WO2025206559A1 - Medical nanofiber membrane with antibacterial and hemostatic functions and hemostatic agent containing same - Google Patents

Abstract

The present invention provides a medical nanofiber membrane, which is produced by electrospinning a polymer solution comprising: polyethylene oxide; at least one selected from hyaluronic acid, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), alginic acid, carrageenan, poloxamer, and cellulose; and at least one selected from chitosan, thrombin, and collagen.

Description

Medical nanofiber membrane with antibacterial and hemostatic functions and hemostatic agent containing the same

The present invention relates to a medical nanofiber film having antibacterial and hemostatic functions and a hemostatic agent comprising the same.

Hemostasis refers to the body's natural defense mechanism, which occurs when bleeding occurs due to trauma or surgery. This process involves constriction of blood vessels, aggregation of platelets, and activation of the blood clotting mechanism. However, hemostatic agents are essential for the survival of patients with blood clotting disorders such as hemophilia and diabetes caused by platelet abnormalities, as well as patients undergoing critical conditions such as laparotomy or open-thoracotomy.

Bleeding during or after surgical procedures is a very serious problem that can lead to higher than expected mortality rates, making hemostatic agents essential during surgery. From minor procedures to laparotomy, hemostatic agents are crucial for preventing unexpected events, such as massive bleeding, in critical cases like laparotomy or thoracotomy, and various types of hemostatic agents are required.

Hemostatic agents can be classified into liquid glues that harden when applied to a wound, mesh-like patches that apply pressure to the wound, and mixed types, which contain hemostatic agents coated on a patch. Of these, mixed types are particularly useful because their hemostatic properties are activated upon contact with blood at the wound site. They are highly effective in vivo, flexible, and move with tissue, making them particularly versatile.

Depending on the type of surgery, various types of hemostatic agents are used, but a hemostatic agent that prevents external infection and adheres well to the site of bleeding is necessary. Representative laparoscopic surgeries such as hernia and colectomy often result in minimal bleeding, and electrocautery is sufficient for hemostasis. However, for patients with blood coagulation disorders such as cirrhosis or liver disease, a patch-type hemostatic agent is essential for reliable hemostasis. However, unlike liquid hemostatic agents, patch-type hemostatic agents are difficult to apply to wide or curved areas of bleeding, and their poor adhesiveness necessitates additional gauze or secondary hemostasis.

The present disclosure aims to provide a medical nanofiber membrane capable of stopping bleeding caused by trauma or surgery and applicable to a wide range of areas, and a hemostatic member including the same.

The present disclosure provides a medical nanofiber membrane manufactured by electrospinning a polymer solution comprising polyethylene oxide; at least one selected from hyaluronic acid, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), alginic acid, carrageenan, poloxamer, and cellulose; and at least one selected from chitosan, thrombin, and collagen.

The above medical nanofiber film may be prepared by mixing 1 to 10 parts by weight of one or more selected from hyaluronic acid, polyvinyl alcohol, polyvinylpyrrolidone, alginic acid, carrageenan, poloxamer, and cellulose with respect to 100 parts by weight of polyethylene oxide.

The above medical nanofiber membrane may be a mixture of 1 to 20 parts by weight of at least one of chitosan, thrombin, and collagen with respect to 100 parts by weight of polyethylene oxide.

The above polymer solution may contain chitosan, thrombin, and collagen.

The above polymer solution may contain 3 to 10 parts by weight of collagen and 1 to 10 parts by weight of chitosan per 1 part by weight of thrombin.

The above polymer solution may include a mixed solvent of water and ethanol.

The above polymer solution may have a weight ratio of polyethylene oxide, water, and ethanol of 10:10 to 30:40 to 70.

The above polyethylene oxide may have a number average molecular weight of 10,000 to 2,000,000 g/mol.

The above medical nanofibers may have an average diameter of 10 to 1000 nm.

The present disclosure provides a hemostatic member comprising a medical nanofiber membrane according to one embodiment of the present disclosure.

The biodegradable medical nanofiber film of the present invention has excellent skin adhesion and absorbency, and has the effect of reducing the sensation of heat in a wound and blocking bacterial infection.

The biodegradable medical nanofiber film of the present invention has antibacterial properties and can effectively protect the wound site from external infectious substances when applied to an open wound.

The hemostatic member of the present invention can be attached to a tissue site to quickly and easily stop the bleeding at the site.

Figure 1 illustrates a conceptual diagram of a method for manufacturing a medical nanofiber membrane using electrospinning of the present disclosure.

FIG. 2 illustrates a medical nanofiber membrane (Example 3) according to one embodiment of the present disclosure.

FIG. 3 illustrates a medical nanofiber membrane (Example 4) according to one embodiment of the present disclosure.

The present invention will be described in detail below. Terms used herein, unless specifically defined, should be interpreted as generally understood by those skilled in the art. The drawings and examples in this specification are intended to facilitate the understanding and practice of the present invention by those skilled in the art. Content that may obscure the gist of the invention may be omitted from the drawings and examples, and the present invention is not limited to the drawings and examples.

The singular forms used in this specification are intended to include the plural forms as well, unless the context clearly indicates otherwise.

Additionally, the numerical range used in the present invention includes lower and upper limits and all values within that range, increments logically derived from the shape and width of the defined range, all doubly defined values, and all possible combinations of upper and lower limits of numerical ranges defined in different shapes. Unless otherwise specifically defined in the specification of the present invention, values outside the numerical range that may arise due to experimental error or rounding of values are also included in the defined numerical range.

In this specification, terms such as include, have, and have mean that a feature or component described in the specification exists, and unless specifically limited, do not preclude the possibility that one or more other features or components may be added.

Traditionally, various types of hemostatic agents have been used depending on the type of surgery. Preventing external infection and maintaining a firm adherence to the site of bleeding are essential. Patch-type hemostatic agents are particularly essential for reliable hemostasis in patients with blood clotting disorders, such as cirrhosis or liver disease. However, their application is difficult in cases where the bleeding area is wide or curved.

The present disclosure aims to provide a medical nanofiber membrane that can be applied to a wide area where bleeding occurs due to trauma or surgery, and that can realize regeneration, hemostasis, and antibacterial effects at the wound site along with excellent adhesiveness and absorbency.

The present disclosure provides a medical nanofiber membrane manufactured by electrospinning a polymer solution comprising polyethylene oxide; at least one selected from hyaluronic acid, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), alginic acid, carrageenan, poloxamer, and cellulose; and at least one selected from chitosan, thrombin, and collagen.

The present disclosure can provide a medical nanofiber membrane that implements excellent hemostatic and antibacterial effects by being manufactured by electrospinning a polymer solution containing the material of the aforementioned composition.

According to one embodiment, the polymer solution may contain 1 to 10 parts by weight of at least one selected from hyaluronic acid, polyvinyl alcohol, polyvinylpyrrolidone, alginic acid, carrageenan, poloxamer, and cellulose, based on 100 parts by weight of polyethylene oxide, and specifically, the lower limit may be 1, 2, 3, or 4 parts by weight, and the upper limit may be 10, 9, 8, 7, or 6 parts by weight.

Specifically, the polymer solution may include at least one of hyaluronic acid, polyvinyl alcohol, polyvinyl pyrrolidone, and alginic acid, may include at least one of polyvinyl alcohol and polyvinyl pyrrolidone, and more specifically may include polyvinyl alcohol.

The viscosity average molecular weight (Mv) of the above hyaluronic acid, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), alginic acid, carrageenan, poloxamer, and cellulose may be, but is not limited to, 100,000 to 1,000,000 g/mol.

The above hyaluronic acid may be extracted from Streptococcus zooepidemicus , and may have a weight average molecular weight selected from 100,000 to 1,600,000 Da, specifically 156,000 to 1,590,000 Da, and may have a terminal group substituted with Na to increase solubility. At this time, if the weight average molecular weight of the hyaluronic acid is less than 100,000 Da, hydrolysis is too rapid and the stability of the final manufactured sheet is low, and if it exceeds 1,600,000 Da, the yield of nanofiber manufacturing is low, which is not preferable.

According to one embodiment, the polymer solution may contain 1 to 20 parts by weight of at least one of chitosan, thrombin, and collagen based on 100 parts by weight of polyethylene oxide, and specifically, the lower limit may be 1, 2, 3, 4, 5, or 5 parts by weight, and the upper limit may be 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, or 10 parts by weight.

According to one embodiment, the polymer solution may include chitosan, thrombin, and collagen.

According to one embodiment, the polymer solution may contain 3 to 10 parts by weight of collagen and 1 to 15 parts by weight of chitosan per 1 part by weight of thrombin, specifically 3 to 7 parts by weight of collagen and 3 to 15 parts by weight of chitosan, and more specifically 4 to 6 parts by weight of collagen and 5 to 12 parts by weight of chitosan.

According to one embodiment, the polymer solution may include a mixed solvent of water and ethanol.

According to one embodiment, the polymer solution may have a weight ratio of polyethylene oxide, water and ethanol of 10:10 to 30:40 to 70, specifically 10:10 to 25:50 to 60, specifically 10:15 to 20:50 to 60.

According to one embodiment, the polyethylene oxide may have a number average molecular weight of 10,000 to 2,000,000 g/mol, the lower limit being 10,000, 20,000, 30,000, 40,000, 50,000, 60,000, 70,000, 80,000, 90,000 or 100,000 g/mol, and the upper limit being 2,000,000, 1,800,000, 1,600,000, 1,400,000, 1,200,000 or 1,000,000 g/mol.

The above polymer solution may be reacted at 40 to 80°C for 0.5 to 5 hours, the lower limit may be 45, 50, or 55°C, and the upper limit may be 75, 70, or 65°C.

The above electrospinning may be performed under conditions of a voltage of 10 to 200 V and a radiation distance of 5 to 50 cm, and specifically, may be performed under conditions of a voltage of 10 to 100 V. In addition, the average voltage of the electrospinning may be 5 to 50 V, 10 to 40 V, or 15 to 30 V.

The polymer solution may contain a variety of substances such as flavoring agents and/or additives, for example, suitable flavoring agents may include menthol, eugenol, spearmint, peppermint, cocoa, vanilla, cinnamon, licorice, citrus or other fruit flavors and combinations thereof, examples of non-flavoring additives may include refrigerants, diluents, aerosol formers and the like.

According to one embodiment, the medical nanofiber membrane may have an average diameter of 10 to 1000 nm, specifically the lower limit may be 10, 50, 100 or 150 nm, and the upper limit may be 1000, 900, 800, 700, 600, 500 or 400 nm.

In one embodiment, the density of the medical nanofiber membrane may be from 1.00 to 2.00 mg/cm ³ , specifically the lower limit may be 1.00, 1.10, 1.20, 1.30, 1.40, 1.50 or 1.60 mg/cm ³ , and the upper limit may be 2.00, 1.80 or 1.70 mg/cm ³ .

The present disclosure provides a hemostatic member comprising the medical nanofiber membrane of the present disclosure. The hemostatic member may be manufactured in the form of a hemostatic patch, and may be specifically a hemostatic foam patch. The hemostatic member may be directly attached to a wound to perform a hemostatic action. As described above, the hemostatic member may include the medical nanofiber membrane of the present disclosure, thereby achieving an effective hemostatic action, as well as regeneration and antibacterial effects at the wound site.

Hereinafter, the medical nanofiber membrane according to the present invention will be described in more detail through specific examples. However, the following examples are merely references for further explanation of the present invention and are not intended to limit the present invention, which may be implemented in various forms. Furthermore, the terminology used in the description of the present invention is merely intended to effectively describe specific embodiments and is not intended to limit the present invention.

[Example 1]

1. Weigh 10 mg of polyethylene oxide (PEO) on a precision scale.

2. Place the Falcon tube on the still water scale, adjust the zero point, and add 18.75 mg of water.

3. Slowly add stirrer 1 powder to solution 2 and mix.

4. Add 5 mg of hyaluronic acid, 0.11 mg of chitosan, 0.53 mg of collagen, and 0.11 mg of thrombin to solution 3.

5. Stir the solution 4 at 60℃ for 1 hour to ensure it is fully dissolved.

6. Slowly add 56.25 mg of alcohol (ethanol) (density: 0.79 g/mL) to solution 5 while rotating it on a stirrer.

7. Stir solution 6 at 60℃ for 1 hour.

8. Prepare a polymer solution by filling the syringe with solution 7 in 1 cc units.

9. The prepared polymer solution was electrospun to produce a nanofiber membrane. At this time, the electrospinning can be performed using the friction electrostatic portable device of Patent No. 10-2353832.

[Examples 1 to 8 and Comparative Examples 1 to 2]

The same procedure as Example 1 was followed, except that the contents in Tables 1 and 2 below were used.

The diameter and density of the nanofiber membranes manufactured through Examples 1 to 8 and Comparative Examples 1 to 2 are shown in Tables 1 and 2 below.

In addition, the appearance of the nanofiber membranes manufactured through Examples 3 and 4 was observed using a scanning electron microscope (SEM) and the results are shown in FIGS. 2 and 3, respectively.

[Evaluation Example 1] Hemostasis and antibacterial evaluation

Hemostasis assessment

The rabbit abdominal aorta was exposed and injured using a 22G needle. Bleeding was checked, and the nanofiber specimens manufactured in Examples 1 to 8 and Comparative Examples 1 and 2 were applied to the bleeding site. Using a timer, the blood coagulation pattern and time were visually observed at 5-second intervals over time, and the time required for hemostasis was measured. The results are shown in Tables 1 and 2 below.

Antibacterial evaluation

The test was conducted according to the KSM 0146 antibacterial test method. 1.0 mL of E. coli was collected and inoculated into a 250 mL sterilized Erlenmeyer flask and cultured at 37°C for 24 hours. The dilution was prepared so that the number of bacteria was 1 × 10 ⁵ CFU/mL as measured by an absorbance photometer, and 1.0 mL was inoculated and mixed. The nanofiber specimens manufactured through Examples 1 to 8 and Comparative Examples 1 to 2 were added to this and shaken for 24 hours. After measuring the absorbance of the culture solution using a absorbance meter to calculate the number of bacteria, the decrease in the number of bacteria (antibacterial activity) compared to the initial number of bacteria was measured. The results are shown in Tables 1 and 2 below.

Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 polyethylene oxide
(mg) 10 10 10 10 10 10 10 (mg) Hyaluronic acid 5 Hyaluronic acid 5 Hyaluronic acid 5 polyvinyl alcohol 5 polyvinyl pyrrolidone 5 Alginic acid 5 Cellulose 5 Chitosan (mg) 0.11 0.53 1.06 0.11 0.11 0.11 0.11 Collagen (mg) 0.53 0.53 0.53 0.53 0.53 0.53 0.53 Thrombin (mg) 0.11 0.11 0.11 0.11 0.11 0.11 0.11 Water (mg) 18.75 18.75 18.75 18.75 18.75 18.75 18.75 Ethanol (mg) 56.25 56.25 56.25 56.25 56.25 56.25 56.25 Whether to produce tribostatic nanofibers O O O O O O O Whether or not hemostasis has occurred (hemorrhage time) 90 seconds 90 seconds 90 seconds 85 seconds 85 seconds 85 seconds 95 seconds Antibacterial activity (%) 90.38 97.25 98.12 99.99 96.15 64.12 98.23 nanofibers
Diameter (nm) 150~350 150~350 150~350 150~350 150~350 150~350 150~350 Density (mg/cm ³ ) 1.61 1.64 1.63 1.61 1.62 1.60 1.61

Example 8 Comparative Example 1 Comparative Example 2 polyethylene oxide
(mg) 10 10 10 (mg) Polyoxamer 5 Hyaluronic acid 5 Hyaluronic acid 5 Chitosan (mg) 0.11 0 2.12 Collagen (mg) 0.53 0 0.53 Thrombin (mg) 0.11 0 0.11 Water (mg) 18.75 18.75 18.75 Ethanol (mg) 56.25 56.25 56.25 Whether to produce tribostatic nanofibers O O X Whether or not hemostasis has occurred (hemorrhage time) 90 seconds 600 seconds - Antibacterial activity (%) 90.38 0 - nanofibers
Diameter (nm) 150~350 150~350 - Density (mg/cm ³ ) 1.65 1.59 -

According to Tables 1 and 2 above, when there is no content of chitosan, collagen, and thrombin as in Comparative Example 1, the hemostasis time takes longer than in the examples, and when the content of chitosan, collagen, and thrombin is added in an amount of 8 parts by weight or more to 100 parts by weight of a mixed solution of polyethylene oxide and water as in Comparative Example 2, it can be confirmed that nanofibers are not formed.

The present invention described above is not limited to the above-described embodiments and the attached drawings, and it will be apparent to a person skilled in the art to which the present invention pertains that various substitutions, modifications, and changes are possible within the scope that does not depart from the technical spirit of the present invention.

Claims (10)

polyethylene oxide; At least one selected from hyaluronic acid, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), alginic acid, carrageenan, poloxamer, and cellulose; and A medical nanofiber membrane manufactured by electrospinning a polymer solution containing at least one selected from chitosan, thrombin, and collagen. In the first paragraph, A medical nanofiber membrane, wherein 1 to 10 parts by weight of at least one selected from hyaluronic acid, polyvinyl alcohol, polyvinylpyrrolidone, alginic acid, carrageenan, poloxamer, and cellulose is mixed with 100 parts by weight of the above polyethylene oxide. In the first paragraph, A medical nanofiber membrane, wherein 1 to 20 parts by weight of at least one of chitosan, thrombin, and collagen is mixed with 100 parts by weight of the above polyethylene oxide. In the first paragraph, A medical nanofiber membrane, wherein the polymer solution comprises chitosan, thrombin and collagen. In paragraph 4, A medical nanofiber membrane, wherein the polymer solution contains 3 to 10 parts by weight of collagen and 1 to 10 parts by weight of chitosan per 1 part by weight of thrombin. In the first paragraph, A medical nanofiber membrane, wherein the polymer solution comprises a mixed solvent of water and ethanol. In paragraph 6, The above polymer solution is a medical nanofiber membrane having a weight ratio of polyethylene oxide, water and ethanol of 10:10~30:40~70. In the first paragraph, The above polyethylene oxide is a medical nanofiber membrane having a number average molecular weight of 10,000 to 2,000,000 g/mol. In the first paragraph, The above medical nanofiber is a medical nanofiber film having an average diameter of 10 to 1000 nm. A hemostatic member comprising a medical nanofiber membrane according to any one of claims 1 to 9.