Dressing for promoting wound healing and method for manufacturing same

Abstract

Provided is a method for manufacturing a dressing for promoting wound healing, comprising at least the following steps: providing a container; adding purified water to the container; adding a first active ingredient to the container, wherein the first active ingredient is low-molecular-weight hyaluronic acid (HA), which has a molecular weight of 500,000 daltons or less and accounts for 0.01 to 1 weight percentage of the entire dressing; and adding a second active ingredient to the container, wherein the second active ingredient is β-nicotinamide mononucleotide (β-NMN), which accounts for 0.01 to 1 weight percentage of the entire dressing, or a plant stem cell extract, which accounts for 0.1 to 5 weight percentage of the entire dressing. (FIG. 1)

Description

A dressing for promoting wound healing and a method for manufacturing the dressing. Technical Field

[0001] This invention relates to a dressing that promotes wound healing and a method for manufacturing the dressing, and particularly to a dressing having effective ingredients such as low molecular weight hyaluronic acid and β-nicotinamide mononucleotide (β-NMN) or plant stem cell extract, and a method for manufacturing the dressing. Background Technology

[0002] The skin, the largest organ in the human body, has multiple functions, including barrier, defense, and regulation. Therefore, damage to the skin not only affects appearance but can also lead to infection and systemic complications. Wound healing is a highly dynamic and multi-stage physiological process, sequentially including hemostasis, inflammation, proliferation, and tissue remodeling, each stage involving the synergistic effects of different cell populations and cytokines. However, in clinical settings such as diabetic foot ulcers, venous ulcers, pressure ulcers, traffic accident wounds, cuts, fall abrasions, and surgical wounds, causing chronic or acute wounds, these processes are often delayed or interrupted. Epidemiological data shows that tens of millions of patients worldwide are affected, placing a heavy burden on the healthcare system.

[0003] Compared to dry dressings, wet dressings offer the ability to provide an ideal microenvironment, thus improving the wound healing process. However, clinical treatment remains challenging, including inadequate infection control in chronic wounds, scar formation, and high heterogeneity among patients. Therefore, the industry continues to strive to improve wet dressings, developing a type that combines protective properties with the ability to promote angiogenesis and regeneration, particularly one that can significantly reduce inflammation, accelerate epidermal coverage, and enhance overall repair in the early and middle stages of the healing process. Summary of the Invention

[0004] As can be understood from the above description, the purpose of this invention is to provide a dressing that uses purified water as a solvent, thus enabling its use as a wet dressing. Thickening or emulsifying agents can also be added to transform the dressing into a gel or emulsion form. The dressing contains at least two active ingredients, one of which is low molecular weight hyaluronic acid (HA), and the other is β-nicotinamide mononucleotide (β-NMN) or a plant stem cell extract. Furthermore, the dressing exhibits anti-inflammatory and accelerated healing effects in the early stages of the healing process.

[0005] Based on one objective of this invention, embodiments of the invention provide a wound-healing dressing, using purified water as a solvent, comprising a first active ingredient and a second active ingredient. The first active ingredient is low molecular weight hyaluronic acid (HA), with a molecular weight below 500,000 Daltons, and accounts for 0.01 to 1% by weight of the total dressing. The second active ingredient is β-nicotinamide mononucleotide (β-NMN), accounting for 0.01 to 1% by weight of the total dressing, or a plant stem cell extract, accounting for 0.1 to 5% by weight of the total dressing.

[0006] Optionally, the wound-healing dressing may also include an osmotic component. The osmotic component is selected from at least one of chloride salts and lactate salts, and comprises 0.4 to 1% by weight of the total dressing.

[0007] Optionally, wound-healing dressings may also include solubilizing and film-forming moisturizing ingredients. The film-forming moisturizing ingredients constitute less than 8% by weight of the total dressing.

[0008] Optionally, the solubilizing and film-forming moisturizing ingredients include at least one of propylene glycol (PG), glycerin, dipropylene glycol (DPG), and 1,3-propanediol (PDO).

[0009] Optionally, the solubilizing and film-forming moisturizing ingredients consist of propylene glycol and glycerin, with propylene glycol accounting for 1 to 3% by weight of the dressing and glycerin accounting for 1 to 3% by weight of the dressing.

[0010] Optionally, wound-healing dressings may also include antiseptic components. The antiseptic components constitute less than 0.3% by weight of the total dressing.

[0011] Optionally, the preservative is methylparaben (MP) or ethylparaben (EP), comprising 0.05 to 0.2% by weight of the total dressing.

[0012] Optionally, the osmotic pressure adjusting component is sodium chloride.

[0013] Optionally, the second active ingredient is β-nicotinamide mononucleotide, and the wound-healing dressing also includes a third active ingredient. The third active ingredient is a plant stem cell extract, which accounts for less than 5% by weight of the total dressing.

[0014] Optionally, the plant stem cell extract is an extract of orchid stem cells.

[0015] Optionally, the wound-healing dressing may also include a pH-adjusting ingredient. This ingredient is used to adjust the pH of the dressing to between 5.5 and 9.

[0016] Optionally, the pH adjusting component is a phosphate.

[0017] Optionally, the dressing is an aqueous liquid.

[0018] Optionally, wound-healing dressings may also include thickening agents. These thickening agents are used to give the dressing a gel-like consistency.

[0019] Optionally, the thickening ingredient is trisaccharide gum, hydroxyethyl cellulose (HEC) gum, high molecular weight hyaluronic acid, or a combination thereof.

[0020] Optionally, wound-healing dressings may also include emulsifying ingredients. These emulsifying ingredients are used to give the dressing an emulsion form.

[0021] Optionally, the emulsifying ingredient is polysorbate (TWEEN), hexanediol laurate (SPAN), mineral oil, vegetable oil, or a combination thereof.

[0022] Based on one objective of the present invention, embodiments of the present invention provide a method for manufacturing a dressing to promote wound healing, comprising: providing a container; adding purified water to the container; adding a first active ingredient to the container, wherein the first active ingredient is low molecular weight hyaluronic acid with a molecular weight of less than 500,000 Daltons and accounting for 0.01 to 1 weight percentage of the total dressing; and adding a second active ingredient to the container, wherein the second active ingredient is β-nicotinamide mononucleotide, accounting for 0.01 to 1 weight percentage of the total dressing, or is a plant stem cell extract, accounting for 0.1 to 5 weight percentage of the total dressing.

[0023] Optionally, the method for manufacturing a dressing that promotes wound healing further includes adding an osmotic pressure adjusting component to a container, wherein the osmotic pressure adjusting component is selected from at least one of chloride salts and lactate salts, and accounts for 0.4 to 1% by weight of the total dressing.

[0024] Optionally, a method for manufacturing a dressing that promotes wound healing further includes adding an antiseptic component to a container, wherein the antiseptic component accounts for less than 0.3% by weight of the total dressing.

[0025] Optionally, the method for manufacturing a dressing that promotes wound healing further includes adding a solubilizing and film-forming moisturizing ingredient to a container, wherein the solubilizing and film-forming moisturizing ingredient accounts for less than 8% by weight of the total dressing.

[0026] Optionally, the dressing is in the form of an aqueous liquid, and the method for manufacturing a dressing to promote wound healing further includes: filtering and sterilizing the liquid in the container before placing it into a canister.

[0027] Optionally, the method for manufacturing a dressing that promotes wound healing further includes adding a pH-adjusting ingredient to the container to adjust the pH of the dressing to between 5.5 and 9.

[0028] Optionally, the second active ingredient is β-nicotinamide mononucleotide, and the method of manufacturing a dressing that promotes wound healing further includes adding a third active ingredient to the container, wherein the third active ingredient is a plant stem cell extract and accounts for less than 5% by weight of the total dressing.

[0029] Alternatively, a method for manufacturing a dressing that promotes wound healing may further include adding a thickening agent to a container to make the dressing gel-like.

[0030] Optionally, the method for manufacturing a dressing that promotes wound healing further includes: adding an emulsifying component to another container; pouring the liquid from one container into another container; and emulsifying the liquid in the other container to make the dressing an emulsion.

[0031] In summary, the present invention provides a dressing and a method for manufacturing the same, wherein the dressing can exhibit anti-inflammatory and healing-accelerating effects in the early stages of the healing process. Attached Figure Description

[0032] Figure 1 is a flowchart of a method for manufacturing a dressing to promote wound healing according to a first embodiment of the present invention.

[0033] Figure 2 is a flowchart of a method for manufacturing a dressing to promote wound healing according to a second embodiment of the present invention.

[0034] Figure 3 is a flowchart of a method for manufacturing a dressing to promote wound healing according to a third embodiment of the present invention.

[0035] Figure 4 is a flowchart of a method for manufacturing a dressing to promote wound healing according to a fourth embodiment of the present invention.

[0036] Figure 5 is a flowchart of a method for manufacturing a dressing to promote wound healing according to a fifth embodiment of the present invention.

[0037] Figure 6 is a histogram of wound healing rate in a cell experiment using the wound-healing dressing provided by the present invention.

[0038] Figure 7A shows sequential wound images from Day 0 to Day 7 of an animal experiment using different dressings for wound healing.

[0039] Figure 7B shows sequential wound images from Day 8 to Day 14 of an animal experiment using different dressings for wound healing.

[0040] Figure 8 is a line graph showing the ratio of wound area from Day 0 to Day 14 in animal experiments using different dressings for wound healing.

[0041] Figure 9 shows microscopic images of skin tissue at 1 and 2 weeks post-surgery in animal experiments using different dressings for wound healing, with a portion magnified to observe inflammation.

[0042] Figure 10 shows microscopic images of skin tissue at 1, 2, and 6 weeks post-surgery in animal experiments using different dressings to promote wound healing.

[0043] [Symbol Explanation] S101~S510: Steps Detailed Implementation

[0044] This invention aims to provide a dressing that exhibits anti-inflammatory and wound-healing-accelerating effects in the early stages of the healing process. The wound-healing-promoting dressing uses purified water as a solvent and includes a first active ingredient and a second active ingredient. The first active ingredient is low molecular weight hyaluronic acid (HA), with a molecular weight below 500,000 Daltons, and accounts for 0.01 to 1% by weight of the total dressing. The second active ingredient is β-nicotinamide mononucleotide (β-NMN), accounting for 0.01 to 1% by weight of the total dressing, or a plant stem cell extract, accounting for 0.1 to 5% by weight of the total dressing. To facilitate understanding of this invention, the following description, in conjunction with the inventive drawings and embodiments, is provided.

[0045] First, please refer to Figure 1, which is a flowchart of a method for manufacturing a wound-healing dressing according to a first embodiment of the present invention. First, in step S101, a container is provided. Then, in step S102, an antiseptic component is added to the container, wherein the antiseptic component accounts for less than 0.3% by weight of the total dressing. For example, but not limited to, 0.01, 0.05, 0.1, 0.15, 0.2, 0.25, or 0.3% by weight of the total dressing. In this embodiment, the antiseptic component is 1.2 grams of methylparaben (MP), but the present invention is not limited thereto, and the aforementioned methylparaben can be replaced with other types of antiseptic components, such as ethylparaben (EP). In other embodiments, the antiseptic component may be methylparaben or ethylparaben, accounting for 0.05 to 0.2% by weight of the total dressing.

[0046] Subsequently, in step S103, a solubilizing and film-forming moisturizing ingredient is added to the container, wherein the solubilizing and film-forming moisturizing ingredient accounts for less than 8% by weight of the total dressing. For example, but not limited to, 1, 2, 3, 4, 5, 6, 7, or 8% by weight of the total dressing. The solubilizing and film-forming moisturizing ingredient may include at least one of propylene glycol (PG), glycerin, dipropylene glycol (DPG), and 1,3-propanediol (PDO). For example, the solubilizing and film-forming moisturizing ingredient is composed of propylene glycol and glycerin, with propylene glycol accounting for 1 to 3% by weight of the total dressing, and glycerin accounting for 1 to 3% by weight of the total dressing.

[0047] In this embodiment, step S103 is detailed as follows: First, 20 grams of propylene glycol as a solubilizing and film-forming moisturizing ingredient are added to the container and stirred appropriately to dissolve the methylparaben preservative ingredient. Then, 30 grams of glycerin as a solubilizing and film-forming moisturizing ingredient are added to the container and stirred evenly. Please note that the stirring action described above is not a limiting requirement of this invention; stirring is merely to accelerate the dissolution and uniform diffusion rate.

[0048] Next, in step S104, purified water is added to the container. Further, in this embodiment, step S104 involves adding 914 grams of reverse osmosis (RO) water to the container and stirring until homogeneous. Please note that the stirring action described above is not a limiting element of the invention; stirring is merely to accelerate dissolution and uniform diffusion.

[0049] Next, in step S105, an osmotic pressure adjusting component is added to the container. This osmotic pressure adjusting component is selected from chloride salts, such as sodium chloride, and constitutes 0.4 to 1% by weight of the entire dressing. Furthermore, the osmotic pressure adjusting component may, for example but not limited to, constitute 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1% by weight of the entire dressing. It should be noted that the present invention is not limited to a single osmotic pressure adjusting component; the osmotic pressure adjusting component may also be selected from at least one of chloride salts and lactate salts. Further, in this embodiment, step S105 involves adding 9 grams of sodium chloride osmotic pressure adjusting component to the container and stirring until homogeneous. Please note that the stirring action described above is not a limiting requirement of the present invention; stirring is merely to accelerate the dissolution and uniform diffusion rate.

[0050] Furthermore, it should be noted that the dressing can be adjusted to low osmotic pressure, isotonic pressure, or high osmotic pressure, and the present invention is not limited to the dressing being low osmotic pressure, isotonic pressure, or high osmotic pressure. Preferably, the dressing is adjusted to high osmotic pressure, thereby allowing the unhealthy tissue fluid in the wound of the organism to escape.

[0051] Next, in step S106, a first active ingredient is added to the container. This first active ingredient is low molecular weight hyaluronic acid, with a molecular weight below 500,000 Daltons, and constitutes 0.01 to 1% by weight of the entire dressing. For example, but not limited to, 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1% by weight of the entire dressing. Further, in this embodiment, step S106 involves adding 2 grams of low molecular weight hyaluronic acid to the container and stirring until homogeneous. It should be noted that the stirring action described above is not a limiting requirement of the invention; stirring is merely to accelerate dissolution and uniform diffusion.

[0052] Then, in step S107, a second active ingredient is added to the container, wherein the second active ingredient is a plant stem cell extract, comprising 0.1 to 5% by weight of the entire dressing. For example, but not limited to, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, or 5% by weight of the entire dressing. Further, in this embodiment, step S107 involves adding 5 grams of the plant stem cell extract to the container and stirring until homogeneous. It should be noted that the stirring action described above is not a limiting requirement of the invention; stirring is merely to accelerate the dissolution and uniform diffusion rate.

[0053] In this embodiment, the plant stem cell extract is an extract of Orchidaceae plant stem cells, but the present invention is not limited thereto. Further, the method for obtaining the plant stem cell extract is described below. Under aseptic conditions, plant tissue is excised, and plant stem cell clusters are cultured using plant tissue culture techniques. A specific weight (e.g., 1 gram) of the aseptic plant stem cell cluster is taken, and an appropriate amount of liquid nitrogen is added to a mortar and rapidly ground into powder. Then, a specific volume (e.g., 1 ml) of sterile distilled water is added to the mortar and ground again. All the material in the mortar is transferred to a centrifuge tube of a specific volume (e.g., 15 ml), rapidly shaken for a specific time (e.g., 3 minutes) to mix thoroughly, and then centrifuged. During centrifugation, the temperature is set to a low temperature (e.g., 4 degrees Celsius), the rotation speed is set to several thousand to tens of thousands of revolutions per minute (e.g., 10,000 revolutions per minute), and the centrifugation time is set to several minutes (e.g., 3 minutes). Next, the supernatant from the centrifuge tube is transferred to another new centrifuge tube. Using a syringe, the supernatant is then passed through a filter membrane with a pore size of 0.1 to 0.5 micrometers (e.g., a 0.22-micrometer filter membrane) and collected in a microcentrifuge tube (Eppendorf Tube) for storage. The liquid in the microcentrifuge tube is a concentrated stock solution of the aforementioned plant stem cell extract. This concentrated stock solution is diluted thousands to tens of thousands of times to avoid damaging the cells. Therefore, the exact weight percentage of the plant stem cell extract dressing will depend on the dilution ratio and, depending on the dilution, will be 0.1 to 5% of the total dressing weight.

[0054] Next, in step S108, since the dressing is an aqueous liquid, the liquid in the container can be filtered and sterilized before being placed into the canister. The filtration can be performed using a filter membrane with a pore size of 0.1 to 0.5 micrometers, for example, but not limited to a filter membrane with a pore size of 0.22 micrometers.

[0055] Please note that the order of steps S102 to S107 is not intended to limit the invention; the order can be adjusted, and at least one of steps S102, S103, S105, and S108 may be removed as needed and in accordance with actual circumstances. Furthermore, between steps S107 and S108, a step of adding purified water to the container to adjust the overall weight of the dressing can be performed, for example, to make the overall weight of the dressing 1,000 grams.

[0056] Please refer to Figure 2, which is a flowchart of a method for manufacturing a wound-healing dressing according to a second embodiment of the present invention. First, in step S201, a container is provided. Then, in step S202, an antiseptic component is added to the container, wherein the antiseptic component accounts for less than 0.3% by weight of the total dressing, and in this embodiment, the antiseptic component is 1.2 grams of methylparaben.

[0057] Then, in step S203, the solubilizing and film-forming moisturizing ingredients are added to the container, wherein the solubilizing and film-forming moisturizing ingredients account for less than 8% by weight of the total dressing. In this embodiment, the details of step S203 are as follows: first, 20 grams of propylene glycol solubilizing and film-forming moisturizing ingredients are added to the container and stirred appropriately to dissolve the methylparaben preservative ingredient; then, 30 grams of glycerin solubilizing and film-forming moisturizing ingredients are added to the container and stirred evenly. Furthermore, as mentioned above, the stirring action is not a limiting requirement of the present invention.

[0058] Next, in step S204, purified water is added to the container. Further, in this embodiment, step S204 involves adding 850 grams of reverse osmosis water to the container and stirring until homogeneous. Additionally, as mentioned above, stirring is not a limiting element of this invention.

[0059] Then, in step S205, an osmotic pressure adjusting component is added to the container, wherein the osmotic pressure adjusting component is selected from at least one of chloride salts and lactate salts, and accounts for 0.4 to 1% by weight of the total dressing. In this embodiment, step S205 is detailed as follows: 9 grams of sodium chloride osmotic pressure adjusting component are added to the container and stirred until homogeneous. Furthermore, as mentioned above, stirring is not a limiting requirement of the present invention.

[0060] Next, in step S206, a first active ingredient is added to the container, wherein the first active ingredient is low molecular weight hyaluronic acid with a molecular weight below 500,000 Daltons and accounts for 0.01 to 1% by weight of the entire dressing. In this embodiment, step S206 is detailed as follows: 3.2 grams of low molecular weight hyaluronic acid are added to the container and stirred until homogeneous. Furthermore, as mentioned above, the stirring action is not a limiting requirement of the present invention.

[0061] Then, in step S207, a second active ingredient is added to the container, wherein the second active ingredient is β-nicotinamide mononucleotide, comprising 0.01 to 1 weight percentage of the total dressing. β-nicotinamide mononucleotide may be, for example, but is not limited to, 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, or 1 weight percentage of the total dressing. Further, in this embodiment, step S207 is detailed as follows: 1 gram of β-nicotinamide mononucleotide is added to the container and stirred until homogeneous. Additionally, as mentioned above, stirring is not a limiting requirement of the present invention.

[0062] Next, in step S208, a pH-adjusting ingredient is added to the container to adjust the pH of the dressing to between 5.5 and 9, for example, to 5.5, 6, 6.5, 7, 7.5, 8, 8.5, or 9. The pH-adjusting ingredient can be a phosphate, such as, but not limited to, at least one of potassium dihydrogen phosphate (KH₂PO₄) and disodium hydrogen phosphate (Na₂HPO₄), but the present invention is not limited to the type of pH-adjusting ingredient. In this embodiment, potassium dihydrogen phosphate and disodium hydrogen phosphate are added to adjust the pH of the dressing to 6.5.

[0063] Next, in step S209, purified water is added to the container to adjust the total weight of the dressing, for example, to a total weight of 1,000 grams. Then, in step S210, since the dressing is an aqueous liquid, the liquid in the container can be filtered and sterilized before being placed into the canister. Filtration can be performed using a filter membrane with a pore size of 0.1 to 0.5 micrometers, for example, but not limited to, a filter membrane with a pore size of 0.22 micrometers.

[0064] Please note that the order of steps S202 to S207 is not intended to limit the present invention. The order can be changed, and at least one of steps S202, S203, S205, S208, S209 and S210 may be removed as needed and in accordance with actual circumstances.

[0065] Please refer to Figure 3, which is a flowchart of a method for manufacturing a wound-healing dressing according to a third embodiment of the present invention. First, in step S301, a container is provided. Then, in step S302, an antiseptic component is added to the container, wherein the antiseptic component accounts for less than 0.3% by weight of the total dressing, and in this embodiment, the antiseptic component is 2 grams of methylparaben.

[0066] Then, in step S303, the solubilizing and film-forming moisturizing ingredients are added to the container, wherein the solubilizing and film-forming moisturizing ingredients account for less than 8% by weight of the total dressing. In this embodiment, the details of step S303 are as follows: first, 30 grams of propylene glycol solubilizing and film-forming moisturizing ingredients are added to the container and stirred appropriately to dissolve the methylparaben preservative ingredient; then, 30 grams of glycerin solubilizing and film-forming moisturizing ingredients are added to the container and stirred evenly. Furthermore, as mentioned above, the stirring action is not a limiting requirement of the present invention.

[0067] Next, in step S304, purified water is added to the container. Further, in this embodiment, step S304 involves adding 800 grams of reverse osmosis water at 40 to 50 degrees Celsius to the container and stirring until homogeneous. Additionally, as mentioned above, the stirring action is not a limiting requirement of this invention.

[0068] Then, in step S305, an osmotic pressure adjusting component is added to the container, wherein the osmotic pressure adjusting component is selected from at least one of chloride salts and lactate salts, and accounts for 0.4 to 1% by weight of the total dressing. In this embodiment, step S305 is detailed as follows: 9 grams of sodium chloride osmotic pressure adjusting component are added to the container and stirred until homogeneous. Furthermore, as mentioned above, stirring is not a limiting requirement of the invention.

[0069] Next, in step S306, a first active ingredient is added to the container, wherein the first active ingredient is low molecular weight hyaluronic acid with a molecular weight below 500,000 Daltons and accounts for 0.01 to 1% by weight of the total dressing. In this embodiment, step S306 is detailed as follows: 1.2 grams of low molecular weight hyaluronic acid is added to the container and stirred until homogeneous. Furthermore, as mentioned above, the stirring action is not a limiting requirement of the present invention.

[0070] Then, in step S307, a second active ingredient is added to the container, wherein the second active ingredient is a plant stem cell extract, which accounts for 0.1 to 5% by weight of the entire dressing. Further, in this embodiment, step S307 involves adding 2 grams of plant stem cell extract to the container and stirring until homogeneous. Additionally, as mentioned above, stirring is not a limiting element of the invention, and the method of obtaining the plant stem cell extract is as described above.

[0071] Next, in step S308, pH adjusting ingredients are added to the container to adjust the pH of the dressing to between 5.5 and 9. In this embodiment, potassium dihydrogen phosphate and disodium hydrogen phosphate are added to adjust the pH of the dressing to 6.6.

[0072] Next, in step S309, a thickening agent is added to the container to make the dressing gel-like. The thickening agent is styrax glue, hydroxyethyl cellulose (HEC) glue, high molecular weight hyaluronic acid, or a combination thereof, and the invention is not limited thereto. In this embodiment, the details of step S309 are as follows: 3.00 grams of hydroxyethyl cellulose glue is added and stirred thoroughly until a gel-like state is achieved, and then the temperature is lowered to room temperature.

[0073] Next, in step S310, purified water is added to the container to adjust the total weight of the dressing, for example, to a total weight of 1,000 grams. After step S310, the gel-type dressing can be placed into a container.

[0074] Please note that the order of steps S302 to S307 is not intended to limit the present invention. The order can be changed, and at least one of steps S302, S303, S305, S308 and S310 may be removed as needed and in accordance with actual circumstances.

[0075] Please refer to Figure 4, which is a flowchart of a method for manufacturing a wound-healing dressing according to a fourth embodiment of the present invention. First, in step S401, a container is provided. Then, in step S402, an antiseptic component is added to the container, wherein the antiseptic component accounts for less than 0.3% by weight of the total dressing, and in this embodiment, the antiseptic component is 2 grams of methylparaben.

[0076] Then, in step S403, the solubilizing and film-forming moisturizing ingredients are added to the container, wherein the solubilizing and film-forming moisturizing ingredients account for less than 8% by weight of the total dressing. In this embodiment, the details of step S403 are as follows: first, 30 grams of propylene glycol solubilizing and film-forming moisturizing ingredients are added to the container and stirred appropriately to dissolve the methylparaben preservative ingredient; then, 30 grams of glycerin solubilizing and film-forming moisturizing ingredients are added to the container and stirred evenly. Furthermore, as mentioned above, the stirring action is not a limiting requirement of the present invention.

[0077] Next, in step S404, purified water is added to the container. Further, in this embodiment, step S404 involves adding 800 grams of reverse osmosis water at 40 to 50 degrees Celsius to the container and stirring until homogeneous. Additionally, as mentioned above, the stirring action is not a limiting requirement of this invention.

[0078] Then, in step S405, an osmotic pressure adjusting component is added to the container, wherein the osmotic pressure adjusting component is selected from at least one of chloride salts and lactate salts, and accounts for 0.4 to 1% by weight of the total dressing. In this embodiment, step S405 is detailed as follows: 6 grams of sodium chloride osmotic pressure adjusting component is added to the container and stirred until homogeneous. Furthermore, as mentioned above, stirring is not a limiting requirement of the invention.

[0079] Next, in step S406, a first active ingredient is added to the container, wherein the first active ingredient is low molecular weight hyaluronic acid with a molecular weight below 500,000 Daltons and accounts for 0.01 to 1% by weight of the entire dressing. In this embodiment, step S406 is detailed as follows: 0.2 grams of low molecular weight hyaluronic acid is added to the container and stirred until homogeneous. Furthermore, as mentioned above, the stirring action is not a limiting requirement of the present invention.

[0080] Then, in step S407, a second active ingredient, β-nicotinamide mononucleotide, is added to the container, comprising 0.01 to 1% by weight of the dressing. Further, in this embodiment, step S407 involves adding 0.2 g of β-nicotinamide mononucleotide to the container and stirring until homogeneous. Additionally, as mentioned above, stirring is not a limiting element of the invention.

[0081] Next, in step S408, pH adjusting ingredients are added to the container to adjust the pH of the dressing to between 5.5 and 9. In this embodiment, potassium dihydrogen phosphate and disodium hydrogen phosphate are added to adjust the pH of the dressing to 6.6.

[0082] Next, in step S409, another container is provided, and an emulsifying agent is added to this container. The emulsifying agent is used to transform the dressing into an emulsion form, wherein the emulsifying agent is polysorbate (TWEEN), hexanediol laurate (SPAN), mineral oil, vegetable oil, or a combination thereof. In this embodiment, the details of step S409 are as follows: 12.6 grams of TWEEN 20 and 17.4 grams of SPAN 20 are first added to another container, followed by 20 grams of mineral oil being added to yet another container.

[0083] Then, in step S410, the liquid in the container of the aqueous solvent-based dressing is transferred to another container containing the emulsifying component, and emulsification is performed to transform the dressing into an emulsion form. Further, in this embodiment, step S410 is detailed as follows: the emulsification device is turned on, and purified water is added to bring the total weight of the emulsion dressing to 1,000 grams. The emulsification speed of the emulsification device is set to 2,000 to 3,000 rpm, and the emulsification time is set to 10 to 15 minutes. After step S410, the emulsion-type dressing can be further processed into containers.

[0084] Please note that the order of steps S402 to S407 is not intended to limit the invention; the order can be adjusted, and at least one of steps S402, S403, S405, and S408 may be removed depending on needs and actual circumstances. Furthermore, in step S410, it is possible to adjust the total weight of the dressing by not adding purified water.

[0085] Please refer to Figure 5, which is a flowchart of a method for manufacturing a wound-healing dressing according to a fifth embodiment of the present invention. First, in step S501, a container is provided. Then, in step S502, an antiseptic component is added to the container, wherein the antiseptic component accounts for less than 0.3% by weight of the total dressing, and in this embodiment, the antiseptic component is 1.2 grams of methylparaben.

[0086] Then, in step S503, the solubilizing and film-forming moisturizing ingredients are added to the container, wherein the solubilizing and film-forming moisturizing ingredients account for less than 8% by weight of the total dressing. In this embodiment, the details of step S503 are as follows: first, 20 grams of propylene glycol solubilizing and film-forming moisturizing ingredients are added to the container and stirred appropriately to dissolve the methylparaben preservative ingredient; then, 30 grams of glycerin solubilizing and film-forming moisturizing ingredients are added to the container and stirred evenly. Furthermore, as mentioned above, the stirring action is not a limiting requirement of the present invention.

[0087] Next, in step S505, purified water is added to the container. Further, in this embodiment, step S405 involves adding 914 grams of reverse osmosis water to the container and stirring until homogeneous. Additionally, as mentioned above, stirring is not a limiting element of this invention.

[0088] Then, in step S505, an osmotic pressure adjusting component is added to the container, wherein the osmotic pressure adjusting component is selected from at least one of chloride salts and lactate salts, and accounts for 0.4 to 1% by weight of the total dressing. In this embodiment, step S505 is detailed as follows: 9 grams of sodium chloride osmotic pressure adjusting component are added to the container and stirred until homogeneous. Furthermore, as mentioned above, stirring is not a limiting requirement of the present invention.

[0089] Next, in step S506, a first active ingredient is added to the container, wherein the first active ingredient is low molecular weight hyaluronic acid with a molecular weight below 500,000 Daltons and accounting for 0.01 to 1% by weight of the entire dressing. In this embodiment, step S406 is detailed as follows: 2 grams of low molecular weight hyaluronic acid are added to the container and stirred evenly. Furthermore, as mentioned above, stirring is not a limiting requirement of this invention.

[0090] Then, in step S507, a second active ingredient, β-nicotinamide mononucleotide, is added to the container, comprising 0.01 to 1% by weight of the dressing. Further, in this embodiment, step S507 involves adding 2 grams of β-nicotinamide mononucleotide to the container and stirring until homogeneous. Additionally, as mentioned above, stirring is not a limiting element of the invention.

[0091] Then, in step S508, a third active ingredient is added to the container, wherein the third active ingredient is a plant stem cell extract, which accounts for less than 5% by weight of the total dressing. Further, in this embodiment, step S307 involves adding 4 grams of plant stem cell extract to the container and stirring until homogeneous. Additionally, as mentioned above, stirring is not a limiting element of the invention, and the method of obtaining the plant stem cell extract is as described above.

[0092] Next, in step S509, purified water is added to the container to adjust the total weight of the dressing and stirred evenly, for example, to make the total weight of the dressing 1,000 grams. Furthermore, as mentioned above, the stirring action is not a limiting requirement of the present invention. Next, in step S510, since the dressing is an aqueous liquid, the liquid in the container can be filtered and sterilized before being placed into the canister. Filtration can be performed using a filter membrane with a pore size of 0.1 to 0.5 micrometers, for example, but not limited to, a filter membrane with a pore size of 0.22 micrometers.

[0093] Please note that the order of steps S502 to S508 is not intended to limit the present invention. The order can be changed, and at least one of steps S502, S503, S505, S509 and S510 may be removed as needed and in accordance with actual circumstances.

[0094] Please refer to Figure 6, which is a histogram of wound healing rate in a cell experiment using the wound-healing dressing provided by this invention. In this cell experiment, wound healing was assessed using the mouse 3T3-L1 cell line. The mouse 3T3-L1 cells are fibroblasts derived from mouse embryos and are a widely used in vitro model for studying adipocyte formation (lipogenesis), obesity, and metabolic diseases. Cells were cultured using DMEM (Durbeco Modified Eagle Medium) supplemented with fetal bovine serum (FBS) and in 6-well cell culture dishes at a density of 1 × 10⁶ cells / well. 6Cells / well were seeded at a density of 1:1 and cultured for 24 hours to facilitate adhesion and monolayer formation. Subsequently, under sterile conditions, linear scratches were made on the cell monolayer using the tip of a 200 μL dropper to simulate wounds. If necessary, the cells were gently rinsed with sterile fetal bovine serum to remove airborne cells and debris generated by the scratches. The culture medium was then replaced with either the test dressing or physiological saline, and the culture dish was returned to the incubator to maintain the culture conditions.

[0095] Example 1 served as the first experimental group, and the culture medium for Example 1 contained the dressing described in the first embodiment. Example 2 served as the second experimental group, and the culture medium for Example 2 contained the dressing described in the second embodiment. The comparative examples served as the comparative group, and their culture medium contained a commercially available aqueous wound dressing that aids in the repair and regeneration of damaged tissue. The dressing contained sterile water, sodium hyaluronate, buffer solution, stabilizer, preservative, microencapsulated polysaccharide, and oligopeptide I. The control examples served as the control group, i.e., the blank control group, and their culture medium was physiological saline, i.e., no dressing was used. After the scratches were established, continuous imaging of the same scratch area was performed using an inverted microscope in a fixed field of view. Image acquisition time points included 0, 12, and 24 hours, and 48 hours of images were also collected for quantitative comparison.

[0096] All images were measured and analyzed using ImageJ software. The wound healing rate at each time point was calculated using the scratch width (or area) at 0 hours as the baseline. The formulas were: (Y_0HR-Y_12HR) / Y_0HR, (Y_12HR-Y_24HR) / Y_0HR, and (Y_24HR-Y_48HR) / Y_0HR, where Y_0HR is the scratch width (or area) at 0 hours, Y_12HR is the scratch width (or area) at 12 hours, Y_24HR is the scratch width (or area) at 24 hours, and Y_48HR is the scratch width (or area) at 48 hours. These calculations represent the healing progress relative to the initial wound size within each time interval.

[0097] As shown in Figure 6, the experimental results show that the quantitative data at 24 hours after the scratch treatment are the most significant. During the 24-hour experimental treatment, all three cases (Example 1, Example 2, and Comparative Case) showed overall healing effects. The wound disclosure rate of Example 1 was 85±8%, that of Example 2 was 80±10%, that of the Comparative Case was 65±8%, and that of the Control Case was 42±6%. Among these, the p-values ​​for Example 1 and Example 2 compared to the Control Case were less than 0.01, indicating that the results were statistically significant.

[0098] Compared to the control, the above cell experiments confirmed that the dressings of Experimental Example 1, Experimental Example 2, and the comparative example all exhibited faster wound closure speeds 24 hours after the cell lines were treated for wound healing. Furthermore, the healing completion rate of Experimental Example 1 and Experimental Example 2 rapidly approached completion within 24 hours, with Experimental Example 1 performing slightly better than Experimental Example 2, and its wound healing rate (or wound healing area) was at least twice that of the control. The dressings of Experimental Example 1 and Experimental Example 2 demonstrated wound healing effects, and showed superior wound healing performance compared to the dressing of the comparative example.

[0099] In addition to the cell experiments described above, related animal experiments were also conducted on the dressing provided by this invention, the details of which are described below.

[0100] Twenty-four thirteen-week-old male Wistar rats were used in the animal experiments. The animals were sourced from LASCO Biotechnology Co., Ltd. (Taiwan). All surgical procedures were performed under sterile conditions. Four 1cm incisions were made on the back of each rat. 2 The patients had full-thickness skin defects and were randomly assigned to control, comparative, experimental case 1, and experimental case 2 according to the experimental design. Immediately after surgery, the wound surface was treated according to each group and covered with the appropriate dressing.

[0101] Furthermore, Experimental Example 1 served as the first experimental group, and the wound was treated with the dressing described in the first embodiment. Experimental Example 2 served as the second experimental group, and the wound was treated with the dressing described in the second embodiment. Comparative Examples served as the comparative group, and the wound was treated with the dressing described in the commercially available form. Control Examples served as the control group, i.e., the blank control group, and the wound was treated with physiological saline.

[0102] In addition, in this invention, after anesthetizing the rat, the wound area is first marked on its back with a marker, and then four 1cm incisions are made at the marked locations. 2 A full-thickness skin defect was used as a skin trauma model. This model can simulate clinical full-thickness trauma and is used to assess the impact of different subsequent treatments on skin repair.

[0103] To compare the healing differences of Experimental Case 1, Experimental Case 2, Comparative Case and Control Case in the skin trauma model, this study followed up continuously from the day of injury (Day 0) to the end of Day 14 (Day 14), and measured and analyzed the wound area ratio (RWA), which was defined as the current wound area divided by the wound area on Day 0.

[0104] The wound area of ​​each group in Experimental Example 1, Experimental Example 2, Comparative Example and Control Example was standardized to 1 on Day 0. The decrease in the wound area ratio over time represents the gradual shrinkage and healing of the wound. The wound area ratios of each group in Experimental Example 1, Experimental Example 2, Comparative Example and Control Example from Day 0 to Day 14 are shown in Table 1 below, and the wound images from Day 0 to Day 14 are shown in Figures 7A and 7B.

[0105] Table 1

[0106] For ease of comparison, the control group was used as the baseline at each time point. If the wound area ratio of Experimental Case 1, Experimental Case 2, and the control group was lower than that of the control group, it indicated better healing speed; conversely, if the wound area ratio of Experimental Case 1, Experimental Case 2, and the control group was higher than that of the control group, it indicated relatively slow healing. The wound area ratios of Experimental Case 1, Experimental Case 2, the control group, and the control group all showed a decreasing trend over time, indicating that all animals entered progressive healing. However, the rate of decrease differed among different dressings, reflecting the different effects of each treatment on early contraction and epithelial coverage.

[0107] Please refer to Figure 8. Figure 8 is a line graph of the wound area ratio from Day 0 to Day 14 in animal experiments using different dressings for wound healing. The lines in Figure 8 are drawn based on Table 1. For ease of observation, the upper and lower error indicators for each point on the lines are omitted from the figure. From Day 1 to Day 3, the wound area ratio of Experimental Example 1 was significantly lower than that of the Comparative and Control examples on Day 1, indicating that the dressing of Experimental Example 1 could promote wound contraction in the early stages of inflammation. The wound area ratios of Experimental Example 2 and the Comparative examples were close to those of the Control example at this stage, showing no significant difference. From Day 1 to Day 3, the wound area ratios of Experimental Example 1, Experimental Example 2, the Comparative example, and the Control example all showed a decreasing trend with each day, indicating that all animals entered progressive healing. However, the rate of decrease differed among the different dressings, reflecting the different effects of each treatment on early contraction, granulation tissue filling, and epithelial coverage.

[0108] From Day 4 to Day 7, the wound area ratios of Experimental Case 1, Experimental Case 2, Comparative Case, and Control Case all continued to decrease. Among them, the wound area ratio of Experimental Case 1 remained at the lowest level, showing a sustained healing advantage. The wound area ratios of Experimental Case 2 and Comparative Case were slightly lower than those of Control Case, but the difference was not as significant as that of Experimental Case 1.

[0109] On Day 8, the wound area ratio of Experimental Example 1 was significantly lower than that of the Comparative and Control Examples. Subsequently, the wound area ratios of Experimental Example 1, Experimental Example 2, Comparative Example, and Control Example all approached stable low values, with the differences gradually narrowing. By Day 14, the wound area ratios of Experimental Example 1, Experimental Example 2, Comparative Example, and Control Example all decreased to around 0.15 to 0.18, indicating that the wounds had almost completely healed. However, the wound area ratio of Experimental Example 1 remained at the lowest value at most time points, indicating that the healing speed was relatively the fastest.

[0110] In summary, the dressing in Experiment 1 showed a sustained advantage over 14 days, especially in significantly accelerating wound healing in the early stages. Although the dressings in Experiment 2 and the control also showed some promoting effect, their effect was more moderate compared to that of the dressing in Experiment 1. The decrease in the wound area ratio in the control mainly reflects the natural healing process.

[0111] In this animal experiment, in order to evaluate the effect of different dressing treatments on skin wound inflammation, the skin tissue in the wound area was stained with hematoxylin-eosin at 1, 2 and 6 weeks after surgery, and the inflammation index was calculated. Hematoxylin-eosin staining is one of the most basic and widely used staining techniques in histology, embryology and pathology, and is also one of the gold standards for pathological diagnosis, so it will not be elaborated on here.

[0112] The interpretation of inflammatory response was based on microscopic histological observation, mainly including the degree of lymphocyte infiltration, vasodilation, and congestion. These indicators were used for evaluation, with 0 points representing no inflammatory response, no significant lymphocyte infiltration or vascular changes; 1 point representing mild inflammation, usually with a small amount of local lymphocyte infiltration accompanied by mild vasodilation; and 2 points representing moderate to severe inflammation, characterized by widespread lymphocyte infiltration and significant vasodilation. In this animal experiment, the above quantitative method was used to compare the effects of different dressing treatments on inflammatory response. Microscopic images of skin tissue at 1 and 2 weeks post-surgery for each case are shown in Figure 9. A portion of each image in Figure 9 was magnified to observe the inflammation. The inflammatory index of skin tissue at 1, 2, and 6 weeks post-surgery for each case is shown in Table 2.

[0113] Table 2

[0114] Table 2 shows the results for week 1. The inflammation indices for experimental case 1, experimental case 2, comparative case, and control case were 1.67±0.33, 1.67±0.33, 1.67±0.33, and 1.33±0.33, respectively. Overall, experimental case 1, experimental case 2, comparative case, and control case all showed high levels of inflammation. Obvious lymphocytic infiltration, vasodilation, and congestion were observed in the tissue sections, indicating that the first week postoperatively was the peak period of acute inflammation. Since all cases were in the pre-inflammatory stage at this time, the differences were not significant and represent part of the normal repair process.

[0115] Table 2 shows that the inflammation index of Case 1 and Case 2 decreased significantly to 0.33±0.33, with only sporadic lymphocyte infiltration observed in their tissue sections. Vascular dilation and congestion were also significantly reduced, indicating that these two dressings could effectively relieve inflammation in the intermediate stage. In contrast, the inflammation index of the comparative and control cases remained at 1.33±0.67, and persistent lymphocyte aggregation and vasodilation were still visible in their tissues, indicating that their inflammation had not been completely relieved.

[0116] Table 2 shows that the inflammation index in all cases decreased to 0.00±0.00, and significant lymphocyte infiltration, vasodilation, and congestion were no longer observed in the tissue sections, indicating that the skin tissue had recovered to a non-inflammatory state at this stage. This also demonstrates that all experimental animals remained healthy under long-term observation and possessed normal self-repair and self-healing abilities.

[0117] In summary, the dressings used in Experiments 1 and 2 significantly reduced inflammation in the middle stage (week 2), which was superior to the dressings used in the control group and the control group without dressings. This indicates that the dressings used in Experiments 1 and 2 have potential application value in promoting inflammation relief in the early and middle stages of wound healing. The small differences among the cases in week 1, which is the peak of inflammation, are normal. By week 6, all cases had recovered to a non-inflammatory state. This also confirms that the experimental animals in this study were healthy and possessed self-healing capabilities, enabling them to complete tissue repair under long-term observation.

[0118] Furthermore, in order to evaluate the impact of different dressing treatments on the epidermal repair of skin wounds, this animal experiment also performed hematoxylin-eosin staining on the skin tissue in the wound area at weeks 1, 2 and 6 postoperatively and calculated the epidermal repair index. The evaluation of epidermal repair mainly observed whether the arrangement of keratinocytes was neat and whether the layers were clearly distinguishable.

[0119] Under a microscope, the basal layer, granulosum, spinosum, and stratum corneum can be completely identified. Ideal repair is characterized by a uniform, intact stratum corneum without fragmentation or cavities, normal cell nuclei morphology, and uniform staining, indicating normal epidermal differentiation and maintenance of intact barrier function. A score of 0 represents no epidermal repair, 1 represents epidermal formation, and 2 represents complete epidermal tissue formation. The epidermal repair index at weeks 1, 2, and 6 post-surgery for the above cases is shown in Table 3.

[0120] Table 3

[0121] Table 3 shows that the epidermal repair index of all the above cases was 0.00±0.00, indicating that no new structures of stratum corneum or cell arrangement were observed in the sections.

[0122] Table 3 shows that the epidermal repair index of Experimental Case 1 and Experimental Case 2 reached 1.67±0.33 and 2.00±0.00 respectively, indicating that the epidermal repair entered an active period and the cell layers were clearly distinguishable. However, the epidermal repair index of the comparative case was only 0.67±0.67, and the epidermal repair index of the control case was 0.00±0.00, indicating that the repair progress was significantly lagging behind.

[0123] Table 3 shows that the epidermal repair index of experimental case 1, experimental case 2 and the control case all reached 2.00±0.00, indicating that the epidermal structure was intact and the stratum corneum was uniform. However, the epidermal repair index of the control case was 1.67±0.33, indicating that although there was recovery, it was slightly lower than that of experimental case 1, experimental case 2 and the control case.

[0124] In summary, the dressings used in Experimental Examples 1 and 2 allowed for noticeable epidermal repair in the skin tissue at the mid-stage. While the comparative and control cases did not show significant epidermal repair at the mid-stage, they ultimately achieved complete repair. Overall, the dressings used in Experimental Examples 1 and 2 effectively accelerated the epidermal reconstruction process.

[0125] Furthermore, to assess the impact of different dressing treatments on the dermal repair of skin wounds, this animal experiment also performed hematoxylin-eosin staining on the skin tissue in the wound area at weeks 1, 2, and 6 post-surgery, and calculated the dermal repair index. Dermal repair was primarily based on the arrangement of fibers and the state of fibroblasts. Under a microscope, when the fibers appeared wavy and evenly arranged, accompanied by a significant increase in the number of plump fibroblasts with clear nuclei, it indicated that the dermal structure was gradually recovering and possessed good tension and support. A score of 0 represented no dermal repair, 1 represented dermal formation, and 2 represented complete dermal tissue formation. The dermal repair indices for the above cases at weeks 1, 2, and 6 post-surgery are shown in Table 4.

[0126] Table 4

[0127] Table 4 shows that the dermal repair index of all the above cases was 0.00±0.00, and no fibers or new dermal tissue were observed in the sections.

[0128] Table 4 shows the results for week 2. The dermal repair index of Experimental Example 1 was 0.33±0.33. A small amount of fiber arrangement was visible in the dermal layer of Experimental Example 1, indicating that the dermal layer had mild repair. The dermal repair index of Experimental Example 2, Comparative Example and Control Example were still 0.00±0.00. No fibers or new dermal growth were observed in the sections.

[0129] Table 4 shows that the dermal repair index of Experimental Example 1, Experimental Example 2 and the control example all reached 1.00±0.00, and the dermal fibers were gradually regularized in the sections. However, the dermal repair index of the control example was 0.67±0.33, indicating that the degree of dermal repair was slightly lower.

[0130] In summary, the results show that the dressing in Experimental Example 1 initiated dermal repair in the middle stage of the skin tissue repair process. The dressings in Experimental Example 1 and the control examples gradually caught up in dermal repair in the later stages. Although the dermal layer of the control group's skin tissue eventually recovered, the extent was smaller. Overall, the dressing intervention in Experimental Example 1 helps to accelerate dermal repair.

[0131] Furthermore, to assess the impact of different dressing treatments on angiogenesis in skin wounds, this animal experiment performed hematoxylin-eosin staining on the skin tissue of the wound area at weeks 1, 2, and 6 post-surgery, and calculated the angiogenesis repair index. The assessment of angiogenesis focused on the number, distribution, and integrity of newly formed microvessels, and observed the regularity of endothelial cell arrangement and the clarity of lumen formation. The formation of new blood vessels represents the gradual recovery of local tissue oxygen and nutrient transport, and also reflects the metabolic demand and regenerative activity. A score of 0 indicates no angiogenesis repair, 1 indicates angiogenesis, and 2 indicates complete vascular tissue formation. The angiogenesis repair indexes for the above cases at weeks 1, 2, and 6 post-surgery are shown in Table 5.

[0132] Table 5

[0133] Table 5 shows that the angiogenesis repair index for all the above cases was 0.00±0.00, indicating that no signs of angiogenesis were observed in the sections.

[0134] Table 5 shows that the angiogenesis repair index of both Experimental Case 1 and Experimental Case 2 reached 1.00±0.00, indicating that new microvessels were visible in the sections. The angiogenesis repair index of the comparative case was 0.67±0.33, indicating that vascular recovery was limited. The angiogenesis repair index of the control case was still 0.00±0.00, indicating that no signs of angiogenesis were seen in the sections.

[0135] Table 5 shows the results of week 6. The angiogenesis repair index of experimental case 1 and the control case was 1.33±0.33, the angiogenesis repair index of experimental case 1 was 1.00±0.00, and the angiogenesis repair index of the control case was 1.33±0.33, indicating that angiogenesis in all cases was significant at this time.

[0136] In summary, the dressings in Experiment 1 and Experiment 2 can promote angiogenesis in the middle stage, while the dressing in the comparative case gradually catches up in the later stage. Although the control case did not show any effect in the early stage, it was eventually able to achieve a similar effect, showing that the self-healing ability can restore angiogenesis in the long term.

[0137] Furthermore, to evaluate the impact of different dressing treatments on hair follicle regeneration in skin wounds, this animal experiment performed hematoxylin-eosin staining on the skin tissue in the wound area at weeks 1, 2, and 6 post-surgery, and calculated the hair follicle repair index. Hair follicle regeneration was assessed based on the structural integrity and layering clarity of the hair matrix, dermal papilla, and hair bulb. When the structures of the hair matrix, dermal papilla, and hair bulb were clearly identifiable, it indicated that the skin appendages possessed regenerative potential, suggesting that the corresponding treatment method had potential value in promoting deep repair and tissue remodeling. A score of 0 represented no hair follicle regeneration, 1 represented hair follicle formation, and 2 represented complete hair follicle tissue formation. The hair follicle repair indices for the above cases at weeks 1, 2, and 6 post-surgery are shown in Table 6.

[0138] Table 6

[0139] Table 6 shows that the hair follicle repair index for all the above cases was 0.00±0.00, indicating that no hair follicle structure was observed in the sections.

[0140] Table 6 shows the results for week 2. The hair follicle repair indices for Experiment 1 and Experiment 2 were 0.33±0.33 and 0.67±0.67, respectively, indicating that hair follicle regeneration had begun. However, the hair follicle repair indices for the comparative and control cases were still 0.00±0.00, indicating that no hair follicle structure was observed in the sections.

[0141] Table 6 shows the results for week 6. The hair follicle repair index of both experimental case 2 and the control case reached 1.33±0.67, and the hair follicle repair index of the control case was also 1.33±0.67, indicating that the structure of the hair follicle, hair papilla and hair bulb area could be seen to be restored in the section. However, the hair follicle repair index of the experimental case was 0.67±0.67, indicating that the degree of hair follicle regeneration was relatively low.

[0142] In summary, the dressings used in Experimental Example 2 and the comparative example showed better results in hair follicle regeneration. The control example, treated without a dressing, also achieved a similar level of regeneration as the dressings used in Experimental Example 2 and the comparative example in the later stages. The dressing in Experimental Example 1 showed slightly lower regeneration, indicating that hair follicle repair occurred in a later stage. In terms of hair follicle repair, the differences between cases were not as significant as those in the other repair indicators mentioned above.

[0143] Furthermore, to assess the impact of different dressing treatments on the overall tissue repair of skin wounds, this animal experiment performed hematoxylin-eosin staining on the skin tissue of the wound area at weeks 1, 2, and 6 post-surgery, and calculated the overall repair index. The overall repair index was calculated by summing scores from four aspects: epidermal restoration and hyperplasia, dermal restoration and hyperplasia, angiogenesis, and hair follicle regeneration. Its significance lies in integrating multi-level repair indicators into a single quantitative value to comprehensively understand the overall healing status of the wound under different dressing treatments. This scoring method eliminates the limitations of judging a single structure, provides a more holistic assessment benchmark, and helps to identify the comprehensive effect of dressings on skin regeneration and tissue reconstruction. Microscopic images of the skin tissue at weeks 1, 2, and 6 post-surgery for the above cases are shown in Figure 10, and the overall repair indices at weeks 1, 2, and 6 post-surgery for the above cases are shown in Table 7.

[0144] Table 7

[0145] Table 7 shows that the overall repair index for all the above cases was 0.00 ± 0.00. The wound area in the sections was mainly composed of inflammatory cell infiltration and tissue loss, and no new epidermal or dermal structures were observed, indicating that the repair response had not yet been initiated.

[0146] Table 7 shows that the overall repair index of Experimental Case 1 and Experimental Case 2 increased to 3.33±0.33 and 3.67±0.67, respectively, indicating that the new epidermis gradually covered the wound and fibroblast proliferation appeared in the dermis. The overall repair index of the comparative case was only 1.33±0.88, indicating that the degree of repair was limited. The overall repair index of the control case remained at 0.00±0.00, indicating that it was still in an inflammatory and defective state.

[0147] Table 7 shows that the results at week 6 indicate that all the above cases achieved significant repair. The overall repair indices for experimental case 1, experimental case 2, comparative case, and control case were 5.00±1.00, 5.33±0.67, 5.67±0.88, and 5.00±1.53, respectively. Intact epidermis and remodeled dermis were visible in the sections, and fibrosis, angiogenesis, and hair follicle regeneration gradually became apparent.

[0148] In summary, the dressings in Test Examples 1 and 2 showed significantly faster healing rates in the intermediate stages. While the dressings in the control examples progressed slowly initially, they eventually reached a similar level of healing. The control examples showed the weakest healing. Over the long term, all the above cases showed healing, but the use of additional dressings accelerated healing, particularly the dressings in Test Examples 1 and 2.

[0149] In summary, compared to prior art, rigorous animal experiments have verified that the dressing provided by this invention is of great significance for improving clinical wound repair strategies. It has been demonstrated that it can significantly reduce the inflammation index, accelerate epidermal coverage and improve overall repair in the early and middle stages of the healing process, showing that it has the ability to promote a smooth transition from the inflammatory phase to the proliferative phase and bring substantial advantages in the healing timeline.

[0150] This invention is disclosed herein only by preferred embodiments. However, it should be understood by any person skilled in the art that the above embodiments are for illustrative purposes only and are not intended to limit the scope of the patent rights claimed by this invention. Any variations or substitutions that are equivalent or equivalent to the above embodiments should be interpreted as being covered within the spirit or scope of this invention. Therefore, the scope of protection of this invention should be based on the claims defined below.

Claims

1. A dressing that promotes wound healing, using purified water as a solvent, characterized in that, The dressing includes: The first active ingredient is low molecular weight hyaluronic acid (HA), with a molecular weight below 500,000 Daltons, and accounts for 0.01 to 1% by weight of the total dressing; and The second active ingredient is β-nicotinamide mononucleotide (β-NMN), which accounts for 0.01 to 1% by weight of the dressing, or is a plant stem cell extract, which accounts for 0.1 to 5% by weight of the dressing.

2. The wound healing promoting dressing as claimed in claim 1, wherein, The dressing also includes: The osmotic pressure adjusting component is selected from at least one of chloride salts and lactate salts, and accounts for 0.4 to 1% by weight of the total dressing.

3. The wound-healing dressing as described in claim 1, characterized in that, The dressing also includes: The solubilizing and film-forming moisturizing ingredients comprise less than 8% by weight of the total dressing.

4. The wound-healing-promoting dressing according to claim 3, characterized in that The solubilizing and film-forming moisturizing ingredients include at least one of propylene glycol (PG), glycerin, dipropylene glycol (DPG), and 1,3-propanediol (PDO).

5. The wound-healing dressing as described in claim 4, characterized in that, The solubilizing and film-forming moisturizing components are composed of propylene glycol and glycerin, with propylene glycol accounting for 1 to 3% by weight of the dressing and glycerin accounting for 1 to 3% by weight of the dressing.

6. The wound-healing dressing as described in claim 1, characterized in that, The dressing also includes: The preservative component comprises less than 0.3% by weight of the total dressing.

7. The wound-healing dressing as described in claim 6, characterized in that, The preservative component is methylparaben (MP) or ethylparaben (EP), which accounts for 0.05 to 0.2% by weight of the total dressing.

8. The wound-healing dressing as described in claim 2, characterized in that, The osmotic pressure adjusting component is sodium chloride.

9. The wound-healing dressing as described in claim 1, characterized in that, The second active ingredient is β-nicotinamide mononucleotide, and the dressing further includes: The third active ingredient is a plant stem cell extract, which accounts for less than 5% by weight of the entire dressing.

10. The wound-healing dressing as described in claim 1, characterized in that, The plant stem cell extract mentioned above is an extract of orchid stem cells.

11. The wound-healing dressing as described in claim 1, characterized in that, The dressing also includes: pH adjusting ingredients are used to adjust the pH value of the dressing to a value between 5.5 and 9.

0.

12. The wound-healing dressing as described in claim 11, characterized in that, The pH-adjusting component is a phosphate.

13. The wound-healing dressing as described in claim 1, characterized in that, The dressing is an aqueous liquid.

14. The wound-healing dressing as described in claim 1, characterized in that, The dressing also includes: A thickening agent is used to make the dressing gel-like.

15. The wound-healing dressing as described in claim 14, characterized in that, The thickening component is styrax, hydroxyethyl cellulose (HEC) gum, high molecular weight hyaluronic acid, or a combination thereof.

16. The wound-healing dressing as described in claim 1, characterized in that, The dressing also includes: An emulsifying component is used to make the dressing an emulsion.

17. The wound-healing dressing as described in claim 16, characterized in that, The emulsifying component is polysorbate (TWEEN), hexanediol laurate (SPAN), mineral oil, vegetable oil, or a combination thereof.

18. A method for manufacturing a dressing to promote wound healing, characterized in that, The method includes: Provide containers; Add purified water to the container; A first active ingredient is added to the container, wherein the first active ingredient is low molecular weight hyaluronic acid with a molecular weight below 500,000 Daltons and accounts for 0.01 to 1% by weight of the total dressing; and A second active ingredient is added to the container, wherein the second active ingredient is β-nicotinamide mononucleotide, which accounts for 0.01 to 1% by weight of the dressing, or is a plant stem cell extract, which accounts for 0.1 to 5% by weight of the dressing.

19. The method for manufacturing a wound-healing dressing as described in claim 18, characterized in that, The method further includes: An osmotic pressure adjusting component is added to the container, wherein the osmotic pressure adjusting component is selected from at least one of chloride salts and lactate salts, and accounts for 0.4 to 1% by weight of the total dressing.

20. The method for manufacturing a wound-healing dressing as claimed in claim 18, characterized in that, The method further includes: A preservative is added to the container, wherein the preservative constitutes less than 0.3% by weight of the total dressing.

21. The method for manufacturing a wound-healing dressing as described in claim 18, characterized in that, The method further includes: A solubilizing and film-forming moisturizing ingredient is added to the container, wherein the solubilizing and film-forming moisturizing ingredient accounts for less than 8% by weight of the total dressing.

22. The method for manufacturing a wound-healing dressing as described in claim 18, characterized in that, The dressing is an aqueous liquid, and the method further includes: The liquid in the container is filtered and sterilized before being transferred to a tank.

23. The method for manufacturing a wound-healing dressing as described in claim 18, characterized in that, The method further includes: A pH-adjusting ingredient is added to the container to adjust the pH of the dressing to between 5.5 and 9.

24. The method for manufacturing a wound-healing dressing as described in claim 18, characterized in that, The second active ingredient is β-nicotinamide mononucleotide, and the method further includes: A third active ingredient is added to the container, wherein the third active ingredient is a plant stem cell extract and accounts for less than 5% by weight of the entire dressing.

25. The method for manufacturing a wound-healing dressing as described in claim 18, characterized in that, The method further includes: A thickening agent is added to the container to make the dressing gel-like.

26. The method for manufacturing a wound-healing dressing as described in claim 18, characterized in that, The method further includes: Add the emulsifying ingredients to another container; Pour the liquid from the container into the other container; and In the other container, the liquid is emulsified to make the dressing an emulsion.

Images