Hyaluronic acid double-network hydrogel and sponge dressing preparation method

Abstract

The invention discloses a preparation method of hyaluronic acid double-network hydrogel and a sponge dressing, and belongs to the technical field of biomedical materials, and the technical key points are that the preparation method comprises the following steps: step 1, dissolving wet squid top bones in a sodium hydroxide solution, then carrying out high-temperature reaction for 120 minutes, then filtering, and washing filtrate with deionized water until the pH value is neutral; step 2, adding diluted hydrochloric acid, reacting at room temperature for 24 hours, filtering, washing, and washing filter residues with deionized water until the pH value is neutral, so as to obtain wet beta-chitin; 3, hyaluronic acid and a cross-linking agent glutaraldehyde are added into the beta-chitin fiber dispersion liquid subjected to ultrasonic dispersion, low-temperature solvent replacement is conducted after cross-linking, freezing and demolding are conducted, and the sponge is obtained through freeze drying; 4, the proportion of glutaraldehyde, beta-chitin and hyaluronic acid is adjusted, the sponge dressings with different properties are obtained, and the sponge dressings have the advantage of being good in tissue growth promoting effect.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical materials, and specifically relates to a preparation method of a hyaluronic acid double-network hydrogel and a sponge dressing. Background Art

[0002] According to the World Health Organization, as of 2015, there were approximately 415 million diabetic patients globally, and more than 15% of them had diabetic ulcers and chronic wound infections. Different from traditional wound formation, chronic diabetic ulcer wounds result from the combined effects of microenvironmental pollution, microcirculation disorders, inflammatory responses, and bacterial infections. The accumulation of high sugar and advanced glycation end products can cause microenvironmental pollution of the wound, resulting in chronic inflammatory reactions, leading to insufficient numbers or low functions of endothelial progenitor cells, causing microcirculation regeneration disorders and tissue inactivation, and then bacterial invasion forms chronic infections, exacerbating the condition. Therefore, it is particularly important and urgent to design and prepare new antibacterial materials with high efficiency and promoting wound epithelialization.

[0003] Chitin is the precursor of chitosan and is a natural high-molecular aminopolysaccharide. The solubility of natural chitin itself is relatively poor, and it generally needs to be modified in applications. Chitin has three basic crystal structure forms: α, β, and γ. The molecular chains of β-chitin are arranged in a parallel manner, and the intermolecular hydrogen bond force is relatively weak, so it is easier to carry out chemical modification and has stronger antioxidant, antibacterial, and depolymerization reactions.

[0004] Hyaluronic acid is a linear acidic mucopolysaccharide composed of D-glucuronic acid and N-acetylglucosamine. It is widely present in the human body and has the functions of regulating cell growth, proliferation, and differentiation. Hyaluronic acid can absorb a large amount of water and has excellent gelling properties, but its own properties are unstable and it can react with chitin under the action of a chemical cross-linking agent to form a stable high-molecular compound.

[0005] Therefore, to solve at least one of the above problems, the present invention provides a preparation method and application of a double-network hydrogel / sponge dressing of β-chitin cross-linked hyaluronic acid. Summary of the Invention

[0006] Aiming at the deficiencies of the existing technology, the purpose of the embodiments of the present invention is to provide a preparation method of a hyaluronic acid double-network hydrogel and a sponge dressing to solve the problems in the above background art.

[0007] To achieve the above purpose, the present invention provides the following technical solutions:

[0008] A preparation method of a hyaluronic acid double-network hydrogel and a sponge dressing, comprising the following steps:

[0009] Step 1: Use wet squid cranial bone, dissolve it with sodium hydroxide solution, then filter after high-temperature reaction for 120 min, and wash the filtrate with deionized water until the pH value is neutral.

[0010] Step 2: Add dilute hydrochloric acid, react at room temperature for 24 hours, filter, wash, and wash the filter residue with deionized water until the pH value is neutral to obtain wet β-chitin.

[0011] Step 3: Add hyaluronic acid and cross-linking agent glutaraldehyde to the ultrasonically dispersed β-chitin fiber dispersion, carry out cross-linking, freezing and demolding, then perform low-temperature solvent replacement, and freeze-dry to obtain the sponge.

[0012] Step 4: Adjust the ratio of glutaraldehyde to β-chitin and hyaluronic acid to obtain sponge dressings with different properties.

[0013] As a further scheme of the present invention, in Step 1, it is dissolved with a sodium hydroxide solution with a concentration of 1 mol / L, filtered after high-temperature reaction at 90 °C for 120 min.

[0014] As a further scheme of the present invention, 0.1 mol / L dilute hydrochloric acid is added in Step 2.

[0015] As a further scheme of the present invention, in Step 3, the concentration of the β-chitin fiber dispersion is 0.5 wt%, and the concentration of hyaluronic acid is 0.5%.

[0016] As a further scheme of the present invention, after uniform dispersion in Step 3, it is injected into a mold for cross-linking for 24 hours, and demolded after freezing at -75 °C.

[0017] As a further scheme of the present invention, after cross-linking, freezing and demolding in Step 3, it is placed in pre-cooled absolute ethanol for low-temperature replacement, and then the ethanol is eluted with deionized water, and freeze-dried to obtain the sponge.

[0018] As a further scheme of the present invention, the molecular formula of the hyaluronic acid is (C 14 H 21 NO 11 )n, where n represents the degree of polymerization, that is, the number of disaccharide units in the hyaluronic acid molecule.

[0019] In summary, the embodiments of the present invention have the following beneficial effects compared with the prior art:

[0020] The double-network hydrogel / foam dressing of β-chitin cross-linked hyaluronic acid provided by the present invention can be used as a new type of biomedical material for promoting wound healing, and various types of dressings, such as liquid dressings, hydrogel dressings, sponge dressings, etc., can be obtained by changing the ratio of hyaluronic acid to glutaraldehyde, so as to be applied to various types of wounds, such as acute wounds like abrasions and burns, and chronic wounds like diabetic ulcers and pressure sores.

[0021] The preparation method of the double-network hydrogel / sponge dressing of β-chitin crosslinked hyaluronic acid provided by the present invention uses β-chitin and hyaluronic acid as the main components. The raw materials are easily obtainable and can be carried out under normal pressure conditions, which is suitable for industrial production.

[0022] The double-network hydrogel / sponge dressing of β-chitin crosslinked hyaluronic acid provided by the present invention is expected to be applied to the field of researching biomedical materials or preparing wound care medical devices.

[0023] To more clearly illustrate the structural features and effects of the present invention, the present invention will be described in detail below with reference to the drawings and specific embodiments. Description of the Drawings

[0024] Figure 1 It is a schematic diagram of the healing condition of the back wound of diabetic rats in the invention embodiment.

[0025] Figure 2 It is a schematic diagram of HE and Masson staining in the invention embodiment. Detailed Embodiments

[0026] In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the drawings and embodiments. 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.

[0027] The following describes the specific implementation of the present invention in detail with specific embodiments.

[0028] In one embodiment, a preparation method of a hyaluronic acid double-network hydrogel and sponge dressing is as follows, see Figures 1 to 2 , including the following steps:

[0029] Step 1: Use wet squid pen, dissolve it in sodium hydroxide solution, then filter after high-temperature reaction for 120 min, and wash the filtrate with deionized water until the pH value is neutral;

[0030] Step 2: Add dilute hydrochloric acid and react at room temperature for 24 hours. After filtration, washing, and washing the filter residue with deionized water until the pH value is neutral, wet β-chitin is obtained;

[0031] Step 3: Add hyaluronic acid and crosslinking agent glutaraldehyde to the ultrasonically dispersed β-chitin fiber dispersion liquid. After crosslinking, freezing, and demolding, low-temperature solvent replacement is carried out, and freeze-drying is performed to obtain a sponge;

[0032] Step 4: Adjust the ratio of glutaraldehyde to β-chitin and hyaluronic acid to obtain sponge dressings with different properties.

[0033] Further, refer to Figures 1 to 2 , in Step 1, dissolve with a sodium hydroxide solution with a concentration of 1 mol / L, react at a high temperature of 90 °C for 120 min, and then filter.

[0034] Further, refer to Figures 1 to 2 , in Step 2, add 0.1 mol / L dilute hydrochloric acid.

[0035] Further, refer to Figures 1 to 2 , in Step 3, the concentration of the β-chitin fiber dispersion is 0.5 wt%, and the concentration of hyaluronic acid is 0.5%.

[0036] Further, refer to Figures 1 to 2 , after being uniformly dispersed in Step 3, inject into a mold for crosslinking for 24 hours, and demold after freezing at -75 °C.

[0037] Further, refer to Figures 1 to 2 , after crosslinking, freezing and demolding in Step 3, place it in pre-cooled absolute ethanol for low-temperature replacement, then wash off the ethanol with deionized water, and freeze-dry to obtain a sponge.

[0038] Further, refer to Figures 1 to 2 , the molecular formula of the hyaluronic acid is (C 14 H 21 NO 11 ), where n represents the degree of polymerization, that is, the number of disaccharide units in the hyaluronic acid molecule.

[0039] In this embodiment, the present invention provides a novel double-network hydrogel / sponge dressing of β-chitin crosslinked hyaluronic acid, where the molecular formula of the hyaluronic acid is (C 14 H 21 NO 11 ), where n represents the degree of polymerization, that is, the number of disaccharide units in the hyaluronic acid molecule, and the molecular structure is shown in Formula (I). The molecular formula of β-chitin is (C 8 H 15 NO 6 ), and the molecular structure is shown in Formula (II). The molecular structure of β-chitin crosslinked hyaluronic acid is shown in Formula (III).

[0040] ;

[0041] Formula (I).

[0042] ;

[0043] Formula (II).

[0044] ;

[0045] Formula (III).

[0046] The present invention provides a method for preparing the above-mentioned β-chitin crosslinked hyaluronic acid double-network hydrogel / sponge dressing, comprising the following steps:

[0047] (1) Preparation of β-chitin: Using wet squid pen, dissolving it with a sodium hydroxide solution with a concentration of 1 mol / L, reacting at a high temperature of 90 °C for 120 min, and then filtering. The filtrate is washed with deionized water until the pH value is neutral. Then, 0.1 mol / L dilute hydrochloric acid is added and reacted at room temperature for 24 hours. Filter, wash, and the filter residue is washed with deionized water until the pH value is neutral to obtain wet β-chitin.

[0048] (2) Crosslinking of β-chitin and hyaluronic acid: Add 0.5% hyaluronic acid and crosslinking agent glutaraldehyde to the ultrasonically dispersed β-chitin fiber dispersion (0.5 wt%). After dispersing evenly, inject it into a mold and crosslink for 24 hours. Demold after freezing at -75 °C, place it in pre-cooled absolute ethanol for low-temperature replacement, and then wash off the ethanol with deionized water and freeze-dry to obtain a sponge.

[0049] (3) By adjusting the ratio of glutaraldehyde to β-chitin and hyaluronic acid, sponge dressings with different properties can be obtained.

[0050] In the third aspect, the present invention provides an application of the above-mentioned β-chitin crosslinked hyaluronic acid hydrogel / foam dressing in the preparation of biomedical materials or medical devices for the care of diabetic ulcers and chronic wounds.

[0051] From Figure 1 it is shown that the β-chitin / hyaluronic acid double-network sponge dressing has an obvious effect of promoting the healing of the back wounds of rats, and the effect is better than that of the drug-loaded group.

[0052] From Figure 1 it is shown that HE and Masson staining confirm that the β-chitin / hyaluronic acid double-network sponge dressing has the effect of promoting the growth of the back wound tissue of diabetic rats and increasing collagen. (1) Establishment of a back wound model of diabetic rats; (2) Blank control in part A, use of β-chitin / hyaluronic acid double-network sponge dressing in part B, use of iodophor disinfection in part C, and use of drug-loaded β-chitin / hyaluronic acid double-network sponge dressing in part D; (3) Healing situation of the back wounds of rats 8 days after surgery

[0053] From Figure 2 it is shown that A is the blank control group, B is the iodophor disinfection group, C is the drug-loaded β-chitin / hyaluronic acid double-network sponge dressing group, and D is the β-chitin / hyaluronic acid double-network sponge dressing group. The results of HE staining and Masson staining suggest the effects of promoting wound healing and collagen hyperplasia (D > C > B > A).

[0054] The working principle of the present invention is:

[0055] A double-network hydrogel / sponge dressing of β-chitin crosslinked hyaluronic acid is provided. The double-network hydrogel / sponge dressing of β-chitin crosslinked hyaluronic acid has a good effect of promoting tissue growth and has broad application prospects as a biomedical antibacterial material.

[0056] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, and improvements made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for preparing a hyaluronic acid double network hydrogel and a sponge dressing, characterized in that: The following steps are involved: Step 1: using a wet squid parietal bone, dissolving it in a sodium hydroxide solution, reacting it at high temperature for 120 minutes, filtering it, and washing the filtrate with deionized water until the pH value is neutral; Step 2: adding dilute hydrochloric acid, reacting at room temperature for 24 hours, filtering, washing and cleaning the residue with deionized water until the pH value is neutral, to obtain wet β-chitosan; Step 3: adding hyaluronic acid and cross-linking agent glutaraldehyde to the ultrasonically dispersed β-chitin fiber dispersion, performing low-temperature solvent replacement after cross-linking, freezing and demoulding, and freeze-drying to obtain a sponge; Step 4: Adjust the ratio of glutaraldehyde to β-chitin and hyaluronic acid to obtain sponge dressings with different properties.

2. The method for preparing the hyaluronic acid double network hydrogel and sponge dressing according to claim 1, characterized in that: In the step 1, a sodium hydroxide solution with a concentration of 1 mol / L is used for dissolving and reacting at a high temperature of 90° C. for 120 min and then filtering.

3. The method for preparing the hyaluronic acid double network hydrogel and sponge dressing according to claim 2, characterized in that: In the step 2, 0.1 mol / L dilute hydrochloric acid is added.

4. The method for preparing the hyaluronic acid double network hydrogel and sponge dressing according to claim 3, characterized in that: In the step 3, the concentration of the β-chitin fiber dispersion is 0.5wt%, and the concentration of the hyaluronic acid is 0.5%.

5. The method for preparing the hyaluronic acid double network hydrogel and sponge dressing according to claim 4, characterized in that: After being evenly dispersed in step 3, the mixture is injected into a mold and cross-linked for 24 hours, and then frozen and demoulded at -75°C.

6. The method for preparing the hyaluronic acid double network hydrogel and sponge dressing according to claim 5, characterized in that: In the step 3, after cross-linking, freezing and demoulding, the sponge is placed in pre-cooled anhydrous ethanol for low-temperature replacement, and then the ethanol is washed out with deionized water, and freeze-dried to obtain a sponge.

7. The method for preparing the hyaluronic acid double network hydrogel and sponge dressing according to claim 6, characterized in that: The hyaluronic acid molecular formula (C 14 H 21 NO 11 )n, where n represents the degree of polymerization, that is, the number of disaccharide units in the hyaluronic acid molecule.

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