阅读说明：本技术 一种基于霍夫迈斯特效应的蛋白质水凝胶及其制备方法 (Protein hydrogel based on Hofmeister effect and preparation method thereof ) 是由 寿旗扬 李圣宇 吴熙锦 张红 金璐 孙洁文 于 2021-09-29 设计创作，主要内容包括：本发明涉及一种基于霍夫迈斯特效应(Hofmeister)的蛋白质水凝胶及其制备方法。霍夫迈斯特效应辅助制备蛋白质水凝胶的制备方法包括：将牛血清蛋白与N-羟基琥珀酰亚胺(NHS)以及交联剂1-乙基-3-[3-二甲基氨基丙基]碳化二亚胺盐酸化物(EDC)制备蛋白质水凝胶,将其浸泡在不同盐溶液中,成功改变其性能。其操作简单可控,本发明专利探讨了Hofmeister系列中几种不同浓度和类型的盐对蛋白质水凝胶形成和性能的影响,可作为改变蛋白质水凝胶性质的有效方法。(The invention relates to a protein hydrogel based on Hofmeister effect (Hofmeister) and a preparation method thereof. The preparation method for preparing the protein hydrogel with the assistance of the Hofmeister effect comprises the following steps: bovine Serum Albumin (BSA), N-hydroxysuccinimide (NHS) and a cross-linking agent 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) are used for preparing protein hydrogel, and the protein hydrogel is soaked in different salt solutions to successfully change the performance of the protein hydrogel. The method is simple and controllable in operation, and the patent of the invention discusses the influence of several salts with different concentrations and types in Hofmeister series on the formation and performance of the protein hydrogel, and can be used as an effective method for changing the property of the protein hydrogel.)

1. A preparation method of protein hydrogel based on Hofmeister effect is characterized by comprising the following steps:

s1, preparation of BSA hydrogel: adding bovine serum albumin, N-hydroxysuccinimide and a cross-linking agent 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride into pure water to obtain BSA hydrogel; the N-hydroxysuccinimide accounts for 0.5 percent of the mass of the pure water; the 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride accounts for 1.5 percent of the mass of the pure water; the concentration of bovine serum albumin BSA in the BSA hydrogel is 10-40 wt%;

s2, preparation of BxAy hydrogel: soaking the BSA hydrogel prepared in the step S1 in a salt solution with the concentration of 5-40 wt% to prepare a BxAy hydrogel; wherein x represents the BSA concentration and y represents the salt solution concentration.

2. The method for preparing a protein hydrogel based on the hofmeister effect according to claim 1, wherein the BSA hydrogel has a BSA content concentration of preferably 15 wt%.

3. The method for preparing a protein hydrogel based on the hofmeister effect according to claim 1, wherein the salt solution is preferably an ammonium sulfate solution, a sodium chloride solution, a sodium iodide solution, or a sodium dihydrogen phosphate solution.

4. The method for preparing a protein hydrogel based on the Hofmeister effect according to claim 3, wherein the salt solution is preferably ammonium sulfate solution.

5. The method for preparing a protein hydrogel based on the hofmeister effect according to claim 1, wherein the concentration of the salt solution is preferably 15 wt%.

6. A protein hydrogel based on the Hofmeister effect, which is prepared by the preparation method of the protein hydrogel based on the Hofmeister effect as claimed in any one of claims 1 to 5.

Technical Field

The invention relates to the field of hydrogel, and particularly relates to a protein hydrogel based on Hofmeister effect and a preparation method thereof.

Background

The hydrogel has the characteristics of softness and water enrichment, is widely applied to drug delivery, tissue engineering and wearable electronic products, can simulate physiological environment, and is beneficial to cell culture and mass transfer in materials. But still has the risk of inflammatory reaction and/or cytotoxicity, compared with the natural polymer extracted from protein, polysaccharide and the like, the matrix has excellent biological safety, and meets the pursuit of researchers. The Hofmeister effect is one of the most important properties of proteins, and the research on the effect is very intensive, and the mechanism of the effect is still one of the most interesting basic problems in the debate. With this method, the problem of low fracture toughness or mechanical strength of protein hydrogels can be solved. More importantly, anions have been shown to have a more pronounced effect than cations, usually in an ordered arrangement: CO 2₃ ^2->SO₄ ^2->S₂O₃ ^2->H₂PO₄ ^->F^->CH₃COO^->Cl^->Br^->NO₃ ^->I^->ClO₄ ^->SCN^-。

Therefore, the method for preparing the natural protein material with good mechanical property by developing a green and feasible method by virtue of the Hofmeister effect has important significance.

Disclosure of Invention

The invention aims to provide a protein hydrogel based on Hofmeister effect and a preparation method thereof, aiming at overcoming the defects in the prior art, the operation is simple and controllable, and the aim of changing the low fracture toughness or mechanical strength of the protein hydrogel can be controlled by the method.

The technical problem to be solved by the invention is realized by adopting the following technical scheme: a preparation method of protein hydrogel based on Hofmeister effect specifically comprises the following steps:

s1, preparation of BSA hydrogel: adding bovine serum albumin, N-hydroxysuccinimide and a cross-linking agent 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride into pure water to obtain BSA hydrogel; the N-hydroxysuccinimide accounts for 0.5 percent of the mass of the pure water; the 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride accounts for 1.5 percent of the mass of the pure water; the concentration of bovine serum albumin BSA in the BSA hydrogel is 10-40 wt%;

s2, preparation of BxAy hydrogel: soaking the BSA hydrogel prepared in the step S1 in a salt solution with the concentration of 5-40 wt% to prepare a BxAy hydrogel; wherein x represents the BSA concentration and y represents the salt solution concentration.

Further, the BSA content concentration of the BSA hydrogel is preferably 15 wt%.

Further, the salt solution is preferably an ammonium sulfate solution, a sodium chloride solution, a sodium iodide solution, or a sodium dihydrogen phosphate solution.

Further, the salt solution is preferably an ammonium sulfate solution.

Further, the concentration of the salt solution is preferably 15 wt%.

The invention discloses a protein hydrogel based on the Hofmeister effect, which is prepared by the preparation method of the protein hydrogel based on the Hofmeister effect.

The invention has the beneficial effects that: the invention provides a preparation method of protein hydrogel based on Hofmeister effect, which comprises a preparation method of BSA hydrogel, and the aim of enhancing the mechanical property of the protein hydrogel is achieved by the influence of a salt solution on the Hofmeister effect; the problems of brittleness, softness and the like of the protein hydrogel are improved.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 is a stress-strain curve of the hydrogel prepared in experimental examples 1 to 3;

FIG. 2 is a graph of a pull-up test of a B15A0 hydrogel with a gradient salt concentration;

FIG. 3 is a stress-strain graph of 10 compression cycle tests on the hydrogel obtained in Experimental example 15;

FIG. 4 is a line graph showing the water content and ammonium sulfate content of BxAy hydrogels prepared in Experimental examples 1-15;

FIG. 5 is a graph comparing water loss of hydrogels prepared in Experimental examples 8-15;

FIG. 6 is a histogram showing Young's modulus of hydrogels obtained in Experimental examples 16, 17, 18, and 11.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The medical super-hydrophobic antibacterial dressing and the preparation method thereof according to the embodiment of the invention are specifically described below.

The invention provides a preparation method of protein hydrogel based on Hofmeister effect, which comprises the following steps:

s1. preparation of BSA hydrogel: preparing a protein hydrogel for BSA by dissolving Bovine Serum Albumin (BSA), N-hydroxysuccinimide (NHS) and a cross-linking agent 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) in pure water; the N-hydroxysuccinimide (NHS) accounts for 0.5 percent of the mass of the pure water; the 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) accounts for 1.5 percent of the mass of pure water; the concentration of bovine serum albumin BSA in the BSA hydrogel is 10-40 wt%.

The BSA hydrogel preferably has a BSA content concentration of 15 wt%.

S2, preparation of BxAy hydrogel: soaking the BSA hydrogel in a salt solution with the concentration of 5-40 wt% to prepare a BxAy hydrogel; wherein x represents the BSA concentration and y represents the salt solution concentration; the salt solution is preferably an ammonium sulfate solution, a sodium chloride solution, a sodium iodide solution or a sodium dihydrogen phosphate solution.

In the preferred embodiment of the present invention, the concentration of ammonium sulfate is 15 wt%;

the features and properties of the present invention are described in further detail below with reference to examples.

Example 1: preparation of B10A0 hydrogel

Protein hydrogels for BSA were prepared by adding 0.5g bovine serum albumin with 0.5% N-hydroxysuccinimide (NHS) and 1.5% 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) as a cross-linking agent to 5 ml of purified water;

example 2: preparation of B15A0 hydrogel

Protein hydrogel for BSA was prepared by adding 0.75g of bovine serum albumin with 0.5% N-hydroxysuccinimide (NHS) and 1.5% 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) as a crosslinking agent to 5 ml of purified water;

example 3: preparation of B20A0 hydrogel

Protein hydrogel for BSA was prepared by adding 1g bovine serum albumin with 0.5% N-hydroxysuccinimide (NHS) and 1.5% 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) as a cross-linking agent to 5 ml of purified water;

example 4: preparation of B25A0 hydrogel

Protein hydrogel for BSA was prepared by adding 1.25g bovine serum albumin with 0.5% N-hydroxysuccinimide (NHS) and 1.5% 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) as a crosslinking agent to 5 ml of purified water;

example 5: preparation of B30A0 hydrogel

Protein hydrogels for BSA were prepared by adding 1.5g bovine serum albumin with 0.5% N-hydroxysuccinimide (NHS) and 1.5% 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) as a cross-linking agent to 5 ml of purified water;

example 6: preparation of B35A0 hydrogel

Protein hydrogel for BSA was prepared by adding 1.75g of bovine serum albumin with 0.5% N-hydroxysuccinimide (NHS) and 1.5% 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) as a crosslinking agent to 5 ml of purified water;

example 7: preparation of B40A0 hydrogel

Protein hydrogel for BSA was prepared by adding 2g of bovine serum albumin with 0.5% N-hydroxysuccinimide (NHS) and 1.5% 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) as a crosslinking agent to 5 ml of purified water;

example 8: preparation of B15A5 hydrogel

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in a5 wt% ammonium sulfate solution to prepare B15A5 hydrogel;

example 9: preparation of B15A10 hydrogel

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in a10 wt% ammonium sulfate solution to prepare B15A10 hydrogel;

example 10: preparation of B15A15 hydrogel

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in a15 wt% ammonium sulfate solution to prepare B15A15 hydrogel;

example 11: preparation of B15A20 hydrogel

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in a20 wt% ammonium sulfate solution to prepare B15A20 hydrogel;

example 12: preparation of B15A25 hydrogel

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in a25 wt% ammonium sulfate solution to prepare B15A25 hydrogel;

example 13: preparation of B15A30 hydrogel

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in a30 wt% ammonium sulfate solution to prepare B15A30 hydrogel;

example 14: preparation of B15A35 hydrogel

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in 35 wt% ammonium sulfate solution to prepare B15A35 hydrogel;

example 15: preparation of B15A40 hydrogel

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in 40 wt% ammonium sulfate solution to prepare B15A40 hydrogel;

example 16

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in a20 wt% sodium iodide solution;

example 17

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in a20 wt% sodium chloride solution;

example 18

S1, adding 0.75g of bovine serum albumin, 0.5% of N-hydroxysuccinimide (NHS) and 1.5% of 1-ethyl-3- [ 3-dimethylaminopropyl ] carbodiimide hydrochloride (EDC) serving as a cross-linking agent into 5 ml of pure water to prepare B15A0 hydrogel;

s2, soaking the BSA hydrogel in 20 wt% sodium dihydrogen phosphate solution;

FIG. 1 is a stress-strain curve of the hydrogel prepared in experimental examples 1 to 3. The hydrogels of examples 1 to 3 were subjected to compression test comparison using a universal tester. As can be seen from FIG. 1, the mechanical properties of the hydrogel are enhanced with increasing BSA concentration, because the amino acid residues in BSA are more bound in the presence of the cross-linking agent EDC/NHS, making the binding more stable. In order to research the influence of Hofmann specific effect on protein hydrogel, the BSA concentration is preferably selected to be 15 wt% of hydrogel.

Figure 2 is a graph of a pull-up test of a B15a0 hydrogel versus a gradient salt concentration. The effect of gradient salt concentration on the lifting of the B15a0 hydrogel was studied using a universal tester. As shown in the figure, the higher the salt solution concentration is, the greater the tensile stress of the B15A0 hydrogel is, because the salt solution concentration in the solution is high, so that the hydrogel has a "dehydration" effect, and based on this, the toughness of the hydrogel is increased, and a better mechanical effect is achieved.

FIG. 3 is a stress-strain curve obtained by performing 10 compression cycle tests on Experimental example 15 using a universal testing machine; as can be seen from FIG. 3, the hydrogel prepared by the embodiment of the invention can show excellent elastic deformation performance after being compressed for 10 times, and the problems of brittleness and softness of the protein hydrogel are improved.

FIG. 4 is a line graph showing the water content and ammonium sulfate content of BxAy hydrogels prepared in Experimental examples 1-15; the greater the ammonium sulfate solution concentration, the less the water content of the hydrogel. This is because the concentration of the salt solution in the external environment is high, so that the permeation between the hydrogel and the solution is poor, and the hydrogel is easy to lose water.

FIG. 5 is a graph comparing water loss of hydrogels prepared in Experimental examples 8-15; the greater the concentration of ammonium sulfate solution, the greater the water loss of the hydrogel.

FIG. 6 is a histogram showing Young's modulus statistics of hydrogels obtained in Experimental examples 16, 17, 18 and 11 measured by a universal tester. As can be seen from fig. 6, the young's modulus of the hydrogel using the ammonium sulfate solution is the highest, the resistance to elastic deformation is the highest, and the rigidity is higher. Therefore, the salt solution based on the hofmeister effect is preferably an ammonium sulfate solution.

In summary, the preparation method of protein hydrogel assisted by hofmeister effect provided by the embodiment of the invention comprises the following steps: the BSA hydrogel preparation method achieves the purpose of enhancing the mechanical properties of the protein hydrogel by the influence of the salt solution on the Hofmeister effect of the BSA hydrogel; the problems of brittleness, softness and the like of the protein hydrogel are improved.

The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.