Mangiferin and polydatin co-assembled hydrogel as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of hydrogel preparation and application, and relates to mangiferin and polydatin co-assembled hydrogel as well as a preparation method and application thereof.Polydatin and mangiferin are taken and dispersed in a mixed solvent respectively, pH is adjusted to 7.4-8.3, a reaction is performed under the heating condition to form a clear solution, then the clear solution is cooled to the room temperature, and the mangiferin and polydatin co-assembled hydrogel is obtained. The mangiferin and polydatin co-assembled hydrogel can be formed; the mass ratio of polydatin to mangiferin is greater than 1: 1. According to the invention, mangiferin and polydatin are co-assembled to form the hydrogel, so that the hemostasis speed can be accelerated, the antibacterial effect is good, and the hydrogel has a very good promotion effect on healing of diabetic wounds and can be used for healing of diabetic wounds.

Description

Technical Field

[0001] The invention belongs to the technical field of hydrogel preparation and application, and relates to a mangiferin and polydatin co-assembled hydrogel and a preparation method and application thereof. Background Art

[0002] Wound healing refers to the healing process after the body is subjected to external forces and the skin and other tissues are severed or damaged. It includes the regeneration of various tissues and the complex combination of granulation tissue proliferation and scar tissue formation, showing the synergistic effect of various processes. The basic process of wound healing: acute inflammatory phase → cell proliferation phase → scar formation phase → epidermis and other tissue regeneration.

[0003] Common methods to promote faster wound healing include: (1) keeping the wound clean, because cleaning the wound helps remove dirt and bacteria, reduces the risk of infection, and provides a good healing environment; (2) applying medication: applying antibiotic ointments, such as erythromycin ointment, mupirocin ointment, etc., helps prevent or treat wound infection and promotes healing; (3) covering the wound with wet compresses or dressings containing moisturizing ingredients to accelerate wound healing, such as hydrogel dressings, which are semi-permeable, can block bacteria, prevent wound infection, and allow oxygen and water to pass through.

[0004] Diabetes is a metabolic disease characterized by increased blood sugar levels. It is a chronic disease. The harm of diabetes mainly comes from complications throughout multiple systems of the body, among which diabetic foot is one of the most common and serious complications. Diabetic wounds are different from ordinary wounds and are chronic wounds (often ulcerated for a long time). This is because diabetic wounds are prone to breeding bacteria due to the high blood sugar environment. Bacterial infection makes it more difficult for wounds to heal, and in severe cases, it may lead to amputation of patients. Especially in patients with diabetic foot, the most common complication is ulcer wounds combined with bacterial infection. The high blood sugar environment of diabetic patients can serve as a bacterial culture medium, providing convenient conditions for the growth of bacteria. The difficulty in healing of diabetic foot wounds is caused by bacterial infection. It can be seen that bacterial infection is an important factor that causes the difficulty in healing of diabetic foot wounds.

[0005] Currently, there are many hydrogel materials available for treating wounds, which have slow hemostasis speed and poor antibacterial properties and are not suitable for healing wounds in diabetic patients. Summary of the invention

[0006] In view of the technical defects of the existing hydrogel materials such as slow hemostasis speed and poor antibacterial performance, the present invention provides a mangiferin and polydatin co-assembled hydrogel and a preparation method and application thereof.

[0007] The invention can accelerate the hemostasis speed by co-assembling mangiferin and polydatin to form a hydrogel, has a good antibacterial effect, has a good promoting effect on diabetic wound healing, and can be used in the healing of diabetic wounds.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is:

[0009] A method for preparing a mangiferin and polydatin co-assembled hydrogel comprises the following steps:

[0010] Polydatin and mangiferin are dispersed in a mixed solvent respectively, the pH value is adjusted to 7.4-8.3, and reacted under heating conditions to form a clear solution, and then cooled to room temperature to form a mangiferin and polydatin co-assembled hydrogel; the mass ratio of polydatin to mangiferin is greater than 1:1.

[0011] It is further defined that the mixed solvent is formed by PBS and ethanol in a volume ratio of (19-30):1.

[0012] It is further defined that the organic solvent is methanol or ethanol.

[0013] It is further defined that the heating conditions are: maintaining the temperature at 55°C to 70°C for 5 minutes to 25 minutes; and the cooling rate is 1°C / min to 3°C / min.

[0014] The mangiferin and polydatin co-assembled hydrogel is prepared by utilizing the method for preparing the mangiferin and polydatin co-assembled hydrogel.

[0015] Application of mangiferin and polydatin co-assembled hydrogel in improving free radical scavenging rate.

[0016] Application of mangiferin and polydatin co-assembled hydrogel in improving hemostasis speed.

[0017] Application of mangiferin and polydatin co-assembled hydrogel in promoting wound healing.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] 1. The present invention forms a hydrogel by co-assembling mangiferin and polydatin, which can accelerate the hemostasis speed, has a good antibacterial effect, and has a good promoting effect on the healing of diabetic wounds; it is used in the healing of diabetic wounds to accelerate the reconstruction of diabetic wounds.

[0020] 2. In the present invention, hydrogen bonds and π-π stacking effects are formed between mangiferin and polydatin, which makes the structure of the molecule larger. Therefore, mangiferin will form a co-assembled hydrogel with polydatin at a low concentration (2 mg / mL of mangiferin); at the same time, the co-assembled hydrogel can enhance the solubility and administration concentration of two small molecules, polydatin and mangiferin, so that polydatin and mangiferin act synergistically, improve the antibacterial activity and biological activity of the co-assembled hydrogel, and thus promote the healing of diabetic infected wounds.

[0021] 3. The mangoside and polydatin co-assembled hydrogel prepared by the present invention has a good scavenging effect on free radicals, and the higher the concentration of the hydrogel, the higher the scavenging rate, indicating that it has good in vitro antioxidant activity.

[0022] 4. The mangiferin and polydatin co-assembled hydrogel prepared by the present invention has no toxicity to cells at lower concentrations and has good safety; in addition, the mangiferin and polydatin co-assembled hydrogel has a good scavenging effect on ROS in cells.

[0023] 5. The mangiferin and polydatin co-assembled hydrogel prepared by the present invention has a hemolytic property of 3.2% and has high biocompatibility.

[0024] 6. The present invention adopts mangiferin and polydatin to co-assemble and form the hydrogel, the preparation method is simple, the operating temperature is mild, and industrial production is easy to realize. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a physical picture of the prepared mangoside and polydatin co-assembled hydrogel;

[0026] Figure 2 This is a SEM image of the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0027] Figure 3 The rheological properties of the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0028] Figure 4 The in vitro antioxidant activity results of the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0029] Figure 5 This is a graph showing the effect of mangiferin and polydatin co-assembled hydrogel prepared in Example 1 on bacterial growth;

[0030] Figure 6 The bacterial staining results of the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0031] Figure 7 The live / dead staining results of HUVECs in the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0032] Figure 8 This is the NIH-3t3 live-death staining result of the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0033] Fig. 9 The hemolytic results of the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0034] Fig.10 The results of mangiferin and polydatin co-assembled hydrogel prepared in Example 1 on removing intracellular ROS;

[0035] Fig.11 The hemostatic results of the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0036] Fig.12 The healing-promoting effect of the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0037] Fig.13 HE staining of wound tissue using the mangiferin and polydatin co-assembled hydrogel prepared in Example 1;

[0038] Fig.14 Masson staining of wound tissue using the mangiferin and polydatin co-assembled hydrogel prepared in Example 1. DETAILED DESCRIPTION

[0039] The technical solution of the present invention is further described below through the accompanying drawings and embodiments.

[0040] Unless otherwise defined, technical or scientific terms used in the present invention shall have the common meanings understood by one having ordinary skills in the field to which the present invention belongs.

[0041] Technologies, methods, and equipment known to ordinary technicians in the relevant field may not be discussed in detail, but where appropriate, the technologies, methods, and equipment should be considered as part of the specification.

[0042] In addition, it should be understood that although this specification is described according to the implementation modes, not every implementation mode includes only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation modes that can be understood by those skilled in the art. These other implementation modes are also covered within the protection scope of the present invention.

[0043] The present invention provides a method for preparing a mangiferin and polydatin co-assembled hydrogel, comprising the following steps:

[0044] Polydatin and mangiferin are dispersed in a mixed solvent respectively, the pH value is adjusted to 7.4-8.3, and reacted under heating conditions to form a clear solution, and then cooled to room temperature to form a mangiferin and polydatin co-assembled hydrogel; the mass ratio of polydatin to mangiferin is greater than 1:1.

[0045] For mangiferin alone, due to its small molecular structure, a higher concentration is required to form a hydrogel. Generally, mangiferin can form a gel at a concentration of at least 4 mg / mL; while polydatin alone cannot form a hydrogel.

[0046] In the present invention, in 1 ml of co-assembled hydrogel, when the load is greater than 2 mg of polydatin and the concentration of mangoside is 2 mg / mL, polydatin and mangoside can form a gel. This is because when mangoside and polydatin are mixed, hydrogen bonds and π-π stacking effects are formed between mangoside and polydatin, making the structure of the molecule larger, so that co-assembled hydrogels can be formed at low concentrations. At the same time, through the synergistic effect of polydatin and mangoside, the antibacterial activity and biological activity of the co-assembled hydrogel can be improved, the hemostasis speed can be accelerated, and the healing of wounds can be promoted.

[0047] Preferably, the mass ratio of polydatin to mangiferin is 3:2. According to this mass ratio, in 1 ml of co-assembled hydrogel, the concentration of mangiferin is 2 mg / mL, and the concentration of polydatin is greater than 2 mg / mL. Mangiferin and polydatin will form a co-assembled hydrogel.

[0048] In the present invention, the mixed solvent is formed by PBS and an organic solvent in a volume ratio of (19-30): 1. The organic solvent is methanol or ethanol. Preferably, the ethanol is anhydrous ethanol.

[0049] In the present invention, the heating conditions are: maintaining the temperature at 55°C to 70°C for 5 to 25 minutes. Preferably, the temperature is 55°C, 60°C, 65°C, 70°C, and the maintaining time is 5 minutes, 10 minutes, 15 minutes, 20 minutes or 25 minutes.

[0050] It is further defined that the cooling rate is 1°C / min to 3°C / min.

[0051] The mangiferin and polydatin co-assembled hydrogel is prepared by utilizing the method for preparing the mangiferin and polydatin co-assembled hydrogel.

[0052] The preparation and performance of the mangiferin and polydatin co-assembled hydrogel of the present invention are explained in detail with several groups of specific examples below.

[0053] It should be noted that, unless otherwise specified, the pharmaceutical agents and the like used in the following examples are conventional commercially available products.

[0054] It should be noted that, unless otherwise specified, the operations used in the following embodiments are all routine operations in the art.

[0055] Example 1

[0056] The method for preparing the mangiferin and polydatin co-assembled hydrogel provided in this embodiment comprises the following steps:

[0057] Take 3 mg of polydatin and 2 mg of mangiferin and disperse them in a mixed solvent respectively, adjust the pH to 7.5, heat to 70°C and keep the reaction for 10 minutes. After a clear solution is formed, cool it to room temperature at a rate of 1°C / min to form a mangiferin and polydatin co-assembled hydrogel.

[0058] In this embodiment, in 1 mL of mangiferin and polydatin co-assembled hydrogel, the concentration of polydatin is 3 mg / mL, and the concentration of mangiferin is 2 mg / mL.

[0059] In this embodiment, the mixed solvent is formed by PBS and anhydrous ethanol in a volume ratio of 19:1.

[0060] Example 2

[0061] The method for preparing the mangiferin and polydatin co-assembled hydrogel provided in this embodiment comprises the following steps:

[0062] Take 3 mg of polydatin and 2 mg of mangiferin and disperse them in a mixed solvent respectively, adjust the pH to 8.0, heat to 55°C and keep the reaction for 25 minutes. After a clear solution is formed, cool it to room temperature at a rate of 1°C / min to form a mangiferin and polydatin co-assembled hydrogel.

[0063] In this embodiment, in 1 mL of mangiferin and polydatin co-assembled hydrogel, the concentration of polydatin is 3 mg / mL, and the concentration of mangiferin is 2 mg / mL.

[0064] In this embodiment, the mixed solvent is formed by PBS and anhydrous ethanol in a volume ratio of 30:1.

[0065] Example 3

[0066] The method for preparing the mangiferin and polydatin co-assembled hydrogel provided in this embodiment comprises the following steps:

[0067] Take 3 mg of polydatin and 2 mg of mangiferin and disperse them in a mixed solvent respectively, adjust the pH to 8.3, heat to 60°C and keep the reaction for 5 minutes. After a clear solution is formed, cool it to room temperature at a rate of 1°C / min to form a mangiferin and polydatin co-assembled hydrogel.

[0068] In this embodiment, in 1 mL of mangiferin and polydatin co-assembled hydrogel, the concentration of polydatin is 3 mg / mL, and the concentration of mangiferin is 2 mg / mL.

[0069] In this embodiment, the mixed solvent is formed by PBS and anhydrous ethanol in a volume ratio of 19:1.

[0070] Taking the mangiferin and polydatin co-assembled hydrogel (hereinafter also referred to as hydrogel) prepared in the above Example 1 as an example, the performance of the mangiferin and polydatin co-assembled hydrogel prepared by the present invention was studied and evaluated.

[0071] Test 1, SEM morphology

[0072] Sample: mangiferin and polydatin co-assembled hydrogel prepared in Example 1.

[0073] The hydrogel was first placed on a clean silicon wafer at room temperature until its water evaporated completely, and then gold was sprayed on its surface. The microstructure of the hydrogel was recorded using SEM. The results are shown in Figure 2. Figure 2 shown.

[0074] from Figure 2 It can be seen that the mangoside and polydatin co-assembled hydrogel has a porous network structure. When the hydrogel with this structure acts on the wound, it does not affect the gas exchange of the wound, and at the same time provides a moist healing environment for the wound.

[0075] Test 2: Rheological properties

[0076] Sample: mangiferin and polydatin co-assembled hydrogel prepared in Example 1.

[0077] The rheological properties of the hydrogel were determined using a rheometer (Thermo HAAKE RS6000, USA; 35 mm parallel plates, 1 mm gap distance) in oscillation mode. Unless otherwise specified, the measurement temperature of the test was 25°C. The rheological properties of the hydrogel were described in this test using the values ​​of the storage modulus (G′) and loss modulus (G″) of the hydrogel and the relationship between the two.

[0078] First, a strain scan was performed on the hydrogel to determine the linear viscoelastic region (LVR) of the hydrogel. The strain scan range was set to 1%-100%, and the G′ and G″ of the hydrogel were measured. The results are shown in Figure 3 shown.

[0079] See also Figure 3 It can be seen that the hydrogel has a good modulus and is not easily deformed when used for wound healing, and can maintain coverage of the wound.

[0080] Test 3. In vitro antioxidant activity

[0081] Sample: mangiferin and polydatin co-assembled hydrogel prepared in Example 1.

[0082] In this study, the antioxidant properties of the hydrogels were evaluated by scavenging DPPH free radicals, ABTS free radicals, oxygen ion free radicals, and hydroxyl free radicals.

[0083] Before the experiment, the hydrogel was ground into a homogenate using a tissue grinder.

[0084] (1) DPPH free radical scavenging test

[0085] 3 mL of DPPH solution (0.1 mmol / L) was incubated with hydrogel samples (1, 3, 5, 10, and 15 mg) in the dark for 20 min and centrifuged. Then, the absorbance at 517 nm was measured by UV-vis, and the DPPH clearance rate was calculated using the formula.

[0086] DPPH clearance rate (%) = (A 0 -A i ) / A 0 ×100%.

[0087] Among them: A 0 is the absorbance of the control group, A i is the absorbance of the hydrogel sample.

[0088] (2) ABTS free radical scavenging test

[0089] ABTS (7.4×10 3 mol / L) solution and potassium persulfate (2.6×10 3 ABTS was prepared by mixing equal volumes of 2 mol / L solutions and reacting them at room temperature in the dark for 16-18 hours. +. Free radical solution. Dilute ABTS +. The free radical solution was incubated with the hydrogel samples (1, 3, 5, 10, 15 mg) in the dark for 20 min, centrifuged, and the absorbance at 734 nm was measured to calculate the ABTS. +. Clearance rate.

[0090] ABTS +. Clearance rate (%) = (A 0 -A i ) / A 0 ×100%.

[0091] Among them: A 0 is the absorbance of the control group, A i is the absorbance of the hydrogel sample.

[0092] (3) Determination of hydroxyl radical scavenging ability

[0093] The hydroxyl radical scavenging ability of each purified polysaccharide was determined by referring to Zhong Ruifang's method. Specifically, the purified polysaccharides of each fraction were dissolved in ultrapure water to prepare concentrations of 1, 2, 4, 6, 8 and 10 mg / mL, and 50 μL of each concentration of purified polysaccharide was added to a 96-well plate, and 50 μL and 9 mmol / L of FeSO 4 and 50 μL of 9 mmol / L salicylic acid anhydrous ethanol solution, mix well and add 50 μL of 8.8 mmol / L H 2 O 2 The solution was reacted at 37°C in the dark for 30 minutes, and the absorbance was measured at 510 nm. Ultrapure water was used as the blank control group, Vc was used as the positive control group, and each concentration was measured three times in parallel. The calculation formula is as follows:

[0094] Hydroxyl radical scavenging rate (%) = [1-(A 样 -A 对照 ) / A 空白 ]×100%

[0095] Among them: A 空白 is the absorbance value without adding sample, A 对照 : absorbance value of blank control group; A 样 is the absorbance value of the added sample.

[0096] The scavenging results of the above three free radicals are as follows Figure 4 shown.

[0097] See also Figure 4 The mangoside and polydatin co-assembled hydrogel had good scavenging effects on the three free radicals, and the higher the concentration of the hydrogel, the higher the scavenging rate, indicating good antioxidant activity in vitro.

[0098] Test 4: Antimicrobial activity

[0099] In this study, methicillin-resistant Staphylococcus aureus was selected for antibacterial activity study.

[0100] Sample: mangiferin and polydatin co-assembled hydrogel prepared in Example 1.

[0101] The test process is as follows: Methicillin Staphylococcus aureus, using LB medium, first put 400 μL of hydrogel in a 1.5 mL centrifuge tube, then take 400 μL of 1×10 7The CFU bacterial solution was placed on top of the hydrogel, and finally incubated on a shaker with a rotation speed of 120r / min and a temperature of 37°C. After the hydrogel and the bacterial solution were incubated for 24 hours, the bacterial solution on top of the hydrogel was diluted 10 times and evenly spread on the solid culture medium. Finally, it was incubated in a constant temperature and humidity incubator (temperature 37°C, humidity 60%) for 18 hours, and the growth of bacteria was observed to determine the bactericidal ability of the hydrogel, and photographed for record. At the same time, after the above bacterial solution was centrifuged, fixed and dehydrated, the bacteria were stained for live or dead using a bacterial live or dead staining kit (live bacteria are green and dead bacteria are red). The test results are as follows Figure 5 and Figure 6 shown.

[0102] See also Figure 5 and Figure 6 , it can be seen that the growth of bacteria in the hydrogel group was inhibited, and the staining results showed that the bacteria in the hydrogel group appeared red, indicating that the material has excellent bactericidal ability.

[0103] Test 5: Biocompatibility

[0104] Sample: mangiferin and polydatin co-assembled hydrogel prepared in Example 1.

[0105] In this experiment, HUVEC and NIH-3T3 cells were used to investigate the biocompatibility of mangiferin and polydatin co-assembled hydrogels, and live and dead cell staining (AM and PI, AM is a green fluorescent dye that can only stain live cells, that is, live cells are green and dead cells are red) was used to determine the cytotoxicity of mangiferin and polydatin co-assembled hydrogels to mammalian cells. This experiment used the Calcein / PI Cell Viability and Cytotoxicity Detection Kit (referred to as the Calein / PI Kit) produced by Bio-Tech, which is a very convenient kit for detecting the live and dead of animal cells based on the Calcein-AM (calcein) and P (propidium iodide) dual fluorescence staining method.

[0106] The test process is as follows: First, cells were plated in a 96-well plate at a dose of 10,000 cells / well and incubated for 12 hours. Then, the supernatant was discarded, and hydrogels of different concentrations were added and incubated in a cell culture incubator for 24 hours. Then, the Bio-Tech Calein / PI kit was added to evaluate the cytotoxicity of the hydrogels. Figure 7 and Figure 8 shown.

[0107] See also Figure 7 (HUVEC live / dead staining) and Figure 8 (NIH-3t3 live-death staining), MTT results showed that mangiferin and polydatin co-assembled hydrogel had no toxicity to both cells at lower concentrations, but showed an inhibitory effect on HUVEC cells when the concentration increased to 5 mg.

[0108] See also Fig. 9 The hemolytic test of mangoside and polydatin co-assembled hydrogel was carried out using human red blood cells. The results showed that the hemolytic activity of mangoside and polydatin co-assembled hydrogel was 3.2%, showing a safety comparable to that of the negative control group. These all indicate that the mangoside and polydatin co-assembled hydrogel prepared by the present invention has high biocompatibility. The morphological picture of red blood cells shows that the hydrogel treatment group has no effect on the morphology of red blood cells, and the red blood cells have a good morphology without rupture.

[0109] Experiment 6: Effect of removing intracellular ROS

[0110] Sample: mangiferin and polydatin co-assembled hydrogel prepared in Example 1.

[0111] The experimental process is as follows: first, cells were plated in a 96-well plate at a dose of 10,000 cells / well and incubated for 12 hours. Then, the supernatant was discarded, and hydrogels of different concentrations were added and incubated in a cell culture incubator for 24 hours. After the incubation, ROS dye solution was added to each well and incubated for another 15 minutes. The supernatant was then discarded and observed under a fluorescence microscope. The results are as follows: Fig.10 shown.

[0112] See also Fig.10 It can be seen that compared with the control group, the mangiferin and polydatin co-assembled hydrogel prepared by the present invention has a better scavenging effect on intracellular ROS and shows a certain concentration dependence. As the concentration of the hydrogel increases, the scavenging effect is the best.

[0113] Test 7: Hemostasis

[0114] In this study, SD rats were used as research subjects to establish a liver bleeding model187 and evaluate the in vivo hemostatic potential of mangiferin and polydatin co-assembled hydrogel.

[0115] Sample: mangiferin and polydatin co-assembled hydrogel prepared in Example 1.

[0116] The experimental process is: first, use a dose of 50 mg / kg of sodium pentobarbital to anesthetize the rat through intraperitoneal injection, then cut the rat's abdomen along the midline of the abdomen, take out a lobe of the liver and place it on a pre-weighed filter paper. Next, use a 21G needle to break the liver, place the hydrogel on the bleeding point, take pictures at different time points after treatment to record the bleeding of the liver, and record the bleeding time of the liver. Finally, record the weight of the filter paper and calculate the amount of liver bleeding. The untreated group and the PBS solution-treated group were used as controls, with at least 5 mice in each group. The tail hemostasis operation is the same as above, and the results are as follows Fig.11 shown.

[0117] See also Fig.11It can be seen that the mangoside and polydatin co-assembled hydrogel has good hemostatic properties and a faster hemostatic speed. At the same time, the blood loss of mice is reduced compared with the PBS control group.

[0118] Experiment 8: Study on the effect of hydrogel in promoting wound healing

[0119] In this study, SD rats (180-220 g) were used as the research subjects. A diabetes model was established by STZ-induced diabetic model to study the healing effect of hydrogel on diabetic wounds infected with methicillin-resistant Staphylococcus aureus.

[0120] Sample: mangiferin and polydatin co-assembled hydrogel prepared in Example 1.

[0121] Modeling method: STZ was intraperitoneally injected into rats at a dose of 35 mg / kg. After 7 days, the fasting blood glucose (FBG) of the rats was tested, and FBG ≥ 11.1 mmol / L was used as the diabetes modeling standard. Rats with successful diabetic model were selected to establish a MRSA-infected diabetic wound model. The day before surgery, the hair on the back of the rats was removed; on the day of surgery, 50 mg / kg of sodium pentobarbital was intraperitoneally injected to anesthetize the rats, and then a circular full-thickness skin excision wound with a diameter of 7 mm was created on the depilated skin, and 100 μL of MRSA (1×10 7 CFU / mL) were incubated at the wound site. The established diabetic MRSA infection wound model was divided into two groups, one group was treated with 300 μL of normal saline every day, and the other group was treated with 300 μL of hydrogel every day.

[0122] See also Fig.12 It can be seen that compared with the control group, the hydrogel has a significant healing-promoting function, and the effect of the high-concentration hydrogel group is more obvious. After 12 days, the wounds of the drug-treated group have healed, while the wound area of ​​the control group is still larger.

[0123] Test 9: Histological examination of wounds

[0124] Sample: mangiferin and polydatin co-assembled hydrogel prepared in Example 1.

[0125] For the mice in Experiment 8, the treated skin tissues were collected at different time points and fixed with 10% formalin solution, and histological examination was performed by HE and Masson staining.

[0126] In this study, HE staining and Masson staining were performed on the wound tissue. The drug-treated group had less inflammation and more obvious collagen deposition. Fig.13 and Fig.14 shown.

[0127] The results showed that after 12 days of hydrogel treatment, the wound tissue showed a complete epithelial structure, indicating that it had completed preliminary wound reconstruction, and a large amount of collagen deposition appeared; while the epithelial reconstruction of the saline treatment group was not yet complete, and a large number of inflammatory cells were present. The granulation tissue, new blood vessels, hair follicles and other skin appendages in the hydrogel treatment group increased significantly, indicating that the reconstruction of the dermal structure was basically completed. The results of skin histology further demonstrated that hydrogels can accelerate the reconstruction of diabetic wounds.

[0128] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.

Claims

1. A method for preparing a mangiferin and polydatin co-assembled hydrogel, characterized in that: The following steps are involved: Polydatin and mangiferin are dispersed in a mixed solvent respectively, the pH value is adjusted to 7.4-8.3, and reacted under heating conditions to form a clear solution, and then cooled to room temperature to form a mangiferin and polydatin co-assembled hydrogel; the mass ratio of polydatin to mangiferin is greater than 1:

1.

2. The method for preparing the mangiferin and polydatin co-assembled hydrogel according to claim 1, characterized in that: The mixed solvent is formed by PBS and an organic solvent in a volume ratio of (19-30):

1.

3. The method for preparing the mangiferin and polydatin co-assembled hydrogel according to claim 2, characterized in that: The organic solvent is methanol or ethanol.

4. The method for preparing the mangiferin and polydatin co-assembled hydrogel according to claim 1, characterized in that: The heating conditions are: maintaining the temperature at 55°C to 70°C for 5min to 25min; and the cooling rate is 1°C / min to 3°C / min.

5. The mangiferin and polydatin co-assembled hydrogel prepared by the method for preparing the mangiferin and polydatin co-assembled hydrogel according to claim 1.

6. Use of the mangiferin and polydatin co-assembled hydrogel as claimed in claim 5 in improving the free radical scavenging rate.

7. Use of the mangiferin and polydatin co-assembled hydrogel as claimed in claim 5 in improving the hemostasis speed.

8. Use of the mangiferin and polydatin co-assembled hydrogel as claimed in claim 5 in promoting wound healing.

Images