Fullerene hydrogel with anti-oxidation and active shrinkage functions as well as preparation method and application of fullerene hydrogel

Abstract

The invention discloses fullerene hydrogel with antioxidant and active shrinkage functions as well as a preparation method and application of the fullerene hydrogel. The fullerene hydrogel is prepared from water-soluble fullerene, poly (N-isopropylacrylamide) (PNIPAM), a high polymer material and the like. According to the invention, the water-soluble fullerene derivative with excellent oxidation resistance, the polymer (PNIPAM) with temperature response characteristic and the Alg with excellent water retention capacity are compounded, so that a tissue redox signal and tissue mechanical properties are regulated and controlled at the same time under the condition of body temperature, thereby promoting the closure of diabetic wounds.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and particularly relates to a fullerene hydrogel having both antioxidant and active contraction functions, a preparation method thereof, and an application thereof. Background Art

[0002] At present, the most widely used clinical treatment methods for diabetic wounds in China can be roughly divided into two categories: physical therapy, biological therapy, and topical dressings.

[0003] 1. Physical Therapy

[0004] 1.1 Laser Therapy

[0005] In laser therapy, the more representative one is He-Ne laser therapy. By directly acting He-Ne on deep ulcer tissues, it causes polarization and oscillation of the tissues around the ulcer, thereby generating a certain thermal effect, improving the surrounding blood circulation, and promoting tissue regeneration. This therapy is simple to operate, but it is limited by the wound area and distribution position, and it is difficult to implement for skin ulcers with large wound areas or scattered positions, and the curative effect is poor.

[0006] 1.2 Vacuum Sealing Drainage (VSD) Technique

[0007] The VSD technique refers to covering and sealing the wound surface with a special material and maintaining it under a negative pressure state for a period of time to achieve the treatment effects of debridement and healing of the wound surface. The VSD-sealed environment hinders the influence of germs on the wound surface. At the same time, the negative pressure environment makes the capillaries in the granulation tissue dilate and the vascular permeability improve, thereby promoting the proliferation of granulation tissue. Although this therapy shows certain treatment effects, it requires special materials and strict aseptic conditions during the operation, the treatment cost is high, and the operation is relatively complex.

[0008] 1.3 Vascular Reconstruction

[0009] Compared with other vascular reconstruction techniques, the vascular bypass grafting technique can more efficiently promote the healing of diabetic wound surfaces, but it may cause damage to vascular endothelial cells, etc. during the operation, and at the same time, it may cause an inflammatory reaction in the body itself. According to statistics, about 30% of patients will have vascular stenosis and dysfunction again after the operation.

[0010] 1.4 Oxygen Therapy

[0011] Oxygen therapy is a commonly used clinical means for treating DFU, which is mainly divided into hyperbaric oxygen therapy and local oxygen therapy. Its specific mechanism is to promote angiogenesis and tissue regeneration by increasing the blood oxygen content of the body or local tissues. This therapy has been clinically applied for decades, but its curative effect is still controversial.

[0012] 2. Biological Therapy

[0013] 2.1 Growth Factor Drugs

[0014] Commonly used growth factors in clinical practice include recombinant epidermal growth factor (rhEGF), vascular endothelial growth factor (VEGF), basic fibroblast growth factor, platelet-derived growth factor (PDGF), etc. They promote tissue repair and accelerate the healing of diabetic wounds by promoting the growth, proliferation, and differentiation of specific cells. Although the above-mentioned growth factors can act on skin wound cells at the molecular level to achieve the purpose of promoting wound healing, they have problems such as high treatment costs, easy inactivation of growth factors, and inability to exert continuous efficacy, which to a certain extent limits their application.

[0015] 2.2 Stem Cell Therapy

[0016] Stem cells refer to cells with unlimited self-renewal ability that can be transformed into at least one highly differentiated daughter cell under certain conditions. At present, stem cell therapy has been widely used in the clinical treatment of diabetic wounds, mainly using the strong differentiation potential of stem cells to induce the generation of local microvessels to promote tissue regeneration. Stem cell therapy has a good effect on promoting the healing of chronic diabetic wounds, but its high treatment costs and potential carcinogenic risks objectively limit its popularization.

[0017] 3 Topical Dressings

[0018] 3.1 Moisturizing Dressings

[0019] With the proposal of the "moist wound healing theory", various moisturizing dressings such as alginate dressings and sponge dressings have emerged one after another. The extremely strong water retention ability of alginate dressings enables them to form a closed moist environment after contacting the wound surface, making them more suitable for dry wound surfaces; while sponge dressings, due to their rich porous structure, can absorb a large amount of wound exudate and at the same time provide a moist but not wet microenvironment for wound healing, thus promoting wound surface healing. Although moisturizing dressings can actively promote wound surface healing, their single function makes it difficult for them to deal with chronic wounds including diabetic wounds.

[0020] 3.2 Antibacterial Dressings

[0021] Repeated bacterial infections are one of the main reasons why chronic diabetic wounds are difficult to heal. Currently, the commonly used antibacterial strategies in clinical practice are mainly drugs such as topical antibiotics, which to a certain extent reduce the growth of bacteria on the wound surface and ensure that the healing process is not interrupted. However, the widespread use of antibiotics has created conditions for the emergence of drug-resistant bacteria. For this reason, new antibacterial dressings represented by silver ion dressings and chitosan dressings have been proposed in recent years, which have solved the above problems to a certain extent, but their single antibacterial function makes them unable to cope with the complex wound environment of ulcerative wounds, thus limiting their further use in chronic wounds.

[0022] Immunomodulatory gel dressings are currently the most popular strategy for treating chronic diabetic wounds. The wound healing process can be roughly divided into the inflammatory phase, the proliferative phase, and the remodeling phase. For acute wounds, these three phases proceed continuously and orderly. However, due to the influence of hyperglycemia, diabetic wound healing is uncoordinated and chaotic, which is manifested in excessive inflammation, delayed re-epithelialization, impaired angiogenesis, and excessive collagen degradation. Immunomodulatory gel dressings load bioactive molecules to regulate the transformation of immune cells from pro-inflammatory (such as M1 macrophages) to repair (such as M2 macrophages), thereby inhibiting excessive inflammatory responses and promoting tissue regeneration (DOI: 10.1002 / adma.202200521).

[0023] The current functional gel dressing strategies mainly focus on antibacterial, growth factor delivery, stem cell delivery, and immune regulation. As mentioned earlier, these strategies have problems such as single function, high cost, easy inactivation of bioactive substances, and inability to exert sustained therapeutic effects, and cannot meet the complex diabetic wound microenvironment. In addition, the current strategy mainly promotes tissue repair by regulating biochemical signals, but mechanical signals and biochemical signals are interdependent, and changes in mechanical properties can directly regulate cell responses by regulating integrins, ion channels, growth factor receptors, G-protein coupled receptors, etc. For this reason, the development of a multifunctional dressing that can simultaneously regulate biochemical signals and tissue mechanical properties is expected to provide a new idea for the healing of chronic diabetic wounds. Summary of the invention

[0024] The purpose of the present invention is to provide a fullerene active contraction gel for treating and / or promoting wound healing, and a preparation method and application thereof.

[0025] In a first aspect, the present invention provides a fullerene active shrinkage gel.

[0026] The fullerene active shrinkage gel provided by the present invention is made of raw materials including the following mass percentages:

[0027]

[0028] Wherein, the cross-linking agent can be selected from common structures such as N,N'-methylenebisacrylamide (MBAA), N,N'-bis(acryl)cystamine, etc.;

[0029] The initiator may be selected from at least one of the following: ammonium persulfate (APS), potassium persulfate, ethyl bromide isobutyrate (EBIB), a hydrophilic photoinitiator (Irgacure 2959), a hydrophobic photoinitiator (Irgacure 651), etc.;

[0030] The polymer material can be selected from at least one of the following: sodium alginate (Alg), hyaluronic acid, chitosan, and polyacrylic acid.

[0031] When the polymer material is sodium alginate (Alg), the raw materials further include substances with the following mass percentages: 1% - 5% of calcium salt.

[0032] The calcium salt can be selected from at least one of the following: CaCl2, CaSO3, CaCO3, etc.

[0033] In a second aspect, the present invention provides a preparation method of the fullerene active contraction gel described in the first aspect of the present invention.

[0034] The fullerene active contraction gel provided by the present invention is prepared through a traditional free radical reaction. The specific preparation method includes the following steps:

[0035] Weigh the raw materials according to the above mass ratio, mix the water-soluble fullerene derivative, NIPAM, polymer material, cross-linking agent, initiator, and catalyst TEMED in deionized water until completely dissolved, and carry out cross-linking to obtain the fullerene active contraction gel.

[0036] In the above method, the conditions for cross-linking are: standing at 0 - 60°C for 0.5 - 6 hours.

[0037] When the polymer material is sodium alginate (Alg), the above method further includes the following step: putting the obtained gel into a calcium salt solution for secondary cross-linking to obtain the fullerene active contraction gel.

[0038] The calcium salt solution can be a CaCl2 solution with a mass concentration of 1% - 5%.

[0039] Furthermore, the above method further includes: dialyzing the above fullerene active contraction gel in deionized water to remove unreacted monomers and then storing it for later use.

[0040] In a third aspect, the present invention provides a composite material for treating and / or promoting wound healing.

[0041] The composite material is composed of a chitosan-based gel layer and the fullerene active contraction gel layer.

[0042] Furthermore, the fullerene active contraction gel layer completely covers the chitosan-based gel layer, that is, the area of the fullerene active contraction gel layer is smaller than the area of the chitosan-based gel layer.

[0043] The preparation method of the chitosan-based gel in the above chitosan-based gel layer is as follows: A chitosan solution with a mass concentration of 0.5%-4%, a 25-100 mg / ml carbodiimide (EDC) solution, and a 25-100 mg / ml N-hydroxysuccinimide ester (NHS) solution are mixed evenly and crosslinked. The mixed liquid will undergo a sol-gel transition to obtain a chitosan-based gel, which can form an adhesion interface with tissues.

[0044] Further, the mass concentration of the chitosan solution is 0.5%-4%, the concentration of the carbodiimide (EDC) solution is 25-100 mg / ml, and the concentration of the N-hydroxysuccinimide ester (NHS) solution is 25-100 mg / ml.

[0045] The solvent of the chitosan solution is water, the solvent of the carbodiimide (EDC) solution is a MES solution with a pH of 6.0, and the solvent of the N-hydroxysuccinimide ester (NHS) solution is a PBS solution with a pH of 7.4.

[0046] Further, the volume ratio of the chitosan solution, the carbodiimide solution, and the N-hydroxysuccinimide ester solution is 1:1:1.

[0047] When the above composite material is used, the fullerene active contraction gel layer is directly contacted with the wound tissue to be healed. After the composite material and the tissue are pressed, they are closely adhered to form an adhesion interface.

[0048] In the present invention, the water-soluble fullerene derivative is selected from hollow fullerenes or metal fullerenes with hydrophilic groups modified on the surface.

[0049] In one aspect of the present invention, the hydrophilic groups in the water-soluble fullerene derivative include one or more of hydroxyl, carboxyl, mercapto, and amino groups.

[0050] In one aspect of the present invention, the hollow fullerene includes one or more cage structures composed of carbon atoms with the general formula C 2m , where 30 ≤ m ≤ 60.

[0051] In one aspect of the present invention, the metal fullerene is an endohedral metal fullerene with a metal fullerene or a metal cluster embedded in a hollow fullerene.

[0052] In one aspect of the present invention, the metal fullerene includes M@C 2n , M2@C 2n , MA@C 2n , M3N@C 2n , M2C2@C 2n , M2S@C 2n , M2O@C 2n and M xA 3-x N@C 2n One or more of them, wherein M and A both represent metal elements and are each selected from any one of Sc, Y and lanthanide metal elements, where 30 ≤ m ≤ 60 and 0 ≤ x ≤ 3. Wherein N represents nitrogen element, C represents carbon element, S represents sulfur element, and lanthanide metal elements include La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu.

[0053] Furthermore, the water-soluble fullerene (or water-soluble fullerene derivative) described in the present invention may have a general structural formula of C 2n (OH) x (NH2) y ; where 30 ≤ n ≤ 60, 10 < x ≤ 30, 0 < y ≤ 15, and n, x, and y are all natural numbers.

[0054] The above general structural formula indicates that both hydroxyl and amino groups are connected to the fullerene.

[0055] In the general structural formula of the water-soluble fullerene, n can specifically be 30 or 35.

[0056] The hydrated particle size of the above water-soluble fullerene is 40 - 100 nm.

[0057] The water-soluble fullerene of the present invention can be prepared according to the methods disclosed in the prior art, such as the methods described in the references (DOI: 10.1021 / acsami.7b08348, DOI: 10.1126 / sciadv.abc1586).

[0058] According to the embodiments of the present invention, for the water-soluble fullerene, its molecular formula is C 70 (OH) ～26 (NH2) ～7 ·14H2O, and the specific preparation method is as follows: React 100 mg of C 70 , 6 - 15 mL of 30% hydrogen peroxide, and 2 - 6 mL of ammonia water at 50 - 70 °C for 3 - 8 hours to obtain a water-soluble fullerene derivative. The obtained product is precipitated with ethanol and then dialyzed with ultrapure water for standby. The average molecular formula of the obtained material is C 70 (OH) ～26 (NH2) ～7 ·14H2O.

[0059] For the synthesis of water-soluble fullerenes, fullerene materials are applicable to related fullerene derivatives such as C 60 、C 70 etc., and similar products can be obtained at reaction temperatures within 30 - 80 °C.

[0060] Fourth aspect, the present invention provides: an application of the fullerene active contraction gel described in the first aspect of the present invention or the composite material described in the third aspect of the present invention in the preparation of a medicament for treating and / or promoting wound healing.

[0061] The wound healing includes acute wound healing and chronic wound healing.

[0062] Further, the chronic wound healing may be chronic diabetic wound healing.

[0063] Fifth aspect, the present invention provides: a method for treating and / or promoting wound healing, comprising administering a therapeutically effective amount of the fullerene active contraction gel described in the first aspect of the present invention or the composite material described in the third aspect of the present invention to a subject in need thereof.

[0064] In the present invention, the term "effective amount" refers to a dose that can achieve treatment, prevention, alleviation, and / or remission of the diseases or disorders described in the present invention in a subject.

[0065] In the present invention, the term "subject" may refer to a patient or other animal that receives the composition of the present invention for treating, preventing, alleviating, and / or remitting the diseases or disorders described in the present invention, particularly mammals, such as humans, dogs, monkeys, cows, horses, etc.

[0066] Redox signaling is a key regulator in the process of wound healing, and reactive oxygen species (ROS) regulate all stages of wound healing. For example, during the proliferation phase, too high ROS levels will inhibit the transformation of macrophages from M1 type (pro-inflammatory type) to M2 type (repair type), resulting in persistent inflammation. During the proliferation / remodeling phase, ROS can regulate angiogenesis in a vascular endothelial growth factor (VEGF)-dependent / independent manner. At the same time, the ROS level also affects the extracellular matrix (ECM), thereby regulating the wound healing process.

[0067] As a new type of nanoplatform, fullerene has gradually become a rising star in the field of nanobiology due to its excellent ROS scavenging ability and good biocompatibility. However, its non-polar structure itself makes it insoluble in water, which to a certain extent limits its application in organisms. Therefore, the inventors of the present invention successfully prepared water-soluble fullerene derivatized with hydroxyl and amino groups by a one-pot method in the presence of hydrogen peroxide and ammonia water, greatly improving its solubility in water while ensuring its ROS scavenging ability, paving the way for its further biological utilization.

[0068] In addition, mechanical signals can promote keratinocyte proliferation through pathways such as MEK1 / 2 and EGFR; continuous mechanical stress can upregulate the expression of collagen in fibroblasts, thereby promoting wound healing. N-isopropylacrylamide (NIPAM), as a temperature-sensitive polymer monomer, has both hydrophilic amide groups and hydrophobic isopropyl groups. After free radical polymerization and crosslinking, poly(N-isopropylacrylamide) (PNIPAM) undergoes a phase transition when the temperature rises to 32 °C, resulting in a sudden volume shrinkage, which creates conditions for achieving temperature-mechanical property responses. In addition, sodium alginate (Alg) is an ideal material basis for realizing the "wet healing theory" due to its excellent water retention ability and good tissue compatibility.

[0069] Compared with the prior art, the fullerene active contraction gel described in the present invention has the following advantages:

[0070] (1) The preparation process of the fullerene active contraction gel of the present invention is simple and can be prepared in large batches;

[0071] (2) The fullerene active contraction gel of the present invention can effectively promote wound re-epithelialization and wound contraction, thereby promoting the closure of diabetic wounds;

[0072] (3) The fullerene active contraction gel described in the present invention can effectively reduce the inflammatory level at the wound site, promote cell proliferation and migration, and promote the repair of cell oxidative damage.

[0073] In summary, by compounding a water-soluble fullerene derivative with excellent antioxidant properties, a polymer with temperature-responsive characteristics (PNIPAM), and Alg with excellent water retention ability, the present invention realizes the simultaneous regulation of tissue redox signals and tissue mechanical properties under body temperature conditions, thereby promoting the closure of diabetic wounds. Description of the Drawings

[0074] Figure 1 It is the hydrated particle size diagram of the water-soluble fullerene used in the examples of the present invention;

[0075] Figure 2 It is the picture of the acute wound healing situation of each treatment group in Example 5;

[0076] Figure 3 It is the acute wound healing rate of each treatment group in Example 5;

[0077] Figure 4 It is the picture of the diabetic wound healing situation of each treatment group in Example 5;

[0078] Figure 5 It is the diabetic wound healing rate of each treatment group in Example 5. Detailed Embodiments

[0079] The present invention will be further described in detail below in conjunction with specific embodiments. The provided embodiments are only for clarifying the present invention and not for limiting the scope of the present invention. The following provided embodiments can be used as a guide for those of ordinary skill in the art to make further improvements and do not constitute any limitation to the present invention in any way.

[0080] In the experimental methods in the following embodiments, unless otherwise specified, they are all conventional methods, carried out according to the techniques or conditions described in the literature in this field or according to the product specifications. The materials, reagents, etc. used in the following embodiments, unless otherwise specified, can all be obtained from commercial channels.

[0081] The present invention provides a fullerene active contraction gel and a preparation method thereof. This material is composed of water-soluble fullerene, NIPAM, Alg, etc. The fullerene active contraction gel prepared by the present invention can simultaneously regulate the redox signal and tissue mechanical properties of tissues, thereby promoting wound re-epithelialization and skin contraction, and promoting wound closure. Most of the existing mainstream technologies only promote wound healing from the regulation of biochemical signals. On this basis, the present invention additionally adds the regulation of mechanical signals. Research shows that the introduction of mechanical signals promotes the proliferation and re-epithelialization of cells at the wound site and can produce a synergistic effect with the regulation of biochemical signals, further accelerating the healing of diabetic wounds.

[0082] The water-soluble used in the following embodiments has the molecular formula C 70 (OH) ～26 (NH2) ～7 ·14H2O, and the preparation method is as follows: React 100 mg of C 70 , 10 mL of 30% hydrogen peroxide, and 4 mL of ammonia water at 60 °C for 5 hours to obtain a water-soluble fullerene derivative. The obtained product is precipitated with ethanol and then dialyzed with ultrapure water for standby. The average molecular formula of the obtained material is C 70 (OH) ～26 (NH2) ～7 ·14H2O. The water hydration particle size diagram of water-soluble fullerene is as Figure 1 shown. As Figure 1 can be seen, the average water hydration particle size of the obtained AHF is 63.19 ± 0.41 nm (PDI: 0.16).

[0083] Example 1, Fullerene Active Contraction Gel and Its Preparation Method

[0084]

[0085] The preparation method is as follows:

[0086] Dissolve 12.68 mg of water-soluble fullerene derivative, 125 mg of N-isopropylacrylamide (NIPAM), 50 mg of sodium alginate (Alg), and 3.4 mg of N,N'-methylenebisacrylamide (MBAA) in deionized water until completely dissolved. Subsequently, add 3.4 mg of APS and 6 μL of N,N,N',N'-tetramethylethylenediamine (TEMED), mix well, and let stand overnight at 4 °C. Place the obtained gel in a 2% (mass concentration) CaCl2 solution for secondary crosslinking, and dialyze the unreacted monomers in deionized water before storing for later use, thus obtaining the AHF@AS Gel.

[0087] Example 2: Fullerene Active Shrinkable Gel and Its Preparation Method

[0088]

[0089]

[0090] The preparation method is as follows:

[0091] Dissolve 6.34 mg of water-soluble fullerene derivative, 125 mg of N-isopropylacrylamide (NIPAM), 50 mg of sodium alginate (Alg), and 3.4 mg of N,N'-methylenebisacrylamide (MBAA) in deionized water until completely dissolved. Subsequently, add 3.4 mg of APS and 6 μL of N,N,N',N'-tetramethylethylenediamine (TEMED), mix well, and let stand overnight at 4 °C. Place the obtained gel in a 2% (mass concentration) CaCl2 solution for secondary crosslinking, and dialyze the unreacted monomers in deionized water before storing for later use.

[0092] Example 3: Fullerene Active Shrinkable Gel and Its Preparation Method

[0093]

[0094] The preparation method is as follows:

[0095] Dissolve 12.68 mg of water-soluble fullerene derivative, 125 mg of N-isopropylacrylamide (NIPAM), 50 mg of hyaluronic acid, and 3.4 mg of N,N'-methylenebisacrylamide (MBAA) in deionized water until completely dissolved. Subsequently, add 3.4 mg of APS and 6 μL of N,N,N',N'-tetramethylethylenediamine (TEMED), mix well, and let stand overnight at 4 °C. Then dialyze the unreacted monomers in deionized water before storing for later use.

[0096] Example 4: Fullerene Active Shrinkable Gel and Its Preparation Method

[0097]

[0098] The preparation method is as follows:

[0099] Mix 12.68 mg of water-soluble fullerene derivative, 125 mg of N-isopropylacrylamide (NIPAM), 50 mg of chitosan (molecular weight 1000 - 5000), and 3.4 mg of N,N'-methylenebisacrylamide (MBAA) in deionized water until completely dissolved. Subsequently, add 3.4 mg of APS and 6 μL of N,N,N',N'-tetramethylethylenediamine (TEMED), mix well, and let it stand overnight at 4 °C. After dialyzing the unreacted monomers in deionized water, store it for later use.

[0100] Comparative Example 1, Shrinking Gel (AS Gel) and Its Preparation Method

[0101]

[0102] The preparation method is as follows:

[0103] Mix 125 mg of N-isopropylacrylamide (NIPAM), 50 mg of Alg, and 3.4 mg of N,N'-methylenebisacrylamide (MBAA) in deionized water until completely dissolved. Subsequently, add 3.4 mg of APS and 6 μL of N,N,N',N'-tetramethylethylenediamine (TEMED), mix well, and let it stand overnight at 4 °C. Place the obtained gel in a 2% (mass concentration) CaCl2 solution for secondary cross-linking, and after dialyzing the unreacted monomers in deionized water, store it for later use to obtain AS Gel.

[0104] Example 5, A Composite Material for Treating and / or Promoting Wound Healing

[0105] The composite material is composed of a chitosan-based gel layer and the fullerene active shrinking gel layer.

[0106] Preparation method of chitosan-based glue:

[0107] Mix an aqueous chitosan solution with a mass concentration of 2%, a 50 mg / mL solution of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (the solvent is a MES solution, pH 6.0), and a 50 mg / mL solution of N-hydroxysuccinimide ester (NHS) (the solvent is a PBS solution, pH 7.4) in a volume ratio of 1:1:1, and perform cross-linking to obtain chitosan-based glue.

[0108] Apply the chitosan-based gel water on the surface of the fullerene active shrinking gel prepared in the above example to obtain the composite material.

[0109] When in use, the fullerene active contraction gel layer in the composite material is directly contacted with the surface of the fresh wound tissue to be healed. After pressing against the tissue and closely adhering, an adhesion interface can be formed.

[0110] In the following pharmacodynamic experiments, the composite materials prepared in this example were used in the dosing groups. It is only necessary to replace the fullerene active contraction gel layer therein with the fullerene active contraction gel prepared in the specific example, or the contraction gel prepared in Comparative Example 1.

[0111] Example 6. Pharmacodynamic evaluation of fullerene active contraction gel

[0112] 1. Acute wound model:

[0113] Twenty female ICE mice (24 - 26 g) were randomly divided into 4 groups: control group, ASGel group (the contraction gel prepared in Comparative Example 1), low-dose fullerene contraction gel group (L-AHF@AS Gel, the fullerene active contraction gel prepared in Example 2), and high-dose fullerene contraction gel group (H-AHF@AS Gel, the fullerene active contraction gel prepared in Example 1), with 5 mice in each group. One day in advance, the back hair was removed with depilatory cream, and then a full-thickness excision model with a diameter of 5 mm was established on the back of the mice. On Day 0 and 5, drug treatment was carried out, and the wound closure was observed on Day 0, 5, and 9, and the healing rate was calculated. Except for the control group, each treatment group was administered on Day 0 and Day 5, and a total of 2 doses were administered during the 9-day treatment process. Each time the drug was administered, the gel samples of each group were pressed against the tissue and closely adhered to form an adhesion interface. The drug delivery object was: a gel sheet with a diameter of 10 mm and a thickness of 2 mm, with a volume of 0.157079 cm 3 , and an area of 0.785398 cm 2 ; the AHF dosage in H-AHF@AS Gel was 0.8 mg / cm 2 , and the AHF dosage in L-AHF@AS Gel was 0.4 mg / cm 2 .

[0114] The wound healing rate was quantified for the wound area using Image J software.

[0115] The formula for calculating the wound healing rate was: 1 - (actual wound size / initial wound size) * 100%

[0116] The healing situation is shown in Figure 2 , and the healing rate is shown in Figure 3 .

[0117] As can be seen from Figure 2 , after 9 days of treatment, the wound areas of the mice in each group were all reduced to a certain extent, and the mice in the H-AHF@AS Gel group had the best healing rate.

[0118] It can be seen from Figure 3 that after 9 days of treatment, the wound healing rate of the control group of mice was about 22%, that of the AS Gel group of mice was 35%, that of the L-AHF@AS Gel group of mice was 40%, and that of the H-AHF@AS Gel group of mice was 59%.

[0119] 2. Diabetic wound model:

[0120] Eighteen male db / db mice (40 - 42 g) were randomly divided into 3 groups: a control group (control), a contractile gel group (AS Gel, the contractile gel prepared in Comparative Example 1), and a fullerene contractile gel group (AHF@AS Gel, the fullerene active contractile gel prepared in Example 1), with 6 mice in each group. One day in advance, the back hair of the mice was removed with depilatory cream, and then a full-thickness excision model with a diameter of 5 mm was established on the back of the mice. On Day 0, 4, 8, and 11, drug treatment was given, and the wound closure was observed on Day 0, 4, 8, 11, and 14, and the healing rate was calculated. Except for the control group, each treatment group was given drugs on Days 0, 4, 8, and 11, and a total of 4 times of drug administration was given during the 14-day treatment process. Each time of drug administration, the gel samples of each group were pressed against the tissue to form a tight adhesion interface. The drug delivery object was: a gel sheet with a diameter of 10 mm and a thickness of 2 mm, with a volume of 0.157079 cm 3 , and an area of 0.785398 cm 2 ; the dosage of AHF in AHF@AS Gel was 0.8 mg / cm 2 .

[0121] The healing situation is shown in Figure 4 , and the healing rate is shown in Figure 5 .

[0122] It can be seen from Figure 4 that after 14 days of treatment, the healing situation of the mice in the AHF@AS gel group was significantly improved compared with that of the control group. After treatment with AS Gel, the wound healing situation of the mice was also improved to a certain extent.

[0123] It can be seen from Figure 5 that after 14 days of treatment, the wound healing rate of the control group of mice was about 46%, that of the AS Gel group of mice was about 66%, and that of the AHF@AS Gel group of mice was about 77%.

[0124] Conclusion: The fullerene active contractile gel prepared by the present invention has a certain therapeutic effect on the skin injury models of large-size acute wound models and diabetic wound models of mice, and can accelerate wound closure compared with the control group.

Claims

1. A fullerene active contraction gel is made from raw materials including the following mass percentages:

2. The fullerene active contraction gel according to claim 1, characterized in that: The crosslinking agent is selected from at least one of N,N'-methylenebisacrylamide and N,N'-bis(acryloyl)cystamine; Or, the initiator is selected from at least one of ammonium persulfate, potassium persulfate, ethyl 2-bromoisobutyrate, hydrophilic photoinitiator, and hydrophobic photoinitiator; Or, the polymer material is selected from at least one of sodium alginate, hyaluronic acid, chitosan, and polyacrylic acid.

3. The fullerene active contraction gel according to claim 2, characterized in that: The polymer material is sodium alginate, and the raw materials further include substances with the following mass percentages: 1% - 5% of calcium salt; Preferably, the calcium salt is selected from at least one of the following: CaCl2, CaSO3, and CaCO3.

4. The fullerene active contraction gel according to any one of claims 1 - 3, characterized in that: The water-soluble fullerene derivative is selected from hollow fullerenes or metal fullerenes surface-modified with hydrophilic groups; Furthermore, the hydrophilic group includes one or more of hydroxyl, carboxyl, mercapto, and amino groups; Further, the hollow fullerene includes one or more cage structures composed of carbon atoms with a general formula of C 2m , where 30 ≤ m ≤ 60; Furthermore, the metal fullerene is an endohedral metal fullerene in a hollow fullerene or an endohedral metal fullerene of a metal cluster; Furthermore, the water-soluble fullerene has a structural general formula of C 2n (OH) x (NH2) y ; where 30 ≤ n ≤ 60, 10 < x ≤ 30, 0 < y ≤ 15, and n, x, and y are all natural numbers.

5. The preparation method of the fullerene active contraction gel according to claim 1 or 2 includes the following steps: Weigh each raw material according to the raw material ratio described in claim 1 or 2. Mix the water-soluble fullerene derivative, N-isopropylacrylamide, polymer material, cross-linking agent, initiator, and N,N,N',N'-tetramethylethylenediamine in deionized water until completely dissolved, and carry out cross-linking to obtain the fullerene active contraction gel.

6. The preparation method according to claim 5, characterized in that: The crosslinking conditions are: standing at 0 - 60°C for 0.5 - 6 hours.

7. The preparation method according to claim 5 or 6, characterized in that: The polymer material is sodium alginate, and the method further includes the following step: putting the obtained gel into a calcium salt solution for secondary crosslinking to obtain a fullerene actively shrinking gel; The calcium salt solution is a 1% - 5% CaCl2 solution by mass concentration.

8. A composite material for treating and / or promoting wound healing is composed of a chitosan-based gel layer and the fullerene active contraction gel layer; Furthermore, the fullerene active contraction gel layer completely covers the chitosan-based gel layer; Furthermore, the preparation method of the chitosan-based gel in the chitosan-based gel layer is as follows: Mix the chitosan solution, carbodiimide solution, and N-hydroxysuccinimide ester solution evenly, and carry out cross-linking. The mixed liquid will undergo a sol-gel transition to obtain the chitosan-based gel, which can form an adhesion interface with tissues.

9. Use of the fullerene active contraction gel according to any one of claims 1-4 or the composite material according to claim 8 in the preparation of a product for treating and / or promoting wound healing; Preferably, the wound healing includes acute wound healing and chronic wound healing; preferably, the chronic wound healing is chronic diabetic wound healing.

10. A method for treating and / or promoting wound healing, comprising administering a therapeutically effective amount of the fullerene active contraction gel according to any one of claims 1-4 or the composite material according to claim 8 to a subject in need thereof.

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