CN116115556A - Physical cross-linked hyaluronic acid hydrogel and application thereof in radioactive skin injury treatment - Google Patents
Physical cross-linked hyaluronic acid hydrogel and application thereof in radioactive skin injury treatment Download PDFInfo
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- CN116115556A CN116115556A CN202211581222.8A CN202211581222A CN116115556A CN 116115556 A CN116115556 A CN 116115556A CN 202211581222 A CN202211581222 A CN 202211581222A CN 116115556 A CN116115556 A CN 116115556A
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- hyaluronic acid
- acid hydrogel
- hydrogel
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Abstract
The invention relates to a physical cross-linked hyaluronic acid hydrogel and application thereof in the treatment of radioactive skin injury. The hydrogel material for promoting radioactive skin injury wound repair, which is prepared by the invention, has good biocompatibility and excellent anti-inflammatory performance, can maintain pharmacological activity of the medicine, and has good promotion effect on wound healing.
Description
Technical Field
The invention relates to the technical field of biomedical materials, in particular to a physical cross-linked hyaluronic acid hydrogel and application thereof in radioactive skin injury treatment.
Background
Wound and wound healing are important problems in the surgical field. Specific types of wounds need targeted treatment to achieve healing of wounds that would otherwise not be healable, or to accelerate healing of wounds that would otherwise require long-term healing. With the progress of production and life, radiation is an indispensable part in the fields of medicine, industry and the like, and damage caused by ionizing radiation to a human body is also a problem to be solved urgently, wherein radioactive skin damage is one of the most frequently occurring body damage after radiation exposure. More than half of malignant tumor patients receiving radiotherapy can have radioactive skin injury at a certain moment, especially in cancers of breast, perineum and head and neck, and the skin is also included in the target range of radiotherapy, so that acute and chronic skin injury generated after treatment is more remarkable. The soft tissue of the irradiated area is fibrosed, the skin gradually loses normal elasticity, the original sebaceous gland secretion function and the skin protection function are damaged, the skin is dry, thinned, the blood vessel and lymphatic vessel wall is thickened and even blocked, and the healing capacity of the skin is reduced. The large amount of active oxygen free radicals and inflammatory mediators generated by radiation make normal cell physiological activities and protein and lipid metabolism difficult to normally progress, such as increased apoptosis of endothelial cells and fibroblasts, neovascular disorders, cell division and differentiation, cell cycle retardation and the like, so that skin injury caused by radiation exposure is often very difficult to heal.
Unlike the general wound healing process, oxidative stress and inflammatory reaction of radiation injury caused by radiation exposure are heavy, a large amount of Reactive Oxygen Species (ROS) are generated in cells in the acute phase after radiation, the ROS accumulated in the cells destroy cell structures, directly cause cell injury and initiate aggregation of inflammatory cells, and further enter the long-term inflammatory phase, the chronic inflammatory reaction and fibrosis process lead to the fact that the radioactive wound is not healed for a long time by activating fibroblasts by transforming factor beta 1 (TGF-beta 1), and in addition, the sensitivity of the radioactive wound to treatment is lower than that of the general wound due to irreversible DNA fracture, excessive extracellular matrix deposition and abnormal cell cycle caused by radiation. At present, no standard guideline for treating radioactive injury is available at home and abroad, and most of treatment methods for radioactive skin injury have poor curative effects, and some of the treatment methods can even produce larger side effects. Common treatment methods are mainly based on anti-inflammatory, antioxidant, anti-infective, angiogenesis promoting, growth factor supplementing, wet healing environment providing and other aspects, but have limited therapeutic effects. Although widely used in wound surfaces, growth factors which can produce an accelerating healing effect in general wound surfaces can not produce a remarkable effect when being subjected to radioactive ulcer, tissues in the wound surfaces of the radioactive ulcer are inert, insensitive to treatment, the course of the disease is repeated, and medicines with anti-inflammatory effects such as retinoic acid and the like are used for treating skin diseases, and although the medicines have a treatment effect on common skin injuries such as white spots, acne vulgaris and the like, the skin surfaces are often additionally damaged due to the skin irritation of the medicines when the radioactive skin injuries are treated. Treatment of radiation skin lesions is a lengthy process, the lesions after irradiation are divided into acute and chronic phases, and in the acute phase within six months, skin pathology may occur from erythema of degree I, hair follicle lesions, to pigmentation, blisters, and even necrotic ulcers of degree IV, whereas long-term lesions over six months appear as atrophic dermatitis, fibrosis, and chronic ulcers. Acute radiodermatitis resembles a burning lesion, and lighter people may regress after several weeks, however if the underlying lesion is not appreciated and treated, the reaction may persist and cause complications that have a significant negative impact on the quality of life of the patient.
In the end of the 20 th century, along with rapid development of tissue engineering, a plurality of biological materials are widely applied in the medical field, and particularly in the aspect of wound healing, the biological materials have the advantages of good cell biocompatibility, degradability, small toxic and side effects, certain bioactivity of promoting repair and the like. Hyaluronic acid is widely used as macromolecular polysaccharide in organisms, is easy for large-scale production due to good biocompatibility, has the functions of regulating osmotic pressure, maintaining tissue morphology, lubricating, buffering stress and the like, and is applied to the biomedical field. In order to meet the practical application, the hyaluronic acid is often required to be constructed into a three-dimensional reticular gel structure in the preparation process, and the hyaluronic acid is often required to be modified by a chemical crosslinking agent during construction so as to be gelled, so that the gel with various mechanical properties and controllable degradation time is obtained. The chemical crosslinking method improves the gel performance of the hyaluronic acid, but the use of the crosslinking agent has potential cytotoxicity, causes local side effects, and reduces the application of the crosslinking agent in the conditions of difficult-to-heal wound surfaces, inflammation and the like. Therefore, development of a green synthetic and non-cytotoxic hydrogel for the treatment of radiation-induced skin lesions has become extremely urgent.
Disclosure of Invention
In order to solve the technical problems, the invention provides a physical crosslinking hyaluronic acid hydrogel of a compound micromolecule drug deferoxamine and retinoic acid, which is used for a tissue repair material for treating radioactive skin injury, and the influence of a chemical crosslinking agent on the biocompatibility of the material is reduced to the greatest extent.
It is a first object of the present invention to provide a physically cross-linked hyaluronic acid hydrogel obtained by dispersing deferoxamine and retinoic acid in a physically cross-linked hyaluronic acid hydrogel by freeze thawing method.
Further, the freezing and thawing method comprises the steps of freezing at the temperature of-25 to-15 ℃ and thawing at the temperature of 1 to 10 ℃.
The second object of the present invention is to provide a method for preparing the above-mentioned physically cross-linked hyaluronic acid hydrogel, comprising the steps of:
s1, preparing hyaluronic acid aqueous solution into gel by a freeze thawing method under an acidic condition;
s2, placing the gel obtained in the step S1 in a buffer system with the pH value of 7-8, then placing the gel in water until the pH value of an aqueous solution rises to 6.8-7.2, preparing neutral gel, adding deferoxamine and retinoic acid into the neutral gel, and obtaining the physical cross-linked hyaluronic acid hydrogel through a freeze thawing method.
Further, the molecular weight of the hyaluronic acid is 89-150 ten thousand.
Further, in step S1, the acidic condition is to adjust the pH to 1-3 with hydrochloric acid.
Further, the concentration of the hydrochloric acid is 0.5-1.5mol/L.
Further, in step S2, the concentration of deferoxamine is 50-70umol/L.
Further, in step S2, the retinoic acid has a concentration of 40-60umol/L.
The third object of the invention is to provide the application of the physical cross-linked hyaluronic acid hydrogel in preparing a medicine for treating radioactive skin injury.
In the invention, the hydrogel for promoting the repair of the radioactive skin injury is prepared by using hyaluronic acid through a freeze thawing method under the condition of no external cross-linking agent. In preliminary experiments, the applicant finds that the combination of retinoic acid and deferoxamine, the preparation of silk fibroin hydrogel by adopting a physical method of temperature concentration regulation, the preparation of drug-loaded hyaluronic acid hydrogel by adopting other physical methods and the like are not ideal in the treatment of radioactive skin injury, and the drug-loaded hyaluronic acid hydrogel prepared by using a freeze thawing method only can effectively treat radioactive skin injury, can maintain pharmacological activity of drugs, does not influence cell activity and has good promotion effect on wound healing.
The hyaluronic acid is selected as a gel matrix, so that not only is the hyaluronic acid a substance naturally existing in a human body, but also the harmful product is not generated, the inflammatory reaction caused by materials and the potential cytotoxicity caused by chemical substance residues can be minimized, the hyaluronic acid has a full water-retaining effect, and the negative ion repulsive effect formed by carboxyl in a disaccharide unit can enable the hyaluronic acid to be combined with more than 100 times of water, so that a good environment is provided for the healing of radioactive wound surfaces. (2) Compared with other physical modes, the freeze thawing method has the advantages of simple preparation process, low cost, safety and stability, small influence on the characteristics of the material, full play of the auxiliary treatment effect of the matrix material, freezing of the solvent in the homogeneous solution through the freeze thawing process, formation of discrete physical crosslinking points by hydrogen bonds between molecular chains of a solute region, formation of three-dimensional reticular gel under the action of non-covalent bonds, good water retention performance, injectability and spreadability. (3) Retinoic acid is a derivative of vitamin a, and can promote skin regeneration and repair by means of anti-inflammatory, fibroblast proliferation promoting, hair follicle regeneration promoting and other skin attachment regeneration promoting. However, retinoic acid has been found to be irritating when applied to wounds, causing localized redness and loss of epidermis, the specific mechanism of which is not yet defined; the effect of deferoxamine on the repair of radioactive skin lesions is not reported. The applicant tests physical hyaluronic acid hydrogel of the composite RA and DFO in a pre-experiment by using a skin drug stimulation experiment on the back of the rabbit, and scores the skin damage degree of the rabbit, and the result shows that no obvious stimulation exists. The hyaluronic acid hydrogel loaded with micromolecule drugs deferoxamine and retinoic acid is prepared in a safe and mild physical crosslinking mode, the three-dimensional network structure of the hydrogel can protect media, the double bonds in the retinoic acid are reduced to be oxidized, the media are uniformly dispersed, the toxicity of local high-concentration drugs and the irritation of pure drugs are reduced, and the hyaluronic acid hydrogel has remarkable effects on the treatment of special wound surfaces.
A fourth object of the present invention is to provide a radioactive skin damage treatment drug comprising the above-mentioned physically crosslinked hyaluronic acid hydrogel.
Further, the physically cross-linked hyaluronic acid hydrogel is in an injectable formulation.
By means of the scheme, the invention has at least the following advantages:
aiming at the key problem of poor treatment effect on radioactive skin injury at present, the invention provides a physical cross-linked hyaluronic acid hydrogel of a compound small molecular drug deferoxamine and retinoic acid, which is used as a tissue repair material for treating radioactive skin injury. The physical crosslinking hyaluronic acid hydrogel has good biocompatibility, is suitable for serving as a carrier of micromolecular drugs deferoxamine and retinoic acid, can promote the healing of radioactive skin ulcers under the condition of maintaining the pharmacological activity of the drugs, and has good promotion effect on wound vascularization and inflammation control. The invention also expands biomedical application of the hyaluronic acid material in the field of special wound surfaces, and provides a new method for preparing the hyaluronic acid material with better biocompatibility.
The foregoing description is only an overview of the present invention, and is presented in terms of preferred embodiments of the present invention and the following detailed description of the invention in conjunction with the accompanying drawings.
Drawings
In order that the contents of the present invention may be more clearly understood, the present invention will be further described in detail with reference to specific embodiments thereof with reference to the accompanying drawings.
FIG. 1 is a schematic illustration of a physically cross-linked hydrogel of hyaluronic acid with complex deferoxamine and retinoic acid;
FIG. 2 is a schematic illustration of a bolus injection of a physically cross-linked hyaluronic acid hydrogel;
FIG. 3 is a diagram of the microscopic morphology of a physically cross-linked hyaluronic acid hydrogel;
FIG. 4 is a Raman spectrum of a physically cross-linked hyaluronic acid hydrogel;
FIG. 5 is a graph showing the effect of deferoxamine and retinoic acid on cell growth after irradiation injury;
FIG. 6 is a fluorescence image of cell proliferation;
FIG. 7 shows the results of a cell clone formation assay;
FIG. 8 is a wound healing image;
FIG. 9 is an evaluation of the vascular function promoting effect of hyaluronic acid hydrogel loaded with deferoxamine and retinoic acid on endothelial cells, and an evaluation of the ability of silk fibroin hydrogel to improve cellular vascular function under the same conditions in comparative example;
FIG. 10 is a functional evaluation of the frozen thawing method hyaluronic acid hydrogel for enhancing the migration ability of cells after irradiation, and an evaluation of the photo-crosslinking method hyaluronic acid hydrogel for promoting the migration ability of cells after irradiation in a comparative example;
wherein p <0.05 represents a ratio compared to the photocrosslinked hydrogel; # represents a comparison with the blank group.
Detailed Description
The present invention will be further described with reference to the accompanying drawings and specific examples, which are not intended to be limiting, so that those skilled in the art will better understand the invention and practice it.
In the following examples, except for the specific description, irradiation was all X-rays, single irradiation was adopted, irradiation intensity unit was Gy, the irradiated area was placed on an irradiation plate, local irradiation was performed with 4Mev electron beam generated by a linear accelerator, the non-irradiated area was shielded with a lead plate, and the absorption dose rate was 598MU/min.
Example 1
(1) Preparation of hyaluronic acid hydrogel under acidic conditions
The Macklin company sodium hyaluronate powder has a molecular weight of about 100 ten thousand, and is dissolved in pure water at room temperature at a concentration of 10mg/ml, magnetically stirred at 300rpm until completely dissolved, pH adjusted to 2 with 1mol/L hydrochloric acid, centrifuged twice at 4000r/min for 20 minutes, and air bubbles removed. The solution was frozen at-20℃for 7 days and thawed at 4℃for 12 hours.
(2) Physical cross-linking type hyaluronic acid hydrogel loaded deferoxamine and retinoic acid
The gel was placed in phosphate buffer at pH7 for 4 hours and then replaced with deionized water until the pH of the aqueous solution increased to 7 to obtain hyaluronic acid hydrogel. Adding deferoxamine with the concentration of 60umol/L and retinoic acid with the concentration of 50umol/L into the hyaluronic acid hydrogel, physically mixing, freezing for 7 days at the temperature of minus 20 ℃ and thawing for 12 hours at the temperature of 4 ℃ to obtain the sample, namely the physical crosslinking hyaluronic acid hydrogel of the compound micromolecular drug deferoxamine and retinoic acid.
Example 2
(1) Preparation of hyaluronic acid hydrogel under acidic conditions
The Macklin company sodium hyaluronate powder has a molecular weight of about 150 ten thousand, and is dissolved in pure water at room temperature at a concentration of 10mg/ml, magnetically stirred at 300rpm until completely dissolved, pH adjusted to 2 with 0.5mol/L hydrochloric acid, and centrifuged twice at 4000r/min for 20 minutes to remove air bubbles. The solution was frozen at-15℃for 5 days and thawed at 10℃for 10 hours.
(2) Physical cross-linking type hyaluronic acid hydrogel loaded deferoxamine and retinoic acid
The gel was placed in phosphate buffer at pH8 for 3 hours and then replaced with deionized water until the pH of the aqueous solution increased to 7 to obtain hyaluronic acid hydrogel. Adding deferoxamine with the concentration of 50umol/L and retinoic acid with the concentration of 60umol/L into the hyaluronic acid hydrogel, physically mixing, freezing at-15 ℃ for 6 days, and thawing at 7 ℃ for 12 hours to obtain a sample, namely the physical crosslinking hyaluronic acid hydrogel of the compound micromolecular drug deferoxamine and retinoic acid.
Example 3
(1) Preparation of hyaluronic acid hydrogel under acidic conditions
The Macklin company sodium hyaluronate powder has a molecular weight of about 89 ten thousand, and is dissolved in pure water at room temperature at a concentration of 10mg/ml, magnetically stirred at 300rpm until completely dissolved, pH adjusted to 1 with 1.5mol/L hydrochloric acid, centrifuged twice at 4000r/min for 20 minutes, and air bubbles removed. The solution was frozen at-25℃for 4 days and thawed at 1℃for 12 hours.
(2) Physical cross-linking type hyaluronic acid hydrogel loaded deferoxamine and retinoic acid
The gel was placed in phosphate buffer at pH7 for 4 hours and then replaced with deionized water until the pH of the aqueous solution increased to 7 to obtain hyaluronic acid hydrogel. Adding deferoxamine with the concentration of 40umol/L and retinoic acid with the concentration of 70umol/L into the hyaluronic acid hydrogel, physically mixing, freezing for 7 days at the temperature of 25 ℃ below zero, and thawing for 12 hours at the temperature of 3 ℃ to obtain a sample, namely the physical crosslinking hyaluronic acid hydrogel of the compound micromolecular drug deferoxamine and retinoic acid.
Example 4
(1) Physical crosslinking type hyaluronic acid hydrogel Performance test of Complex deferoxamine and Retinoic acid (in this example, the gel prepared in example 1 was used for the test)
The hydrogel was tested for injectability before and after drug loading by injecting with a 1ml syringe with and without a needle (23G). The results are shown in FIG. 2.
(2) Physical crosslinking type hyaluronic acid hydrogel scanning electron microscope test of compound deferoxamine and retinoic acid
The hydrogel prepared in example 1 was freeze-dried, and then a cube with a side length of about 10mm was taken, attached to an electronic microscope bench by a conductive adhesive, subjected to platinum spraying treatment (10 ma, 90S), and tested by using an S-4800 type scanning electron microscope with a test voltage of 3kV, and the microstructure of the physical cross-linked hyaluronic acid gel was collected. The result is shown in FIG. 3, and it can be seen that the microscopic morphology of the hyaluronic acid hydrogel prepared by the method is a long fiber network structure formed by mutually crosslinking bundles.
(3) Raman spectrum of physical cross-linked hyaluronic acid hydrogel of composite deferoxamine and retinoic acid
Correction Raman spectrometer at 520.7cm -1 The spectrum peak of the monocrystalline silicon wafer is used as a benchmark, unified parameters are set, HA and DFO-RA-HA are tested, spectra of different substances are collected, the obtained spectra are subjected to statistical treatment by Matlab software, and NGSLabSpec software is used for smoothing and correcting the baselines for further observation and statistics. The results are shown in FIG. 4. The DFO is 1050cm in the Raman spectrum of the composite aqueous gel system -1 Characteristic peak, RA at 1580cm -1 Characteristic peaks at 667 and 945cm -1 The absorption peak typical of HA.
(4) Detection of growth influence of deferoxamine and retinoic acid on irradiation damage type endothelial cells
And testing cells 24h, 48h and 72h after intervention by adopting a cell proliferation experiment, measuring the optical density value of each hole at the wavelength of 450nm by using an enzyme-labeled instrument, and calculating the cell survival condition. Deferoxamine and retinoic acid concentrations suitable for cell growth following radiation injury are measured.
The results are shown in FIGS. 5 and 6. In CCK-8 experiment, free DFO has obvious cytotoxicity to cells damaged by irradiation, the toxicity increases with the increase of the concentration of DFO, when the free DFO is compounded with hyaluronic acid hydrogel, the cytotoxicity of the medicine is relieved under the same condition, and at the concentration of 60uM, endothelial cells have strong angiopoiesis and wound healing promoting capability, so that the concentration of DFO is preferably 60uM, and RA has no obvious cytotoxicity to HUVECs. For screening RA drug concentration, cells after HA-RA intervention irradiation were selected for EdU cell proliferation experiments, the hollow white control in FIG. 6 was PBS group, the HA group was pure HA gel group, and the remaining three groups were HA hydrogels of complex retinoic acid, wherein RA concentrations were different. Under-the-lens EdU positive cells were observed to increase with increasing RA concentration, proliferation capacity was enhanced, and cell proliferation was increased, but at a concentration of 100uM, proliferation was less than 50uM, so 50uM was preferred as RA concentration. Experiments were performed at this preferred concentration as follows.
(5) Influence of physical crosslinking type hyaluronic acid hydrogel of composite deferoxamine and retinoic acid on growth of irradiation damage type HUVECs (human immunodeficiency Virus)
The effect of the fluorescent staining on the biological behavior of the cells was tested using a cell clone formation experiment. The results are shown in FIG. 7. In fig. 7, the control group is a pure HA gel group, and the drug-loaded group is the HA hydrogel of the complex deferoxamine and retinoic acid prepared in example 1.
As can be seen, in the clone formation experiment of FIG. 7, the number of cell clone formation in the low dose irradiation group (2-4 Gy) was not significantly different between the simple HA control group and the drug-loaded group, but the clone formation rate in the high dose irradiation group (6-10 Gy) was higher than that in the control group.
(6) Experiment for promoting healing of radioactive skin ulcer
And constructing a radioactive skin injury model of the rat as a test object, and testing the treatment effect of the physical crosslinking hyaluronic acid hydrogel of the composite deferoxamine and retinoic acid on the actual radioactive skin injury, wherein a control group is a normal saline group and a drug group is independently used. The back shaving area of the rat after anesthesia is irradiated by electron beam of 45Gy, and the irradiation area is about 45mm multiplied by 50mm. In the blank control group, the 60uM DFO-HA group, the 50uM RA-HA group and the DFO-RA-HA group, the skin can be observed to have dehairing and erythema after irradiation for 2 weeks, the skin tissue erosion necrosis manifestations with different degrees appear about 30 days, the radioactive ulcer wound surface is formed, and the healing condition of each wound surface with different degrees is observed at 45 days.
As a result, as shown in fig. 8, early dermatitis symptoms, local hair loss, erythema and desquamation appear gradually in each group at 15 days of irradiation, ulcer wounds appear obviously in the control group at 30 days, the DFO-HA group and RA-HA group have smaller ulcer wounds, healing trend is shown, the combined drug-loaded gel group HAs no hair follicle regeneration, but only forms tiny wounds, the combined treatment group HAs been completely epithelialized at 45 days, and new hair follicles appear, the blank group HAs larger ulcer wounds, slow healing trend is shown, and the other two groups of ulcers heal to different degrees. The combined medicine carrying group HAs better effect of promoting wound healing than the independent medicine carrying group, which proves that the DFO-RA-HA gel can effectively protect against radioactive skin injury, accelerate wound healing and protect skin accessory organs.
Comparative example 1
Silk fibroin nanofiber hydrogel (SF) is prepared by a physical method, specifically a cooling-heating-concentrating method, the concentration of the obtained silk fibroin hydrogel is 1%, the same doses of DFO and RA are compounded, the endothelial cell tube forming capacity of the compound gel is tested, a blank control group is PBS test solution, and the result is shown in figure 9.
As can be seen from fig. 9, the silk fibroin nanofiber hydrogel of the composite DFO and RA HAs a remarkable promotion effect on the vascularization ability of endothelial cells, multiple nodes are formed among endothelial cells, incomplete lumen formation is seen, while the HA hydrogel of the composite DFO and RA forms a complete lumen structure within 8 hours, the tube formation length is longer and the cell proliferation condition is remarkably superior to that of the silk fibroin nanofiber hydrogel.
Comparative example 2
The physical method of photocrosslinking is used for treating hyaluronic acid, in particular to the method that methacrylic acid is crosslinked under ultraviolet light to prepare Hydrogel (HAMA). The effect of the comparative material on the migration ability of HDF-alpha cells was examined by cell scratch experiments.
The results show that for the irradiated HDF-alpha, the physically cross-linked hyaluronic acid hydrogel can generate the capability of promoting cell movement, but compared with HAMA, the scratch area heals more in 8 hours after the gel is used by a freeze thawing method, the migration promoting capability is better, and the capability of promoting the migration of the fibroblast to cover the wound surface is stronger in the wound surface healing and granulation tissue forming processes of the physical gel by the freeze thawing method.
In conclusion, the hyaluronic acid hydrogel prepared by adopting a physical crosslinking method reduces the influence of a chemical crosslinking agent on the biocompatibility of the material to the greatest extent, and simultaneously proposes to apply the hyaluronic acid hydrogel to radioactive skin injury on the basis of moisturizing and preserving water, and compound a small molecular medicine with the effects of promoting angiogenesis and resisting inflammation.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations and modifications of the present invention will be apparent to those of ordinary skill in the art in light of the foregoing description. It is not necessary here nor is it exhaustive of all embodiments. And obvious variations or modifications thereof are contemplated as falling within the scope of the present invention.
Claims (10)
1. A physically cross-linked hyaluronic acid hydrogel, characterized in that: the physical cross-linked hyaluronic acid hydrogel is obtained by dispersing deferoxamine and retinoic acid in hyaluronic acid hydrogel which is cross-linked by a physical mode, wherein the physical mode is a freeze thawing method.
2. The physically cross-linked hyaluronic acid hydrogel according to claim 1, characterized in that: the freezing and thawing method is to freeze at-25 to-15 ℃ and then thaw at 1 to 10 ℃.
3. The physically cross-linked hyaluronic acid hydrogel according to claim 1, characterized in that: the molecular weight of the hyaluronic acid is 89-150 ten thousand.
4. A method for preparing a physically cross-linked hyaluronic acid hydrogel according to any of claims 1-3, comprising the steps of:
s1, preparing hyaluronic acid aqueous solution into gel by a freeze thawing method under an acidic condition;
s2, placing the gel obtained in the step S1 in a buffer system with the pH value of 7-8, then placing the gel in water until the pH value of an aqueous solution rises to 6.8-7.2, preparing neutral gel, adding deferoxamine and retinoic acid into the neutral gel, and obtaining the physical cross-linked hyaluronic acid hydrogel through a freeze thawing method.
5. The method of manufacturing according to claim 4, wherein: in step S1, the acidic condition is to adjust the pH to 1-3 with hydrochloric acid.
6. The method of manufacturing according to claim 4, wherein: in step S2, the concentration of deferoxamine is 50-70umol/L.
7. The method of manufacturing according to claim 4, wherein: in step S2, the concentration of retinoic acid is 40-60umol/L.
8. Use of a physically cross-linked hyaluronic acid hydrogel according to any of claims 1-3 for the manufacture of a medicament for the treatment of radiation-induced skin lesions.
9. The use according to claim 8, characterized in that: the physical cross-linked hyaluronic acid hydrogel is in an injectable formulation.
10. A radioactive skin injury treatment drug, characterized in that: the radioactive skin damage treatment drug comprising the physically cross-linked hyaluronic acid hydrogel of any of claims 1-3.
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| CN110038161A (en) * | 2019-04-29 | 2019-07-23 | 上海交通大学 | A kind of novel transparent matter acid flow-gel and its preparation method and application |
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