respectively, and stirred uniformly to room temperature after dissolution at 100-120°C (the mass ratio of gelatin to pyruvate is 100:10, 100:13, 100:15, 100:13, 100:16, and 100:18). The operation was repeated three to five times.

[0068] B. Preparation of self-powered scaffold by 3D printing by extrusion

[0069] The self-powered bio-ink obtained in step A is subjected to 3D printing, the mixed ink is loaded in the 3D extrusion printer cartridge, the printing parameters are adjusted, and the final 3D structure scaffold is obtained. The prepared is subjected to cell cytotoxicity and proliferation determination, and the CCK-8 results show that the addition amount of 0.5 g sodium pyruvate and 0.5 g calcium pyruvate, 0.5 g sodium pyruvate and 0.8 g calcium pyruvate, 0.5 g sodium pyruvate and 1 g calcium pyruvate, 0.8 g sodium pyruvate and 0.5 g calcium pyruvate groups promote cell survival within 1 day, and promote better cell proliferation at 3 days and 5 days; the addition amount of 0.8 g sodium pyruvate and 0.8 g calcium pyruvate, 0.8 g sodium pyruvate and 1 g calcium pyruvate groups show cytotoxicity due to too high solubility.

[0070] Example 4

[0071] A. Preparation of 3D printing self-powered bio-ink (Mn = 14000) by solvent dispersion method

[0072] First, 10 g of polycaprolactone is placed in a polytetrafluoroethylene beaker, and then 0.1, 0.5, 0.8, 1, 2, and 3 g of calcium pyruvate powder are dissolved in the polymer at 100-120°C, and then stirred uniformly to room temperature (the mass ratio of polycaprolactone to calcium pyruvate is 100:1, 100:5, 100:8, 100:10, 100:20, and 100:30). The operation is repeated for three to five times.

[0073] First, 10 g of polycaprolactone is placed in a beaker, and 100 ml of dichloromethane is added to dissolve and prepare a polymer solution. Then 0.1, 0.5, 0.8, 1, 2, and 3 g of calcium pyruvate powder are dissolved in the polymer at room temperature (the mass ratio of polycaprolactone to calcium pyruvate is 100:1, 100:5, 100:8, 100:10, 100:20, and 100:30). After the dichloromethane is volatilized, a viscous polymer mixed solution is obtained, which is vacuum dried for 24 hours, and then freeze-dried for 24 hours to obtain a 3D printing bio-ink.

[0074] B. Preparation of self-powered scaffold by extrusion 3D printing

[0075] The self-powered bio-ink obtained in step A is subjected to 3D printing, the mixed ink is loaded in the 3D extrusion printer cartridge, the printing parameters are adjusted, and the final 3D structure scaffold is obtained. The prepared scaffold is co-cultured with human bone marrow mesenchymal stem cells (BMSC), and subjected to cell cytotoxicity and proliferation determination, and the CCK-8 results show that the addition amount of 1%, 5%, 8%, and 10% promotes cell survival within 1 day, and promotes better cell proliferation at 3 days and 5 days; the addition amount of 20% and 30% shows cytotoxicity due to too high solubility.

[0076] It can be understood that any combination of the technical features in the above-described embodiments can be made, and for the sake of brevity, not all possible combinations of the technical features in the above-described embodiments are described, however, as long as the combination of the technical features does not exist contradictory, it should be considered within the scope of the present disclosure.

[0077] The above is only a preferred embodiment of the present application, and only the technical principles of the present application are specifically described, and these descriptions are only for the purpose of explaining the principles of the present application, and cannot be interpreted in any way as a limitation on the scope of protection of the present application. Based on the explanation here, any modification, equivalent replacement and improvement made within the spirit and principle of the present application, and other specific embodiments of the present application which can be easily thought by those skilled in the art without creative labor, should be included in the protection scope of the present application.

Claims

1. A method for preparing a self-powered bio 3D printing bio-ink, characterized in that, comprising the following steps: The self-powered biological 3D printing biological ink is prepared by a solvent dispersion method, specifically comprising the following steps: The organic polymer is mixed with an organic solvent to prepare a polymer solution, an energy metabolism promoting substance is dissolved in the polymer solution, the viscous polymer mixed solution is obtained by stirring at room temperature, and the self-powered biological 3D printing biological ink is obtained by vacuum drying the viscous polymer mixed solution; or The self-powered biological 3D printing biological ink is prepared by a solvent dispersion method, specifically comprising the following steps: The organic polymer is mixed with an organic solvent to prepare a polymer solution, an energy metabolism promoting substance is dissolved in the polymer solution, the viscous polymer mixed solution is obtained by stirring at room temperature, and the self-powered biological 3D printing biological ink is obtained by vacuum drying the viscous polymer mixed solution; or 2. The method for preparing the self-powered biological 3D printing bio-ink according to claim 1, wherein: The mass ratio of the organic polymer to the energy metabolism promoting substance is 100:0.1-100:

50.

3. The method for preparing the self-powered biological 3D printing biological ink according to claim 2, wherein: The organic polymer includes at least one of polycaprolactone, gelatin, alginic acid, chitosan, polyethylene glycol, polyurethane, polylactic acid, poly(lactic-co-glycolic acid) and polyhydroxyalkanoate.

4. The method for preparing the self-powered biological 3D printing bio-ink according to claim 2, wherein: The energy metabolism promoting substance includes at least one of pyruvate, citrate, fatty acid, glutamine or glucose.

5. The method for preparing the self-powered biological 3D printing biological ink according to claim 4, characterized in that: The pyruvate includes at least one of sodium pyruvate, potassium pyruvate and calcium pyruvate.

6. The method for preparing the self-powered biological 3D printing bio-ink according to claim 1, wherein: The organic solvent includes dichloromethane, chloroform and acetone.

7. A self-energic bio 3D printing bio-ink, characterized in that, The self-powered biological 3D printing biological ink is prepared by the preparation method of any one of claims 1 to 6.

8. A method of printing a 3D printed bioactive scaffold, characterized in that, The self-powered biological 3D printing biological ink of claim 7 is loaded into a 3D extrusion type printer cartridge, heated and adjusted for printing parameters to obtain the 3D printing biological active scaffold.

9. The printing method of the 3D printing biological active scaffold of claim 8, wherein the printing method adopts single-channel biological printing or multi-channel alternating biological printing.

10. A 3D printed bioactive scaffold, characterized in that, The self-powered biological 3D printing biological ink is prepared by the preparation method of any one of claims 1 to 6.

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