KR102164176B1 - Collagen membrane with excellent physical properties and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a collagen membrane and a method for manufacturing the same, in particular, by optimizing the appropriate concentration of collagen and a crosslinking agent to have a strong tensile force even in a short decomposition period, to a collagen membrane for treatment of periodontal tissue that is easy to perform and a method for manufacturing the same will be.

Description

Collagen membrane with excellent physical properties and its manufacturing method {COLLAGEN MEMBRANE WITH EXCELLENT PHYSICAL PROPERTIES AND MANUFACTURING METHOD THEREOF}

The present invention relates to a collagen membrane and a method for manufacturing the same, and in particular, to a collagen membrane having a short decomposition period and strong tensile strength, and a method for manufacturing the same.

Collagen membrane is a material that is used to induce tissue regeneration (GTR) or to be transplanted to induce bone regeneration (GBR), and requires biocompatibility and tensile strength suitable for the procedure.

For use in the medical field, the collagen membrane must have biocompatible properties that can be adapted to the body and provide sufficient strength to protect regenerative cells from mechanical stress without being damaged. At the same time, it must be flexible enough to be used according to the shape of the treatment area.

In order to satisfy these conditions, conventional inventions prepared a collagen membrane using biocompatible collagen as a raw material, and a crosslinking agent was used to increase strength. Korean Patent Registration No. 10-1434041 (registered on August 25, 2014) discloses an invention in which a membrane is manufactured by pulverizing collagen fibers from a material derived from a biological tissue itself.

However, in this case, there is a problem in that it is difficult to directly suture during a procedure due to low tensile strength, and there is a considerable situation in the industry for a collagen membrane that is biocompatible and easy to manipulate during medical procedures.

Korean Patent Registration No. 10-1434041 (registered on August 25, 2014)

The present invention has been devised to solve the above problems, and an object of the present invention is to provide a collagen membrane having a short decomposition period and easy treatment properties.

In addition, another object of the present invention is to provide a method for producing the collagen membrane.

In addition, another object of the present invention is to provide a filter used to prepare the collagen membrane.

The method for producing a collagen membrane of the present invention in order to achieve the object as described above,

(A) preparing a collagen mixture by adding and dissolving collagen to an aqueous hydrogen chloride solution,

(B) filtering the collagen mixture with hydrated filter paper to obtain a collagen filtration membrane,

(C) 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide [1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide in a mixed solution of ethanol and water; EDC] and N-hydroxysuccinimide; NHS] to prepare a crosslinking solution,

(D) adding the crosslinking solution to a container, submerging the collagen filtration membrane, and crosslinking to prepare a collagen membrane,

(E) freezing the crosslinked collagen membrane, and

(F) characterized in that it comprises the step of lyophilizing the frozen collagen membrane.

In addition, the collagen mixture in step (A)

10000 parts by weight of water,

Collagen 30 to 120 parts by weight, preferably 35 to 110 parts by weight, more preferably 40 to 100 parts by weight, and

Hydrogen chloride 0.003 to 0.02 parts by weight, preferably 0.005 to 0.016 parts by weight, more preferably 0.004 to 0.014 parts by weight may be added.

In addition, some of the collagen in the collagen mixture solution of step (A) may not be dissolved.

In addition, the collagen in the collagen mixture of step (A) may not be dissolved in 40 to 80% by weight, preferably 45 to 70% by weight, and more preferably 50 to 65% by weight of the added collagen.

In addition, the method for producing a collagen membrane of the present invention may further include a step of vibrating the collagen mixture after step (A) and before step (B).

In addition, the vibration may be 500 to 3000 rpm, preferably 1000 to 2500 rpm, more preferably 1500 to 2000 rpm.

In addition, the vibration may be performed for 1 to 30 minutes, preferably 2 to 20 minutes, more preferably 3 to 10 minutes.

In addition, the crosslinking solution of step (C)

100 parts by weight of ethanol,

15 to 70 parts by weight of water, preferably 20 to 65 parts by weight, more preferably 30 to 55 parts by weight,

1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide; EDC] 0.02 to 0.15 parts by weight, preferably 0.03 to 0.12 parts by weight, more preferably 0.05 to 0.1 parts by weight, and

N-hydroxysuccinimide [N-hydroxysuccinimide; NHS] 0.07 to 0.3 parts by weight, preferably 0.09 to 0.28 parts by weight, more preferably 0.12 to 0.25 parts by weight, may be added.

And, the step of preparing the crosslinking solution of step (C) and the step of preparing the collagen membrane of step (D) have a reinforcement index of 70 × 10 ^-6 to 400 × 10 ^-6 , preferably 100 × 10 ^-6 to 350 × 10 ^-6 , more preferably 130 × 10 ^-6 to 300 × 10 ^-6 can be carried out.

And, after the step (D) and before the step (E), it may further include a step of washing the collagen membrane.

In addition, the collagen is characterized in that it is obtained from the tendon of a mammal, preferably a cow, more preferably a cow.

In addition, the lyophilization of step (F) is characterized in that it is carried out at -90 to -50 °C, preferably -80 to -60 °C, more preferably -75 to -65 °C.

In addition, the lyophilization of step (F) is characterized in that it is performed for 8 to 36 hours, preferably 10 to 30 hours, more preferably 12 to 24 hours.

And, it characterized in that it further comprises a step of sterilizing after the step (F).

And, the sterilization is characterized in that sterilization using gamma rays.

In addition, the collagen filtration membrane of step (D) is characterized in that it is immersed in the crosslinking solution in contact with the filter paper.

In addition, after the step (F), it characterized in that it further comprises the step of separating the collagen membrane and the filter paper.

And, the filter paper is characterized in that the square.

And, the filter paper is characterized in that the square.

On the other hand, the filter of the present invention is a filter for manufacturing a collagen filtration membrane, the main body forming the outer wall of the filter; A tray having a filter paper mounted on the inner bottom surface and having a plurality of through holes formed on the bottom surface; A tray holder provided inside the main body so that the tray is seated; A tray rod for transmitting a load to the filter paper seated on the tray; An exhaust pipe for discharging gas inside the filter during filtration; A drain pipe for discharging the liquid inside the filter during filtration; And a lid covering the opening on the upper side of the body to shield the inside of the body from the outside.

In addition, the tray may have a square shape corresponding to the square filter paper.

In addition, the tray may have a square shape corresponding to the square filter paper.

In addition, the filtration of the step (B) is characterized in that the filter is performed.

On the other hand, the collagen membrane of the present invention is characterized in that it is manufactured by the above manufacturing method.

The collagen membrane according to the present invention has an advantage of being completely decomposed so as to provide a barrier for tissue regeneration in the early stage and not to interfere with regeneration in the late stage by having a decomposition rate close to that of normal tissue. In addition, sufficient tensile strength required for suturing can be expressed through adjustment of collagen solubility and crosslinking conditions. In addition, by forming a collagen filtration membrane and a collagen membrane in a square shape, there is an advantage in that the loss in the final production process is significantly reduced compared to the conventional circular filtration membrane.

1 is a plan view, a side view, and a side cross-sectional view showing a main body constituting a filter for manufacturing a collagen filtration membrane of the present invention.
2 is a plan view and a side view showing a tray constituting a filter for manufacturing a collagen filtration membrane.
3 is a plan view and a side view showing a tray rod constituting a filter for manufacturing a collagen filtration membrane.
4 is a plan view and a side view showing a sealing portion and a lid constituting a filter for manufacturing a collagen filtration membrane.
5 is a combined view of a main body, a tray, a tray rod, and a lid constituting a filter for manufacturing a collagen filtration membrane.
6 is a graph showing the results of measuring the tensile strength of the collagen membrane of the present invention and the conventional collagen membrane on the market.
7 is a photograph taken of an example of a collagen membrane prepared by the method of manufacturing a collagen membrane of the present invention.
8 is a photograph taken through H&E staining of the degree of tissue regeneration over time.
9 is a picture taken through MT staining of the degree of tissue regeneration over time.

Hereinafter, preferred embodiments of the present invention will be described in detail. In addition, in the following description, there are many specific matters such as specific elements, etc., which are provided only to help a more general understanding of the present invention, and the present invention can be practiced without these specific matters. It is self-evident to those who have the knowledge of Further, in describing the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the subject matter of the present invention, the detailed description thereof will be omitted.

In this specification, the'reinforcement index' is defined as follows:

Reinforcement index

= (Crosslinking agent concentration / dissolved collagen concentration) × (crosslinking time / target degradation time).

The method for preparing a collagen membrane of the present invention begins with the step of preparing a collagen mixture by first adding and mixing collagen to an aqueous hydrogen chloride solution.

The hydrogen chloride aqueous solution may be added to 10000 parts by weight of water, 0.003 to 0.02 parts by weight of hydrogen chloride, preferably 0.005 to 0.016 parts by weight, more preferably 0.004 to 0.014 parts by weight. When the amount of hydrogen chloride is within the above range, it is possible to obtain a product having a desired property and standard through sufficient dissolution of collagen while preventing side effects during human insertion due to the pH of the product too low.

The collagen mixture is prepared by adding 30 to 120 parts by weight of collagen, preferably 35 to 110 parts by weight, more preferably 40 to 100 parts by weight, based on 10000 parts by weight of water constituting the aqueous hydrogen chloride solution. When the amount of collagen in the collagen mixture is within the above range, it is possible to obtain a collagen membrane having a desired formulation and physical properties, such as tensile strength, while preventing the phenomenon that the formulation manufacturing becomes difficult due to too high viscosity.

The present invention is particularly characterized in that not all of the collagen in the collagen mixture is dissolved, but a part of the collagen is present in the collagen mixture in an undissolved state. That is, the surface of the collagen powder is dissolved and released into collagen fibers, and the central portion of the powder is not dissolved, so that the particle shape is maintained.

This structure of collagen plays a role in expressing strong mechanical properties when formulated into a collagen membrane. Specifically, undissolved collagen particles are connected to each other by collagen fibers, thereby creating a three-dimensional network structure exhibiting strong tensile strength.

As in the case of cement concrete, a coarse aggregate such as gravel plays a major role in the development of strength, and in the collagen membrane of the present invention, collagen in the form of undissolved particles plays the role of such a coarse aggregate. In addition, the dissolved fibrous collagen connects and binds the collagen in the form of particles to each other, and the crosslinking agent to be described later further strengthens the bonding, which will be referred to as a cement functioning as a binder in the cement concrete described above.

The proportion of insoluble collagen among the added collagen may be 40 to 80% by weight, preferably 45 to 70% by weight, and more preferably 50 to 65% by weight. When the amount of undissolved collagen is within the above range, a collagen membrane having a desired formulation and physical properties such as tensile strength can be obtained while preventing the phenomenon that the formulation production becomes difficult due to too high viscosity.

In the method for producing a collagen membrane of the present invention, the collagen can be obtained from a mammal, preferably a cow, more preferably a tendon or ligament of a cow. In addition, the collagen is preferably selected from the group of properties, pH, heavy metals, amino acid analysis, and endotoxin test, or a combination test is performed to use a suitable collagen.

In addition, the method of manufacturing a collagen membrane of the present invention may further include vibrating the collagen mixture before filtering. When vibration is applied to the collagen mixture, undissolved collagen particles and dissolved collagen fibers in the mixture are evenly distributed over the entire area of the mixture, and in particular, the dissolved collagen fibers are sufficiently spread. The collagen membrane prepared through this has an advantage in that the composition is uniform and, as a result, physical properties are evenly expressed over the entire area, and the tensile strength increases due to an increase in the interaction between the sufficiently spread collagen fibers.

The vibration may be 500 to 3000 rpm, preferably 1000 to 2500 rpm, more preferably 1500 to 2000 rpm. When the rpm of the vibration device is within the above range, sufficient economics can be secured, and a problem in that it is difficult to obtain a membrane having low tensile strength and uniform physical properties can be prevented because collagen particles and fibers are not evenly distributed.

In addition, the vibration may be performed for 1 to 30 minutes, preferably 2 to 20 minutes, more preferably 3 to 10 minutes. When the vibration time is within the above range, it is possible to prevent a problem in which it is difficult to obtain a membrane having low tensile strength and uniform physical properties because collagen particles and fibers are not evenly distributed.

The collagen mixture thus prepared is filtered through hydrated filter paper to obtain a collagen filtration membrane.

Here, the filter paper may have a square shape, preferably a square shape, and this is a main feature of the present invention that is distinguished from the conventional collagen membrane.

The filter paper is generally circular, so that the collagen filtration membrane obtained after filtration and the collagen membrane are obtained in a circular shape. However, since the formulation required in the medical field is square, the end product is manufactured by cutting the edge of the circular collagen membrane. The amount of collagen membrane discarded at this stage reaches a significant level.

The present invention is characterized by changing the shape of the filter paper forming the collagen filtration membrane into a square, preferably a square, in order to solve this problem. Through this, it was possible to drastically reduce the amount of the collagen membrane wasted.

In order to convert the collagen membrane into a collagen membrane separately from the preparation of the collagen filtration membrane, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide) in a mixed solution of ethanol and water ; EDC] and N-hydroxysuccinimide; NHS] is added to prepare a crosslinking solution.

A mixed solution of ethanol and water is prepared by mixing 100 parts by weight of ethanol, 15 to 70 parts by weight of water, preferably 20 to 65 parts by weight, and more preferably 30 to 55 parts by weight of water.

To the mixed solution of ethanol and water, 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide [1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide] based on 100 parts by weight of ethanol; EDC] 0.02 to 0.15 parts by weight, preferably 0.03 to 0.12 parts by weight, more preferably 0.05 to 0.1 parts by weight, are added. When the amount of EDC is within the above range, while preventing harmful effects on the human body due to residual crosslinking agent in the final product, the crosslinking reaction completely occurs through a sufficient crosslinking agent, thereby increasing physical properties such as tensile strength in the final product to a desired level.

In addition, in the mixed solution of ethanol and water, N-hydroxysuccinimide [N-hydroxysuccinimide; NHS] 0.07 to 0.3 parts by weight, preferably 0.09 to 0.28 parts by weight, more preferably 0.12 to 0.25 parts by weight, is added. When the amount of NHS is within the above range, the crosslinking reaction is complete, and physical properties such as tensile strength can be raised to a desired level in the final product.

The thus prepared crosslinking solution is put in a container, the prepared collagen filtration membrane is immersed in the crosslinking solution, and then crosslinked to prepare a collagen membrane. Through this crosslinking reaction, the dissolved collagen fibers are further bonded to each other, and as a result, various physical properties such as tensile strength of the final product are improved.

At this time, the collagen filtration membrane may be separated from the filter paper and then immersed in the crosslinking solution, but it is more preferable to immerse it in the crosslinking solution while in contact with the filter paper. This is because separating from the filter paper in the state of the collagen membrane after crosslinking is less likely to be damaged than separating in the state of the collagen filter membrane before crosslinking.

In addition, a separate stirrer may be provided in the container in which the crosslinking proceeds to promote the crosslinking reaction.

The present invention introduces a new dimensionless variable called'reinforcement index' to control the degree of strengthening of the collagen membrane.

The reinforcement index is defined as'(crosslinking agent concentration / dissolved collagen concentration) × (crosslinking time / target degradation time)'.The higher the concentration of the crosslinking agent relative to the collagen concentration, the higher the crosslinking time for the target degradation time. It was designed so that the longer the value, the larger the value. In particular, the strengthening index introduced in the present invention was not based on the total collagen concentration in the mixed solution, but based on the dissolved collagen concentration mainly participating in the crosslinking, so that the actual phenomenon was accurately reflected.

Cross-linking solution and the cross-linking reaction, which constitutes the present invention is the enhanced index 70 × 10 ^-6 to 400 × 10 ^-6, preferably from 100 × 10 ^-6 to 350 × 10 ^-6, and more preferably 130 × 10 ^{- It} is prepared and carried out to have a value of ⁶ to 300 × 10 ^-6 . When the value of the reinforcement index is within the above range, while preventing harmful effects on the human body due to the residual crosslinking agent in the final product, various physical properties such as tensile strength are improved due to a sufficient crosslinking reaction, thereby enabling a procedure such as sealing.

The collagen membrane after the crosslinking reaction is first frozen and then lyophilized. It is more preferable to perform the step of washing the crosslinked collagen membrane before freezing.

The lyophilization may be performed at -90 to -50 °C, preferably -80 to -60 °C, more preferably -75 to -65 °C. When it is within the above range, a liquid state while suppressing excessive economic decline It is possible to secure the desired level of physical properties by preventing problems caused by water.

In addition, the freeze-drying may be performed for 8 to 36 hours, preferably 10 to 30 hours, more preferably 12 to 24 hours, and when it is within the above range, complete drying of the collagen membrane is possible while suppressing excessive economic decline. So that the desired level of physical properties can be secured.

The collagen membrane dried through lyophilization may further include a step of sterilizing afterwards. The sterilization may be used without limitation as long as it is a sterilization method that can be used in the technical field to which the present invention belongs, and may be, for example, sterilization using gamma rays.

Meanwhile, the filter 1 used to obtain a collagen filtration membrane in the method for producing a collagen membrane of the present invention includes a body 10, a tray 20, a tray rod 30, and a lid 40.

The main body 10 forms the outer wall of the filter 1 and has a structure in which the upper part of the cylinder shape is inverted and the lower part of the conical shape.

A tray holder 12 on which the tray 20 is mounted is provided inside the main body to correspond to each corner of the tray 20.

In addition, an exhaust pipe 14 for decompressing the inside of the filter 1 is provided on the conical side of the lower part of the main body 10, and a drain pipe 16 for discharging the filtrate is provided at the conical vertex.

In addition, a leg 18 for separating the filter 1 from the floor and a body handle 19 used to carry the filter 1 are provided outside the main body 10.

On the other hand, the tray 20 has a square, preferably square bottom surface to correspond to a square, preferably square filter paper, and the bottom surface is formed with a plurality of through holes for discharging the filtrate by filtration. .

In addition, the tray 20 may be provided with a tray handle 29 used to carry it.

The filter 1 of the present invention is provided with a tray 20 having a square shape, preferably a square shape, so that a square, preferably square, collagen filtration membrane and a collagen membrane can be obtained after filtration. As described above, the final product It has the advantage of preventing waste.

The filter 1 of the present invention may further include a tray rod 30 for transferring a load to the filter paper and collagen mixture so that filtration can occur evenly in the entire bottom surface of the tray 20. The collagen filtration membrane obtained after filtration by the tray rod 30 can secure a uniform thickness in all areas.

The filter 1 of the present invention further includes a lid 40 covering the opening on the upper side of the main body 10 and shielding the inside of the main body 10 from the outside. It is preferable that a sealing part (not shown) for maintaining a depressurized state is provided at the edge of the lid 40 corresponding to the upper outer circumferential surface of the main body 10, and a lid handle 49 used to transport the lid 40 ) May be provided.

On the other hand, the collagen membrane of the present invention is characterized in that it is manufactured by the above manufacturing method.

Hereinafter, an embodiment of the present invention will be described.

Example

Example 1: Collagen membrane

A collagen mixture was prepared by adding 10 g of collagen (Collagen Solution, New Zealand) to 1500 ml of saline to which 5 ml of 0.01 N hydrogen chloride was added and mixing. In the filter of the present invention, a filter paper moistened with water and hydrated was put in a tray, and the collagen mixture was poured, and then exhausted and filtered for 1 hour to obtain a collagen filtration membrane. A crosslinking solution was prepared by adding 0.2 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 0.5 g of N-hydroxysuccinimide (NHS) to a 500 ml solution of 70 wt% ethanol. . 100 ml of the crosslinking solution was put in a separate container, the collagen filtration membrane was immersed, and crosslinked at 150 rpm for 2 hours and 30 minutes to prepare a collagen membrane. The crosslinked collagen membrane was frozen at -80°C for 5 hours and then lyophilized at -70°C for 24 hours to obtain the collagen membrane of the present invention.

Comparative Example 1: Excessive use of collagen

The same procedure as in Example 1 was performed, but a collagen membrane was prepared using 30 g of the collagen powder.

Comparative Example 2: Use of trace amounts of collagen

The same procedure as in Example 1 was followed, but a collagen membrane was prepared using 1 g of the collagen powder.

Comparative Example 3: EDC overuse

The same procedure as in Example 1 was followed, but a collagen membrane was prepared using 0.5 g of the EDC powder.

Comparative Example 4: Use of trace amounts of EDC

The same procedure as in Example 1 was followed, but a collagen membrane was prepared using 0.01 g of the EDC powder.

Comparative Example 5: NHS excessive use

The same procedure as in Example 1 was followed, but a collagen membrane was prepared using 3 g of the NHS powder.

Comparative Example 6: Use of a trace amount of NHS

The same procedure as in Example 1 was followed, but a collagen membrane was prepared using 0.05 g of the NHS powder.

Test Example 1: Tensile strength

For the collagen membranes of Example 1 and Comparative Examples 1 to 6, the results of measuring tensile strength at a rate of 20 mm/min in a universal testing machine (Testwon, Korea) are shown in Table 1 below. In Table 1, Comparative Examples 7 to 9 are commercially available collagen membranes, Comparative Example 7 is Genos (Korea), Comparative Example 8 is Zimmer Biomet (USA), Comparative Example 9 is a product of Matricel (Germany), and Example 1 And Comparative Examples 7 to 9 were compared with Figure 6.

Tensile strength (N) Remark Example 1 196.5 Comparative Example 1 - Formulation cannot be formed due to high viscosity Comparative Example 2 25.3 Comparative Example 3 182.2 Remaining unreacted EDC Comparative Example 4 54.7 Comparative Example 5 189.6 Remaining unreacted NHS Comparative Example 6 55.1 Comparative Example 7 170.9 Comparative Example 8 142.1 Comparative Example 9 78.4

From Tables 1 and 6, it can be seen that the tensile strength of the collagen membrane of the present invention is improved compared to the prior art, and this high tensile strength makes it suitable for suturing or treatment during the procedure.

Test Example 2: Animal test

The results of animal experiments of the collagen membrane of Example 1 and the collagen membrane of Comparative Example 7 (Genos, Korea) are shown in FIGS. 8 and 9. In the animal experiment, a defect of 8 mm was made in a rabbit's skull (rabbit calvaria), and a collagen membrane was implanted, and bone regeneration was observed after 8 and 16 weeks. The results were confirmed through H&E (Hematoxylin and Eosin) staining (FIG. 8) and MT (Masson's trichrome) staining (FIG. 9) methods. From Figs. 8 and 9, Example 1 is a control example (an example without applying a collagen membrane) as well as bone regeneration through the fact that white bone and circular tissues are further generated compared to Comparative Example 7, which is a commercially available conventional collagen membrane. It can be seen that the effect is improved.

The H&E staining experiment is a method of observing the formation of tissues and bones. Acidic regions of cells, nuclei, and matrix of cartilage are stained blue, and basic sites and collagen fibers are stained pink. In addition, the MT staining experiment is also a method of observing the formation of tissues and bones. The nuclei of cells are stained in dark blue, muscle, keratin and cytoplasm are stained in red, and mucus and collagen are stained in light blue.

Preparation Example 1: All collagen dissolution

Following the same procedure as in Example 1, a collagen membrane was prepared using 5 g of the collagen powder, and the tensile strength was measured at a rate of 20 mm/min in a universal testing machine (Testwon, Korea). It is shown in 2.

Preparation Example 2: Dissolution of trace amounts of collagen

Following the same procedure as in Example 1, a collagen membrane was prepared using 25 g of the collagen powder, and the tensile strength was measured at a rate of 20 mm/min in a universal testing machine (Testwon, Korea). It is shown in 2.

Manufacturing Example 3: Vibration step

The same procedure as in Example 1 was followed, but before filtering the collagen mixture, a collagen membrane was prepared by further comprising the step of vibrating for 5 minutes at 2000 rpm, and 20 mm/in a universal testing machine (Testwon, Korea). The results of measuring the tensile strength at the rate of minutes are shown in Table 2 below.

Preparation Example 4: High Speed RPM

The same procedure as in Preparation Example 3 was carried out, but the vibration was performed at 5000 rpm to prepare a collagen membrane, and the results of measuring the tensile strength at a rate of 20 mm/min in a universal testing machine (Test Institute, Korea) are shown in the following table. It is shown in 2.

Preparation Example 5: Low speed RPM

The same procedure as in Preparation Example 3 was carried out, but the vibration was performed at 100 rpm to prepare a collagen membrane, and the results of measuring the tensile strength at a rate of 20 mm/min in a universal testing machine (Testwon, Korea) are shown in the following table. It is shown in 2.

Manufacturing Example 6: Long vibration time

The same procedure as in Preparation Example 3 was carried out, but the vibration was performed for 1 hour to prepare a collagen membrane, and the results of measuring the tensile strength at a rate of 20 mm/min in a universal testing machine (Testwon, Korea) are shown in the following table. It is shown in 2.

Manufacturing Example 7: Short vibration time

Following the same procedure as in Preparation Example 3, the vibration was performed for 20 seconds to prepare a collagen membrane, and the results of measuring the tensile strength at a rate of 20 mm/min in a universal testing machine (Testwon, Korea) are shown in the following table. It is shown in 2.

Tensile strength (N) Remark Manufacturing Example 1 154.8 Manufacturing Example 2 - Difficulty forming formulation due to high viscosity Manufacturing Example 3 202.3 Manufacturing Example 4 188.6 Manufacturing Example 5 197.2 Manufacturing Example 6 192.7 Manufacturing Example 7 197.1

In the above, preferred embodiments of the present invention have been described, but the present invention is not limited to the specific embodiments described above, and those of ordinary skill in the art can implement various modifications without departing from the gist of the present invention. Of course it is possible. Therefore, the scope of the present invention should not be construed as limited to the above embodiments, and should be defined by the claims and equivalents to the claims described later.

1: filter
10: main body 12: tray holder
14: exhaust pipe 16: drain pipe
18: leg 19: main body handle
20: tray 29: tray handle
30: tray load
40: lid 49: lid handle

Claims (8)

(A) preparing a collagen mixture by adding and dissolving collagen to an aqueous hydrogen chloride solution,
(B) filtering the collagen mixture with hydrated filter paper to obtain a collagen filtration membrane,
(C) 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide [1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide in a mixed solution of ethanol and water; EDC] and N-hydroxysuccinimide; NHS] to prepare a crosslinking solution,
(D) adding the crosslinking solution to a container, submerging the collagen filtration membrane, and crosslinking to prepare a collagen membrane,
(E) freezing the crosslinked collagen membrane, and
(F) freeze-drying the frozen collagen membrane,
In the collagen mixture of step (A), 40 to 80% by weight of collagen in the added collagen is not dissolved,
After the step (A) and before the step (B), further comprising the step of vibrating the collagen mixture at 500 to 3000 rpm for 1 to 30 minutes, a method for producing a collagen membrane. The method according to claim 1,
The collagen mixture in step (A) is
10000 parts by weight of water,
30 to 120 parts by weight of collagen and
A method for producing a collagen membrane, characterized in that 0.003 to 0.02 parts by weight of hydrogen chloride is added. The method according to claim 1,
The crosslinking solution of step (C) is
100 parts by weight of ethanol,
15 to 70 parts by weight of water,
1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide; EDC] 0.02 to 0.15 parts by weight, and
N-hydroxysuccinimide [N-hydroxysuccinimide; NHS], characterized in that the addition of 0.07 to 0.3 parts by weight, a method for producing a collagen membrane. The method according to claim 1,
The collagen is characterized in that obtained from a mammal, a method for producing a collagen membrane. The method according to claim 1,
The method for producing a collagen membrane, characterized in that the collagen filtration membrane of step (D) is immersed in a crosslinking solution in contact with the filter paper. The method of claim 5,
After step (F),
The method of manufacturing a collagen membrane, characterized in that it further comprises the step of separating the collagen membrane and the filter paper. delete delete

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