CN112940293A

CN112940293A - Preparation method of hydrogel based on biological protein

Info

Publication number: CN112940293A
Application number: CN202110213646.8A
Authority: CN
Inventors: 陶虎; 张杨宏; 柳克银; 刘孟玮
Original assignee: Shanghai Institute of Microsystem and Information Technology of CAS
Current assignee: Shanghai Institute of Microsystem and Information Technology of CAS
Priority date: 2021-02-25
Filing date: 2021-02-25
Publication date: 2021-06-11

Abstract

The preparation method of the hydrogel based on the biological protein comprises the steps of adding an inorganic compound containing calcium ions into a biological protein solution, stirring to obtain a first mixed solution, adding an organic compound containing hydroxyl into the first mixed solution, stirring to obtain a second mixed solution, carrying out ultrasonic dispersion treatment on the second mixed solution to obtain a third mixed solution, spin-coating the third mixed solution on a substrate, and carrying out molding treatment on the substrate on which the third mixed solution is spin-coated to obtain the hydrogel. The hydrogel prepared based on the method has good self-healing property, has Young modulus similar to that of human skin, can be well attached to the human skin without falling off, and can provide a moist healing environment for the skin and promote wound healing.

Description

Preparation method of hydrogel based on biological protein

Technical Field

The invention relates to the technical field of material science, in particular to a preparation method of a hydrogel based on biological protein.

Background

Skin damage is one of the most common damages of human body, and unreasonable treatment scheme can cause secondary bed-on and seriously harm human health. Current medical intervention can provide conditions and time for the body to self-heal or to receive a transplant and heal when tissue has regenerative capacity, e.g., fracture fixation, ligament transplantation, and replacement of donor organs of non-regenerative tissue. However, these are passive interventions that result in lost healing, slow healing rates, and even incomplete or no healing at all.

Disclosure of Invention

The embodiment of the application provides a preparation method of hydrogel based on biological protein, which can obtain hydrogel with good self-healing property, has Young modulus similar to that of human skin, can be well attached to the human skin without falling off, and can provide a moist healing environment for the skin to promote the healing of wounds.

The embodiment of the application provides a preparation method of a hydrogel based on biological protein, which comprises the following steps:

adding an inorganic compound containing calcium ions into the biological protein solution, and stirring to obtain a first mixed solution;

adding a hydroxyl-containing organic compound into the first mixed solution, and stirring to obtain a second mixed solution;

carrying out ultrasonic dispersion treatment on the second mixed solution to obtain a third mixed solution;

spin-coating the third mixed solution on the substrate;

and carrying out forming treatment on the substrate coated with the third mixed solution in a spinning mode to obtain the hydrogel.

Further, the ratio of the mass of the inorganic compound containing calcium ions to the mass of the bioprotein solution is within the interval [0.05,0.18 ].

Further, the ratio of the mass of the hydroxyl group-containing organic compound to the mass of the first mixed solution is within the interval [0.05,0.10 ].

Further, the substrate coated with the third mixed solution is subjected to a forming treatment to obtain a hydrogel, which comprises:

and (3) placing the substrate coated with the third mixed solution in a forming device with the temperature within the interval of 20 ℃ and 40 ℃ and the humidity within the interval of 0.2 and 05 for standing for 24-48 hours to obtain the hydrogel.

Further, after the substrate spin-coated with the third mixed solution is subjected to a molding treatment to obtain a hydrogel, the method further includes:

and placing the hydrogel in a laser engraving machine, and cutting the hydrogel by the laser engraving machine based on a preset shape to obtain the cut hydrogel.

Further, the thickness of the hydrogel is within the interval [0.01 μm,100 μm ].

Further, the inorganic compound containing calcium ions includes any one of calcium chloride, calcium sulfate, calcium carbonate, calcium hydroxide, and calcium oxide.

Further, the organic compound includes any one of glycerin and ethylene glycol.

Further, the biological protein solution includes any one of silk fibroin solution, sericin solution, spidroin solution, deer antler protein solution and egg white protein solution.

Further, the second mixed solution is subjected to ultrasonic dispersion treatment to obtain a third mixed solution, which comprises:

and (3) carrying out ultrasonic dispersion treatment on the second mixed solution at room temperature, wherein the treatment time is within an interval of [10min,20min ], so as to obtain a third mixed solution.

Further, after the ultrasonic dispersion treatment is performed on the second mixed solution to obtain a third mixed solution, the method further includes:

and (3) placing the third mixed solution in a storage device with the temperature within the range of [4 ℃ and 10 ℃ for storage treatment.

The embodiment of the application has the following beneficial effects:

Drawings

In order to more clearly illustrate the technical solutions and advantages of the embodiments of the present application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a schematic flow chart of a method for preparing a bioprotein-based hydrogel according to an embodiment of the present disclosure;

FIG. 2 is a schematic view of hydrogel being attached to human skin according to an embodiment of the present disclosure;

FIG. 3 is a schematic diagram of the compression of a hydrogel applied to human skin according to an embodiment of the present disclosure;

FIG. 4 is a schematic diagram of the compression of a hydrogel applied to human skin according to an embodiment of the present disclosure;

FIG. 5 is a comparison graph of an experiment for testing the self-healing performance of a hydrogel provided in the examples of the present application;

FIG. 6 is a graphical representation of the mechanical properties of a hydrogel before and after self-healing provided by an embodiment of the present application;

fig. 7 is a view of an application scenario of a hydrogel provided in an embodiment of the present application.

Detailed Description

To make the objects, technical solutions and advantages of the present application more clear, embodiments of the present application will be described in further detail below with reference to the accompanying drawings. It should be apparent that the described embodiment is only one embodiment of the present application and not all embodiments. 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 application.

An "embodiment" as referred to herein relates to a particular feature, structure, or characteristic that may be included in at least one implementation of the present application. In the description of the embodiments of the present application, it should be understood that the terms "first", "second" and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implying any number of technical features indicated. Thus, features defined as "first", "second" and "third" may explicitly or implicitly include one or more of the features. Moreover, the terms "first," "second," and "third," etc. are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the application described herein are capable of operation in other sequences than described or illustrated herein. Furthermore, the terms "comprising," "having," and "being," as well as any variations thereof, are intended to cover non-exclusive inclusions.

Referring now to specific examples of a method for preparing a bio-protein based hydrogel according to the present application, fig. 1 is a schematic flow chart of a method for preparing a bio-protein based hydrogel according to the present application, and the present specification provides the method steps as shown in the examples or the flow chart, but may include more or less steps based on conventional or non-inventive labor. The sequence of steps recited in the embodiments is only one of many execution sequences, and does not represent the only execution sequence, and in the actual execution, the steps can be executed in the sequence of the method shown in the embodiment or the drawings or executed in parallel. Specifically, as shown in fig. 1, the method includes:

s101: adding an inorganic compound containing calcium ions into the biological protein solution, and stirring to obtain a first mixed solution.

In the present embodiment, the first mixed solution may be obtained by adding the calcium ion-containing inorganic compound to the bioprotein solution and stirring for 5 to 20 minutes, wherein the ratio of the mass of the calcium ion-containing inorganic compound to the mass of the bioprotein solution is within the interval [0.05,0.18 ].

In an alternative embodiment, the bioprotein solution may be a biomaterial that is absorbed by a specific spectrum, for example, the bioprotein solution may be any one of silk fibroin solution, sericin solution, spidroin solution, deer antler protein solution, egg white protein solution.

In an alternative embodiment, the inorganic compound containing calcium ions may be any one of calcium chloride, calcium sulfate, calcium carbonate, calcium hydroxide, and calcium oxide.

S103: adding a hydroxyl-containing organic compound into the first mixed solution, and stirring to obtain a second mixed solution.

In the embodiment of the present application, the hydroxyl group-containing organic compound may be added to the first mixed solution and stirred for 5 to 20 minutes to obtain the second mixed solution, wherein the ratio of the mass of the hydroxyl group-containing organic compound to the mass of the first mixed solution is within the interval [0.05,0.10 ].

In an alternative embodiment, the organic compound comprises any one of glycerol and ethylene glycol.

S105: and carrying out ultrasonic dispersion treatment on the second mixed solution to obtain a third mixed solution.

In the embodiment of the present application, after the second mixed solution is obtained, the second mixed solution may be subjected to ultrasonic dispersion treatment to obtain a third mixed solution, that is, the second mixed solution is subjected to ultrasonic dispersion treatment at room temperature for 10 to 20 minutes to obtain a well-dispersed third mixed solution.

S107: the third mixed solution is spin coated on the substrate.

In the present embodiment, the well-dispersed third mixed solution may be stored in a storage device at a temperature within the range of [4 ℃ C., 10 ℃ C ].

In this embodiment, the third mixed solution may be spin-coated on the substrate, that is, a bio-protein solution to which the calcium ion-containing inorganic compound and the hydroxyl group-containing organic compound are added may be spin-coated on the substrate.

In an alternative embodiment, a third mixed solution may be spin-coated on a substrate based on a rotation speed within a range of [500r/min,8000r/min ], wherein the substrate may be any one of polydimethylsiloxane pdms, glass, a silicon substrate, a piezoelectric thin film ALN, silicon dioxide, and N-type oxide semiconductor-indium tin oxide.

S109: and carrying out forming treatment on the substrate coated with the third mixed solution in a spinning mode to obtain the hydrogel.

In the embodiment of the present application, after the third mixed solution is spin-coated on the substrate, the substrate on which the third mixed solution is spin-coated may be subjected to a molding process to obtain the hydrogel. Specifically, the substrate coated with the third mixture solution by spin coating can be placed in a constant temperature and humidity chamber with a temperature range of [20 ℃,40 ℃ and a humidity range of [0.2,05] for 24-48 hours, and a light and thin hydrogel with a thickness of 0.01-100 μm can be formed. The hydrogel can then be peeled off the substrate with forceps and ready for use.

In the embodiment of the application, after the hydrogel is obtained, the hydrogel can be placed in a laser engraving machine, and the laser engraving machine performs cutting treatment on the hydrogel based on a preset shape to obtain the cut hydrogel. In a specific embodiment, a desired shape of the film can be designed in advance and input into a laser engraving machine, and the film is cut by the laser engraving machine to be cut into the desired shape, specifically, the power of the laser engraving machine can be in the interval [1000mw,3000mw ], and the rotation speed of the laser engraving machine can be in the interval [2000mm/min,3000mm/min ].

By adopting the preparation method of the hydrogel based on the biological protein, provided by the embodiment of the application, the hydrogel with good self-healing property can be obtained, has the Young modulus similar to that of human skin, can be well attached to the human skin without falling off, and can provide a moist healing environment for the skin and promote the healing of wounds.

Based on the preparation methods described hereinbefore, the following examples are given.

Adding calcium chloride into the silk fibroin solution and stirring for 5 minutes to obtain a first mixed solution, wherein the mass of the calcium chloride accounts for 12% of the mass of the silk fibroin solution, and further adding glycerol into the first mixed solution and stirring for 8 minutes to obtain a second mixed solution, wherein the mass of the glycerol accounts for 8% of the mass of the first mixed solution. Then, the second mixed solution was subjected to ultrasonic dispersion treatment for 20 minutes to obtain a third mixed solution, and the third mixed solution was stored in a refrigerator at 4 ℃.

When the hydrogel is used, a layer of modified silk fibroin solution, namely a third mixed solution, can be spin-coated on a pdms substrate at the rotation speed of 500r/min, and is placed in a constant temperature and humidity box at the temperature of 30 ℃ and the humidity of 30% after being spin-coated, the silk fibroin solution is taken out after being placed for 24 hours, and is peeled off by a pair of tweezers, so that hydrogel with the thickness of about 300 mu m can be obtained, and then the silk fibroin solution is cut into a rectangle with the thickness of 3 x 4 cm by a laser engraving machine for later use. The hydrogel with a size of 3 × 4 cm may be attached to the skin of a human body, fig. 2 is a schematic diagram of the hydrogel attached to the skin of a human body according to an embodiment of the present disclosure, and the hydrogel is shown in a dashed box in fig. 2. Fig. 3 is a schematic view of the hydrogel attached to the skin of a human body according to the embodiment of the present invention being compressed, and fig. 4 is a schematic view of the hydrogel attached to the skin of a human body according to the embodiment of the present invention being compressed, and based on fig. 3 and 4, it is clear that the hydrogel has good adhesion to the skin of a human body and does not fall off from the human body by compression and stretching.

In order to test the self-healing performance of the hydrogel, fig. 5 is a comparative graph of an experiment for testing the self-healing performance of the hydrogel provided in the embodiment of the present application, the left graph is a schematic graph of cutting the hydrogel, the other graph is a schematic graph of the hydrogel automatically healing after 3 minutes, fig. 6 is a schematic graph of the mechanical performance of the hydrogel before and after self-healing provided in the embodiment of the present application, in the graph, a solid line represents the stress of the hydrogel before healing, and a dotted line represents the stress of the hydrogel after healing, so that it can be seen that the stress of the hydrogel before and after healing does not change greatly.

The healing performance of the hydrogel is explained by taking the chilblain wound as a wound model. Fig. 7 is an application scene diagram of a hydrogel according to an embodiment of the present application, where a control group is a chilblain wound without any treatment, an experimental group is a chilblain wound with hydrogel attached thereto, and the healing speed of the chilblain wound with hydrogel attached thereto is significantly faster than that of the chilblain wound without any treatment.

The hydrogel obtained by the preparation method of the hydrogel based on the biological protein can automatically heal within 3-5 minutes after being damaged, namely the hydrogel prepared by the method has good self-healing property, has the Young modulus similar to that of human skin, can be well attached to the human skin without falling off, and can provide a moist healing environment for the skin and promote the healing of wounds.

It should be noted that: the foregoing sequence of the embodiments of the present application is for description only and does not represent the superiority and inferiority of the embodiments, and the specific embodiments are described in the specification, and other embodiments are also within the scope of the appended claims. In some cases, the actions or steps recited in the claims can be performed in the order of execution in different embodiments and achieve desirable results. In addition, the processes depicted in the accompanying figures do not necessarily require the particular order shown or connected to enable the desired results to be achieved, and in some embodiments, multitasking and parallel processing may also be possible or may be advantageous.

All the embodiments in the present specification are described in a progressive manner, and the same and similar parts among the embodiments may be referred to each other, and each embodiment is described with emphasis on differences from other embodiments.

The foregoing is a preferred embodiment of the present invention, and it should be noted that it would be apparent to those skilled in the art that various modifications and enhancements can be made without departing from the principles of the invention, and such modifications and enhancements are also considered to be within the scope of the invention.

Claims

1. A method for preparing a bioprotein-based hydrogel, comprising:

spin coating the third mixed solution on a substrate;

and carrying out forming treatment on the substrate coated with the third mixed solution in a spinning mode to obtain hydrogel.

2. The method according to claim 1, wherein the ratio of the mass of the calcium ion-containing inorganic compound to the mass of the bioprotein solution is within the interval [0.05,0.18 ].

3. The production method according to claim 1, wherein a ratio of the mass of the hydroxyl group-containing organic compound to the mass of the first mixed solution is within an interval [0.05,0.10 ].

4. The method according to claim 1, wherein the forming the substrate on which the third mixed solution is spin-coated to obtain a hydrogel comprises:

and placing the substrate spin-coated with the third mixed solution in a forming device with the temperature within the range of [20 ℃,40 ℃ and the humidity within the range of [0.2,05] for standing for 24-48 hours to obtain the hydrogel.

5. The method according to claim 1, further comprising, after the forming process is performed on the substrate on which the third mixed solution is spin-coated to obtain a hydrogel:

and placing the hydrogel in a laser engraving machine, wherein the laser engraving machine cuts the hydrogel based on a preset shape to obtain the cut hydrogel.

6. The method of claim 1, wherein the hydrogel has a thickness within the interval [0.01 μm,100 μm ].

7. The method according to claim 1, wherein the inorganic compound containing calcium ions comprises any one of calcium chloride, calcium sulfate, calcium carbonate, calcium hydroxide, and calcium oxide.

8. The method according to claim 1, wherein the organic compound comprises any one of glycerol and ethylene glycol.

9. The method according to claim 1, wherein the bioprotein solution comprises any one of silk fibroin solution, sericin solution, spidroin solution, deer antler protein solution, and egg white protein solution.

10. The preparation method according to claim 1, wherein the subjecting of the second mixed solution to the ultrasonic dispersion treatment to obtain a third mixed solution comprises:

and carrying out ultrasonic dispersion treatment on the second mixed solution at room temperature, wherein the treatment time is within an interval of [10min,20min ], so as to obtain a third mixed solution.

11. The preparation method according to claim 1, wherein after the ultrasonic dispersion treatment of the second mixed solution to obtain a third mixed solution, the method further comprises: