KR101601674B1 - Surface Modified Nanofibrous GBR membrane and preparation method thereof - Google Patents

Abstract

The present invention relates to a nanofiber shielding film for surface-modified bone tissue induced regeneration, which comprises a nanofiber nonwoven fabric of a biocompatible natural polymer and one surface of the nanofiber nonwoven fabric has a hydrophobic property. According to the present invention, Since the nanoparticles have a high specific surface area and are porous, it is possible to exchange physiologically active substances and can be easily applied to the treatment site. Further, it is not necessary to carry out secondary treatment due to biodegradability, and one side of the natural polymer nanofiber nonwoven fabric The surface modification is made to have a very low water solubility through the plasma treatment, so that the shielding ratio and the regeneration rate can be selectively increased.

Description

[0001] The present invention relates to a nanofiber shielding film for surface-modified bone tissue induction regeneration,

The present invention relates to a nanofiber shielding film made of a biocompatible natural polymer and a method of manufacturing the same, and more particularly, to a shielding film for dental bone tissue induction / regeneration. The present invention relates to a nanofiber shielding film having a high surface area and porosity, which is capable of preventing penetration of cells and foreign matter and imparting hydrophobicity to a surface of a nonwoven fabric through plasma treatment.

The start of Guided Tissue Regeneration (GTR) is based on the concept of blocking the growth of unwanted soft tissues in injured tissues by inducing the regeneration of the desired tissues by placing a barrier in soft tissues with a high regeneration rate. The concept of tissue-induced regeneration (GTR) developed in the periodontal region is intended not only to regenerate bone tissue but also to regenerate periodontal tissue such as periodontal ligament or cementum. However, in implantology, the concept of GTR has changed to the concept of Guided Bone Regeneration (GBR), which means bone regeneration. Techniques to treat shields well for successful implant surgery are also needed, but failures and chaos in many cases are due to insufficient understanding of the shield.

The characteristics of this shielding membrane should prevent intrusion of adjacent fiber connective tissues, prevent the invasion of bacteria even if exposed, and attach or attach well to surrounding tissues. This should stabilize the wound healing, seal between the material and the bone, and prevent the fibrous connective tissue from being exposed to the defect.

In addition, it should be able to bond properly to the shape of the defect with appropriate flexibility and strength without the thickness of the shielding film being too thick, too hard or torn too much, and to provide adequate space for bone regeneration, It must have processability that can be properly shaped.

These dental shields are largely classified into biodegradable and non-degradable. Biodegradable shielding membranes do not require removal of membranes and thus have the advantage of less risk of bacterial infection due to secondary surgery. It is generally easy to manipulate, and when the implant is covered, the shape is kept well in wet condition, so it is not necessary to fix it with a bone tack. However, there is a disadvantage that the effect of regeneration of bone tissue is uncertain due to the limitation of providing space and uncertainty of absorption period and absorption rate.

Biodegradable shields include shields made of collagen. Biogide products are composed of type Ⅰ and type Ⅲ collagen extracted from porcine dermis. Type Ⅰ collagen extracted from bovine tendon And Biomend, a product that is absorbed into tissues within 8 weeks. Biomend Extend, which is a product that increases the absorption time by increasing the amount of cross-linking in this Biomend product, includes Neomem made of bovine collagen, Lyoplant made of bovine collagen, Biosorb, ossix plus and EZ cure.

Another example of a biodegradable polymer is a shielding film made of polylactic acid (PLA) and polyglycolic acid (PGA). As a shielding film using this material, there is a product called BioMesh. And it is known that absorption is almost completed in 8 weeks of experiment (Patent Document 1).

Shielding films of chitosan material and insect origin protein materials have been developed as shielding films for other natural materials (Patent Document 2).

The most widely used shielding membrane is e-PTFE (Expanded-polytetrafluorietylene) shielding membrane, which has been used for a long time. However, since it does not decompose into non-degradable shielding membrane, This is a disadvantage in that the possibility of infection is very high. Product names using e-PTFE include Cytoplast membrane and BioBarrier.

Titanium mesh is a titanium material that is designed to maintain space because conventional shielding membranes have problems in maintaining space. Usually, e-PTFE membranes are used as a reinforcing structure and as a mesh (Patent Document 3). Titanium-based products include Cytoflex®, Ti-Mesh membrane, Osteo-mesh, and Bone Shield. The disadvantage is that secondary surgery is also necessary and there is a risk of infection.

Although the biodegradable shielding membrane is used to reduce the risk of secondary treatment of the non-degradable shielding membrane and thus infection, due to the relatively low shielding property and biodegradation characteristics as described above, the absorbent property and biodegradability There is a disadvantage that the cells can invade according to the speed, and there is a problem that re-operation may be required.

Accordingly, it is an object of the present invention to solve the above-mentioned problems of the prior arts and to provide a biodegradable biodegradable shielding film which can easily be processed and improved in shielding property and space providing, while taking biodegradability characteristics, Is required to be developed.

Patent Document 1: Korean Patent No. 0968231 Patent Document 2: Korean Patent Publication No. 2013-0033826 Patent Document 3: Korean Patent No. 1297814

In order to solve the above problems, the present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide a biocompatible natural polymer by a method of preparing a nanofiber nonwoven fabric sheet by electrospinning and imparting hydrophobicity to one surface of the nonwoven fabric by plasma treatment, The present invention also provides a nanofiber shielding film for surface-modified bone tissue induction and regeneration, which has improved bone regeneration and shielding characteristics from the advantages of nanofibers, and a method for producing the same.

According to an aspect of the present invention, there is provided a nanofiber shielding membrane for surface-modified bone tissue induction and regeneration, which comprises a nanofiber nonwoven fabric of a biocompatible natural polymer, and one surface of the nanofiber nonwoven fabric has a hydrophobic property.

Preferably, the biocompatible natural polymer is at least one selected from the group consisting of silk fibroin, chitosan, collagen, pullulan, gelatin, γ-PGA [poly (γ-glutamic acid)], alginate and mixtures thereof.

Also, preferably, the nanofiber nonwoven fabric has an average diameter of the fiber cross-section of 10 nm to 1000 nm.

Also preferably, the nanofiber nonwoven fabric is selected from the group consisting of penicillin, sephadrine, cephazoline, sepharaxine, cefuroxime, sephacler, sepharose, cephatriaxone, , Cephalosporins, cephalosporins, cephalosporins, cephalosporins, cephalosporins, cephalosporins, cephalosporins, cephalosporins, But are not limited to, tin, ceftazidime, imipenem, meropenem, erthapenem, paropenem, vancomycin, bacitracin, cycloserine, chloramphenicol, tetracycline, erythromycin, azithromycin, clarithromycin, streptomycin, neomycin, But are not limited to, moromycin, sulfamethoxazole, sulfadiazine, kanamycin, daunorubicin, dirubicin, mitomycin, doxorubicin, epirubicin, bleomycin, dactinomycin, Eojin from the group which contains at least one.

Also preferably, the biocompatible natural polymer nanofiber nonwoven fabric comprises reinforced and supported by a Titanium mesh or foil.

According to another aspect of the present invention, there is provided a method for preparing a surface-modified nanofiber shielding layer for bone tissue induction and regeneration, comprising: dissolving silk fibroin in an organic solvent to prepare a silk fibroin solution; Preparing a silk fibroin nonwoven fabric nonwoven fabric by electrospinning the silk fibroin electric spinning solution; Insolubilizing the silk fibroin nanofiber nonwoven fabric to make the silk fibroin nanofiber nonwoven fabric insoluble by using steam; And modifying the insoluble silk nanofiber nonwoven fabric to a hydrophobic surface through plasma treatment.

Preferably, the silk fibroin comprises removing sericin from the silk; Preparing a silk fibroin aqueous solution by using the sericin-removed silk fibroin with a salt and ethanol; Removing the salt and ethanol in the aqueous solution of silk fibroin, and lyophilizing the aqueous solution of silk fibroin from which the salt and ethanol have been removed to obtain a regenerated silk fibroin powder.

Preferably, the organic solvent is at least one selected from the group consisting of 1,1,1,3,3,3-hexafluoroisopropanol, formic acid, and mixtures thereof.

Preferably, the electrospinning is carried out under electrospinning conditions of a solution concentration of 1 to 50 wt%, a voltage of 5 to 100 kV, a discharge rate of 0.1 to 10 ml / h, a radiation distance of 3 to 50 cm and a humidity of 1 to 50% To obtain a nanofiber.

Preferably, the plasma treatment is performed using a gas having a fluorine group under conditions of a treatment time of 0 to 3 hours, a power of 10 to 100 W, and a gas flow rate of 0.1 to 300 cc / min.

Preferably, the gas having the fluorine group is at least one selected from the group consisting of trifluoromethane, tetrafluoromethane, hexafluoroethane, tetrafluoroethylene, octafluoropropane, hexafluoropropylene, perfluorobutane, perfluoropentane, perfluorohexane, At least one of the group consisting of sulfur, phosphorus, sulfur, phosphorus, sulfur, and mixtures thereof.

Preferably, the silk fibroin solution further contains at least one selected from the group consisting of penicillin, sephadrine, cephazoline, sepharaxine, cefuroxime, sephacler, sepharose, cephatriaxone, But are not limited to, dicloxacin, oxacillin, flucloxacin, amoxicillin, ampicillin, piperacillin, ticarcillin, azulocillin, carbenicillin, cephalexin, cephalothin, , Cephacytin, ceftazidime, imipenem, meropenem, erthapenem, paropenem, vancomycin, bacitracin, cycloserine, chloramphenicol, tetracycline, erythromycin, azithromycin, clarithromycin, streptomycin, neo But are not limited to, mitomycin, tobramycin, sulfamethoxazole, sulfadiazine, kanamycin, daunorubicin, darubicin, mitomycin, doxorubicin, epirubicin, bleomycin, Further comprising the step of adding and mixing at least one from the group consisting of a mixture of.

According to another aspect of the present invention, a medical nanofiber sheet includes a surface modified nanofiber shielding layer for regenerating bone tissue.

The nanofiber shielding membrane for surface-modified bone tissue induction regeneration according to the present invention can be manufactured by forming nanoscale non-woven fabrics of a biocompatible natural polymer by an electrospinning method to prevent infiltration of cells and foreign substances according to the nanofiber structure, The effect of the present invention can be remarkably enhanced.

In addition, by producing the nanofiber nonwoven fabric containing the antimicrobial agent easily by adding the antimicrobial agent to the electrospinning aqueous phase, infection and contamination from external germs can be fundamentally blocked.

Further, the natural polymer nanofiber nonwoven fabric produced by the electrospinning method is subjected to the insolubilization treatment using water vapor, whereby the advantages of insolubility and flexibility can be simultaneously improved by using water vapor.

In addition, the hydrophobic surface of the nanofiber shielding film is treated by plasma treatment to prevent the penetration of the soft tissue by the hydrophobic surface, which is not in direct contact with the treatment site. On the other hand, It is possible to simultaneously improve the shielding rate and the bone regeneration efficiency by providing the advantage of biocompatibility and biodegradability.

Accordingly, it is possible to improve the shielding ability and improve the usability of the user by improving adhesion, flexibility, ease of manipulation, biocompatibility and biodegradability to tissues and organs which were problems of the conventional shielding film, To prevent the infiltration of cells and penetration of foreign substances, thereby enhancing shielding ability, promoting healing of wounds, and being able to be manipulated or moved by small surgical tools without being torn or broken even when folded or rolled. This has an easy effect.

1 is a photograph showing an electrospinning device according to an embodiment of the present invention.
2 is an electron micrograph of a silk fibroin nanofiber according to an embodiment of the present invention.
FIG. 3 is an electron micrograph of a silk fibroin nanofiber according to an embodiment of the present invention after the insolubilization treatment. FIG.
4 is a photograph showing a plasma apparatus according to an embodiment of the present invention.
5 is a photomicrograph of the silk fibroin nanofiber after plasma treatment according to an embodiment of the present invention.
FIG. 6 is a graph showing a comparison result of water contact angles before and after the plasma treatment of the silk fibroin nanofibers according to an embodiment of the present invention
7 is a graph showing the results of MTT analysis of nanofibers according to an embodiment of the present invention.
8 is a scanning electron micrograph of a cell proliferation behavior on nanofibers according to an embodiment of the present invention.
FIG. 9 is a photograph showing a H & E staining and a trichrome staining microscope photograph of a bone and a sample according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail with reference to the drawings.

The present invention relates to a nanofiber shielding film for surface-modified bone tissue induced regeneration, which comprises a nanofiber nonwoven fabric of a biocompatible natural polymer and one surface of the nanofiber nonwoven fabric has a hydrophobic property, .

The biocompatible natural polymer used in the present invention refers to a polymeric substance harmless to the human body extracted from a natural product which is decomposed by itself in vivo. The polymer is silk fibroin, chitosan, collagen, pullulan, Gelatin, γ-PGA [poly (γ-glutamic acid)], alginate, and mixtures thereof. Silk fibroin is preferably used.

Unlike cotton or wool fibers, silk is obtained in the form of a long fiber, a fibrous protein made by insects including silkworms and spiders. It is a natural polymer with high toughness, strength and extensibility. The silk is composed of the main component fibroin and the remaining sericin.

Firstly, silk sericin is also called "silk glue", because the rough and hard feeling of silk glue is attached to sericin, and the color of silk cocoon fiber is indicated by the carotenoid pigment contained in sericin , The amino acid composition of sericin is characterized by a remarkably large amount of serine (37 mol%). The physical properties of sericin are dissolved in hot water and gelled when cooled.

Silk fibroin is a kind of light protein. Most of the sericin is melted when it is boiled for about 15 hours in hot water. Only the fibroin is left. In addition, treatment with a dilute alkaline solution dissolves sericin, but fibroin does not dissolve to remove silk sericin. Fibroin, which is removed from sericin in silk fibers, is not dissolved in water, dilute acid, dilute alkali, etc. and has a characteristic of dissolving in an aqueous solution such as lithium bromide, calcium chloride, calcium nitrate, thiocyanic acid and dichloroacetic acid have.

This silk fibroin is a typical fibrous protein and has been mainly used for high-grade garment fibers and surgical sutures. Since silk fibroin has characteristics such as biodegradability, biocompatibility, oxygen and water permeability, and low inflammation reactivity, As an embodiment of the present invention, it can be said that it is a very suitable material to be used as a nanofiber for bone tissue induction regeneration.

The present invention utilizes an electrospinning process for the production of nanofiber nonwoven fabrics, in order to prepare an aqueous solution of silk fibroin for electrical applications. A method for extracting silk fibroin, comprising the steps of: removing sericin from silk as described above; preparing a silk fibroin aqueous solution using said salt and ethanol in said silk fibroin from which said cerasin has been removed; Removing the ethanol, and lyophilizing the aqueous solution of silk fibroin from which the salt and ethanol have been removed to obtain a regenerated silk fibroin powder.

The salt may be at least one selected from the group consisting of zinc chloride, lithium chloride, calcium chloride, zinc nitrate, calcium nitrate, lithium bromide and mixtures thereof. The salt and ethanol may be removed using a dialysis membrane , And a silk fibroin aqueous solution can be prepared by dialyzing distilled water.

The silk fibroin powder prepared by lyophilizing the aqueous solution of silk fibroin is prepared by using an organic solvent to prepare a silk fibroin solution for an electric discharge. The organic solvent used is 1,1,1,3,3,3-hexafluoropropane At least one selected from the group consisting of 1,1,1,3,3,3-hexafluoroisopropanol, HFIP, formic acid and mixtures thereof may be selected and preferably relatively volatile And 1,1,1,3,3,3-hexafluoroisopropanol, which is a solvent having a small residual solvent, is suitable. The concentration of the silk fibroin solution used for the electric discharge is preferably in the range of 1 to 50 wt% Is 3 to 20 wt%.

Silk fibroin nanofibers are obtained through electrospinning from an electric room silk fibroin solution.

FIG. 1 is a photograph of an electrospinning device according to an embodiment of the present invention. In general, the diameter and physical properties of nanofibers vary depending on the concentration, radiation voltage, radiation distance, and flow rate, The higher the voltage and the farther the scatter is, the narrower the diameter of the nanofiber.

The process requirements according to one embodiment of the present invention are as follows: the process conditions of the electrodeposition of the dispersion solution are 1 to 50 wt%, 5 to 100 kV of voltage, 0.1 to 10 ml / h of discharge rate, 3 to 50 cm of radiation distance and 1 to 50% And spinning under spinning conditions to obtain nanofibers

FIG. 2 is an electron micrograph of a silk fibroin nanofiber according to an embodiment of the present invention. In FIG. 2, an average diameter of a fiber cross section according to the present invention is 10 nanometers [nm] to 1000 nanometers [nm] As shown in FIG. 2, since the fine mesh structure having an average diameter of 729.6 nm has a high specific surface area and is porous, it is possible to exchange physiologically active substances and can be easily applied to a treatment site.

A silk fibroin nonwoven fabric having antibacterial activity can be produced by further adding an antimicrobial agent to the solution of the silk fibroin used in the electric room. As antimicrobial agents that can be used herein, antimicrobial agents such as penicillin, cefadrine, caffeolin, cefalexin, cefuroxime, sephacler, cefuroxir, cephatriaxone, But are not limited to, fluoxetine, fluglocracin, amoxicillin, ampicillin, piperacillin, ticarcillin, azlocillin, carbenicillin, cephalexin, cephalotin, Erythromycin, azithromycin, clarithromycin, streptomycin, neomycin, tobramycin, sulfamethoxazole, erythromycin, erythromycin, erythromycin, A group consisting of methoxazole, sulfadiazine, kanamycin, daunorubicin, dirubicin, mitomycin, doxorubicin, epirubicin, bleomycin, dactinomycin and mixtures thereof Since it may contain at least one.

The silk fibroin nanofibers prepared as described above are subjected to a step of insolubilization using water vapor. In general, in the past, a method of insolubilizing by alcohol aqueous solution treatment has been used. However, since this method is hard to apply in a living body since its physical properties are hard after processing, This improved silk fibroin nanofiber could be prepared

After the silk fibroin nanofiber was subjected to insolubilization treatment, the silk fibroin nanofiber was immersed in distilled water for washing to remove residual solvent, and after vacuum drying, the morphology of the insolubilized silk fibroin nanofiber was examined by scanning electron microscope (SEM) 3 shows an electron micrograph of the silk fibroin nanofiber after the insolubilization treatment as shown in Fig. 3. It can be confirmed that even when the insolubilization treatment is performed, no deformation of the fiber shape takes place at all.

The nanofiber sheet is modified into a hydrophobic surface through a plasma treatment using a gas having a fluorine group to produce a nanofiber having both a relatively hydrophobic surface and a hydrophilic surface. In this case, the hydrophobic surface is a layer which does not come into direct contact with the treatment site and serves to prevent infiltration of the soft tissues. On the contrary, the hydrophilic surface is a surface in contact with the treatment site, and has excellent biocompatibility and good biodegradability .

FIG. 4 is a photograph of a plasma apparatus according to an embodiment of the present invention. In the plasma processing used in the present invention, the treatment time is 0 to 3 hours, the electric power is 10 to 100 W, the gas flow rate is 0.1 to 300 cc / min And preferably a power of 10 to 50 W, a treatment time of 100 to 600 S, and a gas flow rate of 100 cc / min.

The gas having the fluorine group used in the present invention may be at least one selected from the group consisting of trifluoromethane, tetrafluoromethane, hexafluoroethane, tetrafluoroethylene, octafluoropropane, hexafluoropropylene, perfluorobutane, perfluoropentane, perfluorohexane, At least one selected from the group consisting of fluorine, sulfur, fluorine, sulfur, and a mixed gas of these can be selected, and tetrafluoromethane is preferably used.

FIG. 5 is an electron micrograph of a silk fibroin nanofiber according to an embodiment of the present invention after plasma treatment, showing that no deformation of the fiber by the plasma treatment occurred at all.

The plasma treatment method using a gas having a fluorine group has been used as an embodiment of the present invention. However, the present invention is not limited to this, but a physical or chemical treatment such as a coating for imparting a low water- And a surface modification process.

Further, as another embodiment according to the present invention, titanium may be used as a reinforcing agent of the nonwoven fabric of the silk fibroin. Although the silk fibroin nanofiber nonwoven fabric according to the present invention can be used alone, when shielding film for guided bone generation (GBR) is required in a stronger strength, excellent corrosion resistance, strength and biocompatibility are required, It is possible to maintain a sufficient space and to prevent the collapse of the soft tissue during the operation. The titanium can be used as a mesh type and a foil type, and it is possible to efficiently reinforce and support a silk fibroin nano nonwoven fabric as a mesh type.

≪ Example 1 > Preparation of silk fibroin nanofiber

Silk fibroin was dissolved in 1,1,1,3,3,3-hexafluoroisopropanol (HFIP) to prepare solutions of 4, 5 and 6 wt / v% . As the process parameters, the voltage was fixed at 10 ~ 20 kV, the radiation distance was 8 ~ 12 cm, the fluid velocity was 1.0 ~ 2.0 ml / h, the drum collector was irradiated for 40 hours and the humidity was kept below 40%. It was then thoroughly dried in a 25 [deg.] C vacuum oven for 24 hours to remove the residual solvent contained in the sheet.

≪ Example 2 > Insolubilization treatment of silk fibroin nanofiber

The silk fibroin nanofibers prepared in the same manner as in Example 1 were insolubilized using steam for 4 hours. After the insolubilization treatment, the silk fibroin nanofibers were immersed in distilled water to remove residual solvent, and then washed and vacuum dried.

≪ Example 3 > The CF of the insolubilized silk fibroin nanofiber ₄  Gas plasma treatment

The insolubilized silk fibroin nanofibers prepared in Example 2 were made into nanofibers having a microphosphate surface through CF ₄ gas plasma treatment. As the process parameters of the plasma treatment, the power was fixed at 10 to 50 W, the treatment time was 100 to 600 S, and the CF ₄ gas flow rate was 100 cc / min.

Experimental Example 1: Measurement of water contact angle

The water contact angle of the plasma treated silk fibroin nanofibers prepared in Example 3 was measured by a sessile drop method (distilled water, 5)) using Contact Angle & Surface Tension Analyzer (Model: Phoenix 300, SEO Co., (Fig. 6).

The water contact angle measurement is the most common method for analyzing the characteristics of a solid surface and is used as an important measure for evaluating wettability or hydrophilicity / hydrophobicity by measuring a contact angle between a solid and a liquid by dropping a certain amount of water droplet on the sample surface .

Before plasma treatment After plasma treatment Contact angle (average) 99.73 DEG 135.8 DEG

Table 1 shows the measurement results of water contact angle.

The water contact angle was measured by dropping water droplets at 10 locations by varying the measurement surface positions of the respective nanofibers for the accuracy of the analysis. As a result, as shown in Table 1, the contact angle significantly increased after the plasma treatment, It can be confirmed that the fibroin surface is modified to have a low water solubility.

<Experimental Example 2> Comparative test of cell proliferation on nanofiber surface

The plasma-treated silk fibroin nanofibers prepared in Example 3 were sterilized by irradiation with UV rays, and the fibroblasts of the silk fibroin nanofibers and the NIH3T3 fibroblasts subcultured on a 12-well cell culture plate (control) Lt; / RTI &gt; At this time, a ring made of glass was placed on the nanofiber, and the cell was inoculated into the ring to induce cell proliferation only on the surface of the nanofiber. As the medium, a medium containing 8% FBS and 0.1% penicillin G-streptomycin was used.

The inoculated cells were cultured for 4 hours, 1 day, 3 days, 5 days and 7 days in an incubator at 37 ° C. and 5% CO 2 atmosphere, and 1 ml of MTT storage solution was added to the inoculated silk fibroin nanofibers , And incubated for 4 hours. DMSO (dimethylsulfoxide) (1 ml) was added to dissolve the cell membrane, and the absorbance at 540 nm was measured using an ELISA reader (enzyme immunoassay) Were measured.

FIGS. 7 and 8 are graphs showing MTT analysis results of nanofibers according to an embodiment of the present invention and scanning electron microscope photographs of cell proliferation behavior on nanofibers. The results of MTT analysis and scanning electron microscopy showed that the adhesion and propagation behavior of the cells on the silk fibroin nanofibers before the plasma treatment was good. On the other hand, in the case of the plasma treated silk fibroin nanofibers, It can be confirmed that the propagation behavior is not relatively active.

In particular, it was confirmed that cell adhesion and propagation behavior on the surface of 12-well cell culture plate is more actively performed, and that the surface of the silk fibroin nanofiber on the surface of the silk fibroin nanofiber before the plasma treatment Confirming that the cell proliferation is relatively inactive, it is confirmed that the shielding property is improved by the plasma treatment.

In the present invention, rabbit was used as a test object in order to evaluate the shielding effect of the surface-modified silk fibroin nanofiber sheet. Five rabbits were weaned at 4 weeks of age (weight: 2.9 ~ 3.1 kg).

Rabbits were punctured at 4 sites. As a control group, one of the 4 punctures was inserted in each rabbit. The shielding membrane was not inserted, and 3 groups of surface-modified silk fibroin nanofiber shields were inserted. After the skull was opened, the perforated skull was taken and analyzed for H & E staining and trichrome staining. As a result, as shown in FIG. 9, it was confirmed that the new bone tissue was formed well without the penetration of the soft tissue into the shielding film.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (13)

A non-woven fabric of a biocompatible natural polymer comprising a hydrophobic surface on a first surface and a hydrophilic surface on a second surface,
Wherein the first surface is modified to a hydrophobic surface by plasma treatment using a gas having a fluorine group under conditions of a treatment time of 100 to 600 sec, a power of 10 to 100 W, and a gas flow rate of 0.1 to 300 cc / min. Coated nanofiber shielding membranes. The method according to claim 1,
Wherein the biocompatible natural polymer is at least one selected from the group consisting of silk fibroin, chitosan, collagen, pullulan, gelatin,? - PGA [poly (? -Glutamic acid)], alginate, Modified nanofiber shielding membrane for induction of bone tissue induction. The method according to claim 1,
Wherein the nanofiber nonwoven fabric has an average diameter of the fiber cross-section of 10 nm to 1000 nm. The method according to claim 1,
The nanofiber nonwoven fabric may be at least one selected from the group consisting of penicillin, cefadrin, cefazolin, cefalexin, cefuroxime, sephacler, cefprozil, cephatriaxone, cytotoxic, Cycloserine, cephadecenone, cephamandol, cephatanthin, cephacytin, ceftazidime, ceftazidime, ceftazidime, ceftazidime, cyclamate, , Imipenem, meropenem, erthapenem, paropenem, vancomycin, bacitracin, cycloserine, chloramphenicol, tetracycline, erythromycin, azithromycin, clarithromycin, streptomycin, neomycin, tobramycin, Or a mixture thereof, from a group consisting of a compound selected from the group consisting of a sulfamoyl group, a sulfamoyl group, a sulfamoyl group, a sulfamoyl group, a sulfamoyl group, a sulfamoyl group, Wherein the nanofibrous shielding layer comprises at least one fiber. delete Dissolving silk fibroin in an organic solvent to prepare a silk fibroin electric spinning solution;
Preparing a silk fibroin nonwoven fabric nonwoven fabric by electrospinning the silk fibroin electric spinning solution;
Insolubilizing the silk fibroin nanofiber nonwoven fabric to make the silk fibroin nanofiber nonwoven fabric insoluble by using steam; And
The first side of the insoluble silk fibroin nanofiber nonwoven fabric is subjected to plasma treatment under the conditions of a treatment time of 100 to 600 sec, a power of 10 to 100 W, and a gas flow rate of 0.1 to 300 cc / min using a gas having a fluorine group Modifying it with a hydrophobic surface; Wherein the surface of the nanofibrous shielding film is in contact with the surface of the nanofiber. The method according to claim 6,
Wherein the silk fibroin comprises: removing sericin from the silk; Preparing a silk fibroin aqueous solution by using the sericin-removed silk fibroin with a salt and ethanol; Removing salt and ethanol in the aqueous silk fibroin solution; And a step of lyophilizing the aqueous solution of silk fibroin from which the salt and ethanol are removed to obtain a regenerated silk fibroin powder. The method according to claim 6,
Wherein the organic solvent is at least one selected from the group consisting of 1,1,1,3,3,3-hexafluoroisopropanol, formic acid, and a mixture thereof. The surface-modified nanofiber shielding film for surface- Gt; The method according to claim 6,
The electrospinning is carried out under an electrospinning condition of a solution concentration of 1 to 50 wt%, a voltage of 5 to 100 kV, a discharge rate of 0.1 to 10 ml / h, a radiation distance of 3 to 50 cm and a humidity of 1 to 50% Wherein the surface of the nanofibrous shielding layer is in contact with the surface of the nanofibers. delete The method according to claim 6,
The gas having the fluorine group may be at least one selected from the group consisting of trifluoromethane, tetrafluoromethane, hexafluoroethane, tetrafluoroethylene, octafluoropropane, hexafluoropropylene, perfluorobutane, perfluoropentane, perfluorohexane, Wherein at least one selected from the group consisting of a mixed gas of barium titanate and a mixed gas of barium titanate is selected. The method according to claim 6,
The silk fibroin solution for electrodialysis is prepared by dissolving or dissolving at least one selected from the group consisting of penicillin, sephadrine, caffeolin, cefalexin, cefuroxin, sepracur, sepharose, cefatriaxone, , Ceftazidime, ceftazidime, ceftazidime, ceftazidime, ceftazidime, ceftazidime, ceftazidime, ceftazidime, ceftazidime, ceftazidime, But are not limited to, thalidomide, tilidem, imipenem, meropenem, erthapenem, paropenem, vancomycin, bacitracin, cycloserine, chloramphenicol, tetracycline, erythromycin, azithromycin, clarithromycin, streptomycin, neomycin, A group consisting of sulfamethoxazole, sulfadiazine, kanamycin, daunorubicin, darubicin, mitomycin, doxorubicin, epirubicin, bleomycin, dactinomycin and mixtures thereof At least the surface-modified bone inductive reproducing method of producing a nanofiber shielding film for which the step of mixing and addition of one, characterized in that it further comprises from. A medical nanofiber sheet comprising the nanofiber shielding membrane for surface-modified bone tissue induction regeneration according to any one of claims 1 to 4.

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