Laminated structure

Abstract

To provide a laminated structure body in which an organic structure body and an inorganic structure body are strongly laminated and integrated by an adhesion layer, and the inorganic structure body is hard to be peeled off.SOLUTION: There is provided a laminated structure body in which an organic structure body and an inorganic structure body are laminated and integrated by an adhesion layer. In the case where the adhesion layer has a layer made of silane coupling agent and a layer of inorganic compound, and the organic structure body, the layer made of silane coupling agent, the layer of inorganic compound and the inorganic structure body are laminated and integrated in this order, that is, in the case where the laminated structure body has a structure that the layer of inorganic compound is laid between the silane coupling agent and the inorganic structure body, the organic structure body and the inorganic structure body are strongly laminated and integrated by the adhesion layer.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a laminated structure in which an organic structure and an inorganic structure are laminated and integrated by an adhesive layer.

Background Art

[0002] Laminated structures in which an organic structure and an inorganic structure are laminated and integrated by an adhesive layer are used in various technical fields, such as cell culture containers in which a cell culture carrier of an inorganic fiber nonwoven fabric is supported in a well of an organic resin well plate, and electronic substrates in which inorganic elements such as electronic components are fixed on an organic film.

[0003] And, in order to laminate and integrate an organic structure and an inorganic structure, a silane coupling agent has been conventionally used as an adhesive forming an adhesive layer as disclosed in JP-A-2003-286289 (Patent Document 1).

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] However, when the applicant of the present application prepared a laminated structure according to the prior art as disclosed in Patent Document 1, an adhesive layer composed only of a silane coupling agent tended to be inferior in the ability to adhere and laminate an organic structure and an inorganic structure. As a result, the problem that the inorganic structure was likely to peel off from the prepared laminated structure occurred.

[0006] Therefore, there has been a demand for realizing a laminated structure in which an organic structure and an inorganic structure are firmly laminated and integrated by an adhesive layer and the inorganic structure is difficult to peel off. [Means for solving the problem]

[0007] The first invention is, “Organic structures and The constituent fibers are silicon oxide fibers. A laminated structure in which fabrics are laminated and integrated by an adhesive layer, The aforementioned adhesive layer comprises a layer made of a silane coupling agent, This is a layer of oligomers or polymers formed by the hydrolysis and / or condensation reactions of the constituent components of a sol containing silicon alkoxide. It has an inorganic compound layer, The inorganic compound layer contains only silicon as a metallic element in its chemical structure. The organic structure, the layer consisting of the silane coupling agent, the layer of the inorganic compound, and the fabric are laminated and integrated in that order. The layer of the inorganic compound fills the gap between the layer made of the silane coupling agent and the fabric. Laminated structure body. " That is the case.

[0008] Furthermore, the second aspect of the present invention is: "A method for manufacturing a laminated structure, (1) an organic structure; Fabrics whose constituent fibers are silicon oxide fibers The process of preparing (2) A step of applying a silane coupling agent onto the organic structure, (3) A step of reacting the organic structure with the silane coupling agent to form a layer consisting of the silane coupling agent on the organic structure. (4) On the layer made of the silane coupling agent, Sol containing silicon alkoxide The process of applying (5) The above Sol containing silicon alkoxide The process of preparing a laminate by laminating the aforementioned fabric on top of, (6) The laminate is included in the Sol containing silicon alkoxide By hydrolyzing and / or condensing the following, The silicon alkoxide-containing sol is formed by hydrolysis and / or condensation of oligomers or polymer layers. A process for forming an inorganic compound layer, A method for manufacturing a laminated structure according to claim 1, comprising the following: an organic structure, a layer consisting of the silane coupling agent, a layer of the inorganic compound, and the fabric, all laminated and integrated in that order. That is the case. [Effects of the Invention]

[0009] As a result of the applicant's consideration, a laminated structure in which an organic structure and an inorganic structure are integrally laminated by an adhesive layer, the adhesive layer having a layer composed of a silane coupling agent and a layer of an inorganic compound, and the organic structure - the layer composed of the silane coupling agent - the layer of the inorganic compound - the inorganic structure being integrally laminated in this order, that is, when it has a structure in which a layer of an inorganic compound is interposed between the silane coupling agent and the inorganic structure, it has been found that the organic structure and the inorganic structure are firmly integrally laminated by the adhesive layer. From the above, the laminated structure according to the present invention and its manufacturing method can provide a laminated structure in which the inorganic structure is difficult to peel off.

Embodiments for Carrying Out the Invention

[0010] In the present invention, various configurations such as the following configurations can be appropriately selected. In addition, unless otherwise specified, all various measurements described in the present invention were measured under atmospheric pressure. Also, the measurements were carried out under the temperature condition of 25°C. And, unless otherwise specified, all various measurement results described in the present invention were measured up to a value one digit smaller than the required value, and the required value was calculated by rounding the said value. As a specific example, when the value up to the first decimal place is the required value, the value up to the second decimal place is obtained by measurement, and the value of the first decimal place is calculated by rounding the obtained value of the second decimal place, and this value was taken as the required value. And, each upper limit value and each lower limit value exemplified in the present invention can be arbitrarily combined.

[0011] The organic structure referred to in the present invention refers to a structure containing organic components such as an organic resin, and its shape can be, for example, plate-like, fabric (fiber web, non-woven fabric, woven fabric, knitted fabric, etc.), non-porous or porous film, foam, etc. In addition, as a specific example of the organic structure, a petri dish or well plate made of polystyrene can be cited.

[0012] The type of the organic resin can be appropriately selected. For example, polyolefin resins (such as polyethylene, polypropylene, polymethylpentene, polyolefin resins with a structure in which a part of hydrocarbons is substituted with a nitrile group or a halogen such as fluorine or chlorine, etc.), styrene resins (such as polystyrene), polyvinyl alcohol resins, polyether resins (such as polyether ether ketone, polyacetal, modified polyphenylene ether, aromatic polyether ketone, etc.), polyester resins (such as polyethylene terephthalate, polytrimethylene terephthalate, polybutylene terephthalate, polyethylene naphthalate, polybutylene naphthalate, polycarbonate, polyarylate, wholly aromatic polyester resin, etc.), polyimide resins, polyamideimide resins, polyamide resins (such as aromatic polyamide resins, aromatic polyetheramide resins, nylon resins, etc.), resins having a nitrile group (such as polyacrylonitrile, etc.), urethane resins, epoxy resins, polysulfone resins (such as polysulfone, polyether sulfone, etc.), fluorine resins (such as polytetrafluoroethylene, polyvinylidene fluoride, etc.), cellulose resins, polybenzimidazole resins, acrylic resins (such as polyacrylonitrile resins copolymerized with acrylic acid esters or methacrylic acid esters, modacrylic resins copolymerized with acrylonitrile and vinyl chloride or vinylidene chloride, etc.), etc., can be configured using known organic resins.

[0013] The organic structure can also be composed of two or more types of organic resins, and two or more types of organic resins may be mixed.

[0014] When the organic structure is a fabric having fibers containing an organic component as constituent fibers, the constituent fibers may generally be fibers called composite fibers. For example, it can be in the forms of core-sheath type, sea-island type, side-by-side type, orange type, bimetal type, etc.

[0015] Furthermore, the material may include organic fibers with irregular cross-sections in addition to approximately circular or elliptical organic fibers. Examples of organic fibers with irregular cross-sections include hollow shapes, polygonal shapes such as triangular shapes, alphabetic shapes such as Y-shapes, irregular shapes, multi-lobed shapes, symbolic shapes such as asterisk shapes, or shapes formed by combining multiple such shapes.

[0016] Furthermore, the surface of the organic structure may be subjected to treatments such as plasma discharge treatment, UV treatment, or corona discharge treatment.

[0017] The physical properties of the organic structure, such as its basis weight and thickness, can be adjusted as appropriate to suit the application of the laminated structure.

[0018] In this invention, an inorganic structure refers to a structure containing inorganic components such as silica, and its shape can be, for example, a plate, a fabric (such as a fiber web, nonwoven fabric, woven fabric, or knitted fabric), a non-porous or porous film, or a foam. A specific example of an inorganic structure is a cell culture carrier made of silica fiber nonwoven fabric.

[0019] The types of inorganic components can be selected as appropriate. Examples include oxides of lithium, beryllium, boron, carbon, sodium, magnesium, aluminum, silicon, phosphorus, sulfur, potassium, calcium, scandium, titanium, vanadium, chromium, manganese, iron, cobalt, nickel, copper, zinc, gallium, germanium, arsenic, selenium, rubidium, strontium, yttrium, zirconium, niobium, molybdenum, cadmium, indium, tin, antimony, tellurium, cesium, barium, lanthanum, hafnium, tantalum, tungsten, mercury, thallium, lead, bismuth, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, or lutetium. Specifically, examples include SiO2, AL2O3, B2O3, TIO2, ZRO2, CEO2, FEO, FE3O4, FE2O3, VO2, V2O5, SNO2, CDO, LIO2, WO3, NB2O5, TA2O5, IN2O3, GEO2, PBTI4O9, LINBO3, BATIO3, PBZRO3, KTAO3, LI2B4O7, NIFE2O4, SRTIO3, hydroxyapatite, carbonate apatite, and zeolite. Note that the inorganic component may consist of an oxide of one element or an oxide containing two or more elements.

[0020] The inorganic structure may be composed of two or more inorganic components, or it may be a mixture of two or more inorganic components. Furthermore, the surface of the inorganic structure may be treated with plasma discharge treatment, UV treatment, corona discharge treatment, etc.

[0021] When an inorganic structure is a fabric having fibers containing inorganic components as constituent fibers, the constituent fibers may be fibers generally referred to as composite fibers. For example, they can be of the core-sheath type, sea-island type, side-by-side type, orange type, bimetal type, etc.

[0022] Furthermore, the material may include inorganic fibers with irregular cross-sections in addition to roughly circular or elliptical fibers. These irregular cross-sections may include hollow shapes, polygonal shapes such as triangular shapes, alphabetic shapes such as Y-shapes, irregular shapes, multi-lobed shapes, symbolic shapes such as asterisks, or shapes formed by combining multiple such shapes.

[0023] The average fiber diameter of inorganic fibers can be 0.01 to 10 μm, 0.02 to 5 μm, 0.05 to 3 μm, or 0.1 to 1.5 μm. The "average fiber diameter" refers to the arithmetic mean of the individual fiber diameters of 50 fibers measured from 5000x electron microscope images of the measurement target area containing the fibers. Furthermore, if the cross-sectional shape of the fiber is non-circular, the diameter of a circle with the same area as the cross-sectional area is considered the fiber diameter.

[0024] Inorganic fibers may contain additives within or on the surface of the fibers, such as flame retardants, fragrances, pigments, antibacterial agents, antifungal agents, photocatalytic particles, emulsifiers, dispersants, surfactants, particles that foam when heated, inorganic particles such as hydroxyapatite, antioxidants, hydrophilic agents, water repellents, surface charge modifiers, and physiologically active substances such as proteins.

[0025] The physical properties of inorganic structures, such as basis weight and thickness, can be adjusted as appropriate to suit the application of the laminated structure. For example, if the inorganic structure is a sheet-like cell culture carrier made of inorganic fibers, its basis weight can range from 0.01 to 100 g / m². 2 It can be 0.05~50g / m 2 It can be 0.1~25g / m 2 It can be. Note that "bare weight" refers to the measurement of 1 m on the main surface of the object being measured. 2This refers to the mass converted to a unit. The thickness can be 1000 μm or less, and can be 500 μm or less. On the other hand, a thickness of 1 μm or more is more practical. Note that "thickness" refers to the length measured using a straight-type tooth thickness micrometer (Mitutoyo Corporation, CLM1-15DK 227-221, measuring force 0.5 N) in a direction perpendicular to the main surface of the object being measured.

[0026] Furthermore, when an inorganic structure has voids, such as in a fabric, its porosity can be between 70% and 99%. This "porosity" refers to the value obtained by the following formula. P = [1 - M / (T × D)] × 100 Here, M is the basis weight of the object (unit: g / m²). 2 ), T is the thickness of the object (unit: mm), and D is the average density of the various components that make up the object (unit: g / cm³). 3 These represent the following meanings, respectively.

[0027] The laminated structure according to the present invention has an adhesive layer that plays a role in bonding and laminating an organic structure and an inorganic structure. The adhesive layer has a layer made of a silane coupling agent and a layer made of an inorganic compound, and the laminated structure according to the present invention is characterized in that the organic structure - silane coupling agent layer - inorganic compound layer - inorganic structure are laminated and integrated in that order.

[0028] In this invention, the silane coupling agent is defined as having the following molecules: • Functional groups that react with and can bond with organic components contained in the organic structure (e.g., amino groups, epoxy groups, methacrylic groups, vinyl groups, mercapto groups, etc.), A precursor of an inorganic compound, and / or a layer of an inorganic compound obtained by hydrolysis and / or condensation of the precursor, and a functional group that can react with and bind to it (e.g., hydrolysis groups such as methoxy groups, ethoxy groups, acetyl groups, etc.), It is a compound that possesses both of these properties, and acts as an intermediary to connect organic and inorganic structures that are normally difficult to bond together.

[0029] The type of silane coupling agent can be selected as appropriate, for example, vinyltrimethoxysilane, vinyltriethoxysilane, 2-(3,4-epoxycyclohexyl)ethyltrimethoxysilane, 3-glycidoxypropylmethyldimethoxysilane, 3-glycidoxypropyltrimethoxysilane, 3-glycidoxypropylmethyldiethoxysilane, 3-glycidoxypropyltriethoxysilane, P-styryltrimethoxysilane, 3-methacryloxypropylmethyldimethoxysilane, 3-methacryloxypropyltrimethoxysilane, 3-methacryloxypropylmethyldiethoxysilane, 3-methacryloxypropyltriethoxysilane, 3-acryloxypropyltrimethoxysilane, N-2-(aminoethyl)-3-aminopropylmethyldimethoxysilane, N- Well-known compounds used as silane coupling agents can be employed, such as 2-(aminoethyl)-3-aminopropyltrimethoxysilane, 3-aminopropyltrimethoxysilane, 3-aminopropyltriethoxysilane, 3-triethoxysilyl-N-(1,3-dimethylbutylidene)propylamine, N-phenyl-3-aminopropyltrimethoxysilane, hydrochloride of N-(vinylbenzyl)-2-aminoethyl-3-aminopropyltrimethoxysilane, tris-(trimethoxysilylpropyl)isocyanurate, 3-ureidopropyltrialkoxysilane, 3-mercaptopropylmethyldimethoxysilane, 3-mercaptopropyltrimethoxysilane, 3-isocyanatetopropyltriethoxysilane, and 3-trimethoxysilylpropyl succinic anhydride.

[0030] Furthermore, the layer consisting of the silane coupling agent refers to a layer derived from the silane coupling agent that is formed on top of the organic structure by the reaction of the functional groups of the silane coupling agent with the organic components contained in the organic structure. In addition, within the layer consisting of the silane coupling agent, the silane coupling agents themselves may be chemically bonded to each other by hydrolysis and / or condensation reactions.

[0031] In this invention, an inorganic compound refers to a compound that is different from the silane coupling agent and contains a metal element in its chemical structure (such as the metal oxides mentioned above). While such inorganic compounds can be selected as appropriate, it is preferable that the inorganic compound contains the same metal element as the metal element constituting the inorganic component of the inorganic structure, in order to minimize the influence of the inorganic compound on the performance that the inorganic structure is intended to exhibit.

[0032] Furthermore, the inorganic compound layer, as will be described in detail later, refers to a layer consisting of compounds derived from the precursor of the inorganic compound, which are obtained by hydrolysis and / or condensation of the precursor of the inorganic compound. Specifically, when a sol containing a metal alkoxide is used as the precursor of the inorganic compound, the inorganic compound layer is a layer of oligomers or polymers obtained by hydrolysis and / or condensation of the constituent components of the sol.

[0033] The layer of inorganic compound can act as an intermediary, connecting the layer of silane coupling agent in the adhesive layer with the inorganic structure.

[0034] The inorganic compound layer is preferably formed by hydrolysis and / or condensation reactions of the constituent components of a sol containing a metal alkoxide (e.g., ethoxysilane). This is because the metal alkoxide contained in the sol, and the compounds formed by hydrolysis and / or condensation reactions of such metal alkoxides, have many reactive groups and therefore have high reactivity with silane coupling agents, thus providing a laminated structure in which the organic and inorganic structures are more strongly laminated and integrated by the adhesive layer.

[0035] Thus, by interposing a layer of inorganic compounds between the silane coupling agent layer and the inorganic structure, it is possible to realize a laminated structure in which the problem of delamination of the inorganic structure is less likely to occur. Although the reason for this is not fully understood, it is thought to be due to the following effects.

[0036] Silane coupling agents are compounds that possess both reactive groups capable of binding to organic components and reactive groups capable of reacting with and binding to precursors or layers of inorganic compounds within their molecular structure. Therefore, when organic and inorganic structures are laminated and integrated using a silane coupling agent, as in conventional techniques, the inorganic structure is laminated and integrated with the organic structure by a layer of silane coupling agents that exists in a two-dimensional arrangement on the organic structure.

[0037] In this case, if voids or irregularities exist on the surface of the inorganic structure, microscopic gaps will form between the layer of silane coupling agent, which is arranged in a two-dimensional state, and the inorganic structure. In other words, these gaps do not contribute to the lamination and integration, so the lamination and integration between the layer of silane coupling agent and the inorganic structure is considered to be weak.

[0038] Therefore, in this invention, a layer of inorganic compound having chemical properties different from the silane coupling agent is formed between the layer made of the silane coupling agent and the inorganic structure. As a result, the aforementioned microscopic gaps can be filled with the inorganic compound. In particular, by employing an inorganic compound precursor that allows molecules to bond three-dimensionally (for example, a sol containing a metal alkoxide (e.g., ethoxysilane)), the microscopic gaps can be effectively filled with the inorganic compound, and the lamination and integration of the silane coupling agent layer and the inorganic structure by the inorganic compound layer can be made stronger.

[0039] As a result of these effects, it is believed that a laminated structure can be realized in which the organic and inorganic structures are firmly laminated and integrated, making it less likely for the inorganic structure to peel off.

[0040] Whether or not a laminated structure satisfies the configuration according to the present invention can be confirmed by the following method. 1. Analyze one of the main surfaces of the laminated structure using methods such as X-ray photoelectron spectroscopy (XPS) to confirm whether that main surface contains an inorganic structure. 2. If the main surface has an inorganic structure, remove the inorganic structure from the laminated structure. 3. The main surface on the side from which the inorganic structure was removed is analyzed using XPS. If the main surface contains an inorganic compound, it is assumed that the inorganic structure and the inorganic compound layer are laminated and integrated in the laminated structure. 4. Remove the layer of inorganic compound from the main surface on the side from which the inorganic structure was removed, using methods such as ion etching. 5. The main surface on the side from which the inorganic compound layer has been removed is analyzed using XPS. If peaks of the element group constituting the silane coupling agent are detected from this main surface, the laminated structure is considered to be a laminated and integrated structure consisting of an inorganic compound layer and a silane coupling agent layer. 6. Remove the layer consisting of the silane coupling agent from the main surface on the side from which the inorganic compound layer was removed, using methods such as ion etching. 7. The main surface on the side from which the silane coupling agent layer has been removed is analyzed using XPS. If the main surface has an organic structure, it is assumed that the laminated structure consists of the silane coupling agent layer and the organic structure laminated together as one. 8. If the above configuration is satisfied, the laminated structure submitted for verification is deemed to satisfy the configuration of the present invention.

[0041] Furthermore, if the manufacturing process of the laminated structure is known, it is possible to determine whether or not the laminated structure satisfies the configuration of the present invention by checking the manufacturing method.

[0042] An example of a laminated structure according to the present invention is a cell culture vessel in which a cell culture carrier made of silica fiber nonwoven fabric is laminated and integrated on the bottom surface of the wells inside a polystyrene well plate, with an adhesive layer in between consisting of a layer of silane coupling agent and a layer of inorganic compound.

[0043] Next, the manufacturing method of the laminated structure according to the present invention will be illustrated and explained. Note that explanations of points that are the same as those already described will be omitted. The manufacturing method of the laminated structure according to the present invention can be appropriately selected, but as an example, (1) A process of preparing organic and inorganic structures, (2) A step of applying a silane coupling agent onto the organic structure, (3) A step of reacting the organic structure with the silane coupling agent to form a layer consisting of the silane coupling agent on the organic structure. (4) A step of applying an inorganic compound precursor onto the layer consisting of the silane coupling agent, (5) A step of preparing a laminate by stacking the inorganic structure on the precursor of the inorganic compound, (6) A step of hydrolyzing and / or condensing the precursor of the inorganic compound contained in the laminate to form a layer of the inorganic compound, A method for manufacturing a laminated structure that includes the above features can be used.

[0044] First, let's explain step (1). As an organic structure, for example, a well plate made of polystyrene can be used. Furthermore, as an inorganic structure, for example, an inorganic fiber nonwoven fabric such as silica glass fiber nonwoven fabric can be used.

[0045] Next, steps (2) and (3) will be explained. The silane coupling agent may contain a solvent or dispersion medium and can be used in the form of a silane coupling agent solution or a silane coupling agent dispersion. The type of solvent or dispersion medium can be selected as appropriate. The mass of the silane coupling agent contained in these can be adjusted as appropriate, but can be 0.00001 to 10% by mass, 0.0001 to 5% by mass, or 0.0005 to 1% by mass. Since the silane coupling agent is also used as a surface modifier, excessive use may cause unintended surface modification of organic or inorganic structures, which may impair the performance of the laminated structure. Therefore, it is preferable to use as little silane coupling agent as possible, as long as the objective of the present invention can be achieved.

[0046] The method for reacting the organic structure with the silane coupling agent can be appropriately selected, but one example is subjecting it to heat treatment. The heating temperature should be adjusted appropriately so that the reaction proceeds smoothly, but it is preferable to adjust it in a way that prevents unintended changes from occurring in the organic structure or other constituent members. The heating method can be appropriately selected, and for example, heating can be done using an oven dryer, far-infrared heater, dry heat dryer, hot air dryer, hot plate, or by irradiating with infrared rays under no pressure.

[0047] Next, I will explain step (4). In the method for manufacturing a laminated structure according to the present invention, an inorganic compound precursor is applied to a layer made of a silane coupling agent, and as described in the following step, the precursor is subjected to hydrolysis and / or condensation reactions by heat treatment, etc., to form a layer of the inorganic compound on top of the layer made of the silane coupling agent. As such an inorganic compound precursor, a sol solution containing an oxide of the same metal element as the inorganic structure can be used.

[0048] The precursor of the inorganic compound may contain a solvent or dispersion medium, and can be used in the form of a precursor solution or a precursor dispersion. The type of solvent or dispersion medium can be selected as appropriate. The mass (solid content) of the precursor contained therein can be adjusted as appropriate, but can be 0.01 to 50% by mass, 0.05 to 25% by mass, or 0.1 to 10% by mass. If the inorganic compound layer and the inorganic structure do not have exactly the same composition, using an excess amount of the inorganic compound precursor may cause unintended alteration of the inorganic structure, impairing its performance. Therefore, it is preferable to use as little of the precursor as possible, as long as the objective of the present invention can be achieved.

[0049] Finally, step (6) will be explained. The method for hydrolyzing and / or condensing the precursor of the inorganic compound can be appropriately selected, but one example is subjecting it to heat treatment. The heating temperature is adjusted appropriately so that the precursor of the inorganic compound undergoes hydrolysis and / or condensation to form a layer of the inorganic compound, but it is preferable to adjust it in a way that prevents unintended changes from occurring in the constituent members such as organic structures. The heating method can be appropriately selected, and the method described above can be used.

[0050] The laminated structure prepared in this manner can be provided and used as is for various industrial applications, but it may also be provided in the form of a product that has undergone sterilization or packaging processes. [Examples]

[0051] The present invention will be specifically described below with reference to examples, but these examples are not intended to limit the scope of the present invention.

[0052] (Preparation of organic structures) A polystyrene well plate (Thermo Fisher Scientific, part number 142475, 24 holes, with each well having a circular bottom surface of 15.5 mm in diameter) was prepared.

[0053] (Preparation of inorganic structures) As an inorganic structure, a high-purity silica fiber nonwoven fabric cell culture carrier is used, which is a glass fiber nonwoven fabric (manufactured by Nippon Vilene Co., Ltd., CB-24CT4, 13mm diameter circular, average fiber diameter: 840nm, basis weight 8g / m²). 2 A material with a thickness of 270 μm and a main component of SiO2 was prepared.

[0054] (Preparation of silane coupling agent) 3-aminopropyltriethoxysilane (Shin-Etsu Chemical Co., Ltd., LS-3150) was diluted with an aqueous ethanol solution to a concentration of 2% by mass. After stirring the prepared dilution at room temperature for 2 hours, ethanol was added to the dilution and mixed until the concentration reached 0.001% by mass to prepare silane coupling agent solution A. Silane coupling agent solution B was also prepared by adding ethanol to the dilution and mixing until the concentration reached 0.005% by mass.

[0055] (Preparation of inorganic compound precursors) Tetraethoxysilane, ethanol, water, and 2 N nitric acid were mixed in a molar ratio of 1:7.2:11:0.0033 and stirred at room temperature for 15 hours to hydrolyze the tetraethoxysilane. Subsequently, ethanol was added and mixed until the solid content (SiO2 in this example) concentration reached 0.5% by mass to prepare sol solution A containing silicon oxide. Sol solution B containing silicon oxide was also prepared by adding ethanol and mixing until the concentration reached 1% by mass.

[0056] (Comparative Example 1) For 12 wells of a well plate, 30 μL of silane coupling agent solution A was applied to the bottom surface of each well. Then, before the silane coupling agent solution dried, a cell culture support made of high-purity silica fiber nonwoven fabric was laminated on top of each well. Subsequently, the silane coupling agent solution was subjected to a 50°C atmosphere for 3 hours to react with the 3-aminopropyltriethoxysilane and remove the solvent. Furthermore, a washing process was performed twice by adding 600 μL of ethanol or pure water to each well (6 wells washed with ethanol, 6 wells washed with pure water). After that, the wells were dried under a 50°C atmosphere to prepare cell culture vessels.

[0057] (Comparative Example 2) For six wells of the well plate, 12 μL of sol solution A was applied to the bottom surface of each well. Then, before the sol solution dried, a cell culture support made of high-purity silica fiber nonwoven fabric was layered on top of each well. Subsequently, the sol solution was subjected to a 50°C atmosphere for 8 hours to hydrolyze and / or condense the tetraethoxysilane contained in the sol solution, and to remove the solvent. Furthermore, a washing process was performed twice by adding 600 μL of pure water to each well (number of wells washed with pure water: 6 wells), and then the solution was dried under a 50°C atmosphere to prepare cell culture vessels.

[0058] (Comparative Example 3) For six wells of the well plate, 12 μL of a mixture consisting of 6 μL each of silane coupling agent solution B and sol solution B was applied to the bottom surface of each well. Then, before the mixture dried, a cell culture support made of high-purity silica fiber nonwoven fabric was laminated on top of each well. Subsequently, the mixture was subjected to a 50°C atmosphere for 8 hours to react the 3-aminopropyltriethoxysilane and hydrolyze and / or condense the tetraethoxysilane, while also removing the solvent. Furthermore, a washing process was performed twice by adding 600 μL of pure water to each well (6 wells washed with pure water), and then the mixture was dried under a 50°C atmosphere. In this way, a cell culture vessel was prepared in which a cell culture support made of high-purity silica fiber nonwoven fabric was laminated and integrated with a layer on the bottom surface inside the well, with a layer in between in which a compound obtained by the reaction of 3-aminopropyltriethoxysilane and an inorganic compound obtained by the hydrolysis and / or condensation reaction of tetraethoxysilane were mixed.

[0059] (Example 1) For six of the wells in the well plate, 30 μL of silane coupling agent solution A was applied to the bottom surface of each well. Subsequently, the well plate was exposed to a 50°C atmosphere for 3 hours to react with the 3-aminopropyltriethoxysilane contained in the silane coupling agent and to remove the solvent. Furthermore, a washing step was performed twice by adding 600 μL of ethanol to each well and removing it, and then the well plate was dried by exposing it to a 50°C atmosphere. In this way, a layer of compound formed by the reaction of 3-aminopropyltriethoxysilane with the polystyrene well plate (a layer consisting of the silane coupling agent) was formed on the bottom surface of the wells inside the well plate. Next, 12 μL of sol solution A was applied to the layer of silane coupling agent formed on the bottom surface of each well. Then, before the sol solution dried, a cell culture carrier made of high-purity silica fiber nonwoven fabric was laminated on top of it. Subsequently, the solution was subjected to a 50°C atmosphere for 8 hours to hydrolyze and / or condense the tetraethoxysilane contained in the sol solution, and to remove the solvent. Furthermore, a washing process was performed twice by adding 600 μL of pure water to each well (number of wells washed with pure water: 6 wells), and then the solution was dried under a 50°C atmosphere. In this way, a cell culture vessel was prepared by laminating and integrating a cell culture support made of high-purity silica fiber nonwoven fabric with a layer of a compound formed by the reaction of 3-aminopropyltriethoxysilane with a polystyrene well plate (a layer consisting of a silane coupling agent) and a layer of an inorganic compound formed by the hydrolysis and / or condensation reaction of tetraethoxysilane, with these layers in between on the bottom surface inside the wells.

[0060] (Method for checking adhesive durability) For each well of each prepared cell culture vessel, we checked whether or not the high-purity silica fiber nonwoven fabric cell culture support had detached. The number of wells in which detachment occurred was summarized in Table 1 as the "number of detachments before 30 pipetting cycles". Next, 600 μL of culture medium was added to each well of each prepared cell culture vessel, and pipetting was performed 30 times using a micropipette set to a 350 μL mark. After pipetting, the presence or absence of detachment of the high-purity silica fiber nonwoven fabric cell culture support was checked in each well. The number of wells in which detachment occurred was summarized in Table 1 as the "number of detachments after 30 pipettings". Note that for wells where the "number of detachments before 30 pipettings" was 6, this pipetting procedure was not performed because there was no subject to check for detachment.

[0061] The results are summarized in Table 1. Furthermore, in the "Number of delaminations after 30 pipettings" column, "-" is indicated in the table for cases where there were no objects to check for delamination. Note that "-" indicates the laminated structure most susceptible to inorganic delamination, while for all other cases, a lower number of delaminations indicates a laminated structure less prone to inorganic delamination.

[0062] [Table 1]

[0063] From the above results, it has been found that the present invention can provide a laminated structure in which an organic structure and an inorganic structure are firmly laminated and integrated by an adhesive layer.

[0064] Furthermore, a cytotoxicity test was performed on cell culture vessels prepared separately using the cell culture vessel manufacturing method according to Example 1.

[0065] (Method for confirming cytotoxicity) In this evaluation, a polystyrene well plate (Thermo Fisher Scientific, 161093, 96 wells, internal well bottom size: circular with a diameter of 6.5 mm) was used; a cell culture carrier made of high-purity silica fiber nonwoven fabric (Vilene Japan Co., Ltd., CB-24CT4, circular with a diameter of 13 mm) was prepared as the inorganic structure and cut into 6.15 mm diameter circles using a laser processing machine; and the amount of adhesive and washing solution per well of the well plate was reduced to one-sixth of the amount used in Example 1. Aside from these differences, the cell culture vessel was prepared in the same manner as in Example 1. Three plates were then prepared and sterilized by irradiation with a 30 kGy electron beam. The cytotoxicity of the cell culture vessels prepared in this manner was evaluated based on "Japanese Pharmacopoeia, 16th Revision, Second Supplement 7.02: Test Methods for Plastic Pharmaceutical Containers" and "Pharmaceutical and Medical Device Notification No. 0301, 20: Basic Concepts Regarding Biological Safety Assessment Required for Applications for Manufacturing and Marketing Approval of Medical Devices." Specifically, 100 μl / well of culture medium (Eagle's MEM medium containing 5% fetal bovine formation) was dispensed into each well of a cell culture vessel, and the mixture was extracted at 37°C under a 5% CO2 atmosphere for 24 hours to obtain a 100% test solution. This solution was then serially diluted (25-100%) with the aforementioned culture medium and exposed to V79 cells to perform a cytotoxicity test using the colony formation method.

[0066] As a result, IC 50 No colony formation inhibition sufficient to calculate the value was observed. Therefore, the cell culture vessel and the adhesion layer of the cell culture vessel according to the present invention were found to be non-cytotoxic. [Industrial applicability]

[0067] The present invention provides a laminated structure in which an organic structure and an inorganic structure are firmly laminated and integrated by an adhesive layer. The laminated structure according to the present invention can be used in various industrial applications such as cell culture vessels and electronic circuit boards.

Claims

1. A laminated structure in which an organic structure and a fabric whose constituent fibers are silicon oxide fibers are laminated and integrated by an adhesive layer, The adhesive layer comprises a layer made of a silane coupling agent and a layer of an inorganic compound which is an oligomer or polymer formed by hydrolysis and / or condensation of the components of a sol containing silicon alkoxide. The inorganic compound layer contains only silicon as a metallic element in its chemical structure. The organic structure, the layer consisting of the silane coupling agent, the layer of the inorganic compound, and the fabric are laminated and integrated in that order. The layer of the inorganic compound fills the gap between the layer made of the silane coupling agent and the fabric. Laminated structure.

2. A method for manufacturing a laminated structure, (1) A step of preparing an organic structure and a fabric whose constituent fibers are silicon oxide fibers, (2) A step of applying a silane coupling agent onto the organic structure, (3) A step of reacting the organic structure with the silane coupling agent to form a layer consisting of the silane coupling agent on the organic structure. (4) A step of applying a sol containing silicon alkoxide onto the layer consisting of the silane coupling agent, (5) A step of preparing a laminate by laminating the fabric onto the sol containing the silicon alkoxide, (6) A step of hydrolyzing and / or condensing the silicon alkoxide-containing sol contained in the laminate to form a layer of inorganic compounds which is an oligomer or polymer layer formed by the hydrolysis and / or condensation of the constituent components of the silicon alkoxide-containing sol, A method for manufacturing a laminated structure according to claim 1, comprising the following: an organic structure, a layer consisting of the silane coupling agent, a layer of the inorganic compound, and the fabric, all laminated and integrated in that order.

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