CN116036358A - A kind of three-dimensional printing antibacterial hydrogel material and its preparation method and application - Google Patents

Abstract

The invention discloses a three-dimensional printing antibacterial hydrogel material, a preparation method and application thereof, wherein the three-dimensional printing antibacterial hydrogel material is prepared from the following components in parts by weight: 10-20 parts of carboxymethyl chitosan, 1-10 parts of polylysine, 1-10 parts of poly glycine, 10-20 parts of gelatin and 20-40 parts of water. The invention comprises a preparation method of the three-dimensional printing antibacterial hydrogel material. The hydrogel material has the advantages of good biocompatibility, small damage to cells, antibacterial effect, and stable performance of the wound dressing obtained by three-dimensional printing, and can be used for clinical work such as wound repair of soft tissues such as skin, ligaments and the like.

Description

A kind of three-dimensional printing antibacterial hydrogel material and its preparation method and application

technical field

The invention relates to a three-dimensional printing antibacterial hydrogel material and its preparation method and application. The hydrogel material and three-dimensional printing technology, specifically, relate to a three-dimensional printing antibacterial hydrogel material and its preparation method and application.

Background technique

Three-dimensional printing, also known as additive manufacturing, is a method of preparing three-dimensional entities by using computer design to control and implement the manufacturing process. The first application field of 3D printing is the industrial field, and then it is used in the biomedical field, which is called 3D bioprinting technology. Hydrogel materials are widely used in 3D bioprinting because of their special three-dimensional network structure that can simulate human tissue, and have the advantages of meeting the morphological requirements of organisms and easy processing. However, the main challenges of hydrogels for tissue engineering are poor mechanical properties, uncontrollable swelling, and inability to customize macroscopic shape and structure. These deficiencies severely limit the practical applications of hydrogels.

The emerging printing technology can efficiently prepare hydrogel tissue engineering materials with complex structures, solve the problem of poor processability of hydrogels, and broaden the application of hydrogels in the field of biomedicine. Currently, they are mainly used for tissue repair, tissue training etc. However, the existing hydrogel materials also have many shortcomings: synthetic materials with high mechanical strength have poor biocompatibility, and some biomaterials from natural sources cannot meet the requirements of printing and mechanical properties during application. .

The hydrogel involved in this patent adopts the cross-linking method of positive and negative charge interactions to form a hydrogel network, and introduces various amino acids on the basis of carboxymethyl chitosan molecules. Due to the use of natural polymer raw materials, its biocompatibility is excellent, and the chemical modification of carboxymethyl chitosan is used to create a 3D printing antibacterial hydrogel material with excellent biocompatibility and strong mechanical properties. .

In summary, suitable hydrogel components for tissue repair are crucial to functionality, and finding suitable components is a key link in the preparation of hydrogels for 3D printing, which will become the basis for the widespread use of hydrogels in the biomedical field in the future. important key points.

Contents of the invention

An object of the present invention is to provide a three-dimensional printing antibacterial hydrogel material.

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

The preparation method of the three-dimensional printing antibacterial hydrogel material provided by the present invention comprises: carboxymethyl chitosan is cross-linked with polyglycine and polylysine to obtain polyamino acid modified carboxymethyl chitosan, and the polyamino acid is modified The antibacterial hydrogel material is obtained by mixing permanent carboxymethyl chitosan, gelatin and water.

Further, the polyamino acid modified carboxymethyl chitosan is obtained by grafting carboxyl groups of carboxymethyl chitosan with amino groups of polyglycine and polylysine.

Furthermore, the molecular weight of the carboxymethyl chitosan is 2×10 ⁴ to 1×10 ⁵ Da, the molecular weight of the polylysine is 3000-4000 Da, and the molecular weight of the polyglycine is 3000-5000 Da.

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

The specific steps are: take the antibacterial hydrogel material and load it in the three-dimensional printing material cylinder, use the three-dimensional bioprinter, adjust the three-dimensional printing parameters, and start printing. After the printing is completed, the sodium β-glycerophosphate solution is used for cross-linking, and the sodium β-glycerophosphate solution is removed to obtain a hydrogel dressing.

Further, the three-dimensional printing parameters are that the temperature of the barrel is 20-25°C, the inner diameter of the needle is 0.20-0.60mm, the printing speed is 1-10mm/s, the distance between the XY axes is 0.5-5mm, and the step height of the Z-axis is 0.2 ~ 0.6mm extrusion air pressure is 1 ~ 6Bar.

Further, the concentration of the sodium β-glycerophosphate is 1-50w/v%.

Beneficial effects: the present invention uses polyamino acid modified carboxymethyl chitosan-gelatin system as a three-dimensional printing material, effectively avoids the use of other chemical cross-linking agents, and has good biocompatibility, gelation, antibacterial properties and processing Type, the hydrogel can maintain stability for a long period of time, meet the conditions for cell proliferation and differentiation, and has excellent cytocompatibility and antibacterial properties. The antibacterial hydrogel obtained through the above technical scheme can be used for tissue construction and repair.

Description of drawings

Fig. 1 is the physical figure of the antibacterial hydrogel prepared in embodiment 1.

Fig. 2-3 is the three-dimensional printed physical map of the antibacterial hydrogel prepared in Example 1.

Fig. 4 is the SEM image of the antibacterial hydrogel prepared in Example 1.

Fig. 5 is the stress-strain curve of the antibacterial hydrogel prepared in Example 1, Example 2 and Example 3. In Fig. 5, the abscissa is the strain Strain (%), and the ordinate is the stress Stress (Pa); Curves A, B, and C represent the antibacterial hydrogels prepared in Example 1, Example 2, and Example 3, respectively.

Fig. 6 is a diagram of swelling properties of antibacterial hydrogels prepared in Example 1, Example 2 and Example 3. In Fig. 6, the abscissa is the time Time (h), and the ordinate is the cell survival rate Swelling Ratio (%); Curves A, B, and C respectively represent the antibacterial hydrogels prepared in Example 1, Example 2, and Example 3 .

Fig. 7 is a biocompatibility diagram of mouse embryonic fibroblast MEF cells with various concentrations of antibacterial hydrogel leachate prepared in Example 1, Example 2 and Example 3. In Fig. 7, the abscissa is the hydrogel extract concentration HydrogelExtract Concentration (%), and the ordinate is the cell viability Cell Viability (%); Columns A, B, and C represent Example 1, Example 2, and Example 3, respectively Prepared antibacterial hydrogels.

Fig. 8 is an antibacterial test diagram containing antibacterial hydrogels prepared in Example 1, Example 2 and Example 3. Antibacterial tests against Staphylococcus aureus and Pseudomonas aeruginosa were performed separately.

Detailed ways

In order to illustrate the present invention more clearly, the present invention will be further described below in combination with preferred embodiments. Those skilled in the art should understand that the content specifically described below is illustrative rather than restrictive, and should not limit the protection scope of the present invention.

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

On the other hand, the present invention provides an application of a three-dimensionally printed antibacterial hydrogel material on a wound dressing, wherein the concentration of the sodium glycerophosphate solution for preparation of the wound dressing is between 1 and 50 w/v%.

The present invention will be described in further detail below in conjunction with specific embodiments and drawings, but the present invention is not limited thereto.

Example 1

(1) At room temperature, add 10 parts of carboxymethyl chitosan to water and stir to fully dissolve, add 1 part of polyglycine and 1 part of polylysine to water and stir to fully dissolve, respectively use NHS EDC to activate for 0 to 30 minutes , was added dropwise into carboxymethyl chitosan aqueous solution, and the polyamino acid was added dropwise in 30 minutes. The resulting mixed solution was placed in a water bath at 25°C for 8 hours, and then the reaction solution was dialyzed for 2 days. After freeze-drying, the material was mixed with 10 parts Gelatin mixed to obtain antibacterial hydrogel material

(2) Using the bioprinter to adjust the printing parameters, the temperature of the barrel is 25°C, the inner diameter of the needle is 0.20mm, the printing speed is 10mm/s, the distance between the XY axes is 1mm, and the step height of the Z axis is 0.2mm. For 2Bar. Finally, a preliminary shaped hydrogel dressing is obtained.

(3) cross-linking the preliminarily shaped hydrogel dressing obtained in step (2) in sodium glycerophosphate solution (1w/v%) to obtain the hydrogel wound dressing.

Example 2

(1) At room temperature, add 12 parts of carboxymethyl chitosan to water and stir to fully dissolve, add 2 parts of polyglycine and 2 parts of polylysine to water and stir to fully dissolve, respectively use NHS EDC to activate for 0 to 30 minutes , added dropwise into carboxymethyl chitosan aqueous solution, the polyamino acid was added dropwise in 30 minutes, the resulting mixed solution was placed in a water bath at 25°C for 8 hours, and then the reaction solution was dialyzed for 2 days, and after freeze-drying, this material was mixed with 12 parts Gelatin mixed to obtain antibacterial hydrogel material

(2) Using the bioprinter to adjust the printing parameters, the temperature of the barrel is 25°C, the inner diameter of the needle is 0.20mm, the printing speed is 10mm/s, the distance between the XY axes is 1mm, and the step height of the Z axis is 0.2mm. For 2Bar. Finally, a preliminary shaped hydrogel dressing is obtained.

(3) cross-linking the preliminarily shaped hydrogel dressing obtained in step (2) in sodium glycerophosphate solution (10w/v%) to obtain the hydrogel wound dressing.

Example 3

(1) At room temperature, add 15 parts of carboxymethyl chitosan to water and stir to fully dissolve, add 5 parts of polyglycine and 5 parts of polylysine to water and stir to fully dissolve, respectively use NHS EDC to activate for 0 to 30 minutes , added dropwise into carboxymethyl chitosan aqueous solution, the polyamino acid was added dropwise within 30 minutes, the resulting mixed solution was placed in a water bath at 25°C for 8 hours, and then the reaction solution was dialyzed for 2 days, and after freeze-drying, this material was mixed with 15 parts Gelatin mixed to obtain antibacterial hydrogel material

(2) Using the bioprinter to adjust the printing parameters, the temperature of the barrel is 25°C, the inner diameter of the needle is 0.20mm, the printing speed is 10mm/s, the distance between the XY axes is 1mm, and the step height of the Z axis is 0.2mm. For 2Bar. Finally, a preliminary shaped hydrogel dressing is obtained.

(3) cross-linking the preliminarily shaped hydrogel dressing obtained in step (2) in sodium glycerophosphate solution (30w/v%) to obtain the hydrogel wound dressing.

After determination, the results of the physical picture of the antibacterial hydrogel prepared in Example 1 are shown in Figure 1. The results of the three-dimensional printing of the antibacterial hydrogel prepared in Example 1 are shown in Figures 2-3. The results of the SEM image of the antibacterial hydrogel prepared in Example 1 are shown in Figure 4. The mechanical test results of the antibacterial hydrogel prepared in Examples 1-3 are shown in Figure 5, from which it can be seen that the maximum compressive fracture stress is 41.76KPa, and the maximum compressive modulus is 0.004MPa. The swelling performance test results of the antibacterial hydrogels prepared in Examples 1-3 are shown in Figure 6, and it can be seen from Figure 6 that the antibacterial hydrogels have excellent water absorption and moisture retention. The biocompatibility results of the antibacterial hydrogels prepared in Examples 1-3 are shown in Figure 7. From Figure 7, it can be known 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 hydrogel prepared in Examples 1-3 are shown in Figure 8, which shows that the antibacterial hydrogel has good antibacterial performance.

The above is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto, and any person familiar with the technical field can easily think of the transformation or replacement within the technical scope disclosed in the present invention, All should be covered within the protection scope of the present invention. Therefore, the protection scope of the present invention should be determined by the protection scope of the claims.

Claims (7)

1. The preparation method of the three-dimensional printing antibacterial hydrogel material is characterized by comprising the following components in parts by weight: 10-20 parts of carboxymethyl chitosan, 1-10 parts of polylysine, 1-10 parts of poly glycine, 10-20 parts of gelatin and 20-40 parts of water. Wherein the molecular weight of the carboxymethyl chitosan is 2 multiplied by 104 to 1 multiplied by 105Da, the molecular weight of the polylysine is 3000 to 4000Da, and the molecular weight of the polyglycine is 3000 to 5000Da.

The preparation method comprises the following steps:

crosslinking the carboxymethyl chitosan with the poly glycine and the polylysine to obtain polyamino acid modified carboxymethyl chitosan;

and mixing the polyamino acid modified carboxymethyl chitosan, gelatin and water to obtain the three-dimensional printing antibacterial hydrogel material.

2. The method for preparing a three-dimensional printed antibacterial hydrogel material according to claim 1, comprising the steps of:

crosslinking the carboxymethyl chitosan with the poly glycine and the polylysine to obtain polyamino acid modified carboxymethyl chitosan;

and mixing the polyamino acid modified carboxymethyl chitosan, gelatin and water to obtain the three-dimensional printing antibacterial hydrogel material.

3. Use of the three-dimensional printing antibacterial hydrogel material according to claim 1 in three-dimensional printing.

4. The use according to claim 4, characterized in that said method of three-dimensional printing comprises the steps of:

loading the three-dimensional printing antibacterial hydrogel material into a printing cylinder, and placing the printing cylinder into a water tank of a printer;

and taking out the printing cylinder from the water tank, placing the printing cylinder in a printer, adjusting parameters of the printer, starting printing, performing cross-linking by using the beta-sodium glycerophosphate solution after printing, and removing the beta-sodium glycerophosphate solution to obtain the hydrogel dressing.

5. The method according to claim 4, wherein the printing parameters comprise a cylinder temperature of 20-25 ℃, a needle inside diameter of 0.20-0.60 mm, a printing speed of 1-10 mm/s, an XY axis spacing of 0.5-5 mm, a Z axis step height of 0.2-0.6 mm, and an extrusion air pressure of 1-6 Bar.

6. The use according to claim 4, wherein the printer tank is maintained at a temperature of 20 ℃ to 25 ℃ for a period of 30 to 60 minutes.

7. The use according to claim 4, wherein the concentration of sodium beta-glycerophosphate is 1-50 w/v%.

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