JPWO2012029971A1 - Composite of fibrin glue and fiber molded body - Google Patents

Abstract

A composite comprising a fiber molded body and a fibrin glue, wherein the fiber molded body is a fiber made of a biodegradable polymer containing phospholipid, the fiber surface has a smoothness of 1.07 or less, and phospholipid It consists of fibers containing 0.1 to 10% by weight with respect to the biodegradable polymer. The present invention provides a composite comprising a fibrin glue having high strength and excellent tissue adhesiveness and a biodegradable fiber molded body.

Description

  The present invention relates to a composite of a fibrin glue and a fiber molded body. Specifically, the present invention relates to a composite of a fiber molded body containing 0.1 to 10% of phospholipid and a fiber having a smooth fiber surface and a fibrin glue. The complex of the present invention is preferably used as an artificial dura mater, an adhesion preventing material, a hemostatic material, etc., as a medical article, particularly a tissue surface or wound site protective material, a covering material, and a sealing material.

In recent years, various studies have been conducted on biodegradable sheet materials as medical materials. Already commercialized products include Neobale (registered trademark), which is a non-woven fabric made of polyglycolic acid for the purpose of reinforcing stitches and preventing air leakage, and periodontal made of a copolymer of lactic acid and glycolic acid. There is GC membrane (registered trademark), which is a tissue-derived regeneration membrane used for diseases. All of these have the property of being decomposed during the tissue regeneration process, and act as a protective material or a sealing material for damaged sites in the regeneration of living tissue. In this way, biodegradable materials have been studied for application in various fields, taking advantage of their characteristics.
Japanese Unexamined Patent Application Publication No. 2002-204826 discloses an artificial substitute biomembrane in which a bioabsorbable and / or biodegradable synthetic fiber cloth is used as a skeleton, and this is coated with fibrin glue. However, although data on the strength of the membrane itself based on a resistance test against water pressure is shown, there is no description about the adhesive force to the biomaterial and the closing force of the biological tissue defect. Also, in the dural defect closure test described in the examples, since the suture with a nylon suture is performed at the time of closure with the substitute biomembrane, it is expected that the tissue adhesion of the biomembrane is not sufficient. Is done.
As a method that does not require suturing, WO 2006/025150 pamphlet describes an intestinal defect occlusion device comprising a polyglycolic acid fiber molded article and fibrin glue. However, it is not sufficient for the strength against pressured tissues such as blood vessels.
An electrospinning method (also referred to as an electrostatic spinning method, an electrospinning method, or an electrospray method) is known as a method for producing a fiber serving as a base material such as an artificial biological membrane as described above. Nanofibers produced by this electrospinning method have the advantage of being able to easily produce yarns with a smaller fiber diameter than conventional molding methods, and are attracting attention as a simple method for obtaining a fiber molded body having a larger surface area. Yes.
As an example using such nanofibers, WO2006 / 022430 pamphlet describes a nanofiber containing a phospholipid. The object is to improve cell adhesion by adding a phospholipid to the fiber surface to have an uneven shape. However, no examination or suggestion has been made about the adhesion to the tissue surface in the complex with fibrin glue.
In US 2004/0013873, there is a description of nanofibers whose fiber surface is porous (having irregularities), and the effect is that the strength of the composite increases when used as a composite with a matrix. is there. However, there has been no suggestion or examination about the smoothness of the fiber surface and the strength of the composite of fibrin glue.

The problem to be solved by the present invention is to provide a composite comprising a fiber molded body of biodegradable fiber excellent in affinity with fibrin glue and fibrin glue, having high strength and excellent tissue adhesiveness. It is.
The inventors diligently studied a composite of a fiber molded body of a biodegradable fiber excellent in adhesiveness with a tissue surface and a fibrin glue. As a result, the present inventors have found that a composite of a biodegradable fiber molded body having a phospholipid-containing fiber having a smooth surface and a fibrin glue is excellent in tissue adhesion, and has completed the present invention.
So far, when the surface of the structure containing phospholipids has irregularities, it was expected that the composite has excellent strength as described in US 2004/0013873, but surprisingly, the fiber surface However, the present inventors have found that a smooth fiber molded article is excellent in the strength of the composite and completed the present invention.
That is, the present invention is a composite comprising a fiber molded body and a fibrin glue, wherein the fiber molded body is a fiber made of a biodegradable polymer containing a phospholipid, and the fiber surface has a smoothness of 1.07 or less. And it is the composite_body | complex characterized by consisting of the fiber which contains a phospholipid 0.1 to 10weight% with respect to a biodegradable polymer.

FIG. 1 is a conceptual diagram of an apparatus for measuring the breaking strength of the fiber composite of the present invention.
FIG. 2 is a photograph of a HE-stained specimen of tissue obtained by autopsy after 1 month when the artificial biological membrane obtained in Example 9 was placed on a beagle cerebrum such that the fibrin gel layer was on the brain parenchyma side.

The composite of the present invention is formed from a fiber molded body and fibrin glue. The fiber molded body used in the present invention contains phospholipid in an amount of 0.1 to 10% by weight based on the biodegradable polymer. When the phospholipid content is less than 0.1% by weight, there is no effect on the affinity with fibrin glue, and when it is more than 10% by weight, the durability of the fiber molded body itself is undesirably lowered. A more preferable content is 0.2 to 5% by weight, still more preferably 0.3 to 1% by weight.
The phospholipid used in the present invention may be extracted from animal tissues or artificially synthesized. Examples of such phospholipids include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylglycerol. One of these may be selected, or a mixture of two or more may be used. Of these, phosphatidylcholine or phosphatidylethanolamine is preferable, and among these, phosphatidylcholine dilauroyl or phosphatidylethanolamine dioleoyl is more preferable.
Specific examples of the biodegradable polymer used in the present invention include polylactic acid, polyglycolic acid, polycaprolactone, polydioxanone, lactic acid-glycolic acid copolymer, lactic acid-caprolactone copolymer, polyglycerol sebacic acid, polyhydroxyalkane. Acids, aliphatic polyesters such as polybutylene succinate, aliphatic polycarbonates such as polymethylene carbonate, polysaccharide derivatives such as cellulose diacetate, cellulose triacetate, methylcellulose, propylcellulose, benzylcellulose, carboxymethylcellulose, fibroin, gelatin, collagen, etc. Proteins and derivatives thereof.
More preferred are aliphatic polyesters such as polylactic acid, polyglycolic acid and lactic acid-glycolic acid copolymer, and most preferred are polylactic acid and lactic acid-glycolic acid copolymer.
At this time, it is preferable that the polylactic acid copolymer has few monomer components imparting stretchability. Here, the monomer component imparting stretchability is, for example, caprolactone monomer, ethylene glycol, 1,2-propylene glycol, 1,3-propylene glycol, 1,2-butanediol, 1,4-butanediol, polycaprolactone diol. , Soft components such as polyalkylene carbonate diol and polyethylene glycol unit. These soft components are preferably less than 20% by weight in polymer ratio. When there are more soft components than this, the polymer tends to lose its self-supporting property and gives a fiber molded product that is too soft and difficult to handle.
As monomers constituting the polymer in polylactic acid, there are L-lactic acid and D-lactic acid, and they are used without particular limitation. There are no particular restrictions on the optical purity or molecular weight of the polymer, the composition ratio of the L-form and the D-form, and the arrangement, but a polymer having many L-forms is preferred. A stereocomplex of poly L lactic acid and poly D lactic acid can also be used.
The weight average molecular weight of the biodegradable polymer is preferably 1 × 10 ^{3 to} 5 × 10 ⁶ , more preferably 1 × 10 ⁴ to 1 × 10 ⁶ , and even more preferably 5 × 10 ^{4 to} 5 × 10 6. ⁵ . Further, the terminal structure of the polymer and the catalyst for polymerizing the polymer can be arbitrarily selected.
The fiber molded body used in the present invention may be used in combination with other polymers and other compounds as long as the purpose is not impaired. For example, it can be used in combination as a polymer copolymer, a polymer blend, or a compound mixture.
The biodegradable polymer used in the present invention is preferably highly pure, and in particular, residues such as additives, plasticizers, residual catalysts, residual monomers, residual solvents used in molding and post-processing are contained in the polymer. Less is preferable. Needless to say, when used for medical treatment, it is necessary to keep it below the safety standard value.
A fiber having a smooth fiber surface refers to a fiber whose surface is not uneven when the fiber surface is observed with a scanning electron microscope or the like. For example, in US 2004/0013873, a fiber molded body having an uneven fiber surface is shown, but it refers to a fiber having no such unevenness. Specifically, the surface morphology of the fiber is evaluated by a surface area ratio (smoothness) in an AFM observation visual field range 1 × 1 μm ² using an atomic force microscope AFM, and the smoothness is 1.07 or less, preferably The fiber which is 1.05 or less, More preferably, it is 1.03 or less.
In Examples described later, Nano Scope IIIa manufactured by Digital Instruments was used as the AFM. The cantilever used was AC-240TS (made of silicon: spring constant 2 N / m). In order to prevent resolution degradation during observation, a new cantilever with no contamination or wear of the probe was used. In addition, the force acting between the probe and the sample surface was set to the minimum necessary force to prevent sample breakage and probe wear during scanning. The observation was performed with an observation field of view of 1 × 1 μm ² so that the probe was in contact with the center of the fiber and the scanning direction of the probe coincided with the fiber axis. The scanning speed was 0.7 Hz. The resolution was 256 × 256 pixels or higher. Since the fiber has a curvature, after the AFM observation, the curvature of the fiber and the undulation on the macro form were canceled using inclination correction software attached to the apparatus. In the tilt correction, when the probe contacted the fiber center completely and the scanning direction coincided with the fiber axis direction, the secondary tilt correction was performed. Otherwise, the tertiary tilt correction processing and flat processing were performed. . After the inclination correction, the surface roughness analysis attached to the apparatus was performed to calculate the surface area ratio. The surface area ratio is the ratio Sratio of the actual surface area S to the area S0 when it is assumed that the observation surface is ideally flat, and is represented by Sratio = S / S0. The evaluation was performed with an average value of 20 observation visual fields performed at random.
The fiber diameter of the fiber molded body used in the present invention is 0.1 to 10 μm. Here, when the average fiber diameter is smaller than 0.1 μm or larger than 10 μm, it is difficult to obtain affinity with a desired fibrin glue. A preferable average fiber diameter is 1.0 to 8.0 μm, and more preferably 2.0 to 7.0 μm. The fiber diameter is the diameter of the fiber cross section. The shape of the fiber cross section is not limited to a circle, and may be an ellipse or an irregular shape. With respect to the fiber diameter in this case, the average of the length in the major axis direction and the length in the minor axis direction of the ellipse is calculated as the fiber diameter. When the fiber cross section is neither circular nor elliptical, the fiber diameter is calculated by approximating a circle or ellipse.
The fiber molded body used in the present invention is composed of long fibers. Specifically, the long fiber refers to a fiber molded body that is formed without adding a step of finely cutting the fiber in the process from spinning to processing of the fiber molded body. It can be formed by a bond method, a melt blow method, or the like. The electrospinning method is preferably used. Here, the electrospinning method is the same in principle as a method called an electrostatic spinning method or an electrospray method, and these are also included in the electrospinning method referred to in the present invention.
The electrospinning method is a method of obtaining a fiber molded body on an electrode by applying a high voltage to a solution in which a polymer is dissolved in a solvent. The steps include a step of producing a solution by dissolving a polymer in a solvent, a step of applying a high voltage to the solution, a step of ejecting the solution, and evaporating the solvent from the ejected solution to form a fiber. A step of forming a body, an optional step of eliminating the charge of the formed fiber molded body, and a step of accumulating the fiber molded body due to charge loss.
Although there is no restriction | limiting in particular in the whole thickness of the fiber molded object used by this invention, Preferably it is 25 micrometers-200 micrometers, More preferably, it is 50-100 micrometers.
The step of producing a solution by dissolving an organic polymer in a solvent in the electrospinning method will be described. The concentration of the biodegradable polymer with respect to the solvent in the solution in the production method of the present invention is preferably 1 to 30% by weight. When the concentration of the biodegradable polymer is less than 1% by weight, it is not preferable because the concentration is too low and it becomes difficult to form a fiber molded body. On the other hand, if it is larger than 30% by weight, the fiber diameter of the obtained fiber molded product is undesirably large. A more preferable concentration of the biodegradable polymer with respect to the solvent in the solution is 2 to 20% by weight.
A solvent may be used individually by 1 type and may combine several solvent. The solvent is not particularly limited as long as it can dissolve the biodegradable polymer and evaporates at the spinning stage to form a fiber. For example, acetone, chloroform, ethanol, 2-propanol, methanol, Toluene, tetrahydrofuran, water, benzene, benzyl alcohol, 1,4-dioxane, 1-propanol, dichloromethane, carbon tetrachloride, cyclohexane, cyclohexanone, phenol, pyridine, trichloroethane, acetic acid, formic acid, hexafluoro-2-propanol, hexafluoro Acetone, N, N-dimethylformamide, N, N-dimethylacetamide, acetonitrile, N-methyl-2-pyrrolidinone, N-methylmorpholine-N-oxide, 1,3-dioxolane, methyl ethyl ketone, mixed solvent of the above solvents And the like. Of these, dichloromethane and ethanol are preferably used in view of handling properties and physical properties.
Next, a step of applying a high voltage to the solution, a step of ejecting the solution, and a step of evaporating the solvent from the ejected solution to form a fiber molded body will be described.
In the method for producing a fiber molded body of the present invention, it is necessary to apply a high voltage to the solution in order to eject a solution in which a biodegradable polymer is dissolved to form a fiber molded body. The method for applying the voltage is not particularly limited as long as a solution in which the biodegradable polymer is dissolved is ejected to form a fiber molded body. There are a method for applying a voltage by inserting an electrode into a solution, a method for applying a voltage to a solution ejection nozzle, and the like.
An auxiliary electrode can be provided separately from the electrode applied to the solution. The value of the applied voltage is not particularly limited as long as the fiber molded body is formed, but is preferably in the range of 5 to 50 kV. When the applied voltage is less than 5 kV, the solution is not ejected and a fiber molded body is not formed, which is not preferable. When the applied voltage is more than 50 kV, discharge is generated from the electrode toward the ground electrode, which is not preferable. More preferably, it is the range of 7-30 kV. The desired potential may be generated by any appropriate method known in the art.
Then, immediately after the solution in which the biodegradable polymer is dissolved is ejected, the solvent in which the biodegradable polymer is dissolved volatilizes to form a fiber molded body. Ordinary spinning is performed at room temperature in the atmosphere, but when volatilization is insufficient, it can be performed under negative pressure or in a high-temperature atmosphere. The spinning temperature depends on the evaporation behavior of the solvent and the viscosity of the spinning solution, but is preferably in the range of 0 to 50 ° C.
Next, the step of accumulating the fiber molded body due to the charge disappearance will be described. The method for accumulating the fiber molded body due to the loss of electric charge is not particularly limited, but a normal method includes a method of losing the electrostatic force of the fiber molded body due to the loss of electric charge, and dropping and accumulating due to its own weight. Moreover, if necessary, a method of sucking and accumulating the fiber molded body from which the electrostatic force has disappeared and accumulating it on the mesh, a method of causing convection of air in the apparatus and accumulating on the mesh, and the like can also be performed.
As a method for producing a fiber having a smooth fiber surface, it can be produced by setting the atmosphere during spinning to low humidity. The relative humidity is preferably 25% or less, more preferably 20% or less.
The processing of laminating a cotton-like fiber structure on the surface of the fiber molded body used in the present invention or forming a sandwich structure with the cotton-like structure sandwiched between the fiber molded bodies is an object of the present invention. Can be arbitrarily implemented within a range that does not impair the process.
The fiber molded body used in the present invention may be subjected to a surface treatment with a chemical such as a surfactant in order to modify the hydrophilicity, hydrophobicity, electrical characteristics and chargeability of the surface. In medical applications, coating treatment for imparting antithrombogenicity and surface coating with an antibody or a physiologically active substance can be optionally performed. The coating method and treatment conditions at this time, and the chemicals used for the treatment can be arbitrarily selected within a range that does not damage the fiber structure and impair the purpose of the present invention.
A drug can be optionally contained inside the fiber of the fiber molded body used in the present invention. In the case of molding by the electrospinning method, the drug used is not particularly limited as long as it is soluble in a volatile solvent and does not impair the physiological activity by dissolution.
Specific examples of such drugs include tacrolimus or its analogs, statin or taxane anticancer agents.
The drug may be a protein preparation or a nucleic acid drug as long as it can maintain activity in a volatile solvent. Moreover, things other than a chemical | medical agent may be included and a metal, polysaccharide, a fatty acid, surfactant, and a volatile solvent tolerance microorganism may be sufficient.
The present invention is a composite comprising the fiber molded body and a fibrin glue. Examples of such fibrin glue include those composed of fibrinogen lyophilized powder, fibrinogen lyophilized solution, thrombin lyophilized powder, and thrombin lysate, which are precursors of fibrin glue. The normal use mode of fibrin glue is to dissolve fibrinogen lyophilized powder with fibrinogen solution to make A solution, thrombin lyophilized powder with thrombin solution to make solution B, and overlay both solutions on the adhesion site. Or mix and apply.
Fibrin glue is a physiological tissue adhesive that utilizes the final stage of blood coagulation, and fibrinogen contained in it becomes a soluble fibrin clot by the action of thrombin, and blood coagulation factor XIII activated by thrombin in the presence of calcium ions To form a stable insoluble urea fibrin clot with physical strength and adhere and close the tissue. In this stabilized fibrin clot, for example, fibroblasts proliferate, collagen fibers and granulation matrix components are produced, and are cured through tissue repair.
The fibrin glue in the present invention is any of those in the above-mentioned soluble fibrin mass state, those in a stable fibrin mass state, or a precursor capable of forming a soluble fibrin mass or a stable fibrin mass and / or a mixture thereof. It may be. However, a fibrin glue precursor is used in the production process of the composite of the present invention.
The fibrin glue used in the present invention can contain an active factor such as a growth factor, a pharmaceutical agent, or a drug, if desired. As the active factor, a water-soluble compound is preferable, and proteins such as antibodies and growth factors can be mentioned as preferable examples.
The method for obtaining the composite of the fibrin glue and the fiber molded body is not particularly limited, and the fiber molded body is preliminarily coated with one of precursors such as fibrinogen and thrombin, coated, and impregnated. Compounding can be performed by mixing other precursors. Preferably, since the fiber molded body of the present invention is excellent in affinity with fibrin glue, a method of mixing and spraying both fibrinogen and thrombin solutions directly onto the fiber molded body by a method such as spraying can easily form a complex. You can do it.
The shape of the fibrin glue and the fiber molded body is not particularly limited, the shape in which the fiber molded body is embedded with fibrin glue, the shape in which the fibrin glue is applied or impregnated on one side of the fiber molded body, a part of the fiber molded body And the like in which fibrin glue is applied.

Hereinafter, although an example explains an embodiment of the present invention, these do not limit the range of the present invention.
1. Average fiber diameter:
Measure the diameter of the fiber by randomly selecting 20 points from the photograph obtained by photographing the surface of the obtained fiber molded body with a scanning electron microscope (Keyence Corporation: trade name “VE8800”) at a magnification of 2000 times. And the average value of all the fiber diameters was calculated | required and it was set as the average fiber diameter. n = 20.
2. Average thickness:
Using a high-precision digital length measuring machine (Mitutoyo Co., Ltd .: trade name “Lightmatic VL-50”), an average value was calculated by measuring the film thickness of the fiber molded body at a length measuring force of 0.01 N and n = 10. . In this measurement, the measurement was performed with the minimum measuring force that can be used by the measuring device.
3. Average apparent density:
The mass of the fiber molded body was measured, and the average apparent density was calculated based on the area and average thickness determined by the above method.
4). Fiber surface smoothness:
The fiber surface 1 × 1 μm ² of the fiber molded body was measured using an atomic force microscope (Digital Instrument Co., Ltd .: trade name “Nano Scope IIIa”), and the smoothness of n = 20 was calculated.
5. Composite strength test of fiber compact and fibrin glue composite:
Rabbit skin was collected as a biological tissue and placed flat on the surface of the apparatus in FIG. Make a hole (5mmΦ) in the center of the rabbit skin 3, fix it with the outer frame 5 and the inner frame 6, apply a fibrinogen solution around it, and place the fiber molding on it so that the hole is blocked Further, a spray of fibrin glue 1 (Bolheel (registered trademark)) was sprayed from above. For the fibrinogen solution and the thrombin solution used for spraying, the Borheel kit was used as it was. After allowing the fibrin glue gel to harden for several minutes, pressure 4 was applied from the outside, and the internal pressure when the composite (fiber molded body + fibrin glue) 2 was broken was measured. When the fibrinogen solution naturally penetrated into the fiber molded body and the composite broke, the internal pressure was 120 mmHg or more.
Example 1
10 parts by weight of polylactic acid (weight average molecular weight 137,000, manufactured by Taki Chemical) to which 1% by weight of phosphatidylcholine dilauroyl was added was dissolved in 90 parts by weight of a dichloromethane solution to obtain a uniform solution. Spinning was performed by electrospinning to obtain a sheet-like fiber molded body. The inner diameter of the ejection nozzle was 0.8 mm, the voltage was 15 kV, and the distance from the ejection nozzle to the flat plate was 15 cm. The flat plate was used as a cathode during spinning. The obtained fiber molded body had an average fiber diameter of 4.4 μm, a thickness of 106 μm, and an average apparent density of 153 mg / cm ³ . The smoothness of the fiber surface was 1.024. The fibrinogen solution naturally penetrated the fiber compact. The internal pressure when the composite broke was 286 mmHg.
Comparative Example 1
A fiber molded body was prepared in the same manner as in Example 1 except that 7.5 parts by weight of polylactic acid (weight average molecular weight 152,000, manufactured by PURAC) was dissolved in 80 parts by weight of a dichloromethane solution and 10 parts by weight of ethanol.
The obtained fiber molded body had an average fiber diameter of 5.0 μm, a thickness of 86 μm, and an average apparent density of 157 mg / cm ³ . The smoothness of the fiber surface was 1.032. The fibrinogen solution did not naturally penetrate into the fiber compact. The internal pressure when the composite broke was 66 mmHg.
Example 2
Example 1 except that 10 parts by weight of polylactic acid (weight average molecular weight 137,000, manufactured by Taki Chemical Co., Ltd.) added with 5% by weight of phosphatidylethanolamine dioleoyl was dissolved in 90 parts by weight of a dichloromethane solution. A fiber molded body was prepared.
The obtained fiber molded body had an average fiber diameter of 4.0 μm, a thickness of 99 μm, and an average apparent density of 165 mg / cm ³ . The smoothness of the fiber surface was 1.014. The fibrinogen solution naturally penetrated the fiber compact. The internal pressure when the composite broke was 294 mmHg.
Example 3
A sample was prepared in the same manner as in Example 1 except that 1% by weight of phosphatidylethanolamine dioleoyl was used. The obtained fiber molded body had an average fiber diameter of 4.3 μm, a thickness of 63 μm, and an average apparent density of 210 mg / cm ³ . The smoothness of the fiber surface was 1.017. The fibrinogen solution naturally penetrated the fiber compact. The internal pressure when the composite broke was 192 mmHg.
Comparative Example 2
A fiber molded article having a concavo-convex structure on the fiber surface made of polylactic acid to which 1% by weight of phosphatidylethanolamine dioleoyl oil is added by the method described in the pamphlet of WO2006 / 022430 (high humidity (relative humidity: 42 to 55%)) Was prepared. The obtained fiber molded body had an average fiber diameter of 3.0 μm, a thickness of 50 μm, and an average apparent density of 136 mg / cm ³ . The smoothness of the fiber surface was 1.074. The fibrinogen solution did not naturally penetrate into the fiber compact. The internal pressure when the composite broke was 68 mmHg.
Example 4
A sample was prepared in the same manner as in Example 1 except that 10% by weight of phosphatidylethanolamine dioleoyl was used. The obtained fiber molded body had an average fiber diameter of 4.3 μm, a thickness of 84 μm, and an average apparent density of 142 mg / cm ³ . The smoothness of the fiber surface was 1.018. The fibrinogen solution naturally penetrated the fiber compact. The internal pressure when the composite broke was 238 mmHg.
Example 5
A sample was prepared in the same manner as in Example 1 except that the concentration of phosphatidylcholine dilauroyl was 0.1% by weight and polylactic acid was made of PURAC having a weight average molecular weight of 152,000. The obtained fiber molded body had an average fiber diameter of 4.1 μm, a thickness of 78 μm, and an average apparent density of 156 mg / cm ³ . The smoothness of the fiber surface was 1.011. The fibrinogen solution naturally penetrated the fiber compact. The internal pressure when the composite broke was 230 mmHg.
Example 6
8 parts by weight of a lactic acid-glycolic acid copolymer (number average molecular weight 204,000, manufactured by PURAC) to which 0.4% by weight of phosphatidylcholine dilauroyl was added was dissolved in 92 parts by weight of dichloromethane to obtain a uniform solution. . Spinning was performed by electrospinning to obtain a sheet-like fiber molded body. The inner diameter of the ejection nozzle was 0.8 mm, the voltage was 8.5 kV, and the distance from the ejection nozzle to the flat plate was 25 cm. The flat plate was used as a cathode during spinning. The obtained fiber molded body had an average fiber diameter of 4.6 μm, a thickness of 82 μm, and an average apparent density of 153 mg / cm ³ . The smoothness of the fiber surface was 1.0088. The fibrinogen solution naturally penetrated the fiber compact. The internal pressure when the composite broke was 223 mmHg.
Example 7
10 parts by weight of polyglycolic acid (weight average molecular weight 100,000, manufactured by Polysciences) to which 0.4% by weight of phosphatidylcholine dilauroyl was added was dissolved in 90 parts by weight of a hexafluoro-2-propanol solution to obtain a uniform solution. . Spinning was performed by electrospinning to obtain a sheet-like fiber molded body. The inner diameter of the ejection nozzle was 0.8 mm, the voltage was 15 kV, and the distance from the ejection nozzle to the flat plate was 25 cm. The flat plate was used as a cathode during spinning. The obtained fiber molded product had an average fiber diameter of 2.5 μm, a thickness of 122 μm, and an average apparent density of 156 mg / cm ³ . The smoothness of the fiber surface was 1.022. The fibrinogen solution naturally penetrated the fiber compact. The internal pressure when the composite broke was 178 mmHg.
Example 8
11 parts by weight of polylactic acid (weight average molecular weight 133,000, manufactured by PURAC) to which 0.4% by weight of phosphatidylcholine dilauroyl was added was dissolved in 79 parts by weight of dichloromethane and 10 parts by weight of ethanol to obtain a uniform solution. It was. Spinning was performed by electrospinning to obtain a sheet-like fiber molded body. The inner diameter of the ejection nozzle was 0.8 mm, the voltage was 15 kV, and the distance from the ejection nozzle to the flat plate was 25 cm. The flat plate was used as a cathode during spinning. The obtained fiber molded body had an average fiber diameter of 3.9 μm, a thickness of 78 μm, and an average apparent density of 147 mg / cm ³ . The smoothness of the fiber surface was 1.004. The fibrinogen solution naturally penetrated the fiber compact. The internal pressure when the composite broke was 256 mmHg.
From the above results, it was shown that the fiber molded body containing a phospholipid having a smooth fiber surface used in the present invention has excellent affinity with fibrin glue.
Example 9
Bolheel (registered trademark), which is a commercially available biological tissue adhesive, was used to produce an artificial biological membrane for implantation in a living body. The fiber molded body produced in Example 8 was cut into a size of ² cm × 2 cm (4 cm ² ) and placed in a 10 cm plastic petri dish. A 1 cm × 1 cm (1 cm ² ) hole was formed in the center of a transparent plastic film having a size of 3 cm × 3 cm (9 cm ² ), and the plastic film was placed on the fiber molded body. Add 3 mL of a solution containing aprotinin (3000KIE) to a vial containing 240 mg of lyophilized fibrinogen in Bolheel and factor XIII 225 units and mix to make 3 mL of fibrinogen solution, then spray 0.2 mL of fibrinogen solution with 1 mL of spray Inhaled into a syringe. Thrombin (750 units) powder was dissolved in 3 mL of a solution containing 17.7 mg of calcium chloride, 0.2 mL of which was drawn into a 1 mL syringe. Each syringe was attached to a Bolheel spray set (Akita Sumitomo Bake Co., Ltd.). Using this spray set, 0.2 mL of each fibrinogen solution and thrombin solution was sprayed evenly from the top of the plastic film overlaid on the fiber molded body. Then, after leaving still for 5 minutes or more, by removing the plastic film, a fibrin gel layer having a size of 1 cm × 1 cm (1 cm ² ) in the center of the fiber molded body and a width of 0.5 cm around the fibrin gel layer. An artificial biofilm with a margin was formed. The artificial biological membrane was peeled from the petri dish and used for animal experiments.
Example 10
Animal experiments with adult beagle dogs were performed using the following method.
(I) Attaching the dura mater A beagle dog was placed under general anesthesia by intubation management, bilateral parietal frontal craniotomy was performed, and a 1-cm square square dural defect part was produced on each of the left and right sides. To one deficient part, 0.1 mL of fibrinogen solution is dropped on the dura mater around the deficient part and rubbed with a finger, and the fibrin gel layer is placed on the brain parenchyma side from the artificial biological membrane described in Example 9 from above. I covered it to become. Next, 0.3 mL of each Borheel solution was sprayed from above the artificial biological membrane using a Bolheel spray set to adhere the artificial biological membrane to the dural defect. After standing for 3 minutes or more, craniotomy was performed.
(Ii) Pathological findings at 1 month after surgery As shown in FIG. 2, in the hematoxylin-eosin stained (HE stained) specimen at 1 month after surgery, the upper part of the artificial biomembrane of the present invention in the dural defect And the layer of self connective tissue was confirmed at the bottom. In addition, connective tissue grew between the fibers of the artificial biological membrane. In addition, the brain tissue showed normal findings. There was no leakage of cerebrospinal fluid from the surgical site from the postoperative period until 1 month at the time of necropsy.
From the above results, it is shown that the fiber molded body containing a phospholipid with a smooth fiber surface used in the present invention has excellent affinity with fibrin glue, adheres to a living tissue, and includes a dura mater. It was confirmed that it can be an artificial biological membrane that promotes regeneration of the layer.
Effect of the Invention According to the present invention, there is provided a composite of a fiber molded body and a fibrin glue having high strength as well as biodegradability and excellent tissue adhesion.

  The composite of the fibrin glue and the fiber molded body of the present invention is useful as an artificial dura mater, an adhesion-preventing material, a hemostatic material, etc. as a medical article, particularly as a tissue surface or wound site protective material, covering material, and sealing material.

Claims (7)

  A composite comprising a fiber molded body and a fibrin glue, wherein the fiber molded body is a fiber made of a biodegradable polymer containing a phospholipid, the fiber surface has a smoothness of 1.07 or less, and produces a phospholipid. A composite comprising fibers containing from 0.1 to 10% by weight based on a degradable polymer.   The composite according to claim 1, wherein the smoothness of the fiber surface is 1.05 or less.   The composite according to claim 1 or 2, wherein the biodegradable polymer is an aliphatic polyester.   The composite according to claim 1 or 2, wherein the biodegradable polymer is polylactic acid, a copolymer of polylactic acid, or polyglycolic acid.   The complex according to any one of claims 1 to 4, wherein the phospholipid is phosphatidylcholine or phosphatidylethanolamine.   The composite according to any one of claims 1 to 5, wherein the fiber molded body is produced by an electrospinning method.   Fiber molded body comprising a biodegradable polymer containing a phospholipid, a fiber surface having a smoothness of 1.07 or less and a fiber containing phospholipid in an amount of 0.1 to 10% by weight based on the biodegradable polymer. A therapeutic kit for biofilm regeneration, which is a combination of a fibrin glue and a fibrin glue.