WO2017171313A1 - Polymer regeneration membrane and preparation method therefor - Google Patents

Abstract

The present invention relates to: a leaf-stacked guided bone regeneration membrane having a membrane layer with a small average pore size at an upper part thereof and having a leaf-stacked structure in the inside and lower part thereof; and a preparation method therefor. The present invention is harmless to the human body since toxic organic materials are not used, and a guided bone regeneration membrane having a leaf-stacked structure can be manufactured by a simple method. In addition, according to the present invention, the leaf-stacked guided bone regeneration membrane allows the sustained release of a bioactive factor only by means of a leaf-stacked structure without using any additive and surface modification method, thereby being very effective in bone regeneration.

Description

Polymer regenerated membrane and preparation method thereof

The present invention relates to a polymer regeneration membrane having a selective permeability and a method for manufacturing the same, and in particular, has a unique internal structure capable of releasing physiologically active substances in a sustained release, and can inhibit the penetration of epithelial tissue and fibrous connective tissue. It relates to a polymer regeneration membrane and a method for producing the same.

In modern society, bone injury occurs more frequently for various reasons such as safety accidents including traffic accidents, fractures caused by sports activities, removal of cancer tissues, and cosmetic surgery. Bone damage is one of the reasons for the deterioration of human life.

When bone damage occurs, the defect is filled with epithelial tissue and fibrous connective tissue prior to bone tissue, thereby preventing regeneration of bone tissue. Physically shielding epithelial and connective tissues that interfere with bone tissue regeneration and accessing the defect to only the cells and materials useful for bone tissue regeneration to guide regeneration to the desired bone tissue. GBR)), and the shielding film used here is called an osteoinductive regeneration membrane (GBR membrane).

In the 1980s, Nyman and Gottlow established the concept of tissue-induced regeneration, and based on this, Dahlin reported that bone regeneration was possible using bone-induced regeneration membranes. Since it is an easy treatment method to cover the bone induction regenerative membrane in bone defects, many researchers are trying to develop a suitable bone induction regenerative membrane.

Materials used for bone induction regeneration membranes include large-scale natural polymers such as collagen and alginate, and synthetic polymers such as polytetrafluoro ethylene (PTFE) and polyglycolic acid (PGA). . Since natural polymers are easily absorbed and disappeared in the body, it is difficult to play a role of bone-induced regenerative membranes.Teflon, a synthetic polymer, has excellent biocompatibility and has been reported to be positive for bone regeneration. The disadvantage is that secondary surgery is required for removal.

Recently, researches using biodegradable synthetic polymers have been actively conducted to overcome the above disadvantages. Osteoinduced regeneration membrane must meet the following conditions in order to perform its function properly. 1) It has biocompatibility, 2) It should have proper shielding to prevent invasion of epithelial tissue and fibrous connective tissue, but 3) It must be easily permeable to nutrients and oxygen, 4) Bone tissue regeneration at bone defect area It should have enough mechanical properties to maintain space, and 5) be able to adhere well to surrounding bone tissue to prevent the penetration of fibrous connective tissue from the edge of the membrane, and 6) to prevent damage and treatment of surrounding normal tissue. It should have flexible characteristics to give ease, and 7) have biodegradability that does not need to be removed through secondary surgery after bone regeneration.

Recently, due to continuous research on biodegradable polymers, products such as Guidor, Vicryl Periodontal Mesh, and Biomesh based on biodegradable polymers have been released.However, the difficulty of the procedure and bone tissue and The low adhesiveness of the membrane (fibrous connective tissue can penetrate the membrane) does not meet the most basic requirements such as bone regeneration membranes. The use was very limited.

In addition, it is possible to permeate nutrients and oxygen essential for bone regeneration, but it is equipped with selective permeable membrane and bioactive factor which have the property of inhibiting the penetration of fibrous connective tissue. Manufacturing is almost never.

On the other hand, in order to solve the above problems, while the permeation of nutrients and oxygen essential for bone regeneration is easy, the permeation of fibrous connective tissue, which is an obstacle to bone regeneration, can be suppressed by the size of the pore and the porous structure of the surface. Porous osteoinductive regeneration membrane having excellent adhesion to bone tissues and very effective in bone regeneration, having an asymmetric structure including an upper layer having an average pore size of 1 to 3,000 nm and a lower layer having an average pore size of 5 to 500 μm. This is presented (see Korean Patent Application No. 10-2008-0124710).

In the above technique, a polymer solution is prepared by warming the polymer so that it is dissolved (about 90 ° C.), casting the polymer solution in a mold at room temperature (25 ° C. to 37 ° C.), and then casting the cast film to water at room temperature. It is added to the process of manufacturing.

That is, since the temperature gradually decreases from the temperature for preparing the polymer solution to the process of casting the polymer membrane and precipitating it in the non-solvent, the polymer maintains its solubility well in the solution. Therefore, the polymer membrane rapidly precipitated by the phase transition between the solvent and the non-solvent at the instant of the casting of the cast membrane into the non-solvent water has a uniform porous structure.

However, the bone regeneration induced membrane according to the above technique has a uniform porosity in the lower layer, so that the bioactive material is released through a plurality of pores in the lower layer. There are some limitations in inducing sustained-release or slow-release releases.

Therefore, the present inventors conducted a study on a polymer regeneration membrane having a new structure capable of seeking sustained release of a bioactive material by improving the structure of the porous osteoinductive regeneration membrane prepared in the above patent.

That is, by changing the various experimental conditions using the same material it is possible to optimize the manufacturing conditions to produce a polymer regenerated film having an improved structure compared to the prior art.

Accordingly, an object of the present invention is to provide a polymer regeneration membrane having an improved structure capable of maximizing the release of physiologically active factors.

In addition, another object of the present invention is to provide a method for producing a polymer regeneration membrane having the above characteristics.

The polymer regeneration membrane according to an embodiment of the present invention is characterized by having a structure including an upper layer having an average pore size of 1 to 5000 nm, and a lower layer having a deciduous laminated internal structure.

According to one embodiment of the invention, the polymer is polycaprolactone, polydioxanone, polylactic acid, polyglycolic acid, polylactic acid-glycolic acid copolymer, polyhydrosibutyrate and polyhydroxybutyric acid-hydroxy At least one biodegradable polymer selected from the group consisting of valeric acid copolymers and polyphosphoesters.

According to another embodiment of the present invention, the polymer is polycaprolactone, polydioxanone, polylactic acid, polyglycolic acid, polylactic acid-glycolic acid copolymer, polyhydrosibutyrate and polyhydroxybutyric acid-hydro At least one biodegradable polymer selected from the group consisting of hydroxyvalic acid copolymers and polyphosphoesters, and

Polyethylene oxide-polypropylene oxide copolymer, polyethylene oxide-polylactic acid copolymer, polyethylene oxide-glycolic acid copolymer, polyethylene oxide-polylactic glycolic acid copolymer, polyethylene oxide-polycaprolactone copolymer, polyethylene oxide-polydioxane It may be a mixture of one or more hydrophilic polymers selected from the group consisting of a copolymer and copolymers thereof.

According to one embodiment of the invention, the hydrophilic polymer may be included in 0.1 to 5 parts by weight based on 100 parts by weight of the biodegradable polymer.

Further, according to another embodiment of the present invention, the upper and lower layers of the polymer regeneration membrane may be mounted with a physiological activator, characterized in that the mounted physiological activator is sustained release from the polymer regeneration membrane .

Such a method for producing a polymer regeneration membrane according to the present invention comprises the steps of preparing a polymer solution; Casting the polymer solution to produce a polymer membrane; And immersing the polymer membrane in a non-solvent.

The concentration of the polymer solution may be 1 to 50% by weight.

The solvent used in the preparation of the polymer solution may be at least one selected from the group consisting of tetraglycol, 1-methyl-2-pyrrolidinone, triacetin, and benzyl alcohol.

According to one embodiment of the invention, the non-solvent is preferably a mixture of water and C ₁ ~ C ₅ lower alcohol in a weight ratio of 30:70 ~ 70:30.

According to one embodiment of the invention, the temperature of the non-solvent is preferably maintained at 10 ~ 20 ℃.

The present invention is to produce a bone-induced regeneration membrane having a deciduous laminated internal structure in a simple process without using a toxic organic solvent using a polymer having biocompatibility, not only performs the function of the bone-induced regeneration membrane properly, but also due to the deciduous laminated structure Since it is possible to release the bioactive factor in a sustained release form without using any additives and surface modification method, it can be very useful as a new osteoinductive regeneration membrane.

1 is a scanning electron micrograph of a bone induction regeneration membrane according to the first embodiment of the present invention,

2 is a scanning electron micrograph of a bone induction regeneration membrane according to Example 2,

3 is a scanning electron micrograph of a bone induction regeneration membrane according to Example 3,

4 is a scanning electron micrograph of a bone induction regeneration membrane prepared according to Patent Document 1 (Comparative Example 1),

5 is a scanning electron micrograph of a bone induction regeneration membrane prepared according to Comparative Example 2,

6 is a scanning electron micrograph of a bone induction regeneration membrane prepared according to Comparative Example 3,

7 is a scanning electron micrograph of OSTEOGUIDE ^™ according to Comparative Example 7,

8 is a schematic diagram showing a process for producing a bone induction regeneration membrane according to the present invention,

9 is a graph comparing the mechanical properties of the bone induction regeneration membrane according to Example 1 and OSTEOGUIDE ^™ according to Comparative Example 7,

10 is a scanning electron micrograph of the bone guided regeneration membrane according to Example 1 (A), Examples 4 to 6 (B to D), and Comparative Examples 4 to 5 (E to F),

11 is a graph of cumulative release behavior of the bioactive factors mounted according to Example 7, Comparative Example 6, and Comparative Example 8. FIG.

Hereinafter, the present invention will be described in more detail.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the singular forms "a", "an" and "the" may include the plural forms as well, unless the context clearly indicates otherwise. Also, as used herein, "comprise" and / or "comprising" specifies the presence of the mentioned shapes, numbers, steps, actions, members, elements and / or groups of these. It is not intended to exclude the presence or addition of one or more other shapes, numbers, acts, members, elements and / or groups.

The present invention relates to a polymer regeneration membrane effective for regeneration of various biological tissues and a method for producing the same by having a unique structure.

Referring to FIG. 1, the structure of the polymer regeneration membrane according to the present invention is characterized by having a structure including an upper layer having an average pore size of 1 to 5000 nm, and a lower layer having a deciduous laminated internal structure. That is, referring to the cross-sectional photograph of the polymer regeneration membrane according to the present invention, the upper layer portion (upper portion of the cross-section photo) having a relatively small pore size and a lower layer portion having an internal structure (bottom photo) such as a plurality of fallen leaves are stacked.

The 'biotissue' of the present invention is meant to include bone, chalky, periodontal ligament and the like, and the polymer regeneration membrane according to the present invention is meant to be used for the regeneration of such biological tissues.

As used throughout the specification of the present invention, the meaning of the 'laminated fallen leaf structure' includes a form in which a plurality of fallen leaves are stacked and stacked as a result of analyzing a cross-sectional photograph of the lower layer of the polymer regeneration membrane according to the present invention. (See bottom view SEM photo of FIG. 1).

The polymer material according to the present invention is a polyester-based polymer having a weight average molecular weight of 1,000 to 1,000,000 g / mol, polycaprolactone [poly (ε-caprolactone)], polydioxanone (polydioxanone), polylactic acid [poly ( lactic acid)], poly (glycolicacid), polylactic acid-glycolic acid copolymer [poly (lactic acid-co-glycolic acid)], polyhydroxybutyrate ([poly (β-hydroxybutyrate)] and poly Hydroxybutyric acid-cohydroxyvalericacid, poly (γ-ethyl glutamate), polyanhydrides, polyethylene oxide-polylactic Polyethylene oxide-polylactic acid, polyethylene oxide polylactic-co-glycolic acid copolymer, and polyethylene oxide-polycaprolacto copolymer One or more biodegradable polymers selected from the group consisting of ne) may be used, but is not limited thereto. In the present invention, polycaprolactone may be most preferably used among the biodegradable polymers.

In addition, according to another embodiment of the present invention, in addition to the biodegradable polymer may further include a hydrophilic polymer in order to improve the role of the polymer regeneration membrane and the ease of introduction of bioactive factors.

The hydrophilic polymer is a polymer containing a large amount of ethylene oxide (ethylen oxide, -CH ₂ CH ₂ O) or hydroxy (hydroxy, -OH) functional group having a weight average molecular weight of 1,000 ~ 1,000,000 g / mol, polyethylene oxide-polypropylene Oxide copolymer (polyethylene oxide-polypropyleneoxide, PEO-PPO), polyethylene oxide-polylactic acid copolymer (PEO-PLA), polyethylene oxide-polylactic glycolic acid copolymer (polyethylene oxide-poly (lactic-co-glycolic acid), PEO-PLGA), polyethylene oxide-polycaprolactone copolymer (polyethylene oxide-polycaprolactone, PEO-PCL) and may be one or more selected from the group consisting of copolymerization thereof, As long as you have it, you can use other materials.

Preferably, the hydrophilic polymer is included in an amount of 0.1 to 5 parts by weight based on 100 parts by weight of the biodegradable polymer, and when the hydrophilic polymer is used in an amount less than 0.1 part by weight, the prepared bone-induced regeneration membrane does not exhibit hydrophilicity and exceeds 5 parts by weight. In this case, there is a problem that the internal structure of the deciduous laminated type is not formed.

Further, according to another embodiment of the present invention, the upper and lower layers of the polymer regeneration membrane may be mounted with a physiological activator, characterized in that the mounted physiological activator is sustained release from the polymer regeneration membrane .

The bioactive factor according to the present invention may be one or more peptides / proteins selected from the group consisting of cytokines, hormones, insulin, and antibodies;

Fibroblast growth factors (FGFs), vascular endothelial growth factor (VEGF), nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) ), Transforming growth factors (TGFs), bone morphogenetic proteins (BMPs), epidermal growth factor (EGF), insulin-like growth factor (IGF) ), One or more growth factors selected from platelet-derived growth factor (PDGF);

gene; And

It may be any one selected from among the vaccines.

In the present invention, the various physiologically active factors listed above are mounted on the upper and lower layers of the polymer regeneration membrane, and the mounted physiologically active factors are sustained-release from the polymer regeneration membrane.

This is because the physiologically active factors mounted on the upper and lower layers of the polymer regenerated membrane exit the complex deciduous laminated structure inside the polymer regenerated membrane, and the physiologically active factors are repeatedly adsorbed and desorbed on the deciduous laminated structure to slow down the release rate. will be.

The sustained release property of the physiologically active factor according to the present invention is different from that of the conventional polymer membrane having a uniform porous structure, and may be attributed to the unique structure of the polymer regeneration membrane of the present invention.

The method for mounting the physiological activator on the polymer regeneration membrane is to put a physiological activator aqueous solution prepared at a constant concentration and a polymer regeneration membrane cut into 12 mm x 12 mm in a syringe and alternating negative pressure and positive pressure into the polymer regeneration membrane. It is introduced into the surface and inside of the polymer regeneration membrane due to the penetration of the aqueous solution containing and adsorption on the surface of the polymer regeneration membrane.

Hereinafter, a method of manufacturing a polymer regeneration membrane according to an embodiment of the present invention will be described.

In the present invention, by having a more improved structure of the polymer structure of the conventional patent document 0001 is to optimize the manufacturing method to induce the sustained release of the bioactive material.

Specifically, as shown in FIG. 8, the polymer solution is prepared through the steps of preparing a polymer solution, casting a polymer solution to prepare a polymer membrane, and immersing the cast polymer membrane in a non-solvent.

First, the polymer material is dissolved in a solvent to prepare a polymer solution. In preparing the polymer solution, 1 is selected from the group consisting of tetraglycol, 1-methyl-2-pyrrolidinone (1-methyl-2-pyrrolidinone (NMP)), triacetin and benzylalcohol. Preference is given to using solvents of species or higher.

The polymer material may be used alone or in combination of one or more biodegradable polymers. The hydrophilic polymer is preferably included 0.1 to 10 parts by weight based on 100 parts by weight of the biodegradable polymer.

In the case of dissolving the polymer in the solvent, the concentration of the polymer solution is preferably 1 to 50% by weight, preferably 10 to 20% by weight. There is a problem that this is not formed or the physical properties are weak, and when the content exceeds 50% by weight, the viscosity of the solution is high, so that it is not easily dissolved or handled.

Preparation of the polymer solution may be carried out by appropriately changing the conditions of maintaining the temperature of room temperature to 100 ℃ depending on the polymer used.

The biocompatible polymer solution is prepared as described above, and then taped to four sides of the glass substrate to a desired size to form a mold having a predetermined thickness, and the polymer solution is coated thereon to prepare a polymer membrane. In the coating, it is convenient to manufacture a general casting method, but is not limited thereto. The thickness of the produced polymer film can be adjusted according to the thickness of the mold used during the casting, preferably 400 to 600㎛.

Next, the cast polymer membrane is immersed in a non-solvent to precipitate a polymer solution.

In the present invention, when the specially cast polymer membrane is immersed in the nonsolvent, the composition of the nonsolvent used and the temperature of the nonsolvent are changed to have the structure of the polymer regeneration membrane according to the present invention.

That is, the non-solvent used at this time is preferably used by mixing water and C ₁ ~ C ₅ lower alcohol in a weight ratio of 30:70 ~ 70:30.

The lower alcohol of C ₁ ~ C ₅ is selected from the group consisting of methanol, ethanol, propanol, t-butyl alcohol, and pentanol. As the non-solvent according to the present invention, alcohols having more carbon atoms than C ₁ to C ₅ lower alcohols (for example, higher alcohols) are not preferable because they may have high molecular weights and high viscosity.

In addition, in the present invention, it is preferable to use a mixture of water and C ₁ to C ₅ lower alcohol as the non-solvent, but when water or alcohol is used as the non-solvent alone, the solvent and phase transition used when preparing the polymer solution are too fast. There is a problem that a deciduous laminated structure such as the present invention can not be obtained only by forming a uniform porous structure. It is possible to obtain a deciduous laminated internal structure such as the present invention because the precipitation rate of the polymer solution is lowered when water and lower alcohol are mixed.

In addition, water and lower alcohols of C ₁ ~ C ₅ can be used by mixing in a weight ratio of 30:70 ~ 70:30, to obtain a deciduous laminated structure like the present invention is used by mixing in a weight ratio of 50:50. Preferably, the optimum non-solvent mixing conditions are essential because the deciduous laminated structure like the present invention cannot be obtained if it is out of the above range.

In addition, according to one embodiment of the present invention, it is necessary to maintain the temperature of the non-solvent at 10 to 20 ℃, preferably 15 to 18 ℃. In this case, since the difference between the temperature in the polymer solution state and the temperature at the time of casting is relatively large, the solubility of the polymer decreases at the moment of placing the casting film in the non-solvent, as well as the precipitation of the polymer in the casting film itself. Since the precipitation occurs at the same time in the medium, the precipitated polymer membrane has a structure having pores of 1 to 5000 nm in the upper layer and at the same time, it is seen that a deciduous laminated structure is formed in the lower layer.

Precipitating the cast membrane in the non-solvent may be performed for 1 minute to 12 hours.

After immersing in the non-solvent as described above, the non-solvent is washed and then freeze-dried to finally obtain a polymer regeneration membrane. The washing method is not particularly limited.

The polymer regenerated membrane thus prepared has a cross-sectional structure in which an upper layer has a small average pore size, and a lower layer has a deciduous laminated structure. The detailed structure of FIG. 1 is shown in FIG. By the presence of pores, penetration of the fibrous connective tissue can be prevented while providing a space for nutrient and oxygen permeation. In addition, the lower layer has a deciduous laminated structure, the effect of easy treatment and adhesion as a variety of tissues.

In addition, due to the deciduous lamination structure of the lower layer, the bioactive factor mounted on the polymer regeneration membrane has a characteristic of exhibiting sustained release behavior. That is, in the present invention, after the physiologically active factors are loaded with only the deciduous layered structure without using any additives or surface modification methods on the polymer regeneration membrane for the physiologically active factor loading and sustained release, the sustained release is possible. This is characterized by repeating the adsorption and desorption while the physiologically active factors mounted on the polymer regeneration membrane pass through the deciduous laminated structure inside the polymer regeneration membrane.

The method of mounting the physiological activator on the polymer regeneration membrane is to inject the aqueous physiological activator solution prepared at a constant concentration and the polymer regeneration membrane into a syringe, and alternating the negative pressure and the positive pressure infiltration of the aqueous solution containing the physiological activator factor into the polymer regeneration membrane. And introduced into the surface and inside of the polymer regeneration membrane due to adsorption on the surface of the polymer regeneration membrane.

Although this invention is demonstrated in detail based on an Example, the following Example is only for illustration of this invention, It should not be interpreted that the scope of the present invention is limited by these Examples. In addition, in the following Examples, although illustrated using a specific compound, it will be apparent to those skilled in the art that even when these equivalents are used, the effects can be similarly similar.

Examples 1-3, Comparative Examples 1-3: Preparation of Osteoinduced Regeneration Membrane

The polycaprolactone showing biocompatibility and biodegradability was dissolved in a tetraglycol solvent at 15 wt% for 15 minutes at 90 ° C. to prepare each polymer solution of polycaprolactone (PCL).

 After casting the prepared polymer solution in a certain mold (50mm x 50mm; thickness 400㎛) that is stored at 37 ℃ and the mold containing the polymer solution while varying the non-solvent composition and non-solvent temperature conditions as shown in Table 1 It was immersed.

After soaking for at least 1 hour, excess ultrapure water was exchanged every hour for 6 hours to remove residual solvent. After the washing was lyophilized to obtain a bone-induced regeneration membrane in the form of a sheet.

sample Nonsolvent composition (weight ratio) Non-solvent temperature (℃) water Alcohol Example 1 50 50 17 Example 2 70 30 15 Example 3 30 70 12 Comparative Example 1 100 - Room temperature (26) Comparative Example 2 - 100 Room temperature (26) Comparative Example 3 50 50 Room temperature (26)

Example  4-6, Comparative example  4 ~ 5: Osteoinduction Regenerative  Produce

Pluronic F127, a PEO-PPO copolymer showing biocompatibility and hydrophilicity, was used as a polymer material with respect to 100 parts by weight of polycaprolactone showing biocompatibility and biodegradability used in Examples 1 to 3 as 1, 3, 5, 10, 20 Example 1, except that the mixture was dissolved in a weight ratio (Examples 4, 5, 6, Comparative Examples 4, 5) and dissolved in tetraglycol at 15 ° C. for 15 minutes at 90 ° C. to prepare a PCL / F127 solution. Osteoinduced regeneration membrane was prepared in the same process.

Examples 4 to 6 The nonsolvent composition and the temperature conditions were maintained to be the same as those of Example 1, respectively.

Example 7, Comparative Example 6: Mounting bioactive factors on bone induction regeneration membrane

The bone induction regenerative membrane prepared in Example 1 and Comparative Example 1 was cut into 12 mm x 12 mm size, and 3 pieces were placed in a 20 ml syringe, and the bone morphogenetic protein (BMP-2) of 1 µg / ml concentration. An aqueous solution was added. The positive and negative pressures were alternately applied to the syringe to allow the BMP-2 aqueous solution to completely penetrate into the bone regeneration membrane. It was then refrigerated and stored at 4 ° C. for 3 hours to induce BMP-2 to be adsorbed on the bone induction regenerative membrane. After 3 hours, the excess BMP-2 aqueous solution remaining in the syringe was removed and lyophilized. The derived bone guide regeneration membrane was obtained.

Comparative example  7: Genoss With OSTEOGUIDE  compare

Among the bone-induced regenerative membranes on the market, OSTEOGUIDE (trademark) of Genoss, a bone-induced regenerative membrane made of polycaprolactone as described in the present invention, was purchased, and the surface and cross-sectional structure thereof were observed by scanning electron microscopy. Shown in

Next, as shown in the SEM image of FIG. 7, it can be seen that commercially available bone-induced regeneration membranes have many pores of 20 to 40 μm or more formed on the entire surface (upper surface), cross section, and lower surface.

Therefore, although the polycaprolactone was prepared using the same material as the present invention, the bone-induced regeneration film according to Comparative Example 7 was confirmed to be different from the structure having the upper layer porous structure and the lower layer deciduous laminated structure. It can be seen as a difference in temperature and nonsolvent composition in the production of bone-induced regeneration membrane.

Comparative example  8: OSTEOGUIDE  Bioactive substance loaded

Mounting of the bioactive factor in OSTEOGUIDE was carried out in the same manner as in Example 7.

Experimental Example 1: Confirming the structure of the bone induction regeneration membrane

The surface of the bone guided regeneration membrane according to Examples 1 to 3 and Comparative Examples 1 to 3 was observed with a scanning electron microscope (SEM), and the results are shown in FIGS. 1 to 6, respectively.

In addition, Examples 4 to 6 (B-D of FIG. 10), and Comparative Examples 4 to 4, which were prepared including the osteoinductive regeneration membrane and the hydrophilic polymer of Example 1 (A of FIG. 10), which did not include a hydrophilic polymer. Scanning electron micrographs of the bone induction regeneration membrane according to 5 (Fig. 10E ~ F) is shown in FIG.

1 to 3, in the case of the bone-induced regeneration membrane prepared in Example 1, Example 2, Example 3, it was confirmed that the upper layer has a small pore size average. In the structure of the lower layer part, the perfect structure of the deciduous laminated type was produced in Example 1, and in the case of Example 2 and Example 3, the lower layer was pressed, but structurally, both of the deciduous laminated structures were produced. It became.

4 manufactured by the method of Comparative Example 1, the upper layer has a small average pore size, but the lower layer has a large average pore size structure rather than a deciduous laminated structure.

In the case of the manufacturing method of Comparative Example 2 and then in Figure 5 it can be seen that the size of the generated pores is 2mm or more, and the lower layer also has no deciduous laminated structure.

6 manufactured by the method of Comparative Example 3, even if the composition ratio of the non-solvent falls within the scope of the present invention, when the temperature conditions of the non-solvent are outside the scope of the present invention, the bottom surface of the bone-induced regeneration membrane falls, It is not so weak that it can not be used as a bone-induced regeneration membrane, it can be confirmed that no deciduous laminated structure was formed in the lower layer.

From these results, it can be seen that the composition and temperature of the non-solvent must be very sensitively controlled in order to produce a bone-induced regeneration membrane having a predetermined size of pores in the upper layer and a deciduous laminated inner structure in the lower layer.

In addition, after confirming the structure of the bone-induced regeneration membrane according to the hydrophilic polymer containing, referring to FIG. 10, even if the hydrophilic polymer is included, the bone-induced regeneration membranes of Examples 4 to 6 according to the present invention (respectively B to D) is a hydrophilic polymer As in the bone induction regeneration membrane (A) of Example 1, which is not included, the upper layer has a small pore size, and the lower layer has a deciduous laminated internal structure.

However, it can be seen that the osteoinductive regeneration membrane according to Comparative Example 4 and Comparative Example 5 containing an excessive amount of hydrophilic polymer is formed a completely different porous structure than the lower layer of the present invention.

Therefore, it can be seen that in order to manufacture a bone-induced regeneration membrane including a lower layer having a deciduous laminated internal structure as in the present invention, a hydrophilic polymer in an appropriate range should be included.

Experimental Example 2: Measurement of mechanical properties of bone induction regeneration membrane

The mechanical properties of the bone guided regeneration membrane according to Example 1 and Comparative Example 7 were analyzed. Mechanical properties can be measured by measuring the tensile and suture pullout strengths of the specimen standard ISO 37-3 in a dry and wet state, using the Universal Testing Machine (UTM, AG-X, SHIMADZU, Japan).

The measured results are shown in Figure 9, the tensile strength of the bone-induced regeneration membrane prepared by the method of Example 1 was superior in mechanical properties than the commercially available OSTEOGUIDE company of Genoss, the suture tensile strength is not a big difference It was confirmed that the bone-induced regeneration membrane prepared by the method of Example 1 has physical properties that can be used in the clinic.

Experimental Example 3: Hydrophilicity Test of Osteoinduced Regeneration

After cutting the bone induction regeneration membrane prepared in Example 1 (sample not containing a hydrophilic polymer) and Examples 4 to 6 to a size of 12mm x 12mm and drop a 20μ volume of water droplets thereon the water droplets It is shown in Table 2 by measuring the time (wetting time) is completely absorbed by the induced regeneration membrane.

sample Wetting time, sec Example 1 No wetting Example 4 234 Example 5 30 Example 6 4

Referring to Table 2, the bone-induced regeneration membrane prepared without adding a hydrophilic polymer as in Example 1 did not absorb water, but as the specific gravity of the hydrophilic polymer was high as in Examples 4 to 6, water was absorbed. You can see that the time is fast.

As a condition of the bone-induced regeneration membrane according to the present invention, invasion of epithelial tissue and fibrous connective tissue should be prevented and nutrients and oxygen should be permeated. Accordingly, as the hydrophilic polymer is added to the biodegradable polymer as in the present invention, the wetting speed of the bone-induced regeneration membrane is increased. Therefore, the higher the specific gravity of the hydrophilic polymer, the easier the permeation of nutrients and oxygen.

Experimental Example 4 Measurement of Release Behavior of Bioactive Factors

Osteoinduced regeneration membrane loaded with the bioactive factor (BMP-2) according to Example 7 and Comparative Example 6, and OSTEOGUIDE ^™ according to Comparative Example 8 was added 1% bovine serum albumin (BSA) The amount of BMP-2 released by sandwich enzyme immunoassay (sandwich Enzyme-Link Immunospecific Assay, sandwich ELISA) was measured daily in phosphate buffer saline (PBS), and the cumulative release amount was measured in FIG. 11. (Example 7 and Comparative Example 8 are A, Comparative Example 6 is B).

Next, referring to Figure 11, it can be seen that in the case of bone induction regeneration membrane (PCL membrane) prepared according to Example 7 of the present invention, the bioactive factor is released in a sustained release form over 40 days.

In addition, in the case of the commercially available OSTEOGUIDE ^™ loaded with the physiologically active factor of Comparative Example 8, it can be confirmed that most of the physiologically active factors are initially released and the rest are released over 5 days.

In addition, in the case of the Asymmetrically PCL membrane prepared according to Comparative Example 6, it can be seen that most of the bioactive factors are released within 7 days.

This difference is because the bone-induced regenerative membrane according to the present invention has a deciduous laminated structure in the lower layer and the inside, and because the bioactive material is mounted on both the surface and the inside of the bone-induced regenerative membrane, Since the physiologically active factors are released while repeating desorption / adsorption while passing through a plurality of deciduous laminated structures, the release of the physiologically active factors can be confirmed to be sustained release.

However, in the case of osteoinduced regeneration membrane according to Comparative Example 6, even if the bioactive material is introduced into the asymmetric structure of porous osteoinduced regeneration membrane comprising two or more layers having different average pore sizes, Since the average pore size (average pore size) is a columnar (column shape) structure of a relatively large 5 to 500 ㎛ can be seen that the bioactive material is quickly desorption is released early.

In addition, in Comparative Example 8, since the internal structure is monotonous, uniform, and has a large average size of pores compared to the bone-induced regeneration membrane prepared according to the present invention, it is released by rapid desorption of the bioactive substance adsorbed on the surface and the inside. It can be seen that.

Claims (10)

An upper layer having an average pore size of 1 to 5000 nm, and The polymer regeneration membrane having a structure including a lower layer having a deciduous laminated internal structure. The method of claim 1, The polymers include polycaprolactone, polydioxanone, polylactic acid, polyglycolic acid, polylactic acid-glycolic acid copolymers, polyhydrocibutyrate and polyhydroxybutyric acid-hydroxybalic acid copolymers and polyphosphoesters. Polymer regeneration membrane that is at least one biodegradable polymer selected from the group consisting of. The method of claim 1, The polymers include polycaprolactone, polydioxanone, polylactic acid, polyglycolic acid, polylactic acid-glycolic acid copolymers, polyhydrocibutyrate and polyhydroxybutyric acid-hydroxybalic acid copolymers and polyphosphoesters. At least one biodegradable polymer selected from the group consisting of, and Polyethylene oxide-polypropylene oxide copolymer, polyethylene oxide-polylactic acid copolymer, polyethylene oxide-glycolic acid copolymer, polyethylene oxide-polylactic glycolic acid copolymer, polyethylene oxide-polycaprolactone copolymer, polyethylene oxide-polydioxane A polymer regeneration membrane, which is a mixture of one or more hydrophilic polymers selected from the group consisting of a copolymer and a copolymer thereof. The method of claim 3, The hydrophilic polymer is a polymer regeneration membrane is included in 0.1 to 5 parts by weight based on 100 parts by weight of the biodegradable polymer. The method of claim 1, The upper and lower layers of the polymer regeneration membrane may be mounted with a physiological activator, wherein the mounted physiological activator is sustained release from the polymer regeneration membrane. Preparing a polymer solution; Casting the polymer solution to produce a polymer membrane; And Method of producing a polymer regeneration membrane comprising the step of immersing the polymer membrane in a non-solvent. The method of claim 6, The concentration of the polymer solution is 1 to 50% by weight of the method for producing a polymer regeneration membrane. The method of claim 6, The solvent used in the production of the polymer solution is tetraglycol, 1-methyl-2-pyrrolidinone, triacetin, and benzyl alcohol, the method for producing a polymer regeneration membrane, characterized in that at least one selected from the group consisting of. The method of claim 6, The non-solvent is a method for producing a polymer regeneration membrane, characterized in that the mixture of water and C ₁ ~ C ₅ lower alcohol in a weight ratio of 30:70 ~ 70:30. The method of claim 6, Temperature of the non-solvent is a method for producing a polymer regeneration membrane, characterized in that maintained at 10 ~ 20 ℃.

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