A baicalein composite hydrogel with fluidity and degradability, and preparation method and application thereof

Abstract

The present invention discloses a baicalein composite hydrogel with fluidity and degradability, a preparation method thereof, and an application thereof in preparing a drug or a scaffold for repairing skin defects. The preparation method comprises the following steps: (1) dissolving baicalein and sodium alginate in a mass ratio of 1:2-20 in deionized water or a PBS solution to obtain a solution A, wherein the mass concentration of baicalein is not greater than 5 mg / ml, and the mass fraction of sodium alginate is about 10 mg / ml; (2) dissolving boric acid in deionized water or a PBS solution to obtain a solution B, wherein the mass concentration of boric acid in the solution B is not less than 5 mg / ml; (3) under ultrasound and continuous stirring, dropwise adding the solution B into the solution A, adjusting the pH value of the final solution to between 6 and 8, and continuing the reaction for 2-4 hours to obtain the baicalein composite hydrogel with fluidity and easy degradation.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to a baicalein composite hydrogel with fluidity and degradability, and a preparation method and application thereof. Background Art

[0002] As the largest organ in the human body, the skin is an important barrier that protects the body from external invasion. Acute skin injuries caused by various accidents and skin defects caused by diseases are common and frequently occurring clinical diseases. If the damaged skin cannot be repaired in time, the wound surface is easily infected and even develops into a chronic wound, which will threaten the patient's health, cause huge psychological pressure and economic burden to the patient, and seriously affect the quality of life.

[0003] Skin wound healing is a complex, continuous and highly coordinated physiological process. People divide this process into three stages: hemostasis and inflammation, new tissue formation and extracellular matrix (ECM) remodeling. The three stages are not isolated from each other, but intertwined. In the process of skin wound healing, a variety of cells and cytokines are involved in this process, which includes multiple cell biological processes such as cell proliferation, cell differentiation, and cell migration. These cells and cytokines are interconnected and influence each other, playing different roles in different stages of skin healing, so that skin wounds can heal better.

[0004] In terms of wound treatment, there are drug therapy, enzyme debridement therapy, electrical stimulation, etc. In the process of drug treatment of trauma, it is very important to choose a suitable drug delivery carrier. Hydrogel has unique advantages as a drug carrier. Hydrogel can absorb wound exudate, create a moist wound environment, provide a good environment for wound healing, and help accelerate wound healing. It is an ideal preparation for wound treatment. Damage to the skin greatly increases the chance of infection by microorganisms, especially bacteria. For a good skin wound repair treatment, it is required to quickly close the wound, promote cell proliferation and tissue growth, and reduce the possibility of infection. Therefore, providing a hydrogel with fluidity and degradability suitable for skin wound repair will be very effective for wound healing. Summary of the invention

[0005] The purpose of the present invention is to provide a method for preparing a baicalein composite hydrogel with fluidity and degradability. The prepared baicalein composite hydrogel has fluidity and degradability, is applied to drugs or scaffolds for repairing skin defects, and is suitable for repairing skin wounds and the like.

[0006] The present invention provides the following technical solutions:

[0007] A method for preparing a baicalein composite hydrogel with fluidity and degradability, the preparation method comprising the following steps:

[0008] (1) dissolving baicalein and sodium alginate in deionized water or PBS solution to obtain solution A;

[0009] (2) dissolving boric acid in deionized water or PBS solution to obtain solution B;

[0010] (3) Under ultrasound and continuous stirring, solution B was added dropwise into solution A to obtain baicalein composite hydrogel.

[0011] Preferably, PBS solution is selected in the preparation method, which can better simulate physiological conditions and be used for skin defect repair more quickly.

[0012] In step (1), the feed ratio of baicalein to sodium alginate is 1:2-20; the mass concentration of baicalein in solution A is not greater than 5 mg / ml, and the mass concentration of sodium alginate is 10 mg / ml.

[0013] In step (2), the mass concentration of boric acid in the solution B is not less than 5 mg / ml.

[0014] In step (3), the volume ratio of solution A to solution B is 1:1. By limiting the mixed volume ratio of solution A and solution B, the concentrations of baicalein and sodium alginate in solution A are indirectly controlled not to be too high. Too high concentrations of baicalein and boric acid will induce precipitation in the subsequent reaction, resulting in the gel being not clear and transparent enough, while too high alginate concentration will cause the fluidity of the baicalein composite hydrogel finally obtained to deteriorate.

[0015] In step (3), solution B is added dropwise to solution A, and the pH value is adjusted to between 6 and 8, and the reaction is continued for 2 to 4 hours to ensure sufficient reaction. Preferably, the pH of the final solution is about neutral (about 7), so that the obtained product will not be toxic due to excessive acidity or alkalinity and cannot be used to prepare skin repair drugs.

[0016] The present invention uses baicalein, sodium alginate and boric acid in a specific ratio as raw materials, strictly controls the concentrations of baicalein and boric acid, and can make fuller use of the reversibility of hydrogen bonds under the above-mentioned specific preparation process (dropwise addition combined with ultrasound, etc.) to prepare a baicalein composite hydrogel with better fluidity and degradability.

[0017] Baicalein is mainly found in Scutellaria baicalensis, and has an anti-platelet aggregation effect and an inhibitory effect on Staphylococcus aureus on the skin surface. Boric acid has lubricity, adhesion and anti-inflammatory effects, and contains a large number of hydroxyl groups. It can form relatively stable hydrogen bonds with the hydroxyl groups in baicalein and the carboxyl groups in sodium alginate, and finally prepare a biogel with fluidity, antibacterial and degradability, which is used for skin wound repair.

[0018] The preparation method provided by the present invention is simple to operate, has readily available raw materials, is low in cost, and is easy to repeat. The obtained hydrogel is a clear and transparent gel-like substance with certain fluidity, antibacterial properties, excellent biodegradability and high repair ability, and is particularly suitable for repairing skin wounds and the like.

[0019] The present invention also provides a baicalein composite hydrogel which is fluid and easily degradable and is obtained according to the above preparation method.

[0020] The baicalein composite hydrogel with fluidity and degradability is clear and transparent, a gel-like substance, and has certain fluidity, adhesion, biocompatibility, degradability and antibacterial properties, etc. Compared with other hydrogel materials, it has better adhesion and is more conducive to absorption.

[0021] The present invention also provides an application of the baicalein composite hydrogel in preparing a drug or a scaffold product for repairing skin defects.

[0022] The baicalein composite hydrogel with fluidity and degradability provided by the present invention adopts small molecules as raw materials with short chain length, small hydrogen bonding effect, strong adhesion effect, and is easier to adhere to and release to the wound area, so as to be absorbed by the wound, so that it can be better used for the repair of wound tissue and achieve the purpose of efficient repair.

[0023] The baicalein composite hydrogel with fluidity and degradability provided by the present invention can be completely degraded and promote the proliferation of bone marrow mesenchymal stem cells (MSC), and has good biocompatibility; it can adhere to the skin surface and be used for wound regeneration; it has an antibacterial effect and can effectively inhibit related bacteria that are not conducive to skin repair; it can effectively achieve complete repair of skin tissue, form epidermis, dermis and subcutaneous tissue, and restore various related functions of the skin.

[0024] Compared with the prior art, the present invention has the following main advantages:

[0025] (1) The present invention utilizes the hydrogen bonding between boric acid and baicalein having hydroxyl groups and sodium alginate having carboxyl groups to obtain the baicalein composite hydrogel having fluidity and degradability: the baicalein composite hydrogel has good fluidity and can be used for skin defects of various shapes; the baicalein composite hydrogel has good degradability and can be quickly degraded on the skin surface for rapid skin defect regeneration; the baicalein composite hydrogel has good antibacterial properties and can effectively inhibit harmful bacteria on the surface of damaged skin, and can be better used for repairing contaminated skin wounds; the hydrogen bonding between boric acid and baicalein and sodium alginate is weak, so a reversible composite gel can be obtained. In view of this reversible effect, the baicalein composite hydrogel provided by the present invention can be quickly degraded and release effective ingredients after acting on skin tissue, thereby achieving the purpose of rapid repair; since the baicalein, boric acid and sodium alginate used in the present invention are all completely absorbable materials, this is also one of the advantages of the present invention and the reasons for its good repair performance.

[0026] (2) The preparation method provided by the present invention is simple to operate, the raw materials are readily available, it is pollution-free, and it is easy to repeat and industrially apply.

[0027] (3) The baicalein composite hydrogel prepared by the present invention is a clear and transparent gel-like substance with good fluidity, adhesion, biocompatibility, degradability and antibacterial properties, and can be used as an efficient drug for repairing skin wounds. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a photo of the fluidity of the baicalein composite hydrogel prepared in Example 1. Scale bar in the figure: 1 cm;

[0029] Figure 2 The degradation ratio diagram of the baicalein composite hydrogel of Application Example 1 after degradation for 3, 6, 12, 18, and 24 hours;

[0030] Figure 3 The biocompatibility test results of the baicalein composite hydrogel of Application Example 2;

[0031] Figure 4 The photos of the baicalein composite hydrogel inhibiting bacterial growth in Application Example 2, with scales of 1 cm (A, C, E, G), 2 μm (B, D, F, H);

[0032] Figure 5 This is a photo of the baicalein composite hydrogel in Application Example 3 promoting skin repair. Scale bar in the figure: 2 mm. DETAILED DESCRIPTION

[0033] The present invention will be further described below in conjunction with the accompanying drawings and specific examples. It should be understood that these examples are only intended to illustrate the present invention and are not intended to limit the scope of the present invention. The operating methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer.

[0034] The baicalein composite hydrogel used in the present invention is prepared and used immediately, and the excess baicalein composite hydrogel is stored at 4°C.

[0035] Example 1

[0036] (1) 100 mg of baicalein and 200 mg of sodium alginate were dissolved in 20 ml of deionized water or PBS solution under stirring to obtain solution A, wherein the mass concentration of baicalein was 5 mg / ml and the mass concentration of sodium alginate was 10 mg / ml.

[0037] (2) Take 100 mg of boric acid and dissolve it in 20 ml of deionized water or PBS solution to obtain solution B, so that the mass concentration of boric acid in the solution is 5 mg / ml.

[0038] (3) Under stirring and ultrasonic vibration conditions, 20 ml of solution B was added dropwise into 20 ml of solution A, and the pH value of the mixed solution was adjusted to about 7.4, and stirring was continued for 3 hours to finally obtain a baicalein composite hydrogel with fluidity and degradability.

[0039] like Figure 1 As shown, the baicalein composite hydrogel prepared in this example is a clear and transparent gel-like substance with good fluidity.

[0040] like Figure 2 As shown, the baicalein composite hydrogel can be almost completely degraded after 24 hours of degradation in PBS solution.

[0041] Example 2

[0042] (1) 50 mg of baicalein and 200 mg of sodium alginate were dissolved in 20 ml of deionized water or PBS solution under stirring to obtain solution A, wherein the mass concentration of baicalein was 2.5 mg / ml and the mass concentration of sodium alginate was 10 mg / ml.

[0043] (2) Take 100 mg of boric acid and dissolve it in 20 ml of deionized water or PBS solution to obtain solution B, so that the mass concentration of boric acid in the solution is 5 mg / ml.

[0044] (3) Under stirring and ultrasonic vibration conditions, 20 ml of solution B was added dropwise into 20 ml of solution A, and the pH value of the mixed solution was adjusted to about 7.4, and stirring was continued for 3 hours to finally obtain a baicalein composite hydrogel with fluidity and degradability.

[0045] Example 3

[0046] (1) 10 mg of baicalein and 200 mg of sodium alginate were dissolved in 20 ml of deionized water or PBS solution under stirring to obtain solution A, wherein the mass concentration of baicalein was 0.5 mg / ml and the mass concentration of sodium alginate was 10 mg / ml.

[0047] (2) Take 100 mg of boric acid and dissolve it in 20 ml of deionized water or PBS solution to obtain solution B, so that the mass concentration of boric acid in the solution is 5 mg / ml.

[0048] (3) Under stirring and ultrasonic vibration conditions, 20 ml of solution B was added dropwise into 20 ml of solution A, and the pH value of the mixed solution was adjusted to about 7.4, and stirring was continued for 3 hours to finally obtain a baicalein composite hydrogel with fluidity and degradability.

[0049] Application Example 1 Biocompatibility of Baicalein Composite Hydrogel on MSC Cells

[0050] 1. 100 μg of the baicalein composite hydrogel of Example 1 was spread on the bottom of a 96-well plate as the experimental group, and a blank 96-well plate was used as the control group. After sterilization by ultraviolet light, the experimental group and the control group were added with culture medium, and MSC cells were planted in the well plate. The culture plate was placed in humidified air containing 5% carbon dioxide at 37°C and cultured, and the culture medium was changed every other day. After 1, 3, and 5 days of culture, the cell growth was analyzed using CCK8. The results are as follows: Figure 3 shown.

[0051] Figure 3 The cell images of the experimental group and the control group after culturing for 1, 3 and 5 days respectively. It can be seen that the number of cells in the experimental group is similar to that in the control group, indicating that the baicalein composite hydrogel prepared in Example 1 has good cell compatibility.

[0052] Application Example 2 Antibacterial effect of baicalein composite hydrogel

[0053] 10 mg of the hydrogel of Example 1 was dispersed in 10 ml of PBS solution and sterilized as the experimental group, and the control group was 10 ml of PBS solution without any material added. 10 μl of bacteria were added to the experimental group and the control group, respectively, and cultured at 37° C. for 12 hours. Afterwards, 100 μl of PBS solution was added to the sample to suspend the surviving bacteria, and after further incubation for 24 hours, the number of surviving bacteria was recorded.

[0054] Figure 4 A in the figure is a photo of E. coli before being treated with materials. Figure 4 C in the figure is a photograph of Escherichia coli after being treated with the material. Figure 4E in the figure is a photo of Staphylococcus aureus before being treated with materials. Figure 4 G in Figure 1 is a photo of Staphylococcus aureus after treatment with the material. Figure 4 It can be seen that the control group has a larger number of bacteria, while the experimental group has a smaller number of bacteria, with almost no bacteria remaining.

[0055] The morphology of Staphylococcus aureus and Escherichia coli and the changes in the number of bacteria before and after material treatment were observed from the microscopic perspective of scanning electron microscope. Figure 4 As shown in B, D, F and H in Figure. Figure 4 As can be seen from B, D, F and H in Figure 1, Escherichia coli is a rod-shaped bacterium and Staphylococcus aureus is a spherical bacterium. The control group has more bacteria, and the scanning electron microscopy results of the experimental group show that there are almost no bacteria in the field of view.

[0056] Application Example 3 Repair of skin defects by baicalein composite hydrogel

[0057] The hydrogel of Example 1 was the experimental group, and the control group was a group without adding any material.

[0058] like Figure 5 As shown in the figure, the defect on the surface of the nude mouse was about 8 mm. After 4 days of repair, the defect in the experimental group was significantly smaller than that in the control group. After 8 days, the wound repair in the experimental and control groups was significantly relieved. After 12 days, the wound defect in the experimental group was basically completely repaired. The defect in the control group was still obvious, indicating that the hydrogel material has a good repair effect.

[0059] In addition, it should be understood that after reading the above description of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A method for preparing a baicalein composite hydrogel with fluidity and degradability, characterized in that: The preparation method comprises the following steps: (1) dissolving baicalein and sodium alginate in deionized water or PBS solution to obtain solution A; (2) dissolving boric acid in deionized water or PBS solution to obtain solution B; (3) under ultrasound and continuous stirring, solution B was added dropwise into solution A to obtain baicalein composite hydrogel; In step (1), the feed ratio of baicalein to sodium alginate is 1:2; the mass concentration of baicalein in solution A is not greater than 5 mg / ml, and the mass concentration of sodium alginate is 10 mg / ml; In step (2), the mass concentration of boric acid in the solution B is 5 mg / ml; In step (3), the volume ratio of solution A to solution B is 1:1; In step (3), solution B is added dropwise to solution A, the pH value is adjusted to between 6 and 8, and the reaction is continued for 2 to 4 hours.

2. A baicalein composite hydrogel having fluidity and being easily degradable obtained by the preparation method according to claim 1.

3. Use of the baicalein composite hydrogel according to claim 2 in the preparation of a drug or scaffold product for repairing skin defects.

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