Method for repairing wound by activating fibroblasts

Abstract

The invention provides a method for repairing a wound surface by activating fibroblasts. Specifically, the invention provides a nanofiber mat, the nanofiber mat continuously delivers curcumin to activate fibroblasts, and the activated fibroblasts further utilize an EMMPRIN and CCL2 target mediated pathway to complete the repair of a steady-state basement membrane. Experimental results show that the delivery mode can effectively activate fibroblasts, and the activated fibroblasts regenerate skin by inhibiting expression of EMMPRIN and CCL2 targets and promoting expression of laminin, so that acute wounds and chronic wounds are treated. The invention also provides a method for preparing the nanofiber mat.

Description

Technical Field

[0001] The present invention relates to the technical field of activated cell wound repair, and in particular to a method and application of utilizing curcumin to activate fibroblasts for basement membrane reconstruction and wound repair. Background Art

[0002] In contrast to scarless healing of fetal skin wounds, where the immune response is known to be weak, wound repair in adults is often imperfect and prone to fibrotic scarring. In chronic diseases such as diabetes, healing is impaired by a dysregulated inflammatory response, represented by prolonged infiltration of inflammatory cells and sustained elevation of proinflammatory cytokines and metalloproteinases (MMPs), which results in tissue destruction (ulcerative skin defects) or proliferation (scars or keloids) with devastating consequences. Therefore, effective wound regeneration occurs more often when the polarized immune response is less, i.e., in steady state, in other words, rapid reestablishment of homeostasis will stop destructive inflammation and result in unconventional healing. Currently, reestablishing homeostasis is difficult to achieve, and effective methods are still lacking to ensure dermal regeneration after injury in adults, or even healing of refractory wounds.

[0003] Although macrophages have long been considered key players in orchestrating the balance of immune responses during wound healing, macrophage-deficient mice unexpectedly exhibit fetal-like scarless repair, in which cellular and matrix debris are engulfed by phagocytic fibroblasts rather than macrophages, suggesting that fibroblasts have a key role in influencing local immune responses, i.e., homeostasis. In fact, fibroblasts have a similar phenotype to mesenchymal stem cells (MSCs) involved in immune regulation; recent studies have shown that dermal fibroblasts can promote the healing of chronic wounds in a state of long-term inflammation. Therefore, this non-immune cell regulation has attracted attention as a potential therapeutic target. In previous studies, in a rat model of acute wounds (normal healing), we found that activation of wound fibroblasts by topical application of curcumin significantly accelerated wound healing. In addition, we noted that the newly formed tissue exhibited some characteristics that are conducive to skin regeneration, suggesting that curcumin-activated fibroblasts (FibroCUR+) can play an active role in regenerative wound repair. However, it remains largely unknown whether or how FibroCUR+ functions to take over the role of macrophages to create a homeostatic environment that includes a dampened immune response that occurs only transiently in the fetus. Therefore, the underlying mechanisms need to be further elucidated before this information can be translated into clinical practice.

[0004] Curcumin (chemical formula: C 21 H 20O6) is the main curcuminoid in the medicinal plant turmeric, which has been identified as an effective anti-wound and scar agent. Its availability as a suitable lead compound remains controversial due to its inherent drawbacks such as instability, insolubility and very low bioavailability, which can interfere with laboratory results that could demonstrate its therapeutic properties due to degradation. To investigate the possible role of FibroCUR+ in regenerating wounds by restoring homeostasis, a suitable carrier system capable of sustained local delivery of therapeutic curcumin has been demonstrated to be functional.

[0005] In summary, there is an urgent need in the art for a method for repairing wounds by using curcumin-activated fibroblasts. Summary of the invention

[0006] The object of the present invention is to provide a method for repairing / rebuilding basement membrane by using curcumin, by activating fibroblasts, and thereby achieving scar-free repair of various wound surfaces through human interference.

[0007] The first aspect of the present invention provides a use of curcumin or a pharmaceutically acceptable salt thereof for preparing a preparation, product or composition, wherein the preparation, product or composition is used for one or more uses selected from the following group:

[0008] (a) for activating fibroblasts;

[0009] (b) used to repair basement membrane;

[0010] (c) Used to repair skin wounds with damaged basement membrane and inhibit scar formation.

[0011] In another preferred embodiment, the preparation, product or composition is used for one or more purposes selected from the following group:

[0012] (d) downregulates the expression of CCL2 in fibroblasts;

[0013] (e) downregulates the expression of EMMPRIN in fibroblasts;

[0014] (f) Promotes the expression of laminin in fibroblasts.

[0015] In another preferred embodiment, the preparation, product or composition is for external use.

[0016] In another preferred embodiment, the preparation, product or composition is applied to the wound surface and / or its surroundings.

[0017] In another preferred embodiment, the preparation, product or composition is applied to a skin wound.

[0018] In another preferred embodiment, the preparation, product or composition inhibits the formation of hypertrophic scars.

[0019] In another preferred embodiment, the skin wound with damaged basement membrane is selected from the following group: a skin wound with mildly damaged basement membrane, a skin wound with severely damaged basement membrane, a skin wound with persistently damaged basement membrane, or a combination thereof.

[0020] In another preferred embodiment, in a skin wound with damaged basement membrane, the ratio of the minimum thickness Y1 of the damaged basement membrane to the average thickness Y0 of the undamaged normal basement membrane (Y1 / Y0) is ≤1 / 2, preferably ≤1 / 3, and more preferably ≤1 / 4.

[0021] In another preferred embodiment, the ratio of the minimum thickness Y1 of the damaged portion of the basement membrane to the average thickness Y0 of the undamaged normal basement membrane is 0.

[0022] In another preferred embodiment, the composition is a pharmaceutical composition.

[0023] In another preferred embodiment, the preparation, product or composition is administered to a subject selected from the following group:

[0024] (Z1) subjects with increased expression of CCL2 in skin wounds;

[0025] (Z2) subjects with increased expression of EMMPRIN in skin wounds;

[0026] (Z3) subjects with decreased expression of laminin in the skin wound surface;

[0027] In another preferred embodiment, the preparation, product or composition is administered to a subject selected from the following group:

[0028] (Z4) Punch biopsy wound of rat;

[0029] (Z5) Inflammatory and non-healing wounds in diabetic mice;

[0030] (Z6) Stress wound in pig.

[0031] In another preferred embodiment, the preparation, product or composition contains curcumin as the only active ingredient.

[0032] In another preferred embodiment, the preparation, product or composition does not contain other active ingredients selected from the following group: anti-inflammatory ingredients and anti-fibrotic ingredients.

[0033] In another preferred embodiment, the preparation, product or composition contains curcumin as the first active ingredient; and a component selected from the following group as the second active ingredient: rhein and ketoprofen.

[0034] In another preferred embodiment, the preparation, product or composition contains curcumin as the first active ingredient and a polymer as an inactive ingredient.

[0035] In another preferred embodiment, the preparation, product or composition contains curcumin as the first active ingredient; and components selected from the following group as inactive ingredients: polyoctyl citrate, gelatin, and methacrylic anhydride gelatin.

[0036] In another preferred embodiment, the basement membrane damaged skin wound is selected from the following group: acute wound, chronic wound, or a combination thereof.

[0037] In another preferred embodiment, the acute wound is selected from the following group: a perforation biopsy wound, a stress wound, or a combination thereof.

[0038] In another preferred embodiment, the diameter of the perforated biopsy wound is 3-9 mm, preferably 6 mm.

[0039] In another preferred embodiment, the stress wound is formed by a method selected from the following group:

[0040] (i) applying tension in the direction opposite to the direction of skin wound healing during the skin wound healing process; and / or

[0041] (ii) In the case of a skin wound with skin and soft tissue defects, the skin on both sides of the skin wound is sutured.

[0042] In another preferred embodiment, the stress wound forms a scar.

[0043] In another preferred embodiment, the chronic wound is an inflammatory refractory wound.

[0044] In another preferred embodiment, the inflammatory refractory wound is a wound in a diabetic patient.

[0045] In another preferred embodiment, the inflammatory refractory wound is a perforation biopsy wound in a diabetic body.

[0046] In another preferred embodiment, the diabetic organism is selected from the following group: diabetic patients, or diabetic mice.

[0047] In another preferred embodiment, the preparation, product or composition comprises curcumin, polycaprylyl citrate, and gelatin.

[0048] In another preferred embodiment, the active ingredient in the preparation, product or composition is curcumin.

[0049] In another preferred embodiment, the mass ratio of curcumin, poly(octyl citrate) and gelatin is (0.15±0.05):(0.5±0.1):(1±0.2).

[0050] In another preferred embodiment, the method for preparing the preparation, product or composition comprises the following steps:

[0051] (S1) dispersing poly(octyl citrate) and curcumin in an organic solvent to obtain a solution 1;

[0052] (S2) dispersing gelatin in an organic solvent to obtain solution 2;

[0053] (S3) Processing solution 1 and solution 2 to obtain the preparation, product or composition.

[0054] In another preferred embodiment, the organic solvent is hexafluoroisopropanol.

[0055] In another preferred embodiment, the processing is electrospinning.

[0056] In another preferred embodiment, the preparation, product or composition is an external preparation, product or composition.

[0057] In another preferred embodiment, the external preparation, product or composition is an external patch.

[0058] In another preferred embodiment, the preparation, product or composition releases curcumin in a controlled manner, and releases about 30% of the contained curcumin cumulatively within 140 hours.

[0059] A second aspect of the present invention provides a method for activating fibroblasts in vitro, comprising the following steps:

[0060] (S1) providing a fibroblast to be activated;

[0061] (S2) Incubating fibroblasts to be activated in the presence of the preparation, product or composition of claim 1.

[0062] In another preferred embodiment, the fibroblasts to be activated are human skin fibroblasts.

[0063] In another preferred embodiment, the preparation, product or composition comprises a cell culture medium containing curcumin.

[0064] In another preferred embodiment, in step (S2), activated fibroblasts are obtained, and compared with the migration rate V0 of the fibroblasts to be activated, the migration rate V1 of the activated fibroblasts satisfies V1 / V0≥2, preferably ≥2.5, and more preferably ≥3.

[0065] In another preferred embodiment, in step (S2), activated fibroblasts are obtained, wherein the expression of CCL2, the expression of EMMPRIN, and the expression of laminin are decreased, and the activated fibroblasts are increased;

[0066] In another preferred embodiment, the method is non-diagnostic and non-therapeutic.

[0067] The third aspect of the present invention provides a product, which contains curcumin or a pharmaceutically acceptable salt thereof, wherein the curcumin or a pharmaceutically acceptable salt thereof is loaded on a fiber membrane, and the fiber membrane comprises polyoctyl citrate and gelatin.

[0068] The fourth aspect of the present invention provides a use of the product described in the third aspect of the present invention for preparing a drug for repairing skin wounds and inhibiting scar formation.

[0069] In another preferred embodiment, the skin wound is a skin wound with intact basement membrane.

[0070] In another preferred embodiment, the skin wound is a skin wound with damaged basement membrane.

[0071] In another preferred embodiment, a method for using the product according to the third aspect of the present invention is provided, comprising the steps of:

[0072] (S1) contacting the preparation, product or composition described in the first aspect of the present invention with the wound surface and / or its surroundings;

[0073] (S2) After a contact period, removing the preparation, product or composition.

[0074] In another preferred embodiment, the wound surface is selected from the following group: a perforation biopsy wound surface, an inflammatory refractory wound surface, a stress wound surface, or a combination thereof.

[0075] In another preferred embodiment, the preparation, product or composition is a fiber membrane loaded with curcumin.

[0076] In another preferred embodiment, the fiber membrane comprises poly(octyl citrate) and gelatin.

[0077] In another preferred embodiment, the time is ≥6 days, preferably ≥11 days, and more preferably ≥15 days.

[0078] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0079] Figure 1 The present invention shows the preparation process of the curcumin-loaded poly(octanediol citrate)-gelatin nanofiber membrane for fibroblast activation. POCA, poly(octanediol citrate).

[0080] Figure 2 The present invention shows the actual object, bright field, fluorescence and electron microscope photos of the curcumin-loaded poly(octyl glycol citrate)-gelatin nanofiber membrane for fibroblast activation. CUR / POCA-Glt NM, curcumin-loaded poly(octyl glycol citrate)-gelatin nanofiber membrane; POCA-Glt NM, non-curcumin-loaded poly(octyl glycol citrate)-gelatin nanofiber membrane.

[0081] Figure 3 The curcumin release curve of the curcumin-loaded poly(octanediol citrate)-gelatin nanofiber membrane for fibroblast activation of the present invention is shown. CUR / POCA-Glt NM, curcumin-loaded poly(octanediol citrate)-gelatin nanofiber membrane.

[0082] Figure 4 Shown are changes in expression of different proteins in activated fibroblasts.

[0083] Figure 5 The expression changes of CCL2 and EMMPRIN at the mRNA and protein levels in activated fibroblasts are shown. β-Actin, a type of actin, was used as a control group.

[0084] Figure 6 Shown are changes in the migration speed of activated fibroblasts.

[0085] Figure 7 Activated fibroblasts were shown to inhibit the chemotactic migration of immune cells.

[0086] Figure 8 The results show that activated fibroblasts inhibit EMMPRIN and promote laminin expression. β-Actin, a type of actin, was used as a control group.

[0087] Fig. 9 Activated fibroblasts were shown to promote basement membrane remodeling in vitro.

[0088] Fig.10 Shows that activated fibroblasts regulate NF-κB signaling. β-Actin is an actin and was used as a control. GAPDH is a housekeeping gene and was used as a control.

[0089] Fig.11 Activated fibroblasts were shown to promote wound healing in punch biopsy.

[0090] Fig.12 It was shown that activated fibroblasts promote the healing of refractory inflammatory wounds and scar-free healing of stress wounds. DETAILED DESCRIPTION

[0091] After extensive and in-depth research and a large number of experiments, the inventors prepared a poly(octanediol) citrate-gelatin nanofiber membrane loaded with curcumin for the first time, and used the carrier to activate fibroblasts. In addition, the inventors unexpectedly discovered for the first time that activated fibroblasts weakened the wound immune response by inhibiting CCL2 expression; and also promoted laminin expression, regulated NK-κB signals to inhibit EMMPRIN expression, thereby repairing / rebuilding the basement membrane, promoting skin regeneration, and completing wound repair of acute wound healing, perforated biopsy wound healing, stress wound healing, etc. On this basis, the present invention was completed.

[0092] Specifically, the present invention aims to provide a nanofiber membrane carrier loaded with curcumin, and also provides a method for using the carrier to slowly release curcumin to activate fibroblasts, thereby repairing / rebuilding the basement membrane, thereby achieving scar-free repair of various wound surfaces through human interference.

[0093] The objective of the present invention is achieved through the following technical solutions:

[0094] The present invention provides a method for wound repair by activating fibroblasts, comprising the following processes: (1) activating fibroblasts by adding drugs or genetic engineering means; (2) the activated fibroblasts inhibit CCL2 expression; (3) the reduced expression of CCL2 leads to a reduction in immune cells migrating to the wound, weakening the wound immune response, reducing the damage of the immune response to the basement membrane, and helping the basement membrane reconstruction; (4) the activated fibroblasts overregulate NF-κB signals to inhibit EMMPRIN expression; (5) EMMPRIN expression is inhibited, promoting basement membrane reconstruction; (6) basement membrane reconstruction promotes wound healing. Preferably, the drug described in step (1) is sustained-release curcumin, such as curcumin continuously released from a polyoctyl citrate-gelatin (POCA-Gel) nanofiber membrane loaded with curcumin. Preferably, the wound in step (6) can be an acute wound, a chronic wound, etc.

[0095] the term

[0096] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0097] Unless otherwise indicated, percentages and parts are by weight.

[0098] As used herein, when used in reference to a specific recited numerical value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 80" includes all values ​​between 79 and 81 (e.g., 79.1, 79.2, 79.3, 79.4, etc.).

[0099] As used herein, the term "pharmaceutically acceptable salt" can improve the stability and solubility of the compound or drug itself to a certain extent, thereby improving its bioavailability. In the present invention, curcumin as the only active ingredient has defects such as low stability, low solubility, and low bioavailability, and the above shortcomings can be improved by loading it on a carrier. Therefore, any curcumin-derived salt having curcumin activity and capable of being loaded on the carrier can further improve the stability, solubility and bioavailability of curcumin.

[0100] As used herein, the term "electrospinning" is a special fiber manufacturing process in which a polymer solution or melt is jet-spun in a strong electric field. This method can produce polymer filaments with nanometer diameters. In the present invention, two polymers, polyoctyl citrate and gelatin, are used for electrospinning to form the nanofiber membrane carrier of the present invention. The materials used to construct the fiber membrane carrier include, but are not limited to, polyoctyl citrate, polycaprolactone, polylactic acid, gelatin, methacrylic anhydride gelatin, and any other molecules that can be used for electrospinning of the matrix component.

[0101] As used herein, the term "active ingredient" refers to a molecule or compound that actually plays a role in a specific area or site. In the present invention, in the polyoctyl citrate-gelatin nanofiber membrane loaded with curcumin, curcumin is the compound that actually plays a role in the wound healing process, while polyoctyl citrate and gelatin are only used to construct the fiber membrane carrier and are inactive ingredients.

[0102] As used herein, the terms "anti-fibrotic ingredients" and "anti-inflammatory ingredients" are collective names for active ingredients classified by function. Anti-fibrotic ingredients refer to molecules or compounds that can alleviate the disordered response during the repair process after tissue damage, while anti-inflammatory ingredients refer to molecules or compounds that can alleviate the symptoms of inflammation. Fibrosis is caused by excessive deposition of extracellular matrix proteins such as collagen and fibronectin by fibroblasts in chronic inflammatory responses, resulting in scarring and tissue hardening.

[0103] Fibroblasts

[0104] Fibroblasts are a type of cell that is widely found in loose connective tissue or fibrous connective tissue. They can secrete collagen and other extracellular matrices to maintain the integrity of the connective tissue structure and play an important role in physiological or pathological processes such as interstitial renewal, wound repair and fibrosis. Existing studies have shown that curcumin-activated fibroblasts migrate faster and can significantly accelerate wound healing.

[0105] Based on the functions and characteristics of the above-mentioned fibroblasts, the present invention activates fibroblasts by curcumin, and finds that the activated fibroblasts inhibit the expression of CCL2 and EMMPRIN and promote the expression of laminin, thereby reducing the chemotactic migration of immune cells at the wound, rebuilding the steady-state basement membrane, and completing the repair of various wound surfaces.

[0106] In the present invention, the method of activating fibroblasts includes in vivo activation and in vitro activation, wherein in vivo activation refers to direct action of curcumin or a topical preparation or composition loaded with curcumin on fibroblasts in vivo to activate fibroblasts, thereby repairing wounds; in vitro activation refers to action of curcumin or a preparation, product or composition loaded with curcumin on fibroblasts cultured in vitro to activate fibroblasts. The method of using in vitro activated fibroblasts for wound repair includes but is not limited to in vitro activation and culture of fibroblasts, preparing them into a preparation with a pharmaceutically acceptable carrier, diluent or excipient, and injecting them into the patient's body.

[0107] CCL2 and EMMPRIN

[0108] As used herein, the term "CCL2" is CC motif chemokine ligand 2, which plays an important role in the inflammatory process. The protein expressed by CCL2 can attract or enhance the expression of other inflammatory factors or inflammatory cells. Through the migration and infiltration of inflammatory cells such as monocytes and macrophages, as well as other cytokines at the site of inflammation, it leads to the progression of various diseases.

[0109] As used herein, the term "EMMPRIN" is an extracellular matrix metalloproteinase inducer, which is a transmembrane glycoprotein that can induce the synthesis of matrix metalloproteinases. EMMPRIN plays an important role in the degradation of extracellular matrix components and the cleavage of cell surface receptors, signaling molecules and other precursor proteins.

[0110] Basement membrane

[0111] The basement membrane (BM) is a thin and dense layer of extracellular matrix between epithelial cells and the connective tissue below. It is mainly composed of proteins and polysaccharides, which play a supporting and separating role, and also have important functions in cell adhesion, differentiation, proliferation and migration. The main components of the basement membrane include: laminin, a macromolecular glycoprotein, is one of the main components of the basement membrane and is responsible for cell adhesion; type IV collagen, the main structural protein, forms a mesh structure and provides mechanical strength for the basement membrane; heparan sulfate proteoglycan: such as dermatan persulfate, contributes to the filtering function of the basement membrane; entactin / nidogen, connects laminin and type IV collagen, and stabilizes the structure of the basement membrane.

[0112] Wound

[0113] As used herein, "wound" and "wound" are used interchangeably and refer to damage caused by physical, chemical or biological factors that results in the rupture and / or loss of skin tissue structure.

[0114] Since the integrity of the basement membrane is very important in the wound healing process, the classification and evaluation of wounds can refer to the degree of damage to the basement membrane. According to the degree of damage to the basement membrane, different types of wounds can be divided into the following categories: acute wounds, chronic wounds, and large surface wounds.

[0115] Acute wounds include superficial acute wounds, such as abrasions and superficial cuts. In these wounds, the basement membrane may be partially damaged or intact. Usually the wounds heal rapidly, and the healing process mainly depends on the proliferation and migration of epithelial cells. Deep acute wounds, such as deep cuts and surgical incisions, are caused by severe damage to the basement membrane and need to be healed through regeneration and reconstruction of the basement membrane and repair of the subcutaneous tissue.

[0116] Chronic wounds include ulcerative chronic wounds, such as diabetic foot ulcers and venous ulcers. In these wounds, the basement membrane is continuously damaged or fails to repair normally, and the wound cannot heal for a long time, often accompanied by inflammatory response and bacterial infection; pressure ulcers are common in patients who are bedridden for a long time or have limited mobility. In these wounds, the basement membrane and subcutaneous tissue are damaged due to continuous inflammatory response and pressure, making healing difficult.

[0117] Large-surface wounds include burns. Depending on the depth of the burn, the basement membrane and underlying tissue may be intact, slightly damaged, or severely damaged; lacerations are common in large-area skin tears caused by trauma. In this type of wound, the basement membrane and dermis may be damaged over a large area, requiring complex surgical repair and a long healing process.

[0118] In clinical treatment, assessing the damage to the basement membrane can help doctors develop appropriate treatment plans and accelerate the wound healing process.

[0119] As used in this article, "punch biopsy wound" is a type of deep acute wound whose basement membrane is severely damaged; "stress wound" is also a type of deep acute wound, which is prone to form scars under the action of external force; "inflammatory non-healing wound" is a type of chronic wound, which is also formed after an acute wound is formed in subjects with specific diseases or limited mobility. The basement membrane is often accompanied by an inflammatory response due to continuous damage, making it difficult to heal.

[0120] Scars and hypertrophic scars

[0121] As used herein, the term "scar" is a general term for the appearance and histopathological changes of normal skin tissue caused by various traumas. Scars include superficial scars, hypertrophic scars, atrophic scars, keloids, etc. Among them, hypertrophic scars are a kind of hypertrophic and raised scars, so they are also called hypertrophic scars, which are abnormal reactions of wound healing. Hypertrophic scars usually occur in tight areas of the skin after skin trauma, burns or surgical incisions.

[0122] The main advantages of the present invention include:

[0123] (1) The present invention imitates the fetal wound healing process and uses the poly(octyl citrate)-gelatin nanofiber membrane loaded with curcumin to regulate a type of non-immune cells at the wound, namely fibroblasts, thereby activating the fibroblasts, allowing the fibroblasts to partially replace immune cells and participate in wound repair.

[0124] (2) The present invention utilizes curcumin to activate fibroblasts, and unexpectedly finds that the activated fibroblasts reconstruct the immune microenvironment of the wound surface by reducing CCL2 expression, reducing EMMPRIN expression, and increasing laminin expression, thereby achieving homeostasis around the wound surface and repairing / reconstructing the basement membrane of the wound tissue.

[0125] (3) The topical preparation or composition containing curcumin of the present invention achieves scar-free healing of acute wounds and chronic wounds.

[0126] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are weight percentages and weight parts.

[0127] Example 1: Preparation of curcumin-loaded poly(octyl citrate)-gelatin nanofiber membrane for fibroblast activation

[0128] The present embodiment relates to a poly(octanediol citrate)-gelatin drug-loaded nanofiber membrane (Cur / POCA-GelNM) for sustained-release curcumin, which is composed of poly(octanediol citrate) (POCA), gelatin, and curcumin.

[0129] like Figure 1 As shown, the specific preparation steps are as follows: 76.8g citric acid, 29.2g 1,8-octanediol are melted at 145°C, and then stirred at 135°C for 1 hour in a nitrogen environment to obtain polyoctanediol citrate. Take 0.5g polyoctanediol citrate and dissolve it in 10mL hexafluoroisopropanol, and add 0.15g curcumin. At the same time, take 1g gelatin and dissolve it in 10mL hexafluoroisopropanol. Then the above two solutions are respectively loaded into the injection mixer for extrusion and processed into nanofiber membranes by electrospinning. The parameters of electrospinning are: flow rate 1ml / h; voltage 15kV; distance: 25cm; needle model: 24G flat-head needle. The nanofiber membrane obtained by electrospinning is placed at 120°C and 2000Pa for light-proof crosslinking for 24 hours to obtain a polyoctanediol citrate-gelatin nanofiber membrane loaded with curcumin.

[0130] Figure 2 Actual photos, bright field microscope photos, fluorescence microscope photos and scanning electron microscope photos were taken to prepare curcumin-loaded poly (octanediol citrate)-gelatin nanofiber membrane (CUR / POCA-Glt-NM) and curcumin-free poly (octanediol citrate)-gelatin nanofiber membrane (POCA-Glt-NM).

[0131] Example 2: Sustained release of curcumin from poly(octylene glycol citrate)-gelatin nanofiber membrane loaded with curcumin for fibroblast activation

[0132] This embodiment relates to the determination of the sustained release of curcumin in a poly(octanediol citrate)-gelatin nanofiber membrane loaded with curcumin for fibroblast activation. The specific steps are as follows: 22 mg of a poly(octanediol citrate)-gelatin nanofiber membrane loaded with curcumin is soaked in 50 ml of 37°C phosphate buffer (PBS, pH 7.4). At each time detection point, 200 μL of solution is taken out for testing, and an equal volume of PBS is added back to maintain a constant volume. The curcumin content in the solution is taken out using high performance liquid chromatography (HPLC) coupled to quadrupole mass spectrometry. Agilent Eclipse XDB-C18 analytical column (4.6×150 mm, 3.5 μm) is used as a separation column and maintained at 45°C. The mobile phase consists of water and methanol, with a flow rate of 300 μL / min in a gradient mode, each containing 0.2% formic acid.

[0133] The obtained curcumin content released into the solution changes with time (release curve) as shown in Figure 3 As shown, compared with pure curcumin, curcumin in CUR / POCA-Glt-NM exhibits a sustained release pattern, so that the concentration of curcumin in the environmental solution can be maintained at a higher concentration for a longer time.

[0134] Example 3: Sustained release of curcumin to activate fibroblasts

[0135] This example involves the use of sustained-release curcumin to activate fibroblasts. The specific steps are as follows: 10 mg of drug-loaded film or empty film was soaked in 50 mL of cell culture medium for 48 hours, and then diluted several times with fetal bovine serum to prepare the drug-loaded film extract. Human skin fibroblasts (HS27) were inoculated in 6-well plates (1×10 5 Cells / well) were added and cultured with drug-loaded membrane extract at 37°C and 5% CO2 until the cells were close to confluence. Then, AIM V serum-free medium (Thermo Fisher Scientific) was replaced and cultured for 48 hours, and the supernatant was collected and semi-quantitative cytokine analysis was performed using the Human XL Protein Chip Kit (R&D Systems).

[0136] The results are as follows Figure 4 As shown in the results, among the 102 different cytokines, chemokines and growth factors tested, monocyte chemoattractant protein 1 (CCL2) and extracellular matrix metalloproteinase inducer (EMMPRIN) in activated fibroblasts were significantly decreased. qPCR and Western blot experiments further confirmed that these two molecules in activated fibroblasts had a significant decrease in both mRNA and protein levels ( Figure 5 ).

[0137] HS27 cells were cultured at 5 × 10 5Cells / ml were seeded in the isolation culture chamber insert (ibidi), activated with the drug-loaded membrane leaching solution, cultured to basic confluence, then the ibidi was removed and photographed every 12 hours under a microscope to record cell migration.

[0138] The results showed that activated fibroblasts migrated faster than unactivated cells ( Figure 6 ).

[0139] Example 4: Activated fibroblasts inhibit chemotactic migration of immune cells

[0140] This example involves studying the effect of activated fibroblasts on the chemotaxis and migration of immune cells. The specific steps are as follows: human skin fibroblasts (HS27) were inoculated in a 6-well plate (1×10 5 cells / well) and activated with drug-loaded membrane extract at 37°C and 5% CO2 until the cells were close to confluence. Then, AIM V serum-free medium (ThermoFisher Scientific) was replaced and cultured for 24 hours. 500 μl of supernatant was added to a 24-well plate, and a transwell chamber was placed on it. Then 300 μl of 1×10 6 Freshly prepared peripheral blood mononuclear cells (PBMC), human peripheral blood mononuclear cell line (THP-1), and human myelomonocytic leukemia cells (MV-4-11) were added to different upper transwell chambers and cultured for 12 hours. For PBMC, after removing non-migrated cells, the transwell chamber was transferred to a clean well; for THP-1 and MV-4-11 cells, cells that migrated to the lower wells were collected by centrifugation and transferred to a clean well. Staining solution was added, and the final reaction solution (100 μl / well) was transferred to a new well of a 96-well plate, and the cell number was measured by a plate reader, and PBMC cells were directly examined using bright field, or THP-1 cells and MV-4-11 cells were examined using a fluorescence microscope.

[0141] The results are as follows Figure 7 As shown, activated fibroblast culture supernatant can effectively inhibit the chemotactic migration of three immune-related cells.

[0142] Example 5: Activated fibroblasts reconstruct basement membrane by inhibiting EMMPRIN

[0143] In this example, the mechanism of activated fibroblasts rebuilding the basement membrane was further studied. The method was as follows: human fibroblasts (HS27) were cultured at an initial density of 10 5Cells / well were inoculated in 6-well plates. When the cell confluence reached about 80%, the culture medium was replaced with fresh culture medium containing 10 μg / mL polyamine, and then EMMPRIN silencing lentivirus (cell: virus = 1: 5) was added and centrifuged (1200RPM, 20min). After overnight culture at 37°C and 5% CO2, fresh culture medium was replaced and cultured for 24 hours. Then, fresh culture medium containing puromycin (2 μg / mL) was replaced and cultured for several days to prepare HS27 cells with EMMPRIN gene silenced.

[0144] Normal HS27 cells and keratinocytes (HaCat) were mixed and cultured at 1×10 5 The cells / well density was seeded in 6-well plates, and the cells were cultured and activated using the drug-loaded membrane extract. Once the cells were fully confluent, the culture was stopped, the cells were fixed, and immunofluorescence staining and western blotting experiments were performed.

[0145] The results are as follows Figure 8 As shown, the expression level of EMMPRIN in activated fibroblasts decreased, while the expression of laminin, an important protein for forming the basement membrane, increased.

[0146] For further experiments, normal or EMMPRIN-silenced HS27 cells and HaCat cells were cultured at 5 × 10 5 / ml concentration was inoculated into two chambers of the separation culture chamber insert (ibidi). After culturing for 24 hours at 37°C and 5% CO2, ibidi was removed and washed 3 times with PBS. The culture medium was replaced with the drug-loaded film extract to continue culturing cells and activate HS27 cells. Once the gap between the two types of cells completely disappeared (direct cell-cell contact was formed), the culture was stopped. The cells were fixed and stained for immunofluorescence analysis.

[0147] The results are as follows Fig. 9 As shown, laminin was highly expressed in the contact interface area between HaCat and activated HS27 cells, forming a basement membrane-like structure, which was similar to the effect after EMMPRIN was silenced, indicating that activated fibroblasts rebuilt the basement membrane by inhibiting EMMPRIN.

[0148] Example 6: Activated fibroblasts reconstruct the homeostatic basement membrane by regulating NF-κB signaling

[0149] This example further explores the mechanism of activated fibroblasts rebuilding the steady-state basement membrane. The method is as follows: human skin fibroblasts (HS27) were seeded in 6-well plates (1×10 5cells / well) and cultured with the drug-loaded membrane extract at 37°C and 5% CO2 until the cell confluence was close to 80%. Then, the pro-inflammatory factor interleukin 1β (IL-1β) was added and cultured for 12 hours, and the cells were fixed for immunofluorescence staining and Western blotting experiments.

[0150] The results are as follows Fig.10 As shown in the figure, IL-1β can significantly increase the expression level of EMMPRIN in unactivated HS27 cells, while the expression level of EMMPRIN in activated HS27 cells is significantly reduced regardless of whether they are stimulated by IL-1β. IL-1β stimulation can significantly increase the expression level of transcription factors (p50 and p65) of the NF-κB family, which play a key role in cellular inflammatory response, immune response and other processes. The expression level of p50 in activated HS27 cells is significantly decreased, indicating that NF-κB signaling is an upstream positive regulator of EMMPRIN in cells, and activated fibroblasts have an inhibitory effect on it.

[0151] This suggests that activated fibroblasts can resist the upstream activation of NF-κB signaling by the pro-inflammatory microenvironment (e.g., cytokines such as IL-1 and TNF-α), thereby protecting basement membrane repair.

[0152] Example 7: Activated fibroblasts promote punch biopsy healing

[0153] This embodiment involves repairing punch biopsy wounds with activated fibroblasts. The specific steps are as follows: 12 Sprague-Dawley male rats with an initial weight of 250-300g were shaved on the back to produce two full-depth punch biopsy wounds (6mm in diameter). Then, CUR / POCA-Glt-NM of the same size was applied to the wound to activate the fibroblasts around the wound. In order to avoid displacement of the drug-loaded film, all wounds were fixed with 3M sterile surgical tape. The changes in the wound were observed. After 15 days, the animals were euthanized and the wound histology was harvested for further analysis. The results are shown in Figure 2. Fig.11 shown.

[0154] like Fig.11 As shown in A, compared with FibroCtrl ivv (without membrane) and FibroCUR-ivv (empty membrane), FibroCUR+ivv (drug-loaded membrane) can significantly accelerate wound closure on days 7, 11, and 15, and can better complete skin repair by causing greater differentiation of the epidermis and better maturation of the dermis.

[0155] like Fig.11 B shows the repaired wound with FibroCUR-ivv ( Fig.11B(b,e,h)) compared with FibroCUR+ivv repair wound ( Fig.11 B(c,f,i)) has complete epithelial re-formation ( Fig.11 B(f,3)) and fully developed epidermis ( Fig.11 B(i,6)), showing a well-differentiated epithelium. In addition to epithelial regeneration, the Fig.11 B(h,8)) or FibroCtrl ivv( Fig.11 B(g,7)) compared with wounds with FibroCUR+ivv ( Fig.11 B(i,9)) also showed significant fibroblast proliferation and abundant collagen deposition.

[0156] Likewise, collagen-specific staining (Masson and Sirius Red) further verified that wounds containing FibroCUR+ivv healed with significantly increased collagen production, thicker and denser collagen bundles ( Fig.11 C(c,f); Fig.11 C(i)).

[0157] On the other hand, wounds containing FibroCUR-ivv or FibroCtrl ivv formed only loose collagen bundles with obvious hollow gaps ( Fig.11 C(b,e); Fig.11 C(a,d), Fig.11 C(h), Fig.11 C(g)). Polarized light microscopy (crossed polarized fields) results showed that the Fig.11 C(k)) or FibroCtrl ivv( Fig.11 In contrast, wounds containing FibroCUR+ivv showed only loosely arranged collagen fibers, while wounds containing FibroCUR+ivv had a more mature collagen network ( Fig.11 C(i)).

[0158] Example 8: Activated fibroblasts promote healing of refractory inflammatory wounds

[0159] In this example, it was further studied whether refractory or difficult-to-heal inflammatory wounds could be rescued by reconstructing a steady-state basement membrane by generating curcumin-activated fibroblasts (FibroCUR+ivv) in vivo. A diabetic mouse model of chronic wounds (db / db mice) was selected, and the curcumin-loaded poly(octyl citrate)-gelatin nanofiber membrane prepared in Example 1 was used to carefully evaluate the healing results from multiple angles.

[0160] This embodiment involves the use of activated fibroblasts to repair difficult-to-heal inflammatory wounds. The specific steps are as follows: 12 type 2 diabetes mouse models (db / db mice) with an initial weight of 250-300g were shaved on the back to produce two full-depth punch biopsy wounds (6mm in diameter). Then, CUR / POCA-Glt-NM of the same size was made into a shape consistent with the wound morphology and applied to the wound to activate the fibroblasts around the wound. In order to avoid displacement of the drug-loaded film, all wounds were fixed with 3M sterile surgical tape. The changes in the wound were observed. After 15 days, the animals were euthanized and the wound histology was harvested for further analysis. The results are as follows. Fig.12 shown.

[0161] like Fig.12 As shown in A, the wounds of wild-type control mice healed on day 13 and re-epithelialization was almost complete; in db / db mice in the FibroCUR-ivv group or the FibroCtrl ivv group, scab formation was observed, but no obvious wound closure was observed. However, the db / db mice in the FibroCUR+ivv group showed significantly accelerated wound healing, with almost complete wound closure and good epithelial regeneration on day 16 ( Fig.12 A, left panel). FibroCUR+ivv mice showed significantly faster wound closure on day 6 and thereafter compared with mice from the FibroCUR-ivv group (***P<0.001) or the FibroCtrl ivv group (***P<0.001). Fig.12 A, right).

[0162] Histological analysis (H&E staining) further demonstrated that tissues from FibroCUR+ivv group db / db mice were histologically more similar to those from healthy wild-type mice ( Fig.12 B(a,e)), showing fully developed epithelial (epidermal) differentiation and abundant collagen deposition in the dermis ( Fig.12 B(d,h)). In contrast, the Fig.12 B(c,g)) or FibroCtrl ivv group ( Fig.12 B (b, f)) Samples from db / db mice showed no or poor re-epithelialization, with significantly reduced collagen deposition and absent or indistinct collagen network organization. The expression levels and localization of relevant markers were investigated by immunohistochemistry.

[0163] Compared with db / db mice in the FibroCUR-ivv group or the FibroCtrl ivv group, the expression of EMMPRIN, CCL2, and CD68 in the FibroCUR+ivv group was significantly decreased, and the expression of laminin and COL4 was significantly increased, which was closer to that of healthy wild-type mice ( Fig.12 C). The significantly enriched laminin and COL4 staining (brown precipitate) is located at the epidermal-dermal junction, indicating successful basement membrane reconstruction.

[0164] These results confirm that the introduction of FibroCUR+ivv into chronic wounds (especially difficult-to-heal inflammatory wounds) can effectively promote regenerative repair through steady-state basement membrane reconstruction.

[0165] Example 9: Activated fibroblasts promote scarless healing of stress wounds

[0166] This embodiment involves scarless repair of stress wounds using activated fibroblasts. The specific steps are as follows: a porcine model of hypertrophic scar (HTS) formation is developed, which increases mechanical stress on the wound to cause hypertrophic scars similar to those in human wound repair failure.

[0167] First, circular full-thickness skin defects were created on the backs of four Chinese Bama miniature pigs. These wounds were randomly divided into three groups (n = 4 / group) and treated with CUR / POCA-Glt NM (FibroCUR+ivv group, Fig.12 D(g)), POCA-Glt NM(FibroCUR-ivv group, i.e., different treatments, Fig.12 D(e)), or untreated (FibroCtrl ivv group, i.e. tension wound healing as experimental control, Fig.12 D(c)). Primary wound closure was then performed. A self-designed, threaded rod-based device capable of providing sustained tension was placed over each closed wound by fixing a metal plate stent to the intact lateral skin ( Fig.12 D, upper panel). Four days after the initial closure, intermittent suture removal was performed (all remaining sutures were removed on day 7), and the mechanical distraction device was activated by carefully tightening the threaded rod by 4 mm (every other day thereafter until the maximum expansion) to the maximum length of the screw (i.e., the maximum tension state), and this maximum tension state was maintained until the measurement endpoint of day 35 (5 weeks) to apply continuous tension on the wound. In addition, two other controls were introduced to verify the usability of the mechanical stress-induced hypertrophic scar formation model: a group of tension-free wound healing as a negative control (linear skin incision with an inactivated traction device, n = 4 / group, Fig.12D(a)), and a group of burn wound healing as positive control, (skin burn with a diameter of 2 cm, n = 4 / group, Fig.12 D(i)).

[0168] The results are as follows Fig.12 As shown in the figure below D, obvious morphological changes occurred after 5 weeks. The wounds healed under mechanical stress (FibroCtrl ivv group) showed a significant increase in proliferation and fibrosis, with the fiber bands arranged parallel to the direction of the applied load, and the structure was the same as that of adult hypertrophic scars ( Fig.12 D(d)). In contrast, wounds healed without tension (negative control) did not show a significant increase in volume and thickness, which was comparable to the surrounding normal skin ( Fig.12 D(b)). Although the creation of burn wounds (positive control) did not cause hypertrophic scars to increase, they shrank significantly into small, dark, and distorted scar tissues ( Fig.12 D(j)).

[0169] The above results suggest that stress wounds under mechanical load are prone to the formation of hypertrophic scars. The experimental results showed that the formation of hypertrophic scars in the wounds of the FibroCUR+ivv group was significantly reduced ( Fig.12 D(h)), while the wounds in the FibroCUR-ivv group still showed a significant increase in the fiber contracture band, with a significant increase in volume and thickness ( Fig.12 D(f)). This indicates that the introduction of curcumin via FibroCUR+ into the stress wound surface (or wound site) can effectively inhibit the formation of hypertrophic scars.

[0170] To provide further evidence, this example continues to evaluate the histological characteristics of newly healed wounds. Fig.12 E shows that the wound healed without tension (negative control) shows well-defined epidermal and dermal layers (including the papillary dermis). In the epidermis, keratinocytes can be seen differentiating and maturing as they pass outward. Meanwhile, the extracellular space is evenly distributed throughout the dermis, as dermal fibroblasts constitute the connective tissue with well-organized collagen fibers in a “basket-weave” orientation ( Fig.12 E(a)).

[0171] In contrast, the histological structure of the burn wound (positive control) was significantly different, with the disappearance of epidermal ridges (flat epidermis), a significant reduction in extracellular space, and the disappearance of the reticular weaving direction of deposited collagen, replaced by very dense and coarse collagen fibers ( Fig.12E(b)). Similar to the positive control, the healed wounds in the FibroCtrl ivv group (tension wound healing) were found to exhibit several typical features that met the histological criteria of hypertrophic scars. These features included a thick epidermal layer without reticular ridges, a high interstitial density in the dermis as it was occupied by stretched but tough and dense collagen bundles parallel to the skin surface ( Fig.12 E(c)). This confirms the success of the porcine model in which refractory wounds terminated with hypertrophic scars.

[0172] On the other hand, the healed wounds in the FibroCUR+ivv group restored a clear epidermal layer (e.g., with reticular ridges) and well-ordered collagen fibers in the dermis arranged in a basket-weave pattern, which was more similar to that found in the negative control ( Fig.12 E(e)). Meanwhile, despite the thinner epidermis and obvious finger-like protrusions, the healed wounds in the FibroCUR-ivv group mainly showed histological features more similar to those in the FibroCtrl ivv group ( Fig.12 E(d)).

[0173] This example also performed PicroSirius red staining and polarized light microscopy (crossed polarized fields) to evaluate the quality of collagen bundles by detecting birefringence. The samples of the FibroCtrl ivv group and the burn group (positive control) showed thick and dense collagen bundles parallel to each other ( Fig.12 F(b), Fig.12 F(e)). In contrast, samples of collagen bundles deposited in tension-free wound healing (negative control) formed a regular interwoven structure that was fine and well-aligned, consistent with the "basket weave" orientation described above in histological studies ( Fig.12 F(a)). Although the healed wounds in the FibroCUR-ivv group showed a collagen bundle structure very similar to that in the FibroCtrl ivv group and the burn group ( Fig.12 F(c)), but the wounds healed in the FibroCUR+ivv group restored the reticular collagen network and regularly interwoven collagen bundles comparable to those in the tension-free wounds ( Fig.12 F(d)). In addition, this example also used synchrotron-based X-ray tomography (SRXTM) to study the three-dimensional structural characteristics of the collagen matrix in the newly developed tissue, and obtained similar results to those of polarized light microscopy. In short, in the repair tissue of the FibroCtrl ivv group and the burn group (positive control), the spatial structure of thick and parallel collagen bundles and even fiber nodules was significantly densified ( Fig.12 G(a, d)), while in the repair tissue of the FibroCUR+ivv group, an anisotropic fibrous structure composed of interwoven thin collagen bundles forming a loose reticular matrix (i.e., a 3D “basket-weave” conformation, Fig.12 G(c)).

[0174] Quantitative assessment of the stiffness level of the repair tissue using atomic force microscopy (AFM) further demonstrated that changes in Young's modulus were consistent with the evolution of scar formation. Similar to the stiffness of burn wound healing tissue (21.0 ± 15.2 kPa), repair tissue from wounds healed under mechanical tension (FibroCtrl ivv group) showed a significantly higher Young's modulus (21.0 ± 15.2 kPa vs. 7.75 ± 4.04 kPa, ***P < 0.001) than wounds healed without tension (negative control). Apparently, topical application of POCA-Glt NM (FibroCUR-ivv group, in which only (un)activated fibroblasts were present in the wound surface) failed to prevent the increase in tissue stiffness, as a higher Young's modulus (18.59±16.61 kPa) could still be detected in the resulting repair tissue, but CUR / POCA-Glt NM treatment (FibroCUR+ivv group, i.e., in which the wound surface contained curcumin-activated fibroblasts) significantly reduced the Young's modulus in the repair tissue (8.39±5.74 kPa), indicating successful prevention of scar formation ( Fig.12 H). In addition, the results of the methenamine silver staining experiment (used to evaluate basement membrane formation) showed a significant increase in dark brown staining, especially at the epidermal-dermal junction to outline the basement membrane structure, which could only be seen in the repaired wounds of the FibroCUR+ivv group or in the tension-free wound healing group (negative control, Figure 7 I(a)) observed ( Fig.12 I(d)). In contrast, in the FibroCtrl ivv group ( Fig.12 I(b)) and FibroCUR-ivv group ( Fig.12 In the repair wounds of I(c)), basement membrane staining was very weak or absent, which was comparable to that of burn wound healing (positive control) ( Fig.12 I(e)).

[0175] These data demonstrate that FibroCUR+ helps heal refractory wounds not only by increasing extracellular matrix deposition but also likely by reestablishing homeostasis through homeostatic basement membrane remodeling. In other words, topical application of CUR / POCA-Glt NM could be an effective means of early intervention in regenerative wound repair.

[0176] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A use of curcumin or a pharmaceutically acceptable salt thereof, characterized in that: For preparing a preparation, product or composition for one or more uses selected from the following group: (a) for activating fibroblasts; (b) used to repair basement membrane; (c) Used to repair skin wounds with damaged basement membrane and inhibit scar formation.

2. The use according to claim 1, characterized in that The preparation, product or composition is used for one or more purposes selected from the following group: (d) downregulates the expression of CCL2 in fibroblasts; (e) downregulates the expression of EMMPRIN in fibroblasts; (f) Promotes the expression of laminin in fibroblasts.

3. The use according to claim 1, characterized in that The basement membrane damaged skin wound is selected from the following group: acute wound, chronic wound, or a combination thereof.

4. The use according to claim 3, characterized in that The acute wound is selected from the group consisting of a perforated biopsy wound, a stress wound, or a combination thereof.

5. The use according to claim 3, characterized in that The chronic wound is an inflammatory wound that is difficult to heal.

6. The use according to claim 1, characterized in that The formulation, preparation or composition comprises curcumin, caprylyl citrate, and gelatin.

7. The use according to claim 6, characterized in that The mass ratio of the curcumin, poly(octanediol citrate) and gelatin is (0.15±0.05):(0.5±0.1):(1±0.2).

8. A method for activating fibroblasts in vitro, characterized in that: The following steps are involved: (S1) providing a fibroblast to be activated; (S2) Incubating fibroblasts to be activated in the presence of the preparation, product or composition of claim 1.

9. A product, characterized in that The products described include: (a) Curcumin or a pharmaceutically acceptable salt thereof; and (b) A fiber membrane loaded with the curcumin or a pharmaceutically acceptable salt thereof, wherein the fiber membrane comprises caprylyl citrate and gelatin.

10. Use of the product according to claim 9, characterized in that: Used to prepare drugs to repair skin wounds and inhibit scar formation.

Images