Resveratrol nanoemulsion gel wound dressing and preparation method thereof
By combining resveratrol nanoemulsion with chitosan hydrogel, a wound dressing with high drug loading, good water solubility and hemostasis properties was prepared, which solved the problem of resveratrol being difficult to dissolve in water, and achieved the rapid healing effect of wound dressing in large-area wound healing.
Patent Information
- Application Number
- CN202311455509.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-11-03
- Publication Date
- 2025-07-08
AI Technical Summary
In the prior art, resveratrol is difficult to dissolve in water and has poor stability, which limits its application in drug carriers, and trauma dressings are not effective in large-area wound healing.
Resveratrol nanoemulsion was combined with chitosan hydrogel to prepare a wound dressing with high drug load, good water solubility and hemostatic properties. Glucose was used as solvent, chitosan hydrochloride solution and hydroxyethyl cellulose solution as auxiliary materials, and resveratrol nanoemulsion gel was prepared by microjet high-pressure homogenization method.
It has achieved high drug loading and good water solubility of resveratrol nanoemulsion gel, has significant hemostasis and ability to promote wound healing, and is suitable for rapid healing of large-area trauma.
Smart Images
Figure CN120267880A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of biological dressings, and particularly to a resveratrol nanoemulsion gel wound dressing and a preparation method thereof. Background Art
[0002] As the largest organ system of the human body, the skin plays a key role in preventing mechanical forces and infections, fluid imbalances, and thermal disorders. The integrity of healthy skin is crucial for maintaining human physiological homeostasis. When the skin is damaged, it poses certain potential risks to human healthy life, and in severe cases, it may even endanger human life and health. The healing of skin wounds is also a relatively complex process, which depends on the interaction of multiple cell types and mediators in a highly complex time sequence. Especially for large-area wounds, it is almost impossible to heal spontaneously.
[0003] Wound dressings are a widely used treatment tool in modern medicine. Their main function is to keep the wound surface moist and prevent infection, thereby promoting wound healing. Nanoemulsion gels are a new type of drug carrier, which have advantages such as high efficiency and low toxicity, are more conducive to drug administration, provide new options for drug delivery systems, and are a dosage form with great development potential.
[0004] Resveratrol (RES) is a natural non-flavonoid polyphenolic compound that widely exists in plants such as grapes, peanuts, blueberries, polygonum cuspidatum, fleece-flower root, and soybeans. Modern research shows that resveratrol has a variety of pharmacological activities, such as inhibiting cell proliferation, anti-inflammatory, antibacterial and antifungal infections, antioxidant, anti-aging, etc., and has great research value; however, due to its poor water solubility and poor stability, its application is greatly limited.
[0005] Chitosan hydrogel has advantages such as low toxicity, high biocompatibility, and high mechanical strength, and has good application prospects in the fields of drug sustained-release materials, medical dressings, etc. Glucose is non-toxic and non-irritating, is an essential component of the body, the energy source and intermediate metabolite of living cells, is a necessary substance in the cell repair process, is a very commonly used excipient in clinical practice, and has higher safety. After retrieval, no report has been found on the preparation of a resveratrol nanoemulsion gel wound dressing using glucose as a solvent and resveratrol raw material and chitosan hydrogel as raw materials. Summary of the Invention
[0006] In view of this, the purpose of the present invention is to provide a resveratrol nanoemulsion gel wound dressing. Based on the anti-inflammatory, antibacterial, angiogenesis-promoting, and cell-protecting effects of resveratrol and the good use of chitosan hydrogel as a wound dressing. The present invention combines resveratrol nanoemulsion with a glucose-based chitosan hydrogel to obtain a new type of wound dressing with high drug loading, good water solubility, and hemostatic properties.
[0007] The technical solution is as follows: A resveratrol nanoemulsion gel wound dressing, comprising the following raw materials in parts by weight: 0.8 - 1.2 parts of resveratrol nanoemulsion, 2 - 4 parts of chitosan hydrochloride solution, and 0.5 - 0.8 parts of hydroxyethyl cellulose solution.
[0008] Furthermore, a resveratrol nanoemulsion gel wound dressing, comprising the following raw materials in parts by weight: 1 part of resveratrol nanoemulsion, 3 parts of chitosan hydrochloride solution, and 0.7 parts of hydroxyethyl cellulose solution.
[0009] Furthermore, the concentration of the chitosan hydrochloride solution is 1% - 5% m / V, and the further preferred concentration is 2% m / V.
[0010] Furthermore, the concentration of the hydroxyethyl cellulose solution is 1% - 5% m / V, and the further preferred concentration is 2.75% m / V.
[0011] Furthermore, the chitosan hydrochloride solution is a D-glucose - sodium bicarbonate (D-glu-NaHCO3) solution, using 0.45% D-glucose (D-glu) as the solvent and 0.3% NaHCO3 as the pH regulator to adjust the pH value of the solution to 6.5 - 7.5.
[0012] Furthermore, the hydroxyethyl cellulose solution is a D-glu-NaHCO3 solution, using 0.45% D-glu as the solvent and 0.6% NaHCO3 as the pH regulator to adjust the pH value of the solution to 6.5 - 7.5.
[0013] Another object of the present invention is to provide a preparation method of the above-mentioned resveratrol nanoemulsion gel wound dressing.
[0014] Step 1, preparation of resveratrol nanoemulsion:
[0015] Precisely weigh triacetin, polyoxyethylene castor oil EL-60, PEG400, and glycerol in a beaker, stir on a magnetic stirrer, then add the resveratrol raw drug, continue stirring until completely dissolved, transfer to a volumetric flask, make up the volume to the scale with water, place in an air bath constant temperature oscillator and keep it at 50 °C for 30 min, and then homogenize by microfluidic high-pressure homogenization method. Homogenization conditions: 28000 - 33000 psi, high-pressure homogenization for 3 - 5 times to obtain the resveratrol nanoemulsion, and place it in a dry vial for storage in the dark.
[0016] Step 2: Preparation of chitosan hydrogel:
[0017] Weigh the prescribed amounts of CSCl (D-glu-NaHCO3) solution and HEC (D-glu-NaHCO3) solution, add them to a dry vial, mix well, and keep at a temperature of 36 - 40 °C until gelation occurs.
[0018] Step 3: Preparation of resveratrol nanoemulsion gel:
[0019] Weigh the prescribed amounts of resveratrol nanoemulsion sample solution and CSCl (D-glu-NaHCO3) solution into a dry vial, mix well, then add the prescribed amount of HEC (D-glu-NaHCO3) solution, mix well again, and keep at a temperature of 36 - 40 °C until gelation occurs, obtaining the resveratrol nanoemulsion gel wound dressing.
[0020] Compared with the prior art, the beneficial effects of the present invention are as follows:
[0021] 1. The present invention uses chitosan hydrochloride as the gel material, HEC as the hardness regulator, NaHCO3 as the pH regulator, and aqueous anhydrous glucose solution as the solvent to prepare an injectable hydrogel wound dressing with self-healing property carrying resveratrol nanoemulsion, namely the resveratrol nanoemulsion gel wound dressing, and the gelation time is 15 min.
[0022] 2. Through experimental determination, it is found that the resveratrol nanoemulsion gel is a strong hydrogel with self-healing property and injectability, having good hemolytic activity and biocompatibility. By constructing liver hemorrhage models and full-thickness skin injury models in animal experiments, the results show that the resveratrol nanoemulsion gel has good hemostatic performance and the ability to promote wound healing, and it is a new type of wound dressing with high drug loading, good water solubility and hemostatic performance. Description of the Drawings
[0023] Figure 1 (Left) is resveratrol nanoemulsion, Figure 1 (Right) is the transmission electron microscope of resveratrol nanoemulsion.
[0024] Figure 2 is the particle size distribution diagram of resveratrol nanoemulsion.
[0025] Figure 3 (Left) is the gelation state of CSCl hydrogel at 0 min, Figure 3 (Right) is the gelation state of CSCl hydrogel at 10 min.
[0026] Figure 4 is the scanning electron microscope morphology of CSCl hydrogel.
[0027] Figure 5 (Left) is the gelation state of resveratrol nanoemulsion gel at 0 min, Figure 5 (Right) is the gelation state of resveratrol nanoemulsion gel at 15 min.
[0028] Figure 6 (Left) shows the gelling state of resveratrol nanoemulsion gel stored at low temperature, room temperature, and high temperature for 0 h. Figure 6 (Right) shows the gelling state of resveratrol nanoemulsion gel stored at low temperature, room temperature, and high temperature for 48 h.
[0029] Figure 7 is the blood loss in the in vivo hemostasis experiment.
[0030] Figure 8 is the blood loss in the liver bleeding model.
[0031] Figure 9 is the wound condition of the mouse.
[0032] Figure 10 is the schematic diagram simulating the mouse wound surface.
[0033] Figure 11 is the area of the mouse wound surface (unit: cm 2 ). Specific implementation manner
[0034] To better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. Those skilled in the art make modifications or equivalent replacements based on the understanding of the technical solution of the present invention, and without departing from the spirit and scope of the technical solution of the present invention, they should all be covered within the protection scope of the present invention.
[0035] Example 1
[0036] A resveratrol nanoemulsion gel wound dressing comprises the following raw materials in parts by weight: 1 part of resveratrol nanoemulsion, 3 parts of 2% m / V chitosan hydrochloride solution, and 0.7 part of 2.75% m / V hydroxyethyl cellulose solution.
[0037] The preparation method comprises the following steps:
[0038] (1) Preparation of resveratrol nanoemulsion:
[0039] Precisely weigh 1.4 g of triacetin, 2 g of polyoxyethylene castor oil EL-60, 0.6 g of PEG400, and 0.65 g of glycerol in a beaker, stir on a magnetic stirrer, then add 0.175 g of resveratrol raw material medicine, continue to stir until completely dissolved, transfer to a 25 mL volumetric flask, make up to the mark with water, place in an air bath constant temperature oscillator and keep at 50 °C for 30 min, then homogenize by microfluidic high-pressure homogenization method. Homogenization conditions: 31500 psi, high-pressure homogenization 4 times to obtain resveratrol nanoemulsion, and store it in a dry vial in the dark.
[0040] (2) Preparation of chitosan hydrogel:
[0041] Weigh the prescription amounts of 2% CSCl (D-glu-NaHCO3) solution and 2.75% HEC (D-glu-NaHCO3) solution, add them to a dry vial, mix well, and place them in a 37°C constant temperature water bath. Observe the gelation situation every 5 minutes until gelation occurs.
[0042] (3) Preparation of resveratrol nanoemulsion gel:
[0043] Weigh the prescription amounts of resveratrol nanoemulsion sample solution and 2% CSCl (D-glu-NaHCO3) solution in a dry vial, mix well, then add the prescription amount of 2.75% HEC (D-glu-NaHCO3) solution, mix well, and place them in a 37°C constant temperature water bath. Observe the gelation situation every 5 minutes until gelation occurs.
[0044] Example 2
[0045] A resveratrol nanoemulsion gel wound dressing, comprising the following raw materials in parts by weight: 0.8 part of resveratrol nanoemulsion, 2 parts of 2% m / V chitosan hydrochloride solution, and 0.5 part of 2.75% m / V hydroxyethyl cellulose solution.
[0046] The preparation method comprises the following steps:
[0047] (1) Preparation of resveratrol nanoemulsion:
[0048] Precisely weigh 1.4 g of glyceryl triacetate, 2 g of polyoxyethylene castor oil EL-60, 0.6 g of PEG400, and 0.65 g of glycerol into a beaker, stir on a magnetic stirrer, then add 0.175 g of resveratrol raw material medicine, continue stirring until completely dissolved, transfer to a 25 mL volumetric flask, make up the volume to the mark with water, place in an air bath constant temperature oscillator and keep at 50°C for 30 minutes, then homogenize by microfluidic high-pressure homogenization method. Homogenization conditions: 31500 psi, high-pressure homogenization 4 times to obtain resveratrol nanoemulsion, and place it in a dry vial for storage in the dark.
[0049] (2) Preparation of chitosan hydrogel:
[0050] Weigh the prescription amounts of 2% CSCl (D-glu-NaHCO3) solution and 2.75% HEC (D-glu-NaHCO3) solution, add them to a dry vial, mix well, and place them in a 37°C constant temperature water bath. Observe the gelation situation every 5 minutes until gelation occurs.
[0051] (3) Preparation of resveratrol nanoemulsion gel:
[0052] Weigh the prescribed amount of resveratrol nanoemulsion sample solution and 2% CSCl (D-glu-NaHCO3) solution into a dry vial, mix well, then add the prescribed amount of 2.75% HEC (D-glu-NaHCO3) solution. After mixing well, place it in a 37°C constant temperature water bath, observe the gelation situation every 5 minutes until gelation occurs.
[0053] Example 3
[0054] A resveratrol nanoemulsion gel wound dressing, comprising the following raw materials in parts by weight: 1.2 parts of resveratrol nanoemulsion, 4 parts of 2% m / V chitosan hydrochloride solution, and 0.8 part of 2.75% m / V hydroxyethyl cellulose solution.
[0055] The preparation method comprises the following steps:
[0056] (1) Preparation of resveratrol nanoemulsion:
[0057] Precisely weigh 1.4 g of glyceryl triacetate, 2 g of polyoxyethylene castor oil EL-60, 0.6 g of PEG400, and 0.65 g of glycerol into a beaker, stir on a magnetic stirrer, then add 0.175 g of resveratrol raw material drug, continue to stir until completely dissolved, transfer to a 25 mL volumetric flask, make up the volume to the mark with water, place it in an air bath constant temperature oscillator and keep it at a constant temperature of 50°C for 30 minutes, then homogenize by microfluidic high-pressure homogenization method. The homogenization conditions are: 31500 psi, high-pressure homogenization 4 times to obtain resveratrol nanoemulsion, and store it in a dry vial in the dark.
[0058] (2) Preparation of chitosan hydrogel:
[0059] Weigh the prescribed amount of 2% CSCl (D-glu-NaHCO3) solution and 2.75% HEC (D-glu-NaHCO3) solution, add them to a dry vial, mix well, then place it in a 37°C constant temperature water bath, observe the gelation situation every 5 minutes until gelation occurs.
[0060] (3) Preparation of resveratrol nanoemulsion gel:
[0061] Weigh the prescribed amount of resveratrol nanoemulsion sample solution and 2% CSCl (D-glu-NaHCO3) solution into a dry vial, mix well, then add the prescribed amount of 2.75% HEC (D-glu-NaHCO3) solution, mix well and place it in a 37°C constant temperature water bath, observe the gelation situation every 5 minutes until gelation occurs.
[0062] Effect Example 1: Preliminary quality evaluation of resveratrol nanoemulsion
[0063] Take 10 μL of the resveratrol nanoemulsion sample solution and place it in a 10 mL volumetric flask. Dilute it to the mark with pure water, filter it through a 0.4 μm microporous filter membrane, and measure the particle size and PDI using a high-sensitivity Zeta potential and particle size analyzer. Take 10 mL of the resveratrol nanoemulsion sample solution in a beaker and measure the pH value using a laboratory pH meter.
[0064] Take the resveratrol nanoemulsion sample solution. Appropriately take the original solution and the sample solutions diluted 10 times and 20 times, respectively. Drop them on a copper plate and let them stand for 5 min. Remove the excess solution, stain them with an appropriate amount of 2% phosphotungstic acid, let them stand for 30 min, remove the excess liquid, let them stand and air-dry, and observe the morphology of the nanoemulsion with a transmission electron microscope at an accelerating voltage of 100.0 kV.
[0065] The resveratrol nanoemulsion (RES-NE) is in a clear and transparent solution state, as Figure 1 (left) shown. The transmission electron microscope photograph of RES-NE is as Figure 1 (right) shown. The nanoemulsion droplets are of uniform size and evenly distributed. The pH value is 6.80, the average particle size is 4.30 nm, the polydispersity index (PDI) is 0.178, and the particle size distribution diagram is as Figure 2 shown.
[0066] Effect Example 2: Preliminary Quality Evaluation of Chitosan Hydrogel
[0067] After freeze-drying the CSCl hydrogel, perform sputtering with gold and study the microscopic morphological characterization of the gel using a scanning electron microscope (SEM). Prepare the CSCl hydrogel, mix it evenly, and measure the pH value of the hydrogel using a wide-range pH test paper.
[0068] The consistency of the CSCl hydrogel under the prescription of Example 1 increased at 5 min and gelled at 10 min, and the pH was 7, as Figure 3 shown. The scanning electron microscope morphology of the CSCl hydrogel is as Figure 4 .
[0069] Effect Example 3: Preliminary Quality Evaluation of Resveratrol Nanoemulsion Gel
[0070] Take three portions of the gelled resveratrol nanoemulsion chitosan hydrogel and place them at 4, 25, and 40 °C respectively. Use the appearance at the 0th and 48th h as the evaluation index to conduct the temperature stability evaluation. Prepare the resveratrol nanoemulsion gel, mix it evenly, and measure the pH value of the hydrogel using a wide-range pH test paper.
[0071] The resveratrol nanoemulsion gel of Example 1 gelled at 15 min and the pH was 6.54, as Figure 5 shown. The temperature stability evaluation is as Figure 6 shown, indicating good stability at low temperatures and showing that the resveratrol nanoemulsion gel is not suitable for storage under high-temperature conditions.
[0072] Effect Example 4: In vivo Hemostatic Performance Evaluation of Resveratrol Nanoemulsion Gel
[0073] In this experiment, an SD rat liver bleeding model was used to analyze the in vivo hemostatic activities of RES-NBGs and CSCl hydrogels. Nine healthy SD rats (all males) were prepared, each rat was marked and weighed, and randomly divided into 3 groups with 3 rats in each group. The SD rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (0.3 mL / 100 g). A scalpel was used to make an incision in the abdomen of the rats to expose the liver. The liquid around the liver was cleaned with filter paper. The pre-weighed filter paper was placed under the liver, and the liver was punctured at 30° with a 20G needle to create a liver bleeding model. The three rats were treated as follows: ① the non-treatment group; ② the blank gel group; ③ the drug-loaded gel group. After 3 min, the weight of the filter paper under the liver was weighed. The increased weight of the filter paper was the blood loss of the rats, and the difference in blood loss was recorded and compared.
[0074] In this experiment, the in vivo hemostatic activities of RES-NBGs and CSCl hydrogels were evaluated through the quantitative results of the blood loss and hemostasis time of the liver bleeding model. In the experiment, the filter paper weights before and after bleeding in the non-treatment group, the blank gel group and the drug-loaded group were compared respectively, and the blood loss of each group was counted. As Figure 7 、 8 shown, it was found that both the blank gel group and the drug-loaded gel group had significant hemostatic functions. Only a small amount of blood stains appeared on the filter papers of these two groups, and the drug-loaded gel group had less blood loss than the blank gel group. A large amount of blood stains remained on the filter paper of the non-treatment group. RES-NBGs and CSCl hydrogels had a rapid hemostatic effect on acute liver bleeding and could effectively absorb the exudate at the wound. The results showed that loading resveratrol nanoemulsion into chitosan hydrogel could effectively improve the in vivo hemostatic ability of chitosan hydrogel.
[0075] Effect Example 5: Wound Healing Experiment of Resveratrol Nanoemulsion Gel
[0076] In this experiment, a full-thickness skin injury model of mice was created to evaluate the wound healing promoting ability of resveratrol nanoemulsion gel. Twenty-four healthy Kunming mice, all males, were weighed and recorded. All the mice were randomly divided into 4 groups with 6 mice in each group. Each group was treated as follows: ① the non-treatment group; ② the commercially available gel group; ③ the blank gel group; ④ the drug-loaded gel group. After anesthesia by intraperitoneal injection of 10% chloral hydrate (0.05 mL / 10 g), the hair on the back of the mice was shaved to prepare for the operation. A skin wound with a diameter of 8 mm was created on the back of each mouse. The dosage of each administration was based on covering the wound. The wound healing conditions of each group were photographed and recorded on the 0th, 3rd, 6th, 9th, 12th, and 15th days after the operation.
[0077] In this experiment, a full-thickness skin injury model of mice was established to investigate the ability of resveratrol nanoemulsion gel dressing to promote wound healing. Through the observation and treatment of the wound surface of mice, the following results were obtained: The wound surfaces of the blank gel group and the drug-loaded gel group showed obvious healing conditions. The wound in the untreated group recovered the slowest, and the commercially available group was the second slowest. The wound healing process is as shown in Figure 9 and 10 . The wound areas at different stages are as shown in Figure 11 . The results indicate that resveratrol nanoemulsion gel has moderate adhesion to the wound surface, can fully contact the wound surface, cover the entire wound surface, accelerate wound healing, and has the potential to become a good dressing.
Claims
1. A resveratrol nanoemulsion gel wound dressing, characterized in that, It comprises the following raw materials in parts by weight: 0.8 - 1.2 parts of resveratrol nanoemulsion, 2 - 4 parts of chitosan hydrochloride solution, and 0.5 - 0.8 parts of hydroxyethyl cellulose solution.
2. The resveratrol nanoemulsion gel wound dressing according to claim 1, characterized in that, The concentration of the chitosan hydrochloride solution is 1% - 5% m / V.
3. The resveratrol nanoemulsion gel wound dressing according to claim 1, characterized in that, The concentration of the chitosan hydrochloride solution is 2% m / V.
4. The resveratrol nanoemulsion gel wound dressing according to claim 1, characterized in that, The concentration of the hydroxyethyl cellulose solution is 1% - 5% m / V.
5. The resveratrol nanoemulsion gel wound dressing according to claim 1, wherein, The concentration of the hydroxyethyl cellulose solution is 2.75% m / V.
6. The resveratrol nanoemulsion gel wound dressing according to claim 1, wherein The chitosan hydrochloride solution and the hydroxyethyl cellulose solution are D-glu-NaHCO3 solutions, using 0.45% D-glu as the solvent and NaHCO3 as the pH regulator.
7. The preparation method of the resveratrol nanoemulsion gel wound dressing according to claim 1, characterized in that It comprises the following steps: Step 1, preparation of resveratrol nanoemulsion: Precisely weigh glyceryl triacetate, polyoxyethylene castor oil EL-60, PEG400, and glycerol in a beaker, stir on a magnetic stirrer, then add the resveratrol raw drug, continue stirring until completely dissolved, transfer to a volumetric flask, make up to the mark with water, place in an air bath constant temperature oscillator and keep at a constant temperature of 50 °C for 30 min, then homogenize using the microfluidic high-pressure homogenization method. The homogenization conditions are: 28000 - 33000 psi, high-pressure homogenize 3 - 5 times to obtain the resveratrol nanoemulsion, and place it in a dry vial for storage in the dark. Step 2: Preparation of chitosan hydrogel: Weigh the prescribed amount of chitosan hydrochloride D-glu-NaHCO3 solution and hydroxyethyl cellulose D-glu-NaHCO3 solution, add them to a dry vial, mix well, and at a temperature of 36 - 40 °C until gelling. Step 3: Preparation of resveratrol nanoemulsion gel: Weigh the prescribed amount of resveratrol nanoemulsion sample solution and chitosan hydrochloride D-glu-NaHCO3 solution in a dry vial, mix well, then add the prescribed amount of hydroxyethyl cellulose D-glu-NaHCO3 solution, mix well, and at a temperature of 36 - 40 °C until gelling, which is the resveratrol nanoemulsion gel wound dressing.