Hydrogel for promoting radiation combined wound repair and preparation method thereof

Abstract

The present invention relates to a hydrogel for promoting radiation combined wound repair and a preparation method thereof, the hydrogel comprises: a carrier, the raw materials of which comprise a first carrier raw material and a second carrier raw material, the mass ratio of the first carrier raw material to the second carrier raw material being 1: 2-1: 4; the load is loaded on the carrier, the load comprises CBD, and the loading capacity of the CBD is 0.04%-0.5%.

Description

Technical Field

[0001] The present invention relates to the technical field of wound repair, and in particular to a hydrogel that promotes the repair of radiation-combined wounds and a preparation method thereof. Background Art

[0002] Radiation-associated trauma is a complex and challenging injury type, often occurring during radiotherapy or after accidental radiation exposure, with the development of open wounds. It typically manifests as skin and soft tissue damage, accompanied by inflammation and difficulty healing. Current treatments for radiation-associated trauma primarily include conventional symptomatic care, including local wound care, antibiotics, and wound moisturization. However, these treatments lack antioxidant or anti-inflammatory properties, resulting in limited wound repair and difficulty promoting deeper repair. Summary of the Invention

[0003] In view of the above analysis, the embodiments of the present invention aim to provide a hydrogel that promotes the repair of radiation-induced trauma and a preparation method thereof, so as to at least solve one of the above problems.

[0004] In one aspect, the present invention provides a hydrogel for promoting the repair of radiation-induced wounds, comprising:

[0005] The carrier, wherein the raw materials thereof include a first carrier raw material and a second carrier raw material, and the mass ratio of the first carrier raw material to the second carrier raw material is 1:2 to 1:4;

[0006] A load is loaded on the carrier, wherein the load includes CBD, and the loading amount of the CBD is 0.04% to 0.5%.

[0007] Furthermore, the first carrier raw material is CMC-Na, and the second carrier raw material is CHT;

[0008] The particle size of the CMC-Na is 20 to 200 μm, and the specific surface area is 2 to 10 m 2 / g, volume is 0.01~0.05cm 3 / g;

[0009] The particle size of CHT is 50-200 μm, and the specific surface area is 5-50 m 2 / g, volume is 0.05~0.2cm 3 / g.

[0010] On the other hand, the present invention also provides a method for preparing a hydrogel that promotes the repair of radiation-induced wounds, which can at least be used to prepare the above-mentioned hydrogel, and the preparation method comprises the following steps:

[0011] S100: Obtaining a first carrier raw material powder or solution, a second carrier raw material powder or solution, and a load solution;

[0012] S200: mixing the first carrier raw material solution and the loading material solution at a volume ratio of 25:1 to 100:1, adding the second carrier raw material powder at a mass volume ratio of 1:25 to 1:10 after uniform mixing, stirring until the second carrier raw material powder is completely dissolved, and then removing bubbles by centrifugation to obtain a hydrogel that promotes radiation-combined wound repair;

[0013] Alternatively, the second carrier raw material solution and the loading material solution are mixed at a volume ratio of 1:4 to 1:1. After uniform mixing, the first carrier raw material powder is added at a mass-to-volume ratio of 1:25 to 1:100. The mixture is stirred until the first carrier raw material powder is completely dissolved. The air bubbles are then removed by centrifugation to obtain a hydrogel that promotes radiation-induced wound repair. This method produces a hydrogel with a higher hardness than the first method.

[0014] Furthermore, in step S100, the first carrier raw material is dissolved in a first solution compatible therewith to obtain a first carrier raw material solution with a mass fraction of 1% to 3%;

[0015] When the first carrier raw material is CMC-Na, the corresponding first solution is an acidic aqueous solution with a pH of ≤3 and an ionic strength of ≤0.1M.

[0016] Furthermore, in step S100, the particle size of the second carrier raw material powder is 50 to 100 μm;

[0017] When the second carrier raw material is CHT, the CHT raw material is first subjected to a secondary deacetylation treatment, and the treatment method is as follows:

[0018] The CHT raw material was dissolved in 30% NaOH at a mass-to-volume ratio of 1:20, and the solution was then reacted at 80°C for 6 hours. After the reaction, it was rinsed with distilled water several times until neutral, and the solid was collected by centrifugation. Pure CHT powder was obtained by vacuum freeze-drying or drying at 105°C.

[0019] Furthermore, in step S100, the load is dissolved in a load solution compatible with the load to obtain a load solution with a mass fraction of 1% to 3%;

[0020] When the loading substance is CBD, CBD is dissolved in 60% ethanol solution, filtered through a 0.22 μm filter membrane to remove plant residues, and then the ethanol is removed by rotary evaporation to obtain CBD crystals with a purity of ≥99%; the CBD crystals are then mixed with 0.5% Tween 80 and a CBD nanosuspension is prepared by a high-pressure homogenizer.

[0021] Furthermore, in step S200,

[0022] Under magnetic stirring, mixing the first carrier raw material solution and the load solution;

[0023] The second carrier raw material powder is added to the first carrier raw material solution and the load mixture, and stirred evenly with a glass rod to completely dissolve the second carrier raw material powder. After complete dissolution, the powder is transferred to a centrifuge tube and the bubbles in the hydrogel are removed by high-speed centrifugation.

[0024] Furthermore, it can be used to prepare the CBD / CMC-Na-CHT hydrogel, comprising the following steps:

[0025] S100: Obtain CMC-Na solution, CHT powder, and CBD nanosuspension;

[0026] S200: CMC-Na solution and CBD nanosuspension are mixed at a volume ratio of 25:1 to 100:1. After uniform mixing, CHT powder is added at a mass volume ratio of 1:25 to 1:10, and stirred until the CHT powder is completely dissolved. Then, bubbles are removed by centrifugation to obtain a CBD / CMC-Na-CHT hydrogel that promotes radiation-combined wound repair.

[0027] Furthermore, the mass ratio of CMC-Na to CHT is 1:3.

[0028] In another aspect, the present invention further provides an application of a hydrogel, wherein the hydrogel is the hydrogel described above, or the hydrogel prepared by the above preparation method, and the hydrogel can at least be used to promote the repair of radiation-induced trauma.

[0029] Compared with the prior art, the present invention can achieve at least one of the following beneficial effects: the present invention prepares a CBD / CMC-Na-CHT hydrogel loaded with cannabidiol (CBD) based on carboxymethylcellulose sodium (CMC-Na) and chitosan (CHT). The CBD / CMC-Na-CHT hydrogel exhibits good healing-promoting function in a mouse model of combined radiotrauma and trauma.

[0030] In the present invention, the above-mentioned technical solutions can be combined with each other to achieve more preferred combinations. Other features and advantages of the present invention will be described in the following description, and some advantages will become apparent from the description or be learned through practice of the present invention. The objectives and other advantages of the present invention can be realized and obtained through the contents particularly pointed out in the description and drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The accompanying drawings are only for the purpose of illustrating particular embodiments and are not to be considered limiting of the present invention. Like reference symbols denote like parts throughout the drawings.

[0032] Figure 1 A bar graph showing the ability to inhibit active oxygen free radicals of Examples 1 to 2 and Comparative Examples 1 to 2 in the specific embodiment;

[0033] Figure 2 The figures are the intuitive diagrams (A) and the statistical diagrams (B) of wound healing rates of Examples 1-2 and Control Examples 1-2 in the specific embodiments;

[0034] Figure 3 Generate visual diagrams for collagen of Examples 1-2 and Control Examples 1-2 in the specific embodiment;

[0035] Figure 4 Graphs showing the fluorescence intensity of M1 (CD86) and M2 (CD206) macrophage markers in Examples 1-2 and Control Examples 1-2 in the specific embodiments;

[0036] Figure 5 The histogram of inflammatory factors and chemokines regulation in Examples 1-2 and Control Examples 1-2 in the specific embodiment;

[0037] Figure 6 It is a bar graph showing the regulation of AGTR2 levels in Examples 1-2 and Control Examples 1-2 in a specific embodiment. DETAILED DESCRIPTION

[0038] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings, wherein the accompanying drawings constitute a part of the present invention and are used together with the embodiments of the present invention to illustrate the principles of the present invention, but are not used to limit the scope of the present invention.

[0039] In the description of the embodiments of the present invention, it should be noted that, unless otherwise specified or limited, the term "connected" should be understood in a broad sense. For example, it can mean a fixed connection, a detachable connection, or an integral connection. It can be a mechanical connection, an electrical connection, a direct connection, or an indirect connection through an intermediate medium. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0040] The terms "top," "bottom," "above," "below," and "on" used throughout the description refer to relative positions of components of a device, such as the relative positions of top and bottom substrates within a device. It will be understood that devices are multifunctional regardless of their orientation in space.

[0041] The working surface of the present invention can be a plane or a curved surface, can be inclined, or can be horizontal. For the convenience of description, the embodiment of the present invention is placed on a horizontal surface and used on the horizontal surface, and "high and low" and "up and down" are defined in this way.

[0042] A specific embodiment of the present invention discloses a hydrogel for promoting the repair of radiation-induced trauma, comprising:

[0043] The carrier raw materials include a first carrier raw material and a second carrier raw material. For example, the first carrier raw material is sodium carboxymethyl cellulose (CMC-Na), the second carrier raw material is chitosan (CHT), and the mass ratio of the first carrier raw material to the second carrier raw material is 1:2 to 1:4. CMC-Na is a white or slightly yellow powder or granule with a particle size of 20 to 200 μm and a specific surface area of 2 to 10 m 2 / g, volume is 0.01~0.05cm 3 / g. CHT is a white or slightly yellow powder with a particle size of 50 to 100 μm and a specific surface area of 5 to 50 m 2 / g, volume is 0.05~0.2cm 3 / g.

[0044] A load is loaded on the carrier, wherein the load includes cannabidiol (CBD), and the loading amount of the cannabidiol is 0.04% to 0.5%.

[0045] The carrier formed by sodium carboxymethylcellulose (CMC-Na) and chitosan (CHT) was designated as CMC-Na-CHT. The hydrogel formed by CMC-Na-CHT loaded with cannabidiol (CBD) and promoting the repair of radiation-induced wounds was designated as CBD / CMC-Na-CHT.

[0046] The present invention also provides a method for preparing a hydrogel that promotes the repair of radiation-induced trauma, which can be used to prepare at least the aforementioned hydrogel, namely, at least CBD / CMC-Na-CHT. The preparation method comprises the following steps:

[0047] S100: Obtaining a first carrier raw material powder or solution, a second carrier raw material powder or solution, and a load solution;

[0048] S200: mixing the first carrier raw material solution and the loading material solution at a volume ratio of 25:1 to 100:1, and after uniform mixing, adding the second carrier raw material powder at a mass volume ratio of 1:25 to 1:10 (i.e., the mass volume ratio of the second carrier raw material powder to the mixed solution is 1:25 to 1:10), stirring until the second carrier raw material powder is completely dissolved, and then removing bubbles by centrifugation to obtain a hydrogel that promotes radiation-combined wound repair;

[0049] Alternatively, the second carrier raw material solution and the loading material solution are mixed at a volume ratio of 1:4 to 1:1. After uniform mixing, the first carrier raw material powder is added at a mass-to-volume ratio of 1:25 to 1:100 (i.e., the mass-to-volume ratio of the first carrier raw material powder to the mixed solution is 1:25 to 1:100). The mixture is stirred until the first carrier raw material powder is completely dissolved, and then centrifuged to remove air bubbles to obtain a hydrogel that promotes radiation-induced wound repair. The hydrogel obtained in this way is harder than the first method.

[0050] In step S100:

[0051] The first carrier raw material is dissolved in a first solution compatible with it to obtain a first carrier raw material solution with a mass fraction of 1% to 3%. For example, when the first carrier raw material is CMC-Na, CMC-Na powder is dissolved in 1% (v / v) hydrochloric acid to form a 2% (m / v) CMC-Na solution. When the first carrier raw material is CMC-Na, the compatible first solution is an acidic aqueous solution with a pH ≤ 3 and an ionic strength ≤ 0.1M. The acidic environment is used to inhibit the premature agglomeration of the CMC-Na molecular chain, improve the uniformity of the solution (viscosity RSD < 5%), and partially hydrolyze the β-1,4 glycosidic bond of CMC-Na to form a shorter chain molecular structure (molecular weight reduced from 200kDa to 80-120kDa), thereby enhancing the swelling rate of the hydrogel (increased by 20%) and the drug diffusion rate.

[0052] Preferably, the first carrier raw material needs to be pretreated. Specifically, CMC-Na powder with a degree of substitution of 0.8 to 1.2 (which can be purchased directly, illustratively, the manufacturer is Shanghai Aladdin Biochemical Technology Co., Ltd.) is selected and sieved with a 200-mesh sieve to remove lumps and ensure uniform distribution of the carboxylmethyl groups in the molecular chains.

[0053] The particle size range of the second carrier raw material powder is 50-100 μm. For example, when the second carrier raw material is CHT, the particle size range of the CHT powder is 50-100 μm. This particle size range is increased by increasing the specific surface area (15m 2 / g) to accelerate dissolution while ensuring the uniformity of electrostatic cross-linking, thereby increasing the mechanical strength of the hydrogel (storage modulus G') by 30% to 650±50Pa.

[0054] Preferably, the second carrier raw material needs to be pretreated. Specifically, CHT raw material with a degree of deacetylation (DD) ≥ 85% is selected (which can be directly purchased, exemplified by the manufacturer Shanghai Aladdin Biochemical Technology Co., Ltd.) and subjected to a secondary deacetylation treatment. Specifically, the CHT raw material is dissolved in 30% NaOH at a mass-to-volume ratio of 1:20. The solution is then reacted at 80°C for 6 hours. After the reaction, it is rinsed with distilled water multiple times until neutral (pH ≈ 7). The solid is collected by centrifugation (4°C, 4900 rpm, 15 min) and vacuum freeze-dried or dried at 105°C to obtain a pure CHT powder. This can increase the degree of deacetylation and ensure sufficient amino content. The amino density is ≥ 6.5 mmol / g as determined by potentiometric titration. The resulting CHT powder is pulverized by air flow to 50-100 μm to increase the specific surface area, accelerate dissolution, enhance electrostatic interaction with CMC-Na, and shorten the hydrogel crosslinking time.

[0055] The load is dissolved in a load solution suitable for it to obtain a load solution with a mass fraction of 1% to 3%. Exemplarily, when the load is CBD, a CBD nanosuspension with a mass fraction of 2% is made by 0.5% Tween 80 solution. Specifically, the purchased CBD is dissolved in 60% ethanol solution, filtered through a 0.22μm filter membrane to remove plant residues, and then ethanol is removed by rotary evaporation to obtain CBD crystals with a purity of ≥99%. The CBD crystals are mixed with 0.5% Tween 80 (polysorbate-80) and a CBD nanosuspension (particle size of 100±20nm, PDI<0.2, PDI is the polydispersity index) is prepared by a high-pressure homogenizer (1500bar×3 times) to improve its dispersion stability in the hydrogel. The specific surface area of nano-sized CBD increases, and the antioxidant activity (DPPH scavenging rate) is enhanced from 80% to 95%.

[0056] In step S200:

[0057] The first carrier raw material solution and the loading substance solution were mixed under magnetic stirring (300-500 rpm).

[0058] The second carrier raw material powder was added to the first carrier raw material solution and the load mixture, and stirred evenly with a glass rod to completely dissolve the second carrier raw material powder. After complete dissolution, the mixture was transferred to a centrifuge tube and removed from the hydrogel by high-speed centrifugation (6000 rpm×10 min).

[0059] Preferably, under magnetic stirring, a CBD solution and a CMC-Na solution are mixed at a volume ratio of 1:50. CHT powder (3:1, m / v) is added to the mixed solution and stirred evenly with a glass rod to completely dissolve the CHT powder. The solution is then transferred to a 50ml centrifuge tube and centrifuged at high speed to remove air bubbles from the hydrogel, thereby obtaining a CBD / CMC-Na-CHT hydrogel that promotes radiation-induced wound repair.

[0060] The hydrogel for promoting the repair of radiation-combined wounds prepared by the above preparation method is stored in a refrigerator at 2-8°C.

[0061] The preparation method of the present invention first fully mixes the first carrier raw material solution and the load solution, and then adds the second carrier raw material to form a hydrogel. Compared with first mixing the first carrier raw material and the second carrier raw material and then loading the load, the hydrogel can fully dissolve the load, so that the final hydrogel is evenly loaded with drugs, with better drug release and higher drug efficacy.

[0062] Illustratively, the present invention discloses at least a method for preparing a CBD / CMC-Na-CHT hydrogel that promotes the repair of radiation-induced trauma, comprising the following steps:

[0063] S100: Obtain CMC-Na solution, CHT powder, and CBD solution;

[0064] S200: CMC-Na solution and CBD solution are mixed in a volume ratio of 25:1 to 100:1. After uniform mixing, CHT powder is added in a mass volume ratio of 1:25 to 1:10 (i.e., the mass volume ratio of CHT powder to the mixed solution is 1:25 to 1:10). The mixture is stirred until the CHT powder is completely dissolved, and then bubbles are removed by centrifugation to obtain a CBD / CMC-Na-CHT hydrogel that promotes radiation-combined wound repair.

[0065] More preferably, CMC-Na powder is dissolved in 1% (v / v) hydrochloric acid to form a 2% (m / v) CMC-Na solution. CBD powder is then prepared into a CBD nanosuspension (i.e., CBD solution). The CHT powder has a particle size range of 50 to 200 μm. Under magnetic stirring, the CBD solution and CMC-Na solution are mixed at a volume ratio of 1:50. CHT powder (3:1, m / v) is added to the mixed solution and stirred with a glass rod to completely dissolve the CHT powder. The mixture is then transferred to a 50 ml centrifuge tube and centrifuged at high speed to remove air bubbles from the hydrogel, yielding a CBD / CMC-Na-CHT hydrogel that promotes radiation-induced wound repair.

[0066] The CBD / CMC-Na-CHT hydrogel prepared by the present invention has an optimal mass ratio of CMC-Na to CHT of 1:3. The obtained CBD / CMC-Na-CHT hydrogel has moderate hardness and is very suitable as an auxiliary material for repairing skin damage.

[0067] Since CMC-Na forms a hydrogel within approximately 10 seconds after contact with CHT, adding the CBD drug later makes it difficult to fully dissolve the CBD. This results in uneven drug loading in the prepared hydrogel, affecting drug release and efficacy. The present invention thoroughly mixes the CMC-Na and CBD first, leaving them in a solution, and then adds CHT to form the hydrogel. This ensures thorough mixing of the CBD, facilitating drug release.

[0068] The present invention also provides a hydrogel for promoting the repair of radiation-induced wounds. Specifically, the prepared hydrogel is applied to the affected area, covered with sterile gauze, and secured with medical tape. The drug is changed daily. CMC-Na, CHT, and CBD can synergistically exert anti-inflammatory effects and promote wound healing.

[0069] The hydrogel provided by the present invention that promotes the repair of radiation-induced trauma has at least the following beneficial effects:

[0070] (1) Excellent antioxidant function. Cannabidiol is a known effective antioxidant that can reduce radiation-induced oxidative stress by scavenging reactive oxygen species. Both CMC-Na and CHT have certain antioxidant capacity, which can reduce the oxidative stress caused by radiation and enhance the repair ability of tissues. CBD / CMC-Na-CHT hydrogel can significantly reduce oxidative stress, thereby protecting tissues from further damage;

[0071] (2) It has strong anti-inflammatory properties. CBD has strong anti-inflammatory properties and can reduce the inflammatory response caused by radiation damage by inhibiting the release of pro-inflammatory factors (such as TNF-α, IL-1β and IL-6). CBD / CMC-Na-CHT hydrogel can continuously inhibit the inflammatory response by slowly releasing CBD, significantly reducing inflammation in the trauma area caused by radiation. CHT naturally has antibacterial and anti-inflammatory properties, which can effectively reduce the possibility of bacterial infection in the wound, while regulating the local inflammatory response and producing a synergistic effect with CBD, thereby promoting wound repair;

[0072] (3) Promotes skin collagen deposition. Collagen is the core component of the extracellular matrix, and its deposition directly determines the mechanical strength and integrity of wound repair. Hydrogel promotes collagen deposition in skin tissue, inhibits the activity of matrix metalloproteinases, maintains the stability of the collagen network, and thus promotes skin wound healing;

[0073] (4) Regulating macrophage polarization. During the wound healing process, the polarization state of macrophages is critical for the different stages of repair. CBD has the ability to regulate macrophage polarization, which can promote the transformation of macrophages from M1 type (pro-inflammatory type) to M2 type (repair type), thereby reducing inflammation and promoting tissue repair and reconstruction. The CBD / CMC-Na-CHT hydrogel of the present invention can continuously release CBD to regulate the response of macrophages at different stages of the healing process.

[0074] (5) No crosslinking agent is used for hydrogel crosslinking. CBD / CMC-Na-CHT hydrogel is formed by physical crosslinking, which avoids the use of traditional chemical crosslinkers and reduces potential toxicity to cells and tissues. The hydrogel network is formed by hydrogen bonding and electrostatic interaction between CMC-Na and CHT, avoiding the residue of harmful crosslinkers, making the material safer and more biocompatible. The preparation process of hydrogel without chemical crosslinkers is relatively simple and does not involve complex chemical reactions. It is more convenient to prepare and more suitable for large-scale production and application.

[0075] (6) High biosafety, low cytotoxicity, and low hemolytic activity. Both CMC-Na and CHT are naturally derived polymer materials with good biocompatibility, low irritation to human tissue, and low toxicity, making them suitable for wound repair and other biomedical applications. CBD / CMC-Na-CHT hydrogels exhibited low cytotoxicity and hemolytic activity in both in vitro and in vivo tests, ensuring their safety. This results in the hydrogel having fewer side effects on surrounding tissues and blood during use, making it particularly suitable for wound repair that requires frequent application. The sustained-release properties of CBD avoid irritation or toxic reactions caused by high-concentration local exposure, ensuring that its anti-inflammatory and antioxidant effects can exert their effects for a long time, while reducing the local concentration of the drug and the occurrence of adverse reactions.

[0076] (7) Targeted inhibition of the immunomodulatory effects of Th-17-related inflammatory factors and chemokines. Proinflammatory factors such as IL-17 and IL-23 secreted by Th-17 cells can aggravate the chronic inflammatory state of radiation trauma. The CBD loaded in the present invention can target and inhibit Th-17-related chemokines, which helps to alleviate radiation skin damage;

[0077] (8) Targeted promotion of AGTR2 gene expression. AGTR2 plays an important role in tissue repair. AGTR2 stimulates endothelial cell proliferation, increases blood supply to wounds, and reduces abnormal activation of fibroblasts. Transcriptome sequencing revealed that CBD / CMC-Na-CHT can significantly promote AGTR2 expression.

[0078] (9) It can promote wound healing. The hydrogel CBD / CMC-Na-CHT forms a good moist environment, which is crucial for wound healing. The moist environment can accelerate the migration of epidermal cells, promote wound closure, and thus shorten the healing time;

[0079] In summary, the CBD / CMC-Na-CHT hydrogel of the present invention demonstrates multiple advantages in the prevention and treatment of radiation-induced trauma, particularly in terms of antioxidant and anti-inflammatory properties, wound healing promotion, and immune response regulation, demonstrating significant efficacy. Furthermore, its preparation via physical crosslinking avoids the use of harmful crosslinking agents, ensuring the material's high biosafety and low toxicity. These properties make the CBD / CMC-Na-CHT hydrogel a highly promising topical formulation for the treatment of radiation-induced injury and complex trauma, with broad potential for clinical application.

[0080] [Example 1] A hydrogel for promoting the repair of radiation-induced trauma (CBD-loaded hydrogel with a CMC-Na to CHT ratio of 1:3, denoted as CBD / CMC-Na-3CHT) is provided. The preparation method thereof comprises the following steps:

[0081] S100: Obtain CMC-Na solution, CHT powder, and CBD nanosuspension;

[0082] Specifically, CMC-Na powder with a degree of substitution of 0.8 to 1.2 is selected, the CMC-Na is sieved through a 200-mesh sieve to remove lumps and ensure uniform distribution of carboxymethyl groups in the molecular chain, and then the CMC-Na powder is dispersed in a 1% (v / v) hydrochloric acid solution to obtain a 2% (m / v) CMC-Na solution;

[0083] CHT raw material with a degree of deacetylation (DD) ≥ 85% was selected and subjected to a secondary deacetylation treatment using a 30% NaOH solution at 80°C for 6 hours to ensure sufficient amino content. The CHT powder was then pulverized by air flow to 50–100 μm to obtain CHT powder.

[0084] Crude CBD was dissolved in 60% ethanol, filtered through a 0.22 μm filter to remove plant debris, and then the ethanol was removed by rotary evaporation. CBD crystals were mixed with 0.5% Tween 80 and passed through a high-pressure homogenizer (1500 bar × 3 times) to prepare a CBD nanosuspension (particle size 100 ± 20 nm, PDI < 0.2).

[0085] S200: Take the prepared CBD nanosuspension and CMC-Na solution in a volume ratio of 1:50 and mix them separately, and stir them evenly under magnetic stirring; add CHT powder to the mixed solution (in this example, the mass ratio of CMC-Na to CHT is 1:3), that is, the ratio of CHT powder to the mixed solution is 1:16.7. Stir magnetically (300-500rpm) until the CHT powder is completely dissolved to form a uniform hydrogel system. The mixed solution is transferred to a 50mL centrifuge tube and the bubbles generated are removed by high-speed centrifugation (6000rpm×10min). Finally, the hydrogel CBD / CMC-Na-3CHT for promoting radiation-combined wound repair is obtained.

[0086] [Example 2] provides a hydrogel that promotes the repair of radiation-combined trauma (CBD-loaded hydrogel with a CMC-Na to CHT ratio of 1:2, denoted as CBD / CMC-Na-2CHT). The difference between its preparation method and [Example 1] is that the ratio of CMC-Na to CHT is 1:2.

[0087] [Control Example 1] Drug-free hydrogel with a CMC-Na to CHT ratio of 1:3 (CMC-Na-3CHT). The difference between Control Example 1 and Example 1 is that CBD is not added.

[0088] [Control Example 2] The commercialized positive drug Biafine (TEA, triethanolamine ointment) which promotes the repair of radiation-induced wounds was used.

[0089] [Mouse model or clinical trial sample] BALB / c mice (6-8 weeks old, male, SPF, 20±2g) were used as experimental animals. In order to ensure that the BALB / c mice received approximately equal irradiation doses, we used a special irradiation box to fix each mouse. The irradiation scheme was whole-body irradiation, with a total irradiation dose of 5Gy 60Coγ rays (dose rate of 0.62Gy / min) and an irradiation distance of 2.5m. The preparation of the traumatic wound was carried out after the irradiation, and a special skin punch was used to ensure that the wound size was basically consistent. Specifically, the hair on the back of the mouse was removed with a depilatory cream before irradiation, and then the cobalt source was irradiated. After the irradiation, the mouse was anesthetized by intraperitoneal injection of 1% sodium pentobarbital (50mg / kg). Then, a skin punch was used to create a circular traumatic wound with a diameter of about 1cm on the skin of the back of the mouse. After establishing the (radiation and trauma) model, the free radical scavenging ability was tested at the cellular level (detected by a ·OH free radical scavenging kit), and the wound healing rate, skin tissue collagen content, skin tissue M1 / M2 macrophage marker levels, inflammatory factors, and chemokine levels were tested at the animal level for 1 to 21 days.

[0090] Radiation damage can trigger a large number of reactive oxygen species (ROS) bursts, among which hydroxyl radicals (·OH) are the main mediators of oxidative stress, which can cause DNA damage, lipid peroxidation and protein denaturation, seriously hindering wound repair. Evaluating the antioxidant capacity of materials is a key indicator to verify their ability to reduce radiation damage. Figure 1 The results show that CBD / CMC-Na-3CHT had a 52% ·OH scavenging rate; CBD / CMC-Na-2CHT had a 43% ·OH scavenging rate; CMC-Na-3CHT, relying solely on CHT's amino radical neutralization, had a ·OH scavenging rate of 15.2%; and TEA had a ·OH scavenging rate of 39.8%. The ·OH scavenging rate of control 1 was significantly lower than that of the other three groups, indicating that CBD and CHT exert a synergistic effect in scavenging ·OH.

[0091] Radiation-induced traumatic wounds heal 2-3 times slower than normal wounds due to vascular endothelial damage and chronic inflammation. After radiation, apply CBD / CMC-Na-3CHT, CBD / CMC-Na-2CHT, CMC-Na-3CHT, or TEA to the wound and change the dressing once a day for 10 consecutive days. Figure 2 It can be seen that the wound healing rate of CBD / CMC-Na-3CHT was 25.4% on the 7th day and 94.2% on the 14th day, which were significantly higher than 6.3% and 76.9% of CMC-Na-3CHT and 7.1% and 91.5% of TEA. The excellent wound healing effect proves that the CBD-loaded hydrogel can improve the therapeutic effect of radiation-combined trauma at the animal level. By continuously releasing CBD and hydrogel to form a good moist environment, it can promote epithelial cell migration and wound closure, thereby shortening recovery time and improving tissue regeneration quality.

[0092] Radiation damage leads to imbalance in collagen metabolism and increases the risk of scar formation. Masson staining is the gold standard for evaluating collagen deposition. Figure 3 It can be seen that CBD / CMC-Na-3CHT and CBD / CMC-Na-2CHT significantly promoted collagen deposition in skin wounds. Higher collagen deposition means that the extracellular matrix is more complete during tissue repair, and wound contraction and mechanical strength are significantly improved, which helps to form structurally stable and beautiful healing tissue, thereby ensuring long-term wound healing effects.

[0093] During wound healing, the polarization state of macrophages plays a key role. M1 macrophages mainly secrete pro-inflammatory factors, while M2 macrophages contribute to tissue repair and regeneration. Promoting the transition from M1 to M2 can help reduce inflammatory responses and enhance the repair process. Skin tissue was extracted and the expression of markers in the skin tissue was detected by immunofluorescence staining. Figure 4CBD / CMC-Na-3CHT and CBD / CMC-Na-2CHT significantly promoted the transformation of local wound macrophages from M1 to M2, with a far greater effect than CMC-Na-3CHT. Promoting the M1 to M2 transformation can effectively reduce local inflammation and stimulate cell repair and angiogenesis, providing a more favorable wound healing microenvironment, thereby shortening healing time and improving repair quality.

[0094] Inflammatory factors are released in large quantities in radiation-induced trauma, which will aggravate local inflammatory reactions and delay wound healing; chemokines attract inflammatory cells to accumulate in large numbers. Reducing the levels of these factors is of great significance for inhibiting inflammation at the wound site, reducing tissue damage and promoting regeneration. The local skin tissue after radiation was extracted and the expression of inflammatory factors and chemokines in the skin tissue was detected using a multi-factor detection kit. Figure 5 It can be seen that compared with CMC-Na-3CHT, CBD / CMC-Na-3CHT and CBD / CMC-Na-2CHT significantly inhibited the expression of IL-1beta, IL-6, IL17A, IL22, TNF-alpha, GM-CSF, CCL3, CCL4, CCL5, and CCL11. Among them, IL-6, IL17A, and IL22 are Th17-related cytokines.

[0095] The AGTR2 (angiotensin II receptor type 2) gene plays an important role in tissue repair. Its expression can promote endothelial cell proliferation, improve blood supply, and regulate local inflammatory responses. Upregulation of AGTR2 helps promote wound healing and tissue reconstruction and is an important molecular indicator for detecting wound repair effects. RT-PCR detection of AGTR2 mRNA levels revealed that CBD / CMC-Na-3CHT and CBD / CMC-Na-2CHT significantly promoted AGTR2 expression. Figure 6 shown.

[0096] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by any technician familiar with this technical field within the technical scope disclosed by the present invention should be covered by the scope of protection of the present invention.

Claims

1. A hydrogel for promoting the repair of radiation-induced trauma, characterized in that: include: The carrier, wherein the raw materials thereof include a first carrier raw material and a second carrier raw material, and the mass ratio of the first carrier raw material to the second carrier raw material is 1:2 to 1:4; A load is loaded on the carrier, wherein the load includes CBD, and the loading amount of the CBD is 0.04% to 0.5%.

2. The hydrogel according to claim 1, wherein The first carrier raw material is CMC-Na, and the second carrier raw material is CHT; The particle size of the CMC-Na is 20 to 200 μm, and the specific surface area is 2 to 10 m 2 / g, volume is 0.01~0.05cm 3 / g; The particle size of CHT is 50-200 μm, and the specific surface area is 5-50 m 2 / g, volume is 0.05~0.2cm 3 / g.

3. A method for preparing a hydrogel for promoting the repair of radiation-induced trauma, characterized in that: At least it can be used to prepare the hydrogel according to claim 1 or 2, and the preparation method comprises the following steps: S100: Obtaining a first carrier raw material powder or solution, a second carrier raw material powder or solution, and a load solution; S200: mixing the first carrier raw material solution and the loading material solution at a volume ratio of 25:1 to 100:1, adding the second carrier raw material powder at a mass volume ratio of 1:25 to 1:10 after uniform mixing, stirring until the second carrier raw material powder is completely dissolved, and then removing bubbles by centrifugation to obtain a hydrogel that promotes radiation-combined wound repair; Alternatively, the second carrier raw material solution and the load solution are mixed in a volume ratio of 1:4 to 1:1, and after uniform mixing, the first carrier raw material powder is added in a mass volume ratio of 1:25 to 1:100, and stirred until the first carrier raw material powder is completely dissolved. Then, bubbles are removed by centrifugation to obtain a hydrogel that promotes radiation-combined wound repair. The hardness of the hydrogel obtained in this way is stronger than that of the first method.

4. The preparation method according to claim 3, characterized in that In step S100, the first carrier raw material is dissolved in a first solution compatible therewith to obtain a first carrier raw material solution with a mass fraction of 1% to 3%; When the first carrier raw material is CMC-Na, the corresponding first solution is an acidic aqueous solution with a pH of ≤3 and an ionic strength of ≤0.1M.

5. The preparation method according to claim 3, characterized in that In step S100, the particle size of the second carrier raw material powder is 50 to 100 μm; When the second carrier raw material is CHT, the CHT raw material is first subjected to a secondary deacetylation treatment, and the treatment method is as follows: The CHT raw material was dissolved in 30% NaOH at a mass-to-volume ratio of 1:20, and the solution was then reacted at 80°C for 6 hours. After the reaction, it was rinsed with distilled water several times until neutral, and the solid was collected by centrifugation. Pure CHT powder was obtained by vacuum freeze-drying or drying at 105°C.

6. The preparation method according to claim 3, characterized in that In step S100, the load is dissolved in a load solution compatible with the load to obtain a load solution with a mass fraction of 1% to 3%; When the loading substance is CBD, CBD is dissolved in 60% ethanol solution, filtered through a 0.22 μm filter membrane to remove plant residues, and then the ethanol is removed by rotary evaporation to obtain CBD crystals with a purity of ≥99%; the CBD crystals are then mixed with 0.5% Tween 80 and a CBD nanosuspension is prepared by a high-pressure homogenizer.

7. The preparation method according to claim 3, characterized in that In step S200, Under magnetic stirring, mixing the first carrier raw material solution and the load solution; The second carrier raw material powder is added to the first carrier raw material solution and the load mixture, and stirred evenly with a glass rod to completely dissolve the second carrier raw material powder. After complete dissolution, the powder is transferred to a centrifuge tube and the bubbles in the hydrogel are removed by high-speed centrifugation.

8. The preparation method according to claim 3, characterized in that It can be used to prepare the CBD / CMC-Na-CHT hydrogel, comprising the following steps: S100: Obtain CMC-Na solution, CHT powder, and CBD nanosuspension; S200: CMC-Na solution and CBD nanosuspension are mixed at a volume ratio of 25:1 to 100:

1. After uniform mixing, CHT powder is added at a mass volume ratio of 1:25 to 1:10, and stirred until the CHT powder is completely dissolved. Then, bubbles are removed by centrifugation to obtain a CBD / CMC-Na-CHT hydrogel that promotes radiation-combined wound repair.

9. The preparation method according to claim 8, characterized in that The mass ratio of CMC-Na to CHT is 1:

3.

10. An application of a hydrogel, characterized in that: The hydrogel is the hydrogel according to claim 1 or 2, or the hydrogel prepared by the preparation method according to any one of claims 3 to 9, and the hydrogel can at least be used to promote the repair of radiation-induced trauma.

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