KR20150125881A - Nano fiber sheet for anti-adhesion using water soluble chitosan and poly ethylene oxide, manufacturing method thereof - Google Patents

Abstract

The present invention relates to a nanofiber sheet which can effectively prevent adhesion among bio-tissues, and to a manufacturing method thereof. More specifically, the present invention relates to a nanofiber sheet in a non-woven fabric form, which is made up of uniform nanofiber and has large surface area without toxicity by being fabricated through electrospinning using water-soluble chitosan and polyethylene oxide (PEO) which is a hydrophilic polymer. The present invention further relates to a manufacturing method thereof. The manufacturing method of the present invention comprises steps of: acquiring a polymer solution by dissolving the water-soluble chitosan and polyethylene oxide in water; and electrospinning the polymer solution thereafter.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a nanofiber sheet for preventing adhesion, which uses water-soluble chitosan and polyethylene oxide, and a manufacturing method thereof,

The present invention relates to a nanofiber sheet capable of effectively preventing adhesion between living tissues and a method for producing the nanofiber sheet. More particularly, the present invention relates to a nanofiber sheet capable of effectively preventing adherence between biotissues, and more particularly, to a nanofiber sheet formed by uniformly spinning a water-soluble chitosan and a hydrophilic polymer polyethylene oxide (PEO) To a nanofiber sheet in the form of a nonwoven fabric having a large surface area and no toxicity, and a method for producing the same.

Adhesion usually occurs in surgery and refers to a phenomenon in which tissues or organs of a human body, which should be separated from each other, form a fibrous tissue by a wound and are not separated from each other.

If tissue damage occurs after surgical operation or due to inflammation, foreign body, hemorrhage, infection, wound, or friction, blood coagulates in the healing process of the wound and coagulates, resulting in abnormal adhesion with peripheral tissues. When the cells penetrate into these adhesion sites, stronger adhesion occurs.

In particular, tissue adhesion is a major cause of intestinal obstruction (80% to 90%) and causes long-term sequelae and pelvic pain after gynecologic procedures (Eur.J. Surg. 1997, Supp1 577: 32-39).

As a known anti-adhesion method to date, there are the following: 1) a micro-surgical technique that minimizes a wound in an open surgery; and 2) a chemical method in which an anti-inflammatory agent, a fibrin formation inhibitor, an anticoagulant, (Eg, t-PA). However, some of the drugs have the disadvantage of causing side effects that delay the wound healing rate as compared with the anti-adhesion effect. Third, apart from this, recently, a method of using a physical barrier and wrapping a wound after surgery or inserting it between tissues or mesenteric membranes to isolate it from surrounding tissues has been used.

As the most frequently used physical barrier, a method of inserting an anti-adhesion agent is mainly used.

Recently, nanofibers obtained by electrospinning a polymer solution have been proposed as physical barriers to prevent tissue adhesion.

Electrospinning is a technique for fabricating micrometer or nanometer-sized fibers by emitting a large potential difference between a charged polymer and a grounded collecting plate by applying an electrostatic force to the polymer solution or melt, , It is possible to spin at a high spinning speed and a small amount and to obtain a nonwoven form at the same time as spinning, which is a useful method for producing an anti-adhesion agent.

Korean Patent Laid-Open No. 10-2011-0066615 discloses a method for producing a nanofiber sheet by electrospinning a chitosan in which a hydrophobic polymer PLGA (polylactic-co-glycolic acid) and a hydrophilic polymer polyethylene oxide are mixed and dissolved in an organic solvent Is disclosed.

However, the above-mentioned patented technique produces a nanofiber sheet by dissolving it in an organic solvent such as hexafluoroisopropanol (HFIT) and trifluoroethanol (TFE) for the dissolution of the water-insoluble chitosan and the hydrophobic polymer. The fabricated nanofiber sheet showed lower survival rate compared to the group containing only PLGA in cytotoxicity test (MTT assay). It seems that the removal of the organic solvent to dissolve the chitosan did not work well and it affected the toxicity.

The present invention has been made in order to overcome the above problems, and it is an object of the present invention to provide a water-soluble chitosan and a polyethylene which can reduce the toxicity by use in an organic solvent by converting the chemical structure of chitosan, And an object of the present invention is to provide a nanofiber sheet for preventing adhesion using oxide.

In order to accomplish the above object, the present invention provides a nanofiber sheet for adhesion prevention using water-soluble chitosan and polyethylene oxide, which is formed by electrospinning a polymer solution containing water-soluble chitosan and polyethylene oxide.

The water-soluble chitosan is N-Carboxyethyl Chitosan.

The water-soluble chitosan and the polyethylene oxide are contained in a weight ratio of 1: 0.5 to 2.

In order to accomplish the above object, the present invention provides a method for producing a nanofiber sheet for preventing adhesion using water-soluble chitosan and polyethylene oxide, comprising the steps of: dissolving water-soluble chitosan and polyethylene oxide in water to obtain a polymer solution; And electrospinning the polymer solution.

The polymer solution is characterized in that the water-soluble chitosan and the polyethylene oxide are mixed and dissolved in a weight ratio of 1: 0.5 to 2.

Wherein the water-soluble chitosan is prepared by a method comprising the steps of: a) stirring a chitosan solution in which non-water-soluble chitosan is dissolved in a mixed solvent of acrylic acid and distilled water; b) dialyzing after adding sodium hydroxide to the chitosan solution; c) And N-Carboxyethyl Chitosan obtained by performing a step of lyophilizing the obtained solution.

The electrospinning is performed by spinning the polymer solution at a temperature of 22 to 28 DEG C, a relative humidity of 27 to 33%, a voltage of 10 to 40 kV, a radiation distance of 10 to 20 cm, and a fluid velocity of 0.02 to 0.1 mL / min do.

As described above, according to the present invention, the chemical structure of chitosan, which is an antibacterial material and natural polymer material, is changed and converted to water-soluble chitosan, thereby reducing the toxicity of organic solvents.

The nanofiber sheet of the present invention is a physical barrier capable of effectively preventing the adhesion of serous tissue with a wound site or hemorrhage, and is excellent in the property as an anti-adhesion agent by being decomposed and absorbed in vivo.

1 is a view of a nanofiber sheet having a composition of CEC: PEO = 3: 3 prepared according to an embodiment,
FIGS. 2 to 4 are electron micrographs of a nanofiber sheet having a composition of CEC: PEO = (2: 4, 3: 3, 4: 2)
5 is an FT-IR graph using an infrared spectrophotometer of water-soluble chitosan having a carboxyethyl group formed on chitosan,
6 is an FT-IR graph using an infrared spectrophotometer of a nanofiber sheet having a composition of CEC: PEO = 3: 3,
7 is a thermal analysis graph of a nanofiber sheet having a composition of CEC: PEO = 3: 3,
8 is a graph showing the cytotoxicity results of a nanofiber sheet having a composition of CEC: PEO = 3: 3,
FIG. 9 is a photograph showing the naked eye observation of the adhesion of the tissue on the 7th day after suturing,
Figs. 10 to 12 are photographs taken by microscope after staining of tissue at 7 days after suturing.

Hereinafter, a nanofiber sheet for adhesion prevention using water-soluble chitosan and polyethylene oxide according to a preferred embodiment of the present invention and a method for producing the same will be described in detail.

The nanofiber sheet for preventing adhesion according to an embodiment of the present invention is formed by electrospinning a polymer solution containing water-soluble chitosan and polyethylene oxide.

Chitosan is a natural polymer material deacetylated by treatment with chitin at high temperature and strong alkali in shells of crabs such as crabs and shrimps. It has no toxicity, is biodegradable and has excellent biocompatibility. It has been well-known as a physiologically active substance such as wound healing agent, artificial skin, blood coagulant, immunity enhancer, antibacterial and antioxidant since it can be applied for medical use since the age of onset. In addition, they are used as scaffolds for tissue engineering such as drug or cell delivery carriers and artificial skin, artificial cartilage, artificial bone, and the like.

Conventional chitosan obtained by deacetylating chitin has an acetylamino group having a very strong hydrogen bond in the molecule, so that it is not soluble in water. Chitosan dissolved in water has low molecular weight chitosan or chitooligosaccharide, but low molecular weight chitosan has low efficacy. Chitosan has been reported to have higher efficacy as the molecular weight increases (Jung B. O. et al., J. Chitin Chitosan, 6 (1), 12-17, 2004).

Therefore, in the present invention, instead of a chemical organic solvent having toxicity as in the prior art, a high molecular weight chitosan which is soluble in water and is excellent in water-soluble chitosan is applied. One example of such water-soluble chitosan is carboxyethyl chitosan (CEC), which is a combination of a common chitosan (non-water-soluble chitosan) obtained by deacetylating chitin and a carboxyethyl group.

For example, acrylic acid may be added to the chitosan to render it water-soluble.

The water-soluble chitosan used as a main material in the present invention is difficult to be electrospun by itself, and therefore it is essential to select a polymer material to be mixed with water-soluble chitosan for electrospinning.

In the present invention, polyethylene oxide (PEO) is used as a polymer material to be mixed with water-soluble chitosan. Polyethylene oxide has excellent hydrophilicity and solubility as well as excellent decomposability due to the characteristics of the ethylene oxide (EO) group present in the repeating unit.

Water-soluble chitosan and polyethylene oxide must be dissolved in water in order to electrospray the water-soluble chitosan and polyethylene oxide. A solution of water soluble chitosan and polyethylene oxide in water is used for electrospinning.

Electrospinning is a technology to produce micrometer or nanometer fiber by radiating electrostatic force to a polymer solution by electrostatic force. It is cheap and simple, and it is possible to spin at high spinning speed and small amount, At the same time, nonwoven forms can be obtained, which is a useful method for producing nanofiber sheets.

Electrospinning is a technique for producing ultrafine fibers with a diameter of several tens to several hundreds of nanometers. When an electric force is applied to a polymer solution dissolved or dissolved in a solvent, charge is induced to the liquid surface of the polymer solution, And the force due to the mutual repulsive force of the induced charges is generated in the direction opposite to the surface tension. When the applied voltage exceeds the threshold voltage of the polymer solution droplet, the polymer solution jet discharged by the electrical repulsive force is released. The jet is finely torn and fibrillated while the air is blown, and the solvent is volatilized, ), A nanofiber sheet in the form of a nonwoven fabric in which microfine fibers are laminated is produced.

Hereinafter, a method for producing a nanofiber sheet will be described in detail.

First, water-soluble chitosan and polyethylene oxide are dissolved in water to obtain a polymer solution for electrospinning.

As an example of the water-soluble chitosan, Carboxyethyl Chitosan (CEC) described above is used.

Carboxyl ethyl chitosan can be used by purchasing a commercialized product or by directly producing it.

For example, 1 to 10 parts by weight of chitosan is dissolved in 100 parts by weight of a solvent in which acrylic acid and distilled water are mixed to obtain a chitosan solution. Then, the chitosan solution is stirred for 1 to 5 days while maintaining the temperature at 45 to 55 ° C. Then, a 1 N sodium hydroxide solution in which sodium hydroxide is dissolved in distilled water is added to the chitosan solution, and the chitosan solution is adjusted to pH 10-12. Then, the chitosan solution is purified and then lyophilized to prepare pure N-carboxyethylchitosan. The dialysis membrane can be used as a purification method. For example, the chitosan solution can be dialyzed in the dialysis membrane for one to three days in the third distilled water.

Next, the prepared water-soluble chitosan and polyethylene oxide are dissolved in water to obtain a polymer solution. At this time, the water-soluble chitosan to polyethylene oxide is mixed and dissolved in a weight ratio of 1: 0.5 to 2.

If the content ratio of polyethylene oxide to water-soluble chitosan is less than 0.5, there is a problem that the diameter of the fibers is increased and the beads are formed in the middle of the fibers. When the content ratio of polyethylene oxide is more than 2, the diameter of the fibers is irregular and the fibers are aggregated.

The polymer solution having a concentration of about 2 to 10% can be obtained by dissolving 2 to 10 g in total weight of the water-soluble chitosan and polyethylene oxide in 100 ml of distilled water.

Next, a prepared polymer solution is electrospun to produce a nonwoven fabric-shaped nanofiber sheet.

A typical electrospinning apparatus for electrospinning can be broadly classified into a polymer solution supply unit, a high voltage supply unit, and a collector unit in which nanofibers are formed. There is no particular limitation to the use of the electrospinning device so long as it can produce fibers having micrometers or nanometers in diameter. However, since the size and characteristics of fibers produced vary according to the spinning conditions of the electrospinning apparatus, the spinning conditions need to be kept constant.

In the present invention, the polymer solution is spun at a temperature of 22 to 28 DEG C, a relative humidity of 27 to 33%, a voltage of 10 to 40 kV, a radiation distance of 10 to 20 cm, and a fluid velocity of 0.02 to 0.1 mL / min to produce a nanofiber sheet .

Hereinafter, the present invention will be described with reference to the following experimental examples. However, the following experimental examples are intended to illustrate the present invention in detail, and the scope of the present invention is not limited to the following experimental examples.

≪ Preparation of nanofiber sheet >

1. Preparation of N-carboxyethyl chitosan (CEC)

The chitosan solution in which 2 g of chitosan (Mw: 190,000 ~ 310,000, viscosity (20 ° C): 20,000 cps) was dissolved in a solvent mixed with acrylic acid (1.68 mL) and distilled water (50 mL) Respectively. The chitosan was purchased from Sigma Aldrich Co., Ltd. (St. Louis, USA).

Then, a 1N NaOH aqueous solution in which sodium hydroxide was dissolved in distilled water was added to the chitosan solution to adjust the pH to 10-12. Then, the mixed solution was dialyzed in a dialysis tube (Mw: cut-off: 3500 g / mol) and dialyzed in the third distilled water for 2 days. The solution obtained was lyophilized to obtain pure N-Carboxyethyl Chitosan (CEC).

2. Fabrication of Nanofiber Sheet

Polyethylene oxide (PEO, Mw: 600,000) was purchased from Sigma Aldrich Co., Ltd. (St. Louis, USA). Other reagents were used without purification and tertiary sterile water was used for distilled water.

To make a polymer solution, the total amount of N-carboxy chitosan (CEC) and polyethylene oxide (PEO) was fixed at 6 g and dissolved in 100 mL of distilled water to prepare a polymer solution having a concentration of 6%. The polymer solution was prepared according to the mixing ratio of N-carboxy chitosan and polyethylene oxide.

Each of the polymer solutions was subjected to measurement under the conditions of a temperature of 25 ± 3 ° C. and a relative humidity of 30 ± 3% using an electrospinning device (HM300-60K-SR-MR, ZEFA.Co., LTD, Korea), a voltage of 25 kV, , And a fluid velocity of 0.05 ml / min, for 3 hours for 5 hours to prepare a nanofiber sheet. FIG. 1 shows a second nanofiber sheet prepared by electrospinning a polymer solution having a composition of CEC: PEO = 3: 3, for example, as a nanofiber sheet.

The weight ratios of N-carboxy chitosan and polyethylene oxide, the viscosity of the polymer solution, and the average diameter of the fibers of each of the polymer solutions used in the three types of nanofiber sheets produced are shown in Table 1 below.

division The weight ratio of CEC to PEO in the polymer solution Viscosity of polymer solution
(20 < 0 > C, CPS) Fiber Diameter (nm) The first nanofiber sheet CEC: PEO = 2: 4 8800 ± 100 230 ± 27.4 The second nanofiber sheet CEC: PEO = 3: 3 3515 ± 327.6 134.2 ± 9.33 The third nanofiber sheet CEC: PEO = 4: 2 4633 + - 57.74 224 ± 89.13

<Experimental Example 1: Measurement of morphology of nanofiber sheet>

The nanofiber sheet was observed to examine the surface properties and morphological structure of the nanofibers obtained by electrospinning. The morphology of the nanofiber sheet was observed through a scanning electron microscope (SEM, S-4100, Hitachi, Japan) and dried in a freeze dryer to remove moisture in the sample and then measured by gold coating.

FIG. 2 is an electron micrograph of a first nanofiber sheet prepared by electrospinning a polymer solution having a composition of CEC: PEO = 2: 4, and FIG. 3 is an electron micrograph of a first nanofiber sheet prepared by electrospinning a polymer solution having a composition of CEC: PEO = 3: FIG. 4 is an electron micrograph of a second nanofiber sheet prepared by electrospinning a polymer solution having a composition of CEC: PEO = 4: 2. FIG.

Referring to FIG. 2, the first nanofiber sheet was found to have an average diameter of 230 ± 27.4 nm. It was observed that the fibers were not uniform in shape and the beads were formed in the middle.

Referring to FIG. 3, the average diameter of the second nanofiber sheet was 134.2 ± 9.33 nm, and uniform and consistent fiber morphology was observed compared to other nanofiber sheets.

Referring to FIG. 4, the third nanofiber sheet was found to have an average diameter of 224 ± 89.13 nm, and the shape of the fiber was observed to be highly non-uniform.

Through the above experiment, it was confirmed that a nanofiber sheet can be prepared in a weight ratio of N-carboxylate chitosan and polyethylene oxide within a range of 1: 0.5 to 2. In particular, the nanofiber has excellent properties at a weight ratio of 1: 1 appear.

Experimental Example 2: Infrared spectroscopy (FT-IR)

To investigate the chemical structure of the N-carboxy chitosan and the second nanofiber sheet, an infrared spectrophotometer (FTIR spectroscopy, Shimadzu-IR Prestige-21, Japan) was used. Creating a KBr pellet was prepared in the specimen, the sample and KBr parts by weight 1: After preparing a pellet by mixing well 100, and removing water by using a vacuum-dried at 60 ℃ for 12 hours 4000~500cm ^- was measured from the ^first region.

FIG. 5 is an FT-IR graph of N-carboxy chitosan, and FIG. 6 is an FT-IR graph of a second nanofiber sheet prepared by electrospinning a polymer solution having a composition of CEC: PEO = 3: 3.

Referring to FIG. 5, in the case of N-carboxy chitosan, peaks of carboxyl groups were observed in comparison with water-insoluble chitosan.

6, peaks of the 1553 cm- ¹ amide (NH, amide) peak of N-carboxy chitosan and 1098 cm ^-1 ether group (CO, ether) of polyethylene oxide were observed in the nanofiber sheet, Are combined together. &Lt; tb &gt;&lt; TABLE &gt;

<Experimental Example 3> Differential Scanning Calorimetry (DSC)>

The glass transition temperature was measured under a nitrogen atmosphere using Mettler Toledo DSC (DSC821) for thermal analysis of a second nanofiber sheet prepared by electrospinning a polymer solution having a composition of CEC: PEO = 3: 3. Fig. 7 shows the thermal change of nanofibers using differential calorimetry. Lt; RTI ID = 0.0 &gt; 400 C &lt; / RTI &gt; under a nitrogen flow of 50 ml per minute.

Referring to FIG. 7, it can be seen that the characteristics of PEO and CEC are integrated in the nanofiber sheet similarly to the FT-IR results. The peak down to the lower end between 60 and 80 ℃ is the melting state of the crystallized polymer, and the endothermic process can be observed. The peak at about 270 ° C is due to the exothermic reaction and can be seen as a characteristic when heated in the presence of air or oxygen.

<Experimental Example 4: Cytotoxicity Test>

Cytotoxicity tests were conducted using PEO, CEC, and second nanofiber sheet as samples. Cell viability was measured by MTT assay.

Each sample was added to the cell culture medium (DMEM, Dulbecco's modified Eagle's medium, Gibco) at concentrations of 10, 100, 1000 and 10000 ppm, and dissolved at 37 ° C for more than 24 hours to prepare an eluate. 96 RAW 264.7 per each well in the well plate cell 5 × 10 ⁴ cells / well for dispensing and after 24 hours 37 ℃ for 24 h gave after transfer to a DMEM culture solution containing a leaching solution prepared at different concentrations, 5% CO ₂ Lt; / RTI &gt; 20 μL of MTT solution (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) per well was added and reacted at 37 ° C and 5% CO ₂ for 4 hours. And formazan was dissolved by adding 100 μL of DMSO. The solution was measured at 570 nm using an ELISA measuring device (ELX 808, Biotek Instruments, Vermont, USA) and is shown in FIG.

Referring to FIG. 8, it was observed that the eluted solution of the second nanofiber sheet obtained after electrospinning had no cytotoxicity.

Experimental Example 5: Animal Experiment [

A 6-week-old male Sprague-Dawley (SD) male rats weighing 180-220 g were purchased from Seongnam, Korea and maintained at 25 ± 1 ° C and a humidity of 50 ± 10% Respectively. Feed and water were also freely consumed, and all experimental animals were adapted to the animal room environment for one week before being used in the experiment.

(1) induction of adhesion

Zoletil (2.5 mg per 100 g body weight) and xylazine (0.35 mg per 100 g body weight) were injected intraperitoneally with zoletil (5 mg per 100 g body weight) and xylazine (0.7 mg per 100 g body weight) Lt; / RTI &gt; At the time of anesthesia, the abdomen was shaved and disinfected with povidone and alcohol, followed by laparotomy along the midline at 4 to 5 cm. The cecum was removed and the 1 × 2 cm ² sachets were used to damage the membranes enough to cause bleeding.

The experiment was divided into PBS treatment group (negative control group), positive control group and experimental group. In the PBS treatment group and the positive control group, 3 animals were placed and 4 animals were placed in the experimental group. In the positive control group, 5 ml of hyaluronic acid-containing injectable antiadhesive agent (Gadix, Genewell, Korea) was applied to the injured area, and the peritoneal membrane was closed by a continuous suturing method. In the PBS treatment group, PBS (Phosphate buffered saline) solution was applied at 5 ml to the wound area and then sutured. In the experimental group, the second nanofiber sheet was inserted between the peritoneal membrane and the serosa and then sutured.

The animals were sacrificed for 1 week with enough water and food to evaluate the degree of adhesion.

(2) Evaluation of adhesion

The degree of adhesion was assessed by Vlahos et al. (Milligan, DW, and Raftery, AT: Observations on the pathogenesis of peritoneal adhesions: a light and electron microscopical study. , 274-280 Harris, ES, Morgan, RF, and Rodeheaver, GT: Analysis of the kinetics of peritoneal adhesion formation in the rat and evaluation of potential antiadhesive agents, Surgery, 1995, 117, 663-669). The degree of adhesion was classified into six grades as shown in Table 2 below.

Rating Justice 0 No adhesion One One thin film-type adhesion 2 Two or more thin film-type adhesives 3 Centralized thick adhesion of the points 4 Centralized thick adhesion of plate 5 Very thick adhesion with blood vessels

The degree of adhesion and the strength of adhesion in each group were evaluated according to the criteria of Table 2 and evaluated as shown in Table 2 below. When evaluating the adhesion, the evaluator made it impossible to know the experimental group so that the objectivity can be improved. FIG. 9 is a photograph showing the naked eye observation of the adhesion of the tissue on the 7th day after suturing.

Rating
PBS treated group Positive control group
(Positive control) Experimental group (n = 3) (n = 3) (n = 4) 0 0 0 4 One 0 0 0 2 0 0 0 3 0 2 0 4 One One 0 5 2 0 0 Mean of score 4.7 3.33 0 Total with adhesion 3 (100%) 3 (100%) 0 (0%)

Referring to Table 3 and FIG. 9, in the PBS-treated group, the degree of adhesion was mostly 4, 5 and the average degree of adhesion formation was 4.6. In the positive control group treated with injectable antiadhesive agents, adhesion grades 3 and 4 were also exhibited. The average degree of adhesion formation was 3.33.

However, the degree of adhesion was 0 in the experimental group, and it was observed that the biocompatibility and hydrophilicity of the water - soluble chitosan improved the wound healing effect. The uniform nanofibers formed at the optimal concentration of CEC or PEO inhibit the formation of adhesion, especially inhibiting the cell proliferation and angiogenesis, and decomposing and absorbing repeatedly, so that the degradation products are harmless to the human body and appear to be low.

Visual observations also showed that the PBS treated group and the positive control group showed severe adhesion in the tissue, but the treated group with the 2-nanofiber sheet showed a cleaned abdominal wall.

Therefore, the nanofiber sheet of the present invention is expected to have an excellent effect as an anti-adhesion agent after surgery.

Experimental Example 6: Histological examination Experiment 6:

For histological evaluation, the surgical site was removed with the muscle layer and dura mater, and then placed in a 10% neutral formalin solution for 24 hours. Subsequently, the tumor was placed in a decolorization solution for 48 hours. made. This block was cut to a thickness of 6-8 μm and stained with hematoxylin-eosin and histologically evaluated through a microscope. Observation results are shown in Figs. 10 to 12, respectively.

Adhesion is formed by the release of kinin and histamine, which are vasoactive factors in the injured mucosa, and capillary permeability is increased to accumulate inflammatory cells and promote cell matrix formation, leading to formation of fibrinous blood clots. After that, it is known that fibrous breakdown does not occur smoothly in damaged mucosal layer, but fibroblasts, macrophages, giant cells and the like are infiltrated into blood vessel granulation tissues, and collagen production and connective tissue are structured and adhesion is organized.

10 to 12, adhesion to the cecum and abdominal wall was observed in the PBS-treated group, and adhesion was observed in the positive control group.

On the other hand, adhesion did not occur in the experimental group, and wound healing was also natural in the wound restoration aspect. In particular, in the case of physical barriers, it encapsulates or covers the wound area, thereby blocking the contact with the surrounding tissue, inducing regeneration of the tissue, promoting the mutual activation of immune cells during the regeneration process after adhesion of cells, Promoting the function of immune cells such as promoting further. 12, it was observed that the mesothelial layer was regenerated during the treatment of the second nanofiber sheet prepared by spinning the polymer solution having the composition of CEC / PEO = 3: 3, and it was confirmed that it could be used as an adhesion inhibitor .

While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. Accordingly, the true scope of protection of the present invention should be determined only by the appended claims.

Claims (7)

A nanofiber sheet for adhesion prevention using water-soluble chitosan and polyethylene oxide, which is formed by electrospinning a polymer solution containing water-soluble chitosan and polyethylene oxide. The nanofiber sheet for preventing adhesion according to claim 1, wherein the water-soluble chitosan is N-Carboxyethyl Chitosan, and the water-soluble chitosan is polyethylene oxide. The nanofiber sheet according to claim 1, wherein the water-soluble chitosan and the polyethylene oxide are contained in a weight ratio of 1: 0.5 to 2, based on the weight of the water-soluble chitosan and the polyethylene oxide. Dissolving water-soluble chitosan and polyethylene oxide in water to obtain a polymer solution;
And a step of electrospinning the polymer solution. The method for producing a nanofiber sheet for adhesion prevention using water-soluble chitosan and polyethylene oxide. The method according to claim 4, wherein the polymer solution is prepared by dissolving the water-soluble chitosan and the polyethylene oxide in a weight ratio of 1: 0.5 to 2, and dissolving the water-soluble chitosan and the polyethylene oxide. The method of claim 4, wherein the water-soluble chitosan is prepared by a method comprising the steps of: a) stirring a chitosan solution in which non-water-soluble chitosan is dissolved in a mixed solvent of acrylic acid and distilled water; b) dialyzing after adding sodium hydroxide to the chitosan solution , and c) N-Carboxyethyl Chitosan obtained by performing the step of lyophilizing the solution obtained after the dialysis. Production of a nanofiber sheet for adhesion prevention using water-soluble chitosan and polyethylene oxide Way. 5. The method according to claim 4, wherein the electrospinning is performed by irradiating the polymer solution under conditions of a temperature of 22 to 28 DEG C, a relative humidity of 27 to 33%, a voltage of 10 to 40 kV, a radiation distance of 10 to 20 cm and a fluid velocity of 0.02 to 0.1 mL / The method for producing a nanofiber sheet for adhesion prevention using water-soluble chitosan and polyethylene oxide.

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