3D printing ultraviolet light crosslinking sesbania glue hydrogel stent as well as preparation method and application of 3D printing ultraviolet light crosslinking sesbania glue hydrogel stent
By preparing 3D-printed hydrogel scaffolds using modified guar gum, the problems of insufficient biocompatibility and mechanical properties of existing materials in tissue engineering are solved, enabling efficient and low-cost tissue repair applications.
Patent Information
- Application Number
- CN202410487768.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-04-22
- Publication Date
- 2025-10-24
AI Technical Summary
Existing 3D printing materials are difficult to simultaneously satisfy good biocompatibility, cell adhesion and mechanical properties in tissue engineering. Furthermore, synthetic polymers have long degradation cycles and poor biocompatibility, while natural polymers degrade quickly and have poor printing and processing performance.
A 3D-printed hydrogel scaffold was formed by cross-linking methacrylic anhydride-modified guar gum with ultraviolet light and combined with bone marrow mesenchymal stem cells to prepare a guar gum hydrogel scaffold with a porous structure, achieving rapid cross-linking and good biocompatibility.
High-fidelity printing of scaffolds has been achieved, exhibiting good biocompatibility, cell adhesion, and mechanical properties, making them suitable for tissue repair. They are also low in cost and widely available.
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Figure CN120827641A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of biological composite materials, in particular to a 3D printing ultraviolet cross-linking sesbania gum hydrogel scaffold, a preparation method and application thereof. BACKGROUND
[0002] The development of tissue engineering provides a technical means for tissue regeneration and repair, which will change the traditional treatment mode of "repairing trauma with trauma" and enter a new stage of non-traumatic repair. The three elements of tissue engineering mainly include seed cells, scaffold materials and growth factors. The currently studied scaffold materials include the following: fiber scaffold, porous scaffold, microsphere aggregation and 3D printing technology. The former several kinds provide a two-dimensional microenvironment for seed cells, while the 3D printing technology can provide a three-dimensional microenvironment required for tissue repair. The 3D printing technology can control the position and geometry of biological micropores, and the three-dimensional porous structure can promote cell adhesion and proliferation, allow tissue fluid, cells and growth factors to enter the material interior, and is beneficial to the ingrowth of new tissue and the remodeling of complex physiological microenvironment of tissue.
[0003] At present, hydrogel is a commonly used biological ink. Hydrogel has a three-dimensional cross-linked network structure and also contains a large amount of water. Hydrogel has adjustable strength, degradability, functional modification and other properties, and can be used as a soft material to simulate the microenvironment of extracellular matrix. It includes three types: one type is prepared from natural polymers such as agar, gelatin, cellulose, collagen, silk fibroin and hyaluronic acid; one type is prepared from synthetic polymers such as polyacrylamide, polyurethane and polyethylene glycol; and the other type is a composite hydrogel composed of synthetic polymers and natural polymers. Synthetic polymers have poor biocompatibility, long degradation period and accumulated degradation products such as lactic acid, which can cause local inflammation, and these shortcomings limit the application of synthetic polymers. Natural polymers have good biocompatibility and low immunogenicity, but they cannot meet the requirements of 3D printing due to their fast degradation rate, poor printability and poor mechanical properties. Functional groups such as tyramine, thiol and vinyl sulfone are used to modify natural hydrogels to achieve printability and improve material mechanical properties, which is crucial for 3D biological printing. Although many printable materials have their own advantages, finding a biologic material that can meet the requirements of 3D biological printing mechanical properties, cell adhesion and good cell compatibility is the key to the development of 3D printing technology in the direction of tissue engineering. SUMMARY
[0004] The present application discloses a 3D printing ultraviolet cross-linking sesbania gum hydrogel scaffold, a preparation method and application thereof, and the preparation method comprises the following steps:
[0005] Step S1, the pannic gum raw material is dissolved in ultrapure water, and is dissolved at 60°C under sufficient stirring to obtain a pannic gum aqueous solution;
[0006] Step S2, slowly add methacrylic anhydride at 60°C, slowly add methacrylic anhydride under 3-5°C ice bath, and continue stirring at 50°C for 1 hour;
[0007] Step S3, drop sodium hydroxide to maintain the pH of the reaction system at 7.0, continue the reaction for 5-7h, and different methacrylate pannic gum derivatives with different grafting degrees are obtained according to the amount of methacrylic anhydride added; the obtained product is dialyzed with a 3500-4000kd dialysis bag, and after freeze-drying, a porous sponge-like modified pannic gum is obtained;
[0008] Step S4, prepare a pannic gum hydrogel by completely dissolving in PBS containing a light initiator (irgacure2959) with a concentration of 0.8-0.12% w / v;
[0009] Step S5, mix the obtained pannic gum hydrogel with bone marrow mesenchymal stem cells, and extrude and print through a 3D bioprinting system; through the sprayed fiber filaments, the pannic gum hydrogel scaffold is formed by layer-by-layer accumulation, and the printed scaffold is crosslinked under ultraviolet light 365nm, 200mW / cm2, to obtain a 3D printed pannic gum scaffold loaded with mesenchymal stem cells.
[0010] Further, when the pannic gum is modified, the concentration of the pannic gum aqueous solution is 0.8-0.12% w / v.
[0011] Further, the modifier is methacrylic anhydride.
[0012] Further, the concentration of the methacrylic anhydride is 1mL / g.
[0013] Further, the concentration of the pannic gum hydrogel for printing is 3.5-4.5%.
[0014] Further, the ultraviolet light crosslinking time of the pannic gum hydrogel scaffold is 55-60s.
[0015] Further, the concentration of the bone marrow mesenchymal cell solution is 1×10 7 cell / ml.
[0016] A 3D printed hydrogel scaffold prepared according to the above method, and application of the hydrogel scaffold in preparing a human tissue engineering scaffold.
[0017] The advantages and effects of the present application are as follows:
[0018] 1. The reaction conditions in the preparation method are mild, and the required conditions are not harsh; the scaffold is crosslinked by ultraviolet light in a short time, so that the fidelity of the printed scaffold can be better realized, and the scaffold can be used for tissue repair.
[0019] 2. The excellent performance of the amaranth gum scaffold material is fully exhibited, and the scaffold material has good biocompatibility, injectability, and cell adhesion, etc. Compared with other materials, the scaffold material has simple design, controllable shape and microstructure, good mechanical properties, low cost, wide source, and can be used for bone marrow mesenchymal stem cells, and provides a preliminary research basis for discovering new tissue engineering scaffolds.
[0020] The above description is only a summary of the technical solutions of the present application. In order to more clearly understand the technical means of the present application, the contents of the specification can be implemented, and in order to make the above and other purposes, characteristics and advantages of the present application more obvious and easy to understand, the following will be described in detail with the preferred embodiments of the present application and the accompanying drawings.
[0021] According to the detailed description of the specific embodiments of the present application in the following text combined with the drawings, those skilled in the art will more clearly understand the above and other purposes, advantages and characteristics of the present application. BRIEF DESCRIPTION OF DRAWINGS
[0022] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiment or the prior art description. Obviously, the drawings in the following description are some embodiments of the present application, and those skilled in the art can also obtain other drawings according to these drawings without creating any creative labor. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, each element or part is not necessarily drawn according to the actual proportion.
[0023] Among them,
[0024] Figure 1 are respectively the Fourier infrared spectrum detection result graphs of amaranth gum and modified amaranth gum;
[0025] Figure 2 is a 3D printed amaranth gum hydrogel scaffold;
[0026] Figure 3 is a result graph of mechanical property test;
[0027] Figure 4 is a result graph of cell activity after culturing amaranth gum composite chondrocytes for 7 days, 14 days and 21 days; DETAILED DESCRIPTION
[0028] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the following will be combined with the accompanying drawings for the embodiments of the present application to make a clear and complete description of the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. In the following description, specific details such as specific configurations and components are provided only to help a comprehensive understanding of the embodiments of the present application. Therefore, those skilled in the art should understand that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of the present application. In addition, in order to be clear and concise, the description of known functions and structures is omitted in the embodiments.
[0029] It should be understood that the "one embodiment" or "the embodiment" mentioned throughout the specification means that the specific features, structures or characteristics related to the embodiment are included in at least one embodiment of the present application. Therefore, "one embodiment" or "the embodiment" appearing throughout the specification does not necessarily refer to the same embodiment. In addition, these specific features, structures or characteristics can be combined in one or more embodiments in any suitable manner.
[0030] In addition, reference numerals and / or letters can be repeated in different examples in the present application. Such repetition is for the purpose of simplification and clarity, and does not indicate the relationship between the various embodiments and / or arrangements discussed.
[0031] The term "and / or" herein is only a description of the association relationship of the associated objects, which means that there can be three relationships, for example, A and / or B can mean that A exists alone, B exists alone, and A and B exist together. The term "and" herein is a description of another association relationship of the associated objects, which means that there can be two relationships, for example, A and B can mean that A exists alone and A and B exist together. In addition, the character " / " herein generally represents an "or" relationship between the associated objects before and after it.
[0032] The term "at least one" herein is only a description of the association relationship of the associated objects, which means that there can be three relationships, for example, at least one of A and B can mean that A exists alone, A and B exist together, and B exists alone.
[0033] It should also be noted that the relationship terms such as first and second in the present document are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between the entities or operations. Moreover, the terms "include", "contain" or any other variants thereof are intended to cover non-exclusive inclusion.
[0034] Embodiment 1
[0035] This embodiment discloses a 3D-printed UV-crosslinked sesbania gum hydrogel scaffold, preparation method, and application. In experiments with methacrylic anhydride-modified sesbania gum, 5M sodium hydroxide was added dropwise to maintain a certain pH for reaction, preferably 7-8 or 8-9. Methacrylated sesbania gum was prepared as follows: a 1% (w / v) sesbania gum solution was prepared at 60°C, methacrylic anhydride was added at the same temperature, and after magnetic stirring, 5M sodium hydroxide was added dropwise to maintain the pH at 7-8 or 8-9. After the reaction was continued at 50°C for 6 hours, the solution was dialyzed against ultrapure water at room temperature for one week and freeze-dried for one week to obtain the product.
[0036] By comparing the grafting degree of methacrylic anhydride of the product, it was found that the efficiency of grafting methacrylic anhydride of the product was higher at pH 7.0. Figure 1 The following are the Fourier transform infrared spectra of sesbania gum and modified sesbania gum. SGMA has a higher peak at 1710 cm-1 than SG. -1 The emergence of a new peak at , indicates that methacrylic anhydride is successfully grafted onto the sesbania gum structure.
[0037] Example 2
[0038] This embodiment discloses a 3D printed UV-crosslinked sesbania glue hydrogel scaffold and its preparation method and application.
[0039] Freeze-dried methacrylated sesbania gum was completely dissolved in PBS containing a photoinitiator (0.1% w / v) to prepare a hydrogel. The resulting hydrogel was then composited with bone marrow mesenchymal stem cells (1×107 cells / ml) and extrusion-printed using a 3D-Bioplotter system. The ejected fibers were stacked layer by layer to form a scaffold. The printed scaffold was then cross-linked with ultraviolet light at 365 nm and 200 mW / cm2 for 58 s to obtain the product.
[0040] like Figure 2 As shown in the figure, it is a 3D printed sesbania glue hydrogel scaffold of the present invention. It is found that the number of printed layers can be more than 20 layers. The printing concentration is 4%. The printed cube has a length × width × height of 16.9 mm × 16.9 mm × 10 mm. It can be seen that the spindles are continuously and evenly stacked. After printing is completed, the spindle texture on the scaffold is clearly visible.
[0041] Example 3
[0042] The embodiment discloses a 3D printing ultraviolet cross-linking sesbania gum hydrogel scaffold and a preparation method and application thereof, the concentration of a sesbania gum raw material during modification can be 1%-2% (w / v), and the preparation method is as follows: a 3D printing sesbania gum hydrogel scaffold is prepared by the following method: a sesbania gum raw material is used to prepare a 1%-2% (w / v) sesbania gum aqueous solution at 50 DEG C, different grafting degree derivatives are prepared by adding different concentrations of methacrylic anhydride, the obtained product is dialyzed, freeze-dried, and modified sesbania gum is obtained.
[0043] It is found through comparison that the grafting degree of methacrylic anhydride in the solution with a concentration of 1% is higher than that of other concentrations.
[0044] Embodiment 4
[0045] The embodiment discloses a 3D printing ultraviolet cross-linking sesbania gum hydrogel scaffold and a preparation method and application thereof, the method comprises the following steps: a sesbania gum raw material is dissolved in ultrapure water at a concentration of 1% w / v, is fully stirred and dissolved at 60 DEG C to obtain a sesbania gum aqueous solution, methacrylic anhydride is slowly added at a speed of 500 uL / min at 60 DEG C, stirring is continued for 1 hour at 60 DEG C, then sodium hydroxide is added dropwise to maintain the pH of the reaction system at 7.0, and the reaction is continued for 6h at 50 DEG C, different grafting degree methacrylated sesbania gum derivatives are obtained according to the amount of methacrylic anhydride added, the obtained product is dialyzed with 3500-4000 kd, freeze-dried for one week, and porous sponge-like modified sesbania gum is obtained; the modified sesbania gum is completely dissolved in PBS containing a concentration of 0.1% W / v of a photoinitiator (irgacure2959) to prepare a sesbania gum hydrogel; then the obtained sesbania gum hydrogel or is fully mixed with bone marrow mesenchymal stem cells, extrusion printing is carried out through a 3D-Bioplotter system, fiber filaments are sprayed to be stacked layer by layer, a sesbania gum hydrogel scaffold is formed, and the scaffold after printing is cross-linked through ultraviolet light 365 nm, 200 mW / cm2, so that a 3D printing sesbania gum scaffold loaded with mesenchymal stem cells is obtained.
[0046] Embodiment 5
[0047] The embodiment discloses a 3D printing ultraviolet cross-linking sesbania gum hydrogel scaffold and a preparation method and application thereof, as shown in Figure 3 The results of the mechanical property test of the application show that the Young's modulus of the modified sesbania gum is higher than that of the unmodified sesbania gum, and the addition of methacrylic anhydride can affect the rigidity of the material.
[0048] Embodiment 6
[0049] The embodiment discloses a 3D printing ultraviolet cross-linking sesbania gum hydrogel scaffold and a preparation method and application thereof, as shown in Figure 4As shown, the cell activity result diagram of the modified sesbania gum composite chondrocyte after 7 days, 14 days and 21 days of culture, shows that most of the cells are dyed green (live cells) and the survival rate of the cells after printing is greater than 85%, indicating that the material is suitable for bioprinting and has good biological activity. The application potential in tissue repair is increased.
[0050] The above only describes the preferred embodiments of the present application and does not limit the protection scope of the present application. For those skilled in the art, the present application can have various changes and variations. Any changes, modifications, replacements, integrations and parameter changes of these embodiments within the spirit and principles of the present application, which can realize the same functions without departing from the principles and spirit of the present application, fall within the protection scope of the present application.
Claims
1. A 3D printed UV-crosslinked sesbania glue hydrogel scaffold and its preparation method and application, characterized in that: The preparation method comprises the following steps: Step S1, the sesbania gum raw material is dissolved in ultrapure water at a concentration of 1% w / v, and is dissolved at 60 DEG C under sufficient stirring to obtain a sesbania gum aqueous solution; Step S2, slowly add methacrylic anhydride at a speed of 500 uL / min at 60 DEG C, slowly add methacrylic anhydride under 3-5 DEG C ice bath, and continue stirring at 50 DEG C for 1 hour; Step S3, drop sodium hydroxide to maintain the pH of the reaction system at 7.0, continue the reaction for 6h, and obtain a methacrylated sesbania gum derivative with different grafting degrees according to the amount of methacrylic anhydride added; the obtained product is dialyzed by a 3500-4000 kd dialysis bag, and after freeze-drying, a porous sponge-like modified sesbania gum is obtained; Step S4, completely dissolve in PBS containing a concentration of 0.8-0.12% w / v of a photoinitiator (irgacure2959) to prepare a sesbania gum hydrogel; Step S5, the obtained sesbania gum hydrogel is mixed with bone marrow mesenchymal stem cells, and is extruded and printed by a 3D Bioprint system; the fiber filaments are sprayed layer by layer to form a sesbania gum hydrogel scaffold, and the printed scaffold is crosslinked by ultraviolet light 365nm, 200mW / cm2 to obtain a 3D printed sesbania gum scaffold loaded with mesenchymal stem cells.
2. The 3D printed UV-crosslinked amorphous hydrogel scaffold of Sesbania gum according to claim 1, characterized in that, When the sesbania gum is modified, the concentration of the sesbania gum aqueous solution is 0.8-0.12% w / v.
3. The 3D printed UV-crosslinked amorphous hydrogel scaffold of Sesbania gum according to claim 1, wherein, The modifier is methacrylic anhydride.
4. The 3D printed UV-crosslinked amorphous hydrogel scaffold of Sesbania gum according to claim 3, characterized in that, The concentration of the methacrylic anhydride is 1mL / g.
5. The 3D printed UV-crosslinked amorphous hydrogel scaffold of Sesbania gum according to claim 1, wherein, The concentration of the sesbania gum hydrogel for printing is 3.5-4.5%.
6. The 3D printed UV-crosslinked amorphous hydrogel scaffold of Sesbania gum according to claim 1, wherein, The time for ultraviolet light crosslinking of the sesbania gum hydrogel scaffold is 55-60s.
7. The 3D printed UV-crosslinked amorphous hydrogel of Sesbania gum scaffold according to claim 1, wherein, The bone marrow mesenchymal cell solution concentration is 1 x 10 7 cells / ml.
8. A 3D printed UV light crosslinked amorphous hydrogel of the quince seed gum and a method of preparation and use thereof, characterized in that, Prepared according to the method of any one of claims 1-7.
9. The 3D printed ultraviolet crosslinked sesbania gum hydrogel scaffold, the preparation method and the application according to claim 8, wherein the application is in the preparation of human tissue engineering scaffolds.