Three-dimensional printing antibacterial hydrogel material and preparation method and application thereof

Abstract

The application discloses a kind of three-dimensional printing antibacterial hydrogel materials and preparation method and application thereof, which are made of the following components by weight fraction: carboxymethyl chitosan 10-20 parts, polylysine 1-10 parts, polyglycine 1-10 parts, gelatin 10-20 parts, water 20-40 parts.The application includes the preparation method of three-dimensional printing antibacterial hydrogel materials.The hydrogel material has the advantages of good biocompatibility, little damage to cells, antibacterial effect, stable performance of wound dressing obtained by three-dimensional printing, and can be used for skin, ligament and other soft tissue wound repair and other clinical work.

Description

Technical Field

[0001] This invention relates to a 3D-printed antibacterial hydrogel material, its preparation method, and its applications. Specifically, it relates to a 3D-printed antibacterial hydrogel material, its preparation method, and its applications. Background Technology

[0002] 3D printing, also known as additive manufacturing, is a method for creating three-dimensional entities using computer-aided design and control of the manufacturing process. Initially applied in the industrial sector, 3D printing has since found use in the biomedical field, known as 3D bioprinting. Hydrogel materials are widely used in 3D bioprinting due to their unique three-dimensional network structure, which can mimic human tissue, and their ability to meet biological morphological requirements and ease of processing. However, the main challenges for using hydrogels in tissue engineering lie in their poor mechanical properties, uncontrollable swelling, and inability to customize macroscopic shapes and structures. These shortcomings severely limit the practical applications of hydrogels.

[0003] Emerging printing technologies can efficiently prepare hydrogel tissue engineering materials with complex structures, solving the problem of poor processability of hydrogels and broadening their applications in the biomedical field. Currently, they are mainly used for tissue repair and tissue culture. However, existing hydrogel materials also have many shortcomings: synthetic materials with high mechanical strength have poor biocompatibility, while some naturally derived biomaterials cannot meet the mechanical performance requirements for printing and application.

[0004] The hydrogel involved in this patent is formed by a cross-linking method involving positive and negative charge interactions to create a hydrogel network. Multiple amino acids are introduced into the carboxymethyl chitosan molecule. Due to the use of natural polymer raw materials, it exhibits excellent biocompatibility. Furthermore, the flexibility of carboxymethyl chitosan in chemical modification allows for the creation of a three-dimensional printing antibacterial hydrogel material with excellent biocompatibility and strong mechanical properties.

[0005] In conclusion, the use of suitable hydrogel components for tissue repair is crucial for functionality, and finding suitable components is a key step in the preparation of hydrogels for 3D printing, which will become an important key point for the widespread use of hydrogels in the biomedical field in the future. Summary of the Invention

[0006] One object of the present invention is to provide a three-dimensional printing antibacterial hydrogel material.

[0007] Another object of the present invention is to provide the application of the above-mentioned three-dimensional printed antibacterial hydrogel.

[0008] The method for preparing a three-dimensional printed antibacterial hydrogel material provided by the present invention includes: crosslinking carboxymethyl chitosan with polyglycine and polylysine to obtain polyamino acid modified carboxymethyl chitosan; mixing polyamino acid modified carboxymethyl chitosan, gelatin and water to obtain an antibacterial hydrogel material.

[0009] Furthermore, the polyamino acid-modified carboxymethyl chitosan is obtained by grafting the carboxyl groups of carboxymethyl chitosan with the amino groups of polyglycine and polylysine.

[0010] Furthermore, the molecular weight of the carboxymethyl chitosan is 2 × 10⁻⁶. 4 ~1×10 5 The molecular weight of polylysine is between 3000 and 4000 Da, and that of polyglycine is between 3000 and 5000 Da.

[0011] The present invention also provides an application of three-dimensional printed antibacterial hydrogel, characterized in that the antibacterial hydrogel material is three-dimensionally printed into a hydrogel dressing for tissue repair.

[0012] The specific steps are as follows: Antibacterial hydrogel material is loaded into a 3D printing cylinder. Using a 3D bioprinter, the 3D printing parameters are adjusted, and printing begins. After printing, cross-linking is performed using a sodium β-glycerophosphate solution. The sodium β-glycerophosphate solution is then removed to obtain the hydrogel dressing.

[0013] Furthermore, the 3D printing parameters are as follows: barrel temperature of 20-25℃, needle inner diameter of 0.20-0.60mm, printing speed of 1-10mm / s, XY axis spacing of 0.5-5mm, Z-axis step height of 0.2-0.6mm, and extrusion air pressure of 1-6Bar.

[0014] Furthermore, the concentration of the β-glycerophosphate sodium is 1–50 w / v.

[0015] Beneficial effects: This invention uses a polyamino acid-modified carboxymethyl chitosan-gelatin system as a 3D printing material, effectively avoiding the use of other chemical crosslinking agents. It has good biocompatibility, gelling properties, antibacterial properties, and processability. The hydrogel can maintain stability for a long time, meet the conditions for cell proliferation and differentiation, and has excellent cell compatibility and antibacterial properties. The antibacterial hydrogel obtained through the above technical solution can be used for tissue construction and repair. Attached Figure Description

[0016] Figure 1 This is a photograph of the antibacterial hydrogel prepared in Example 1.

[0017] Figure 2-3 The image shown is a 3D printed image of the antibacterial hydrogel prepared in Example 1.

[0018] Figure 4The image shows a SEM image of the antibacterial hydrogel prepared in Example 1.

[0019] Figure 5 The stress-strain curves are for the antibacterial hydrogels prepared in Examples 1, 2, and 3. Figure 5 In the figure, the horizontal axis represents strain (%) and the vertical axis represents stress (Pa); curves A, B, and C represent the antibacterial hydrogels prepared in Examples 1, 2, and 3, respectively.

[0020] Figure 6 The swelling properties of the antibacterial hydrogels prepared in Examples 1, 2, and 3 are shown in the graph. Figure 6 In the figure, the horizontal axis represents time (h), and the vertical axis represents cell survival rate (%); curves A, B, and C represent the antibacterial hydrogels prepared in Examples 1, 2, and 3, respectively.

[0021] Figure 7 Biocompatibility diagrams of mouse embryonic fibroblasts (MEF) cells at various concentrations of the antibacterial hydrogels prepared in Examples 1, 2, and 3. Figure 7 In the figure, the horizontal axis represents the concentration of hydrogel extract (%), and the vertical axis represents the cell viability (%). Columns A, B, and C represent the antibacterial hydrogels prepared in Examples 1, 2, and 3, respectively.

[0022] Figure 8 The image shows antibacterial test results for the antibacterial hydrogels prepared in Examples 1, 2, and 3. Antibacterial tests were performed against Staphylococcus aureus and Pseudomonas aeruginosa, respectively. Detailed Implementation

[0023] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive, and should not be construed as limiting the scope of protection of the present invention.

[0024] On one hand, the present invention provides a three-dimensional printing antibacterial hydrogel material, which is made from the following raw materials in parts by weight: 10-20 parts carboxymethyl chitosan, 1-10 parts polylysine, 1-10 parts polyglycine, 10-20 parts gelatin, and 20-40 parts water. Preferably, the three-dimensional printing antibacterial hydrogel material is made from the following components in parts by weight: 10-15 parts carboxymethyl chitosan, 1-5 parts polylysine, 1-5 parts polyglycine, 10-15 parts gelatin, and 20-40 parts water.

[0025] On the other hand, the present invention provides an application of a three-dimensional printed antibacterial hydrogel material in wound dressings, wherein the concentration of the sodium glycerophosphate solution used in the preparation of the wound dressing is between 1 and 50 w / v%.

[0026] The present invention will be further described in detail below with reference to specific embodiments and accompanying drawings, but the present invention is not limited thereto.

[0027] Example 1

[0028] (1) At room temperature, 10 parts of carboxymethyl chitosan were added to water and stirred until fully dissolved. 1 part of polyglycine and 1 part of polylysine were added to water and stirred until fully dissolved. The mixtures were then activated with NHS EDC for 0–30 min and added dropwise to the carboxymethyl chitosan aqueous solution. The polyamino acids were added dropwise over 30 min. The resulting mixed solution was placed in a 25°C water bath for 8 hours. The reaction solution was then dialyzed for 2 days. After freeze-drying, this material was mixed with 10 parts of gelatin to obtain the antibacterial hydrogel material.

[0029] (2) The printing parameters were adjusted using a bioprinter as follows: barrel temperature 25℃, needle inner diameter 0.20mm, printing speed 10mm / s, XY axis spacing 1mm, Z axis step height 0.2mm, and extrusion pressure 2Bar. Finally, a preliminary hydrogel dressing was obtained.

[0030] (3) Crosslink the preliminary hydrogel dressing obtained in step (2) in a sodium glycerophosphate solution (1 w / v%) to obtain the hydrogel wound dressing.

[0031] Example 2

[0032] (1) At room temperature, 12 parts of carboxymethyl chitosan were added to water and stirred until fully dissolved. 2 parts of polyglycine and 2 parts of polylysine were added to water and stirred until fully dissolved. The polyamino acids were activated with NHS EDC for 0–30 min and then added dropwise to the carboxymethyl chitosan aqueous solution. The polyamino acids were added dropwise over 30 min. The resulting mixed solution was placed in a 25°C water bath for 8 hours. The reaction solution was then dialyzed for 2 days. After freeze-drying, this material was mixed with 12 parts of gelatin to obtain the antibacterial hydrogel material.

[0033] (2) The printing parameters were adjusted using a bioprinter as follows: barrel temperature 25℃, needle inner diameter 0.20mm, printing speed 10mm / s, XY axis spacing 1mm, Z axis step height 0.2mm, and extrusion pressure 2Bar. Finally, a preliminary hydrogel dressing was obtained.

[0034] (3) Crosslink the preliminary hydrogel dressing obtained in step (2) in a sodium glycerophosphate solution (10 w / v%) to obtain the hydrogel wound dressing.

[0035] Example 3

[0036] (1) At room temperature, 15 parts of carboxymethyl chitosan were added to water and stirred until fully dissolved. 5 parts of polyglycine and 5 parts of polylysine were added to water and stirred until fully dissolved. The solutions were then activated with NHS EDC for 0–30 min and added dropwise to the carboxymethyl chitosan aqueous solution. The polyamino acids were added dropwise over 30 min. The resulting mixed solution was placed in a 25°C water bath for 8 hours. The reaction solution was then dialyzed for 2 days. After freeze-drying, this material was mixed with 15 parts of gelatin to obtain the antibacterial hydrogel material.

[0037] (2) The printing parameters were adjusted using a bioprinter as follows: barrel temperature 25℃, needle inner diameter 0.20mm, printing speed 10mm / s, XY axis spacing 1mm, Z axis step height 0.2mm, and extrusion pressure 2Bar. Finally, a preliminary hydrogel dressing was obtained.

[0038] (3) Crosslink the preliminary hydrogel dressing obtained in step (2) in a sodium glycerophosphate solution (30 w / v%) to obtain the hydrogel wound dressing.

[0039] The results of the measurement of the antibacterial hydrogel prepared in Example 1 are as follows: Figure 1 As shown in the figure. The result of the 3D printing of the antibacterial hydrogel prepared in Example 1 is shown in the figure. Figure 2-3 As shown in the figure. The SEM image results of the antibacterial hydrogel prepared in Example 1 are as follows. Figure 4 As shown. The mechanical test results of the antibacterial hydrogels prepared in Examples 1-3 are as follows. Figure 5 ,Depend on Figure 5 It can be seen that the maximum compressive fracture stress is 41.76 kPa, and the maximum compressive modulus is 0.004 MPa. The swelling performance test results of the antibacterial hydrogels prepared in Examples 1-3 are as follows: Figure 6 ,Depend on Figure 6 It can be seen that the antibacterial hydrogel has excellent water absorption and moisturizing properties. The biocompatibility results of the antibacterial hydrogels prepared in Examples 1-3 are as follows: Figure 7 As shown, by Figure 7 It can be seen that the antibacterial hydrogels prepared in Examples 1-3 have good biocompatibility and are harmless to cells and tissues. The antibacterial test results of the antibacterial hydrogels prepared in Examples 1-3 are as follows: Figure 8 , Figure 8 This indicates that the antibacterial hydrogel has good antibacterial properties.

[0040] The above description is merely a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A method for preparing a three-dimensional printed antibacterial hydrogel material, characterized in that, The steps are as follows: (1) 10-15 parts of carboxymethyl chitosan is added into water and stirred to be fully dissolved at room temperature, 1-5 parts of polyglycine and 1-5 parts of polylysine are respectively added into water and stirred to be fully dissolved, and then activated by using NHS EDC for 0-30 min, and then added dropwise into the carboxymethyl chitosan aqueous solution, the polyamino acid is added dropwise for 30 min, the obtained mixed solution is placed in a 25℃ water bath for 8 hours, and then the reaction solution is dialyzed for 2 days, and then freeze-dried, and then the material is mixed with 10-15 parts of gelatin to obtain an antibacterial hydrogel material; (2) the printing parameters of a biological printer are adjusted as follows: barrel temperature is 25℃, inner diameter of needle is 0.20 mm, printing speed is 10 mm / s, XY axis distance is 1 mm, Z axis step height is 0.2 mm, and extrusion air pressure is 2 Bar, and finally a preliminarily shaped hydrogel dressing is obtained; (3) the preliminarily shaped hydrogel dressing obtained in step (2) is crosslinked in a glycerol sodium phosphate solution to obtain a hydrogel wound dressing.

2. The method for preparing the three-dimensional printed antibacterial hydrogel material according to claim 1, characterized in that, The concentration of the glycerol sodium phosphate solution is 1-30 w / v%.

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