A thiolated hyaluronic acid hydrogel crosslinked by polydopamine mesoporous nanoparticles, and a preparation method and application thereof
By crosslinking polydopamine mesoporous nanoparticles with thiolized hyaluronic acid, a hydrogel with a mesoporous structure was prepared, which solved the problems of adhesion and drug loss in the treatment of skin wounds by hyaluronic acid hydrogels, and achieved the effects of stable adhesion and promoting wound healing.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- TIANJIN UNIV
- Filing Date
- 2023-01-05
- Publication Date
- 2026-06-09
AI Technical Summary
Existing hyaluronic acid hydrogels have insufficient adhesion in the treatment of skin wounds, cannot remain stably for a long time, and have excessively large pore sizes that cause small molecule drugs to be lost rapidly, making it impossible to effectively load and sustain release them.
By introducing polydopamine mesoporous nanoparticles and crosslinking them with thiolized hyaluronic acid, a hydrogel with a mesoporous structure is formed. The porous physical structure and chemical functional groups of polydopamine nanoparticles are used as crosslinking points to prepare hydrogels with micron and nanopores.
It achieves stable adhesion and long-lasting moisturization of hydrogels on skin wounds, improves the loading capacity and sustained-release effect of small molecule drugs, and promotes the healing of diabetic wounds.
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Figure CN117820858B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of multifunctional hydrogel technology, and more specifically, to a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles, its preparation method, and its application. Background Technology
[0002] Dressings act as a physical barrier by forming a protective layer on the skin surface, promoting the healing of superficial skin wounds. Hydrogels, due to their high water content, can maintain the humidity required for wound healing while isolating external pathogens, thus facilitating wound healing. Hyaluronic acid, a viscous polysaccharide found in the natural extracellular matrix, possesses good tissue compatibility and promotes tissue regeneration, making it widely used in clinical wound treatment. However, pure hyaluronic acid has limited viscosity, failing to effectively adhere to wounds or remain stably on the skin wound site, thus hindering its long-term physical barrier and tissue repair effects. Therefore, chemical cross-linking techniques are being used to prepare hyaluronic acid-based hydrogels that adhere to wounds and provide long-lasting hydration, potentially promoting the healing of refractory wounds such as those from diabetes. Hydrogels typically have interconnected porous structures ranging from tens to hundreds of micrometers, allowing for the loading of various drugs. However, for small molecule drugs, the large pore size of hydrogels leads to rapid drug loss. Therefore, introducing nanoporous structures into hydrogels is expected to improve the "burst release" problem associated with loading small molecule drugs onto existing hydrogels. Summary of the Invention
[0003] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles, its preparation method, and its applications. This invention obtains mesoporous polydopamine nanoparticles by adding dopamine and 4-formylphenylboronic acid as monomers, and thiol and poloxamer as template agents. Polydopamine nanoparticles with a mesoporous structure (dmPDA, i.e., degradable mesoporous polydopamine) are prepared through an oxidative self-polymerization reaction between monomers. Then, an aqueous dispersion of the polydopamine nanoparticles is mixed with a thiolized hyaluronic acid solution in a certain proportion, and the mixture forms a hydrogel through physical and chemical crosslinking.
[0004] The technical objective of this invention is achieved through the following technical solution.
[0005] A thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles and its preparation method thereof, comprising the following steps:
[0006] A PBS solution containing uniformly dispersed biodegradable mesoporous polydopamine and an aqueous solution containing uniformly dispersed thiol-modified hyaluronic acid were mixed and shaken to obtain a thiol-modified hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles, wherein:
[0007] The shaking should be performed at room temperature (20-30 degrees Celsius) for 1-10 minutes, preferably 3-5 minutes.
[0008] The hydrogel of the present invention is composed of biodegradable mesoporous polydopamine and mercaptohyaluronic acid, wherein the mass ratio of biodegradable mesoporous polydopamine to mercaptohyaluronic acid is (1:2) to (2:1), preferably (1:2) to (1:1).
[0009] The hydrogel of the present invention, after freeze-drying, has a porous structure with a diameter of tens to hundreds of micrometers (μm).
[0010] During preparation, thiol hyaluronic acid is uniformly dispersed in water, with a mass fraction of 1%-2% (ratio of mass of thiol hyaluronic acid in g to volume of deionized water in ml × 100%); biodegradable mesoporous polydopamine is uniformly dispersed in PBS solution, with a mass fraction of 1%-2% (ratio of mass of biodegradable mesoporous polydopamine in g to volume of PBS solution in ml × 100%), and the pH of the PBS solution is 7-8, preferably 7.4-7.5.
[0011] The biodegradable mesoporous polydopamine is a nanosphere with a mesoporous structure, a diameter of 200-300 nm, and an average pore size of 10-20 nm. In its preparation, dopamine and 4-formylphenylboronic acid are uniformly dispersed in a solvent. The polydopamine microspheres are prepared according to the reference "Versatile Nanoemulsion Assembly Approach to Synthesize Functional Mesoporous Carbon Nanospheres with Tunable Pore Sizes and Architectures" (J.Am.Chem.Soc.2019,141,7073-7080). The mass ratio of dopamine to 4-formylphenylboronic acid is (1-3):1, preferably (1.5-2):1.
[0012] Compared with existing technologies, this invention discloses a method for preparing a thiolized hyaluronic acid hydrogel crosslinked from polydopamine mesoporous nanoparticles. The main characteristics of dmPDA nanoparticles are their porous physical structure, larger specific surface area than ordinary polydopamine nanoparticles, and abundant surface chemical functional groups. Therefore, they can serve as crosslinking points for thiolized hyaluronic acid, enabling the preparation of a thiolized hyaluronic acid hydrogel crosslinked from dmPDA nanoparticles for the treatment of diabetic wounds (e.g., in SD rats). This hydrogel possesses a storage modulus matching that of skin tissue and exhibits both micron and nanopore structures, making it a promising candidate as a skin dressing for the treatment of diabetic wounds. Attached Figure Description
[0013] Figure 1 This is a characterization diagram of the physical properties of dmPDA nanoparticles.
[0014] Figure 2 This is the infrared spectrum of dmPDA nanoparticles.
[0015] Figure 3 This is the X-ray photoelectron spectrum of dmPDA nanoparticles.
[0016] Figure 4 This is a rheological property characterization diagram of thiolized hyaluronic acid hydrogel crosslinked with dmPDA nanoparticles.
[0017] Figure 5 These are macroscopic morphology and microstructure images of dmPDA nanoparticles crosslinked with thiolized hyaluronic acid hydrogels.
[0018] Figure 6 This image shows the repair effect of using dmPDA nanoparticles to crosslink thiolized hyaluronic acid hydrogel to repair diabetic wounds. Detailed Implementation
[0019] The technical solution of the present invention will be further described below with reference to specific examples and accompanying drawings, but these examples are not intended to limit the present invention.
[0020] First, dmPDA nanoparticles were prepared according to the description on page 7078 of the reference "Versatile Nanoemulsion Assembly Approach to Synthesize Functional Mesoporous Carbon Nanospheres with Tunable Pore Sizes and Architectures" (J.Am.Chem.Soc.2019,141,7073-7080). Specifically, PDA nanoparticles were prepared using the process conditions prior to high-temperature carbonization, with 4-formylphenylboronic acid added during the preparation process, while keeping the other process conditions unchanged, to form dmPDA nanoparticles.
[0021] Weigh 100 mg of dopamine and 50 mg of 4-formylphenylboronic acid using a balance. Transfer the mixture to a 50 mL round-bottom flask. Weigh 5 mL each of water and ethanol using a graduated cylinder, mix them thoroughly in a beaker, and add them to the round-bottom flask. Place a magnet and stir at 37°C and 1000 rpm for 30 minutes. Weigh 100 mg of poloxamer using a balance and transfer it to the round-bottom flask. Continue stirring until the solution is clear and transparent. Under the aforementioned stirring conditions, slowly and dropwise add 160 μL of trimethylbenzene. Seal the round-bottom flask with a glass stopper and transfer it to a 480 W ultrasonic cleaner for sonication for 15 minutes. Subsequently, add 375 μL of 25% ammonia solution dropwise. Transfer the round-bottom flask to a container and stir open at 37°C and 1000 rpm for 12 hours. Centrifuge at 14000 rpm to separate the contents. Wash three times alternately with water and ethanol. The product was dispersed in 10 mL of PBS solution (pH = 7.4) and stored at 4 °C for later use.
[0022] Figure 1 The images show the physical characteristics of the dmPDA nanoparticles, where a is a transmission electron microscope (TEM) image and b is the BET test result. The prepared dmPDA nanoparticle dispersion was dropped onto a 400-mesh copper grid and allowed to dry naturally, followed by observation using an electron microscope. The electron microscopy results showed that the morphology consisted of mesoporous nanospheres with a diameter of 200–300 nm. Figure 1 (a). The results of the nitrogen desorption experiment (BET) showed that ( Figure 1 In b), the average pore size of dmPDA is 10 nm. These results demonstrate that dmPDA is a nanoparticle with a mesoporous structure. Figure 2 This is the infrared spectrum of dmPDA nanoparticles. The prepared dmPDA nanoparticle dispersion was thoroughly dried in a vacuum drying oven, and the infrared spectrum of the dmPDA nanoparticles was obtained using a Fourier transform infrared spectrometer, thus successfully preparing dmPDA nanoparticles. Figure 3 This is the X-ray photoelectron spectrum of dmPDA nanoparticles. The prepared dmPDA nanoparticle dispersion was thoroughly dried in a vacuum drying oven and analyzed using X-ray photoelectron spectroscopy. The results show a significant peak at a binding energy of 399 eV, corresponding to the N1s characteristic peak. Figure 3 (a) A distinct peak is observed at a binding energy of 188 eV, corresponding to the characteristic peak of B1s. Figure 3 b) proves that dmPDA nanoparticles contain N and B elements.
[0023] Weigh 50-100 mg of thiol hyaluronic acid (HA-SH) (degree of substitution = 25%-100%) using a balance, transfer it to a 50 mL beaker, add 5 mL of deionized water, seal the beaker, and heat in a 60°C oven for 10 minutes. Remove and stir with a glass rod until completely dissolved to obtain a 1%-2% thiol hyaluronic acid solution (ratio of thiol hyaluronic acid mass (g) to deionized water volume (ml) × 100%). Aliquot the above thiol hyaluronic acid solution into 1 mL centrifuge tubes and store at -80°C for later use. Take the previously prepared dmPDA nanoparticle PBS dispersion, centrifuge to obtain a 1%-2% dmPDA dispersion (ratio of degradable mesoporous polydopamine mass (g) to PBS solution volume (ml) × 100%). Take 1 mL of dmPDA dispersion with a mass fraction of 1%-2% and 1 mL of thiol hyaluronic acid solution with a mass fraction of 1%-2%, add them to a 5 mL centrifuge tube, and shake and mix for 5 minutes at room temperature to prepare a series of thiol hyaluronic acid hydrogels crosslinked with dmPDA nanoparticles.
[0024] Figure 4 This is a rheological characterization diagram of thiolized hyaluronic acid hydrogels crosslinked with dmPDA nanoparticles. A series of hydrogels were prepared using different concentrations of dmPDA and HA-SH solutions (dispersions), including dmPDA (1% dmPDA nanoparticle PBS dispersion) + HA-SH (1% thiolized hyaluronic acid solution), dmPDA (2%) + HA-SH (1%), dmPDA (1%) + HA-SH (2%), and dmPDA (2%) + HA-SH (2%). Time-scanning rheometers were used to characterize the hydrogels. Figure 4 Part a) and stress-strain scan ( Figure 4 (Part b). The results showed that the storage modulus (G') of the product obtained according to the above ratio was higher than its loss modulus (G”) within 5 minutes, which rheologically confirmed that the product was a hydrogel structure.
[0025] In the above formulations, the hydrogel products G' of dmPDA(1%) + HA-SH(2%) and dmPDA(2%) + HA-SH(2%) were stable within 5 minutes without a significant upward trend, demonstrating that this formulation can rapidly provide mechanical support and form a hydrogel network, which is beneficial for implementing efficient and rapid clinical treatment. Considering the economic benefits, the hydrogel with the dmPDA(1%) + HA-SH(2%) formulation was selected for further research.
[0026] Figure 5These are images showing the macroscopic morphology and microstructure of thiolized hyaluronic acid crosslinked with dmPDA nanoparticles after hydrogelation. The thiolized hyaluronic acid crosslinked with dmPDA nanoparticles exhibits a clear gelation phenomenon in the tilting experiment. Figure 5 Part a). The prepared hydrogel was freeze-dried and observed using a scanning electron microscope. The results showed that the hydrogel contained a porous structure ranging from tens to hundreds of micrometers (μm). Figure 5 Part b).
[0027] Male SD rats, aged 4 to 6 weeks and weighing 180g to 200g, were selected to study the effect of hydrogel on the repair of diabetic wounds. All rats were fasted for 12 hours and then intraperitoneally injected with 55mg / kg streptozotocin (STZ). Fasting blood glucose was continuously monitored for 7-14 days post-injection, and 10 rats with a consistently high blood glucose level above 16.7mmol / L were selected for subsequent experiments. A full-thickness skin defect was created by punching a 10mm diameter hole in the rat's back, establishing a diabetic wound model. The 10 model rats were randomly divided into a control group (PBS) and a treatment group (Gel). The control group received 200μL of sterile PBS in the wound via syringe, while the treatment group received 200μL of hydrogel in the wound. All wounds were then covered with a 3M medical dressing (Tegaderm). TM To minimize the loss of liquid or hydrogel. Starting after treatment, wounds were photographed every three days, and the wound area was measured using ImageJ software. Rats were euthanized 12 days after treatment, and tissue from the wound site was dissected, fixed in 4% paraformaldehyde solution for 24 hours, and then sectioned and stained with H&E.
[0028] Figure 6 This study reflects the efficacy of hydrogels in repairing diabetic wounds. PBS served as the control group, and Gel represented the treatment group using the hydrogel of this invention. Through gross observation and statistical analysis, the wound closure speed in the treatment group was higher than that in the control group. Figure 6 Parts a and b). Observation by H&E staining of paraffin sections showed that the treatment group had completed epithelialization by 12 days; while the control group still had a large gap in the epithelial tissue. Figure 6 (Part c) The above results consistently demonstrate the wound-healing effect of hydrogel.
[0029] Adjusting the process parameters according to the present invention can achieve the preparation of the hydrogel of the present invention, and testing has shown that it exhibits performance substantially consistent with that of the present invention. The present invention has been described above as exemplary. It should be noted that any simple modifications, alterations, or other equivalent substitutions that can be made by those skilled in the art without creative effort, without departing from the core of the present invention, fall within the protection scope of the present invention.
Claims
1. A thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles, characterized in that, The hydrogel is composed of biodegradable mesoporous polydopamine and mercaptohyaluronic acid, with a mass ratio of biodegradable mesoporous polydopamine to mercaptohyaluronic acid of (1:2)-(2:1). The biodegradable mesoporous polydopamine is a nanosphere with a mesoporous structure, a diameter of 200-300 nm, and an average pore size of 10-20 nm. In preparation, dopamine and 4-formylphenylboronic acid are uniformly dispersed in a solvent to prepare biodegradable mesoporous polydopamine microspheres, with a mass ratio of dopamine to 4-formylphenylboronic acid of (1-3):
1.
2. The thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles according to claim 1, characterized in that, The mass ratio of biodegradable mesoporous polydopamine to thiol hyaluronic acid is (1:2)-(1:1).
3. A thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles according to claim 1 or 2, characterized in that, The degree of substitution of thiolated hyaluronic acid is 25%-100%.
4. A thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles according to claim 1 or 2, characterized in that, The mass ratio of dopamine to 4-formylphenylboronic acid is (1.5-2):
1.
5. A method for preparing a thiolized hyaluronic acid hydrogel crosslinked from polydopamine mesoporous nanoparticles as described in claim 1, characterized in that, The preparation was carried out according to the following steps: a PBS solution in which biodegradable mesoporous polydopamine was uniformly dispersed and an aqueous solution in which thiol-modified hyaluronic acid was uniformly dispersed were mixed and shaken to obtain a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles. Specifically: thiol-modified hyaluronic acid was uniformly dispersed in water, with a mass fraction of 1%-2%. The mass fraction of thiol-modified hyaluronic acid was defined as the ratio of the mass of thiol-modified hyaluronic acid to the volume of deionized water, multiplied by 100%. The mass of thiol-modified hyaluronic acid was expressed in g, and the volume of deionized water was expressed in mL. Biodegradable mesoporous polydopamine was uniformly dispersed in PBS solution, with a mass fraction of 1%-2%. The mass fraction of biodegradable mesoporous polydopamine was defined as the ratio of the mass of biodegradable mesoporous polydopamine to the volume of PBS solution, multiplied by 100%. The mass of biodegradable mesoporous polydopamine was expressed in g, and the volume of PBS solution was expressed in mL. The pH of the PBS solution was 7-8.
6. The method for preparing a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles according to claim 5, characterized in that, The pH of the PBS solution is 7.4-7.
5.
7. The method for preparing a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles according to claim 5, characterized in that, Choose to oscillate at 20-30 degrees Celsius for 1-10 minutes.
8. The method for preparing a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles according to claim 7, characterized in that, Choose to oscillate at 20-30 degrees Celsius for 3-5 minutes.
9. The method for preparing a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles according to claim 5, characterized in that, The biodegradable mesoporous polydopamine is a nanosphere with a mesoporous structure, a diameter of 200-300 nm, and an average pore size of 10-20 nm. In the preparation, dopamine and 4-formylphenylboronic acid are uniformly dispersed in a solvent to prepare biodegradable mesoporous polydopamine microspheres. The mass ratio of dopamine to 4-formylphenylboronic acid is (1-3):
1.
10. The method for preparing a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles according to claim 9, characterized in that, The mass ratio of dopamine to 4-formylphenylboronic acid is (1.5-2):
1.
11. The use of a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles as described in any one of claims 1-4 in the preparation of a wound treatment drug.
12. The application of a thiolized hyaluronic acid hydrogel crosslinked with polydopamine mesoporous nanoparticles as described in any one of claims 1-4 in the preparation of skin wound dressings.