KR102864293B1 - Manufacturing Method of collagen fiber wet-laid non-woven fabric - Google Patents

Abstract

The present invention relates to a method for manufacturing a nonwoven fabric composed of pure collagen polymer fibers, and according to the present invention, a method for manufacturing a nonwoven fabric manufactured only with pure collagen without adding a bonding fiber or other fibers after single-fiberizing a multifilament fiber manufactured using fish skin-extracted atelocollagen is provided, thereby providing a nonwoven fabric applicable to medical products such as locally absorbable hemostatic agents requiring flexibility, strength, and biocompatibility.

Description

Manufacturing Method of Collagen Fiber Wet-Laid Non-Woven Fabric

The present invention relates to a method for manufacturing a nonwoven fabric composed of pure collagen polymer fibers.

Collagen, also known as collagen fibers, is an essential protein that constitutes the extracellular matrix (ECM) of living organisms and plays a role in binding and supporting living tissues. Collagen is a fiber bundle composed of three twisted strands of amino acid linkages with a molecular weight of approximately 100,000 daltons. Collagen that maintains its triple helix structure has a molecular weight of approximately 300,000 daltons.

Collagen exists in over 20 types, varying in amino acid composition and three-dimensional structure. The most representative types include Type I collagen, found in skin; Type II, abundant in cartilage; and Type III, primarily found in blood vessels and the dermis. Of these, Type I collagen, which can be obtained from food byproducts such as livestock and marine products, is being extensively studied due to its superior properties as a medical material.

Products that utilize collagen's excellent cell affinity, mechanical properties, hemostatic properties, and biocompatibility include cell culture supports (SCAFFOLD), artificial dermis, hemostatic agents, and wound dressings. Most of them are used in the form of a sponge or single fibers obtained by freeze-drying collagen solutions. Methods for manufacturing fibers for utilizing the collagen as a medical material are being developed, such as a method of manufacturing fibers through wet spinning after solubilizing collagen in an acid solution, and a method using electrospinning. In the production of these fibers, atelocollagen is used, in which the telopeptide at the end of the triple helix structure, which does not cause rejection reactions in the human body, is removed.

Meanwhile, topically absorbable hemostasis is a product that has rapid hemostatic properties and biodegradability when applied to the bleeding site after surgical operations. Currently, the most widely used product is a freeze-dried collagen sponge type product.

However, sponge-type freeze-dried collagen hemostatic agents have the disadvantages of low flexibility, being brittle, and being difficult to manufacture in various weights and sizes. To overcome these shortcomings, the development of collagen products in the form of non-woven fabrics is required, but a method for producing non-woven fabrics composed solely of 100% collagen fibers that can maintain flexibility and strength has not yet been developed.

Accordingly, methods such as mixing PVA, a binder fiber, or mixing biocompatible fibers such as chitin or chitosan have been proposed to supplement the properties of relatively weak collagen fibers and to provide binding strength to nonwoven fabrics. However, compared to nonwoven fabrics composed only of pure collagen fibers, they have the disadvantage of lower hemostatic performance and bioabsorbability.

Additionally, because medical collagen products currently use collagen extracted from mammals such as cows and pigs, there are concerns about the transmission of zoonotic viruses.

Republic of Korea Patent Registration No. 10-2470501 (announced on November 25, 2022)

Accordingly, the present invention provides a nonwoven fabric composed only of pure collagen fibers having excellent hemostatic performance and bioabsorbability by not adding any bonding fibers or other fibers, and a nonwoven fabric composed only of fish-derived atelocollagen fibers that can reduce concerns about viral cross-infection by using collagen extracted from fish rather than collagen extracted from mammals such as cows and pigs, which are at risk of zoonotic virus transmission.

Therefore, according to the present invention, there is provided a step of preparing a spinning solution comprising 4 to 12 wt% of atelocollagen extracted from fish skin having a molecular weight of 100,000 to 300,000 Da and the remainder being an acetic acid solution;

A step of forming collagen multifilaments by discharging a predetermined amount of the above-mentioned radiation solution through a nozzle as a coagulant consisting of a mixture of ethanol and ammonia aqueous solution through a gear pump;

A step of passing the coagulated collagen multifilament through an alcohol bath containing 1.0 wt% of pluronic, a nonionic polymer surfactant, drying the same, and then winding the same on a winder;

A step of cutting the above collagen multifilament into 1 to 10 mm using a cutter;

A step of dispersing the cut collagen fibers in a dispersion solution containing an organic acid solution and ethanol;

A step of injecting the above collagen fiber-containing dispersion solution into a starting machine and then de-dissolving it to make a collagen fiber sheet;

A step of immersing the above collagen fiber sheet in ethanol to remove residual organic acid;

A method for manufacturing a collagen fiber wet nonwoven fabric is provided, characterized by comprising a step of compressing and drying to induce bonding between collagen fibers using some of the collagen fibers available on the sheet to create a wet nonwoven fabric.

The present invention will be described in more detail.

The present invention relates to a technology for manufacturing a sheet-shaped pure collagen wet nonwoven fabric having flexibility and strength through a process of spinning multifilament fibers using atelocollagen extracted from fish skin, cutting them into a predetermined length, uniformly dispersing them in a mixed solution of an acid solution and alcohol at a predetermined ratio, and then desolvating, compressing, and drying them.

The method for manufacturing a collagen fiber nonwoven fabric of the present invention begins with manufacturing a multifilament fiber from pure atelocollagen polymer extracted from fish skin.

The spinning solution is prepared by dissolving atelocollagen, an acid-soluble fish skin-extracted polymer with a molecular weight of 100,000 to 300,000 daltons (Da), in a weakly acidic solution. Collagen, the main component of the spinning solution, is also called collagen fiber and is an essential protein that constitutes the extracellular matrix (ECM) of living organisms and plays a role in binding and supporting living tissues. In the present invention, the acid-soluble polymer atelocollagen is preferably TYPE I collagen extracted from the skin of tilapia, a tropical fish species that has a higher hydroxyproline content among the collagen amino acid composition components than other fish, because the hydroxyproline content is 10 wt% or more of the total composition amino acids and the heat denaturation temperature is 35°C or higher, which can improve the strength of the manufactured fiber and reduce the possibility of heat denaturation during the nonwoven fabric manufacturing process.

The above-mentioned atelocollagen extracted from fish with a molecular weight of 100,000 to 300,000 daltons is dissolved in a weak acid solution of one of acetic acid, citric acid, formic acid, and lactic acid at a concentration of 4 to 12 wt%, and then defoamed to produce a spinning solution.

The above spinning solution is discharged through a nozzle with a diameter of 0.1 to 0.3 mm and an odd number of holes of 100 to 300 using a gear pump, and then spun in a coagulating solution consisting of a mixture of ethanol and ammonia solution (50%) to cause a dehydration reaction, coagulate, and dry to form a multifilament. At this time, it is preferable that the ethanol and ammonia solution be mixed in a volume ratio of 90:10, and the physical properties can be improved by adding a cross-linking agent such as genipin or glutaraldehyde, if necessary.

The physical properties of the atelocollagen multifilament manufactured in this way are suitable for manufacturing wet nonwoven fabrics, such as a strength of 2 to 4 g/denier, an elongation of 10 to 15%, and a single yarn fineness of 2 to 10 denier.

Afterwards, the coagulated collagen multifilament is passed through an alcohol bath containing 1.0 wt% of Pluronic, a nonionic polymer surfactant, and then dried and wound on a winder. The reason for using the Pluronic surfactant is that it prevents the multifilament collagen fibers from clumping together, thereby producing a nonwoven fabric with a uniform surface and solving the problem of biological safety (cytotoxicity, etc.) that is a problem when used as a medical material. The Pluronic is a nonionic surfactant developed by Gibco and is a surfactant used to control shear force in suspension, and is also commonly referred to as Pluronic F-68.

Afterwards, the collagen multifilament is cut into 1 to 10 mm lengths using a cutter to create a length suitable for non-woven fabric production.

A step of dispersing the cut collagen fibers in a dispersion solution containing a mixture of an organic acid solution and ethanol is performed. The reason for using a dispersion solution containing a mixture of an organic acid solution and ethanol as the dispersion solution in the present invention is to solubilize a portion of the collagen fibers, thereby inducing bonding between collagen fibers during compression for forming a nonwoven fabric, thereby improving strength.

The above dispersion solution is preferably a dispersion solution in which an organic acid solution and ethanol are mixed in a volume ratio of 5:95 to 30:70. However, when the ratio of the organic acid solution is less than 5, solubilization of collagen fibers is insufficient, and when the ratio of the organic acid solution exceeds 30, a phenomenon in which collagen fibers dissolve and form a film is observed, and there is a disadvantage in that the nonwoven fabric becomes stiff.

The organic acid of the above organic acid solution is any one of acetic acid, lactic acid, and citric acid, and the concentration is preferably 0.1 to 0.5 M to ensure both bonding and flexibility of the nonwoven fabric.

Afterwards, a step is performed in which the collagen fiber-containing dispersion solution is injected into a paper machine and then desolved to make a collagen fiber sheet, and then a step is performed in which the collagen fiber sheet is immersed in ethanol to remove any residual organic acid present in the fiber sheet.

Next, some of the collagen fibers on the sheet are compressed to induce bonding between the collagen fibers, and then dried to provide the wet nonwoven fabric of the present invention.

According to the present invention, a pure collagen fiber nonwoven fabric having excellent hemostatic performance and bioabsorbability is provided by manufacturing fibers from high molecular weight atelocollagen extracted from fish skin and manufacturing a wet-laid nonwoven fabric composed solely of the fibers, and, by using collagen extracted from fish rather than collagen extracted from mammals such as cows and pigs, which are at risk of zoonotic virus transmission, a nonwoven fabric composed solely of atelocollagen fibers extracted from fish skin is provided, which can reduce concerns about viral cross-infection, and can be utilized as a medical biocompatible material such as a locally absorbable hemostatic agent.

Figure 1 is a surface photograph of a collagen fiber nonwoven fabric manufactured by Example 1 of the present invention.
Figure 2 is a photograph of the fibers that make up Comparative Example 2.

The following examples provide non-limiting examples of the method for manufacturing a collagen fiber nonwoven fabric of the present invention.

[Example 1]

Atelocollagen with a molecular weight of 300,000 daltons extracted from tilapia skin was dissolved in a 0.5 M acetic acid solution to make 5 wt%, and then defoamed to prepare a spinning solution. Then, it was discharged through a nozzle with a hole diameter of 0.1 mm and an odd number of 300 using a gear pump, and then coagulated in a coagulation bath composed of ethanol and ammonia aqueous solution (50%) in a volume ratio of 90:10, and then wound. The wound fiber was passed through an ethanol treatment tank containing 1.0 wt% of Pluronic, a nonionic polymer surfactant, for 10 seconds, and then dried at 35°C for 3 hours to produce a collagen multifilament fiber.

After cutting the fibers manufactured in this way into 5mm lengths, the fibers are dispersed by slowly stirring in a dispersion solution prepared by mixing 0.3M acetic acid solution and ethanol in a volume ratio of 20:80. Then, the solution in which the fibers are dispersed is poured into a storage tank of a wet nonwoven fabric machine. After the dispersion of the fibers is stabilized for a certain period of time, the solution is drained, and a sheet in which single-fiber collagen fibers are evenly distributed is removed on a strainer, and then immersed in ethanol for 30 minutes to remove residual organic acids. After that, the sheet is pressed to induce inter-fiber bonding using partially solubilized collagen fibers, and then dried in a hot air dryer at 38℃ for 3 minutes. At this time, the basis weight (g/㎡) of the nonwoven fabric can be adjusted by controlling the amount of fibers dispersed in the mixed solvent. In the example, the nonwoven fabric was manufactured to have a basis weight of 50g/㎡.

[Comparative Example 1]

After the collagen multifilament fibers manufactured in the same manner as in Example 1 were cut in the same manner, they were stirred in a dispersion solution made of pure ethanol to disperse the fibers, and a nonwoven fabric was manufactured in the same manner.

[Comparative Example 2]

After manufacturing collagen multifilaments in Example 1, the coagulated collagen multifilaments were cut directly without passing them through an alcohol bath containing 1.0 wt% of pluronic acid, a nonionic polymer surfactant, and then a nonwoven fabric was manufactured using the same method as in Example 1 (acetic acid-ethanol mixed solution). The physical properties of the manufactured nonwoven fabric are as shown in Table 1 below.

division Example Comparative Example 1 Comparative Example 2 robbery 0.2kgf/5cm 0.1kgf/5cm 0.13kgf/5cm believer 5% 3% 4% weight 51G/M ² 45G/M ² 55G/M ² Uniformity excellence Good error

As shown in Table 1, in Comparative Example 1, since no organic acid that can solubilize collagen is added due to the nature of collagen not soluble in water, fiber-to-fiber bonding does not occur, resulting in very weak physical properties. In addition, in Comparative Example 2, a large amount of fiber clumping occurred, resulting in a very poor uniformity of the nonwoven fabric, and consequently, a phenomenon of deterioration in physical properties was observed. Fig. 1 shows that some of the collagen fibers manufactured into a wet nonwoven fabric by Example 1 were solubilized in an acid and then bonded to each other, and Fig. 2 shows that the fibers constituting Comparative Example 2 are clumped together.

Claims (4)

A step for preparing a spinning solution comprising 4 to 12 wt% of atelocollagen extracted from fish skin having a molecular weight of 100,000 to 300,000 Da and the remainder being an acetic acid solution;
A step of forming collagen multifilaments by discharging a predetermined amount of the above-mentioned radiation solution through a nozzle as a coagulant consisting of a mixture of ethanol and ammonia aqueous solution through a gear pump;
A step of passing the coagulated collagen multifilament through an alcohol bath containing 1.0 wt% of pluronic, a nonionic polymer surfactant, drying the same, and then winding the same on a winder;
A step of cutting the above collagen multifilament into 1 to 10 mm using a cutter;
A step of dispersing the cut collagen fibers in a dispersion solution containing an organic acid solution and ethanol;
A step of injecting the above collagen fiber-containing dispersion solution into a starting machine and then de-dissolving it to make a collagen fiber sheet;
A step of immersing the above collagen fiber sheet in ethanol to remove residual organic acid;
A method for manufacturing a collagen fiber wet nonwoven fabric, characterized by comprising a step of compressing and drying to induce bonding between collagen fibers using some of the collagen fibers available on the sheet to create a wet nonwoven fabric. In paragraph 1,
A method for manufacturing a collagen fiber wet-laid nonwoven fabric, characterized in that the atelocollagen extracted from the above-mentioned fish skin is atelocollagen extracted from tilapia fish skin. In paragraph 1,
A method for manufacturing a collagen fiber wet nonwoven fabric, characterized in that the above dispersion solution is a dispersion solution in which an organic acid solution and ethanol are mixed in a volume ratio of 5:95 to 30:70. In the second paragraph,
A method for manufacturing a collagen fiber wet nonwoven fabric, characterized in that the organic acid of the organic acid solution is any one of acetic acid, lactic acid, and citric acid, and the concentration is 0.1 to 0.5 M.