Streamlined nano zinc oxide reinforced gelma hydrogel and preparation method thereof

Abstract

The application discloses streamline nano zinc oxide reinforced GelMA hydrogel and a preparation method thereof, and the preparation method is as follows: 1) synthesis of streamline nano zinc oxide, streamline nano zinc oxide is abbreviated as str-ZnO NPs; 2) synthesis of methacrylated gelatin, methacrylated gelatin is abbreviated as GelMA; 3) preparation of GelMA aqueous solution, addition of a photoinitiator and str-ZnO NPs, stirring until uniform; irradiation with ultraviolet light, and the streamline nano zinc oxide is obtained; after zinc oxide is formed into streamline nano zinc oxide, the size and morphology of the zinc oxide change, the surface area of the nano zinc oxide is increased, and therefore the mechanical property of the hydrogel is improved. The hydrogel disclosed by the application has a storage modulus which is 1.8 times that of the GelMA hydrogel, has good biocompatibility, and has the characteristics of injectability and degradability; and can meet the requirements of the biomedical field such as cell culture and tissue engineering on the performance of the hydrogel.

Description

Technical Field

[0001] This invention belongs to the field of biomaterials technology, specifically relating to a streamlined nano-zinc oxide reinforced GelMA hydrogel and its preparation method. Background Technology

[0002] Hydrogels are polymer systems with a three-dimensional network structure that can absorb large amounts of water and have both solid and liquid properties. They have a wide range of applications in drug delivery, wound dressings, tissue engineering, and hygiene products, and are a rapidly developing polymer material.

[0003] Gelatin is a natural material, a hydrolysis product of collagen. By modifying gelatin with methacrylic anhydride, methacrylamide gelatin (GelMA) hydrogels with photocrosslinking properties can be obtained. GelMA hydrogels combine the advantages of synthetic and natural hydrogels, exhibiting excellent biocompatibility and adjustable mechanical strength.

[0004] However, when GelMA hydrogels are used in the biomedical field, they need to meet certain mechanical properties. Traditional GelMA hydrogels cannot withstand large strains or stresses, which greatly limits their applications. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide a streamlined nano-zinc oxide reinforced GelMA hydrogel.

[0006] The second objective of this invention is to provide a method for preparing streamlined nano-zinc oxide reinforced GelMA hydrogel.

[0007] The technical solution of this invention is summarized as follows:

[0008] A method for preparing streamlined nano-zinc oxide reinforced GelMA hydrogel includes the following steps:

[0009] 1) Synthesis of streamlined nano-zinc oxide:

[0010] According to the proportion, 1.83g of anhydrous zinc acetate was dissolved in 100ml of dibenzyl ether, and 10.7g of benzylamine was added. The mixture was reacted at 210-220℃ for 5h, cooled to room temperature, centrifuged, and washed to obtain streamlined nano zinc oxide, which is abbreviated as str-ZnO NPs.

[0011] 2) Synthesis of methacrylamide gelatin:

[0012] Dissolve 10g of gelatin in distilled water according to the proportion, add 5mL of methacrylic anhydride, react at 40-60℃ for 1-3h, dialyze with distilled water, and freeze dry at -80℃ to obtain methacrylamide gelatin, which is abbreviated as GelMA.

[0013] 3) Synthesis of streamlined nano-zinc oxide-reinforced GelMA hydrogels:

[0014] A 10% (w / w) aqueous solution of GelMA was prepared, and 0.3%-0.5% (w / w) of lithium phenyl (2,4,6-trimethylbenzoyl)phosphate photoinitiator was added. 0.5% (w / w) of str-ZnO NPs was added, and the mixture was stirred until homogeneous under water bath heating at 37-40℃. After irradiation with ultraviolet light, a streamlined nano-zinc oxide-reinforced GelMA hydrogel was obtained.

[0015] The distilled water dialysis is performed at 40-50℃ with a molecular weight cutoff of 3500 for 4-7 days.

[0016] A streamlined nano-zinc oxide reinforced GelMA hydrogel was prepared by the above preparation method.

[0017] The beneficial effects of this invention are as follows:

[0018] After zinc oxide forms streamlined nano-zinc oxide, the changes in size and morphology lead to an increase in the surface area of ​​the nano-zinc oxide, thereby improving the mechanical properties of the hydrogel. The streamlined nano-zinc oxide-reinforced GelMA hydrogel prepared in this invention exhibits a storage modulus 1.8 times that of the original GelMA hydrogel.

[0019] The preparation method of this invention is simple and the raw materials are readily available. The resulting streamlined nano-zinc oxide reinforced GelMA hydrogel is a light white and transparent gel-like substance with excellent biocompatibility, as well as injectability and degradability. The preparation method of this invention can adjust the dynamic properties of the hydrogel according to the needs of cells or tissues, which can meet the requirements of hydrogel performance in biomedical fields such as cell culture and tissue engineering, and has broad application prospects. Attached Figure Description

[0020] Figure 1 This is a transmission electron microscope image of the streamlined nano-zinc oxide prepared in Example 1.

[0021] Figure 2 Appearance images of GelMA hydrogel and GelMA / str-ZnO hydrogel prepared in Example 1;

[0022] Figure 3 The rheological properties of the GelMA / str-ZnO hydrogel prepared in Example 1 are shown in the figure.

[0023] Figure 4 The strain recovery diagram of the GelMA / str-ZnO hydrogel prepared in Example 1;

[0024] Figure 5 Viscosity (A) and injectability evaluation diagram (B) of the GelMA / str-ZnO hydrogel prepared in Example 1. Detailed Implementation

[0025] The present invention will be further described below with reference to the accompanying drawings and embodiments:

[0026] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0027] Example 1

[0028] A method for preparing streamlined nano-zinc oxide reinforced GelMA hydrogel includes the following steps:

[0029] 1) Synthesis of streamlined nano-zinc oxide:

[0030] 1.83 g of anhydrous zinc acetate was dissolved in 100 ml of dibenzyl ether, and 10.7 g of benzylamine was added. The mixture was reacted at 210 °C for 5 h, cooled to room temperature, centrifuged (8000 rpm, 10 min), and washed successively with acetone and ethanol to obtain streamlined nano-zinc oxide, which is abbreviated as str-ZnO NPs.

[0031] The prepared streamlined nano-zinc oxide was dropped onto a copper grid, dried, and its morphology was observed under a transmission electron microscope. (See...) Figure 1 .from Figure 1 As can be seen, the streamlined nano zinc oxide is a unique, dispersed streamlined structure with an average width and length of approximately 30 nanometers and 150 nanometers, respectively.

[0032] 2) Synthesis of methacrylamide gelatin:

[0033] Dissolve 10g of gelatin in 100mL of distilled water, add 5mL of methacrylic anhydride, react at 50℃ for 2h, dialyze with distilled water (40℃, molecular weight cutoff of 3500, dialyze for 5 days), freeze dry at -80℃ to obtain methacrylamide gelatin, which is abbreviated as GelMA.

[0034] 3) Synthesis of streamlined nano-zinc oxide-reinforced GelMA hydrogels:

[0035] A 10% (w / w) aqueous solution of GelMA was prepared, and 0.5% (w / w) of lithium phenyl (2,4,6-trimethylbenzoyl)phosphate photoinitiator was added. 0.5% (w / w) of str-ZnO NPs was added, and the mixture was stirred evenly under water bath heating at 37°C. The mixture was then irradiated with ultraviolet light (405 nm) to obtain a streamlined nano-zinc oxide-reinforced GelMA hydrogel (GelMA / str-ZnO hydrogel).

[0036] Example 2

[0037] A method for preparing streamlined nano-zinc oxide reinforced GelMA hydrogel includes the following steps:

[0038] 1) Synthesis of streamlined nano-zinc oxide:

[0039] 1.83 g of anhydrous zinc acetate was dissolved in 100 ml of dibenzyl ether, and 10.7 g of benzylamine was added. The mixture was reacted at 215 °C for 5 h, cooled to room temperature, centrifuged (8000 rpm, 10 min), and washed successively with acetone and ethanol to obtain streamlined nano-zinc oxide, which is abbreviated as str-ZnO NPs.

[0040] The streamlined nano-zinc oxide prepared in this step has a unique dispersed streamlined structure with an average width and length of approximately 30 nm and 150 nm, respectively.

[0041] 2) Synthesis of methacrylamide gelatin:

[0042] Dissolve 10g of gelatin in 100mL of distilled water, add 5mL of methacrylic anhydride, react at 40℃ for 3h, dialyze with distilled water (40℃, molecular weight cutoff of 3500, dialyze for 7 days), freeze dry at -80℃ to obtain methacrylamide gelatin, which is abbreviated as GelMA.

[0043] 3) Synthesis of streamlined nano-zinc oxide-reinforced GelMA hydrogels:

[0044] A 10% (w / w) aqueous solution of GelMA was prepared, and a photoinitiator (lithium phenyl (2,4,6-trimethylbenzoyl)phosphate) with a final mass content of 0.3% was added; str-ZnO NPs with a final mass content of 0.5% were added, and the mixture was stirred evenly under water bath heating at 37°C; and then irradiated with ultraviolet light (405 nm) to obtain a streamlined nano-zinc oxide-reinforced GelMA hydrogel (GelMA / str-ZnO hydrogel).

[0045] Example 3

[0046] A method for preparing streamlined nano-zinc oxide reinforced GelMA hydrogel includes the following steps:

[0047] 1) Synthesis of streamlined nano-zinc oxide:

[0048] 1.83 g of anhydrous zinc acetate was dissolved in 100 ml of dibenzyl ether, and 10.7 g of benzylamine was added. The mixture was reacted at 220 °C for 5 h, cooled to room temperature, centrifuged (8000 rpm, 10 min), and washed successively with acetone and ethanol to obtain streamlined nano-zinc oxide, which is abbreviated as str-ZnO NPs.

[0049] The streamlined nano-zinc oxide prepared in this step has a unique dispersed streamlined structure with an average width and length of approximately 30 nm and 150 nm, respectively.

[0050] 2) Synthesis of methacrylamide gelatin:

[0051] Dissolve 10g of gelatin in 100mL of distilled water, add 5mL of methacrylic anhydride, react at 60℃ for 1h, dialyze with distilled water (50℃, molecular weight cutoff of 3500, dialyze for 4 days), freeze dry at -80℃ to obtain methacrylamide gelatin, which is abbreviated as GelMA.

[0052] 3) Synthesis of streamlined nano-zinc oxide-reinforced GelMA hydrogels:

[0053] A 10% (w / w) aqueous solution of GelMA was prepared, and a photoinitiator (lithium phenyl (2,4,6-trimethylbenzoyl)phosphate) with a final mass content of 0.4% was added; str-ZnO NPs with a final mass content of 0.5% were added, and the mixture was stirred evenly under water bath heating at 40°C; and then irradiated with ultraviolet light (405 nm) to obtain a streamlined nano-zinc oxide reinforced GelMA hydrogel (GelMA / str-ZnO hydrogel).

[0054] Example 4

[0055] Preparation of GelMA hydrogel:

[0056] A 10% (w / w) aqueous solution of GelMA was prepared, and a photoinitiator (lithium phenyl (2,4,6-trimethylbenzoyl)phosphate) with a final mass content of 0.5% was added. The solution was heated in a water bath at 37°C and stirred until homogeneous. The solution was then irradiated with ultraviolet light (405 nm) to obtain a GelMA hydrogel.

[0057] Example 5

[0058] Images of the appearance of the GelMA hydrogel and the GelMA / str-ZnO hydrogel prepared in Example 1 are shown below. Figure 2 The result is from Figure 2As can be seen, the prepared GelMA / str-ZnO hydrogel is a light white, transparent gel-like substance. The introduction of streamlined nano-zinc oxide caused the gel to turn white, its transparency to decrease, and its color distribution to become more uniform, indicating that the streamlined nano-zinc oxide is distributed relatively evenly within the GelMA hydrogel.

[0059] Example 6

[0060] Using a rheometer (DHR-2, Waters) TM The dynamic viscoelasticity of the prepared GelMA hydrogel and the GelMA / str-ZnO hydrogel prepared in Example 1 was characterized by TA Instruments. The storage modulus (G') and loss modulus (G”) as a function of frequency were recorded. The test temperature was room temperature, the strain was 1%, and the frequency range was 0.01-10 Hz. (See [reference needed]) Figure 3 .

[0061] The result is from Figure 3 As can be seen, the storage modulus of the GelMA / str-ZnO hydrogel increases, indicating its enhanced resistance to external forces. The reasons for this are twofold: firstly, the streamlined zinc oxide nanoparticles possess rigidity and can form cross-linking bonds with collagen; secondly, the high aspect ratio of the streamlined zinc oxide nanoparticles increases the specific surface area, thereby increasing the contact area for interaction with the GelMA hydrogel, thus achieving the enhanced effect.

[0062] Example 7

[0063] GelMA / str-ZnO hydrogel was analyzed using a rheometer (DHR-2, Waters). TM The hydrogel was tested using TA Instruments. It was subjected to strain cycles at a frequency of 1 Hz, with high strain (200%, 1 min) and low strain (1%, 1 min) to obtain the storage modulus (G') and loss modulus (G”). See [link to TA Instruments]. Figure 4 .

[0064] The result is from Figure 4 As can be seen, even after being subjected to multiple large strains of 200%, which disrupts the hydrogel structure in the nonlinear region, the rheological viscoelastic behavior of the hydrogel remains unchanged when a small strain of 1% (i.e., in the linear region) is applied again. This observation indicates that the GelMA / str-ZnO hydrogel exhibits strong shear recovery.

[0065] Example 8

[0066] Using a rheometer (DHR-2, Waters) TM TA Instruments) at room temperature for 0.1-100s -1The shear rate range was used to record the shear stress of the hydrogel samples. The injectability of the GelMA / str-ZnO hydrogel prepared in Example 1 was evaluated by injecting it with a syringe at room temperature, see [reference needed]. Figure 5 .

[0067] The result is from Figure 5 As can be seen in (A), the GelMA / str-ZnO hydrogel prepared in Example 1 exhibits obvious shear-thinning properties. Figure 5 (B) It can be seen that at room temperature, the GelMA / str-ZnO hydrogel can be injected with a syringe and has good injectability.

[0068] Experiments showed that the GelMA / str-ZnO hydrogels prepared in Examples 2 and 3 had similar appearance, rheological properties, strain recovery, viscosity (A), and injectability (B) to the GelMA / str-ZnO hydrogel prepared in Example 1.

[0069] It should be understood that the specific embodiments described above are merely illustrative or explanatory of the principles of the present invention and do not constitute a limitation thereof. Therefore, any modifications, equivalent substitutions, improvements, etc., made without departing from the spirit and scope of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing streamlined nano-zinc oxide reinforced GelMA hydrogel, characterized in that... Includes the following steps: 1) Synthesis of streamlined nano-zinc oxide: According to the proportion, 1.83g of anhydrous zinc acetate was dissolved in 100ml of dibenzyl ether, and 10.7g of benzylamine was added. The mixture was reacted at 210-220℃ for 5h, cooled to room temperature, centrifuged, and washed to obtain streamlined nano zinc oxide, which is abbreviated as str-ZnO NPs. 2) Synthesis of methacrylamide gelatin: Dissolve 10g of gelatin in distilled water according to the proportion, add 5mL of methacrylic anhydride, react at 40-60℃ for 1-3h, dialyze with distilled water, and freeze dry at -80℃ to obtain methacrylamide gelatin, which is abbreviated as GelMA. 3) Synthesis of streamlined nano-zinc oxide-reinforced GelMA hydrogels: A 10% (w / w) aqueous solution of GelMA was prepared, and 0.3%-0.5% (w / w) of lithium phenyl (2,4,6-trimethylbenzoyl)phosphate photoinitiator was added. 0.5% (w / w) of str-ZnO NPs was added, and the mixture was stirred until homogeneous under water bath heating at 37-40℃. After irradiation with ultraviolet light, a streamlined nano-zinc oxide-reinforced GelMA hydrogel was obtained.

2. The preparation method according to claim 1, characterized in that: The distilled water dialysis is performed at 40-50℃ with a molecular weight cutoff of 3500 for 4-7 days.

3. A streamlined nano-zinc oxide reinforced GelMA hydrogel prepared by the preparation method of claim 1 or 2.

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