Polymer-coated glass substrate

Abstract

Provided is a polymer-coated glass substrate in which the unevenness of the surface is controlled and the elastic modulus of the surface is low. The present invention relates to a polymer-coated glass substrate in which a hydrophilic polymer layer comprising a mixture of hydrophilic polymers having different molecular weights is formed on the surface of a glass substrate.

Description

Technical Field

[0001] The present invention relates to a glass substrate coated with a polymer. Background Art

[0002] In order to produce an instrument for adsorbing specific cells (blood cells, cancer cells present in blood / body fluid, stem cells, T cells, etc.) in blood and body fluid, a technique of coating a special polymer on the surface of a glass substrate has been proposed.

[0003] However, among special polymers, there are substances that are difficult to produce a smooth surface by coating. Since the concavity and convexity of the surface affect the adsorption performance of specific cells, it is desired to provide a substrate having a surface with controlled concavity and convexity and excellent adsorption performance for specific cells such as cancer cells, stem cells, and T cells (see Patent Document 1, etc.). Further, when the surface has large concavity and convexity, cloudiness of the coating surface is likely to occur, and it is desired to suppress the cloudiness of the coating surface by controlling the concavity and convexity of the surface. In addition, by suppressing the cloudiness of the coating surface, an improvement in the adsorption performance of specific cells such as cancer cells, stem cells, and T cells is expected.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Problems to be Solved by the Invention

[0005] An object of the present invention is to solve the above problems and provide a glass substrate coated with a polymer, which can control the surface concavity and convexity and has a low surface elastic modulus.

Means for Solving the Problems

[0006] The present invention relates to a glass substrate coated with a polymer, on the surface of which a hydrophilic polymer layer composed of a mixture of hydrophilic polymers having different molecular weights is formed.

Advantages of the Invention

[0007] According to the present invention, since it is a glass substrate coated with a polymer, on the surface of which a hydrophilic polymer layer composed of a mixture of hydrophilic polymers having different molecular weights is formed, it is possible to provide a glass substrate coated with a polymer having controlled surface concavity and convexity and a low surface elastic modulus. Therefore, based on the above glass substrate coated with a polymer, an effect of improving the adsorption performance of specific cells such as cancer cells, stem cells, and T cells can be obtained. Detailed Description

[0008] The glass substrate of the coated polymer is a substrate having a hydrophilic polymer layer formed on the surface of the glass substrate and composed of a mixture of hydrophilic polymers with different molecular weights. By forming the hydrophilic polymer layer on the surface of the glass substrate with a mixture of hydrophilic polymers having different molecular weights, the unevenness of the substrate surface can be controlled, and a glass substrate of a coated polymer with a hydrophilic polymer layer having a small surface roughness, high smoothness, and a low elastic modulus on the surface can be provided.

[0009] The number of tumor cells (such as cancer cells) generated in body fluids such as circulating tumor cells in blood (several to hundreds per 1 mL of blood) is very small. For examination, it is considered important to adsorb as many tumor cells present in the sampled body fluid as possible. The glass substrate of the coated polymer can form a hydrophilic polymer layer with controlled surface unevenness, small surface roughness, and high smoothness by forming a hydrophilic polymer layer composed of a mixture of hydrophilic polymers with different molecular weights on the glass substrate. In addition, the surface of the formed hydrophilic polymer layer has a low elastic modulus. Moreover, the unevenness of the hydrophilic polymer layer affects the adsorption of specific cells such as cancer cells, stem cells, and T cells. By controlling the surface unevenness and improving the smoothness, excellent adsorption performance of specific cells can be obtained. In addition, the elastic modulus of the surface also affects the adsorption of specific cells. The lower the elastic modulus, the higher the adsorption performance. On the other hand, if the elastic modulus of the surface is too low, the adsorption will decrease instead. That is, there is an optimal value for the elastic modulus of the surface. In addition, for example, tumor cells are adsorbed on the hydrophilic polymer layer composed of a mixture of hydrophilic polymers with different molecular weights formed on the surface of the glass substrate. By measuring its value, the number of tumor cells in the body fluid can be determined, which can be used for the confirmation of the treatment effect of cancer, etc. In addition, by culturing the adsorbed tumor cells and confirming the effect of anticancer agents, etc. with the cultured cells, the effect of anticancer agents, etc. can be confirmed in vitro before administering anticancer agents, etc., and it can also be used for the selection of anticancer agents, etc. In addition, gene analysis is performed on the adsorbed or cultured tumor cells to investigate the mutation state and expression state of the tumor cells, which can be used for the selection of anticancer agents, etc. and can also contribute to the analysis of the mechanism of cancer.

[0010] The type of glass of the glass substrate constituting the above-mentioned coated polymer is not particularly limited. For example, soda-lime glass, alkali-free glass, borosilicate glass (such as SiO2-B2O3-ZnO-based glass, SiO2-B2O3-Bi2O3-based glass, etc.), potash glass, crystal glass (glass containing PbO, for example, SiO2-PbO-based glass, SiO2-PbO-B2O3-based glass, SiO2-B2O3-PbO-based glass, etc.), titanium crystal glass, barium glass, borate glass (such as B2O3-ZnO-PbO-based glass, B2O3-ZnO-Bi2O3-based glass, B2O3-Bi2O3-based glass, B2O3-ZnO-based glass, etc.), strontium glass, aluminosilicate glass, soda zinc glass, soda barium glass (such as BaO-SiO2-based glass, etc.) can be cited. These glasses can be used alone or in combination of two or more.

[0011] In the above-mentioned glass substrate, the contact angle of the bubbles in water is preferably 100 degrees or more and 120 degrees or less. The above-mentioned contact angle is more preferably 103 degrees or more, and furthermore, more preferably 117 degrees or less. In addition, in this specification, the "contact angle of the bubbles in water" means: in water, bubbles (air) are attached to the surface of the substrate, and the contact angle between the bubbles and the surface of the substrate is measured.

[0012] The thickness of the above-mentioned glass substrate is not particularly limited. As the average thickness, it is preferably 100 μm or more and 4000 μm or less, and more preferably 100 μm or more and 3000 μm or less. In addition, the average thickness is the value obtained by measuring the thickness at any 10 places using a micrometer and taking the average of the measured values.

[0013] The glass substrate of the above-mentioned coated polymer is a substrate on the surface of which a hydrophilic polymer layer (hereinafter also referred to as "mixed hydrophilic polymer layer") composed of a mixture of hydrophilic polymers with different molecular weights is formed. The hydrophilic polymers with different molecular weights can be a substance containing two polymers of the same hydrophilic polymer but with different molecular weights (for example, a mixture of hydrophilic polymers containing poly(2-methoxyethyl acrylate) with a number average molecular weight of 16,000 and poly(2-methoxyethyl acrylate) with a number average molecular weight of 80,000), or a substance containing different hydrophilic polymers (hydrophilic polymers with different structures) and polymers with different molecular weights (for example, a mixture of hydrophilic polymers containing poly(2-methoxyethyl acrylate) with a number average molecular weight of 16,000 and polyacrylmorpholine with a number average molecular weight of 80,000). Among them, from the viewpoint of obtaining better effects, the above-mentioned mixed hydrophilic polymer layer is preferably a layer formed by polymers of the same hydrophilic polymer and with different molecular weights.

[0014] The hydrophilic polymer constituting the above-mentioned mixed hydrophilic polymer layer is not particularly limited. For example, a publicly known hydrophilic polymer can be appropriately selected. The above-mentioned hydrophilic polymer can be used alone or in combination of two or more.

[0015] The above-mentioned hydrophilic polymer can be produced by a publicly known method. For example, a solution of a hydrophilic monomer constituting the hydrophilic polymer can be used, and the hydrophilic monomer can be polymerized by a publicly known method to synthesize it. The solvent of the hydrophilic monomer solution is not particularly limited. For example, the following solvents can be used. Among them, toluene and methanol are preferred.

[0016] As the above-mentioned hydrophilic polymer, for example, homopolymers and copolymers of one or more hydrophilic monomers, copolymers of one or more hydrophilic monomers and one or more other monomers, etc. can be cited. On the glass substrate of the above-mentioned coating polymer, a mixed hydrophilic polymer layer containing such a mixture of hydrophilic polymers with different molecular weights is formed.

[0017] The above-mentioned hydrophilic monomer is not particularly limited. For example, various monomers having a hydrophilic group can be used. As the hydrophilic group, for example, publicly known hydrophilic groups such as an amide group, a sulfate group, a sulfonic acid group, a carboxyl group, a hydroxyl group, an amino group, an amide group, and an oxyethylene group can be cited.

[0018] As specific examples of the above-mentioned hydrophilic monomer, (meth)acrylic acid; (meth)acrylate; (meth)acrylic acid alkoxyalkyl esters such as (meth)acrylic acid 2-methoxyethyl ester; (meth)acrylic acid hydroxyalkyl esters such as (meth)acrylic acid 2-hydroxyethyl ester; (meth)acrylamide; (meth)acrylamide derivatives having a cyclic group such as (meth)acryloylmorpholine, etc. can be cited. Among them, (meth)acrylic acid, (meth)acrylate, (meth)acrylic acid alkoxyalkyl esters, and (meth)acrylamide derivatives having a cyclic group are preferred, (meth)acrylic acid alkoxyalkyl esters and (meth)acryloylmorpholine are more preferred, (meth)acrylic acid alkoxyalkyl esters are further preferred, and 2-methoxyethyl acrylate is particularly preferred. These can be used alone or in combination of two or more.

[0019] The above-mentioned other monomers can be appropriately selected within the range that does not hinder the effect of the hydrophilic polymer. As specific examples of the above-mentioned other monomers, for example, aromatic monomers such as styrene, vinyl acetate, N-isopropylacrylamide capable of imparting temperature responsiveness, etc. can be cited. These can be used alone or in combination of two or more.

[0020] As the above-mentioned homopolymers and copolymers, specifically, polyacrylic acid, polyacrylate, polymethacrylic acid, polymethacrylate, polyacrylmorpholine, polymethacrylmorpholine, polyacrylamide, polymethacrylamide, polyalkoxyalkyl acrylate, polyalkoxyalkyl methacrylate, etc., which are homopolymers composed of one kind of hydrophilic monomer; copolymers composed of two or more of the above-mentioned hydrophilic monomers; copolymers composed of one or more of the above-mentioned hydrophilic monomers and one or more of the other monomers mentioned above, etc. can be cited.

[0021] Among them, the hydrophilic polymer constituting the above-mentioned mixed hydrophilic polymer layer preferably contains a hydrophilic polymer represented by the following chemical formula (I) with different molecular weights. Here, the hydrophilic polymer represented by the following formula (I) with different molecular weights can be a substance containing two or more hydrophilic polymers composed of the same structural unit but with different molecular weights (for example, a polymer containing poly(2-methoxyethyl acrylate) with a number-average molecular weight of 16,000 and poly(2-methoxyethyl acrylate) with a number-average molecular weight of 80,000), or a substance containing two or more hydrophilic polymers composed of different structural units and with different molecular weights (for example, a polymer containing poly(2-methoxyethyl acrylate) with a number-average molecular weight of 16,000 and poly(2-ethoxyethyl methyl acrylate) with a number-average molecular weight of 80,000). The hydrophilic polymer represented by the following formula (I) can be any one of one kind or two or more kinds.

Chemical formula 1

[0022] As the hydrophilic polymer represented by the above formula (I), for example, a hydrophilic polymer represented by the following formula (I-1) is preferably used. The hydrophilic polymer represented by the following formula (I-1) can be any one of one kind or two or more kinds.

Chemical formula 2

[0023] As the above hydrophilic polymer, a copolymer of a hydrophilic monomer represented by the following formula (II) and other monomers with different molecular weights is also preferably used. Here, the copolymer of a hydrophilic monomer represented by the following formula (II) with different molecular weights and other monomers may be a substance containing two or more hydrophilic polymers composed of the same structural unit but having different molecular weights (for example, a polymer containing a copolymer of 2-methoxyethyl acrylate with a number-average molecular weight of 16,000 and styrene and a copolymer of 2-methoxyethyl acrylate with a number-average molecular weight of 80,000 and styrene), or a substance containing two or more hydrophilic polymers composed of different structural units and having different molecular weights (for example, a polymer containing a copolymer of 2-methoxyethyl acrylate with a number-average molecular weight of 16,000 and styrene and a copolymer of 2-methoxyethyl arylate with a number-average molecular weight of 80,000 and vinyl acetate). The hydrophilic polymer represented by the following formula (II) may be any one of one kind or two or more kinds. The other monomers may be any one of one kind or two or more kinds.

[0024]

Chemical formula 3

[0025] As the compound (hydrophilic monomer) represented by the above formula (II), for example, the compound represented by the following formula (II-1) is preferably used. The compound represented by the following formula (II-1) may be any one of one kind or two or more kinds.

Chemical formula 4

[0026] In the above formulas (I), (I-1), (II), and (II-1), the alkyl group of R 52 preferably has 1 to 10 carbon atoms, more preferably 1 to 5 carbon atoms. Among them, R 52 is particularly preferably a methyl group or an ethyl group. p is preferably 1 to 5, more preferably 1 to 3. m is preferably 1 to 3. n (number of repeating units) is preferably 15 to 1500, more preferably 40 to 1200.

[0027] Among the foregoing hydrophilic polymers, from the viewpoints of controlling the concavity and convexity of the substrate surface and being able to form a hydrophilic polymer layer with a small surface roughness and high smoothness, the hydrophilic polymer represented by the above formula (I) is preferred, and the hydrophilic polymer represented by the above formula (I-1) is particularly preferred.

[0028] Among the hydrophilic polymers with different molecular weights described above, the molecular weight is not particularly limited, and examples include number average molecular weight (Mn), weight average molecular weight (Mw), etc. Among them, a mixture of hydrophilic polymers with different Mn is preferably used.

[0029] When using a mixture of the above hydrophilic polymers with different Mn, from the viewpoint of obtaining better effects, it is preferable to contain hydrophilic polymers with a difference in Mn of 30,000 or more, more preferably to contain hydrophilic polymers with a difference in Mn of 50,000 or more. In addition, it is preferable to contain hydrophilic polymers with a difference in Mn of 195,000 or less.

[0030] From the viewpoint of obtaining better effects, the mixture of the above hydrophilic polymers with different molecular weights preferably contains a low molecular weight hydrophilic polymer with a number average molecular weight (Mn) of 5,000 or more and 25,000 or less. The Mn of the above low molecular weight hydrophilic polymer is preferably 10,000 or more, more preferably 14,000 or more, and in addition, preferably 23,000 or less. In addition, from the viewpoint of obtaining better effects, the mixture of the above hydrophilic polymers with different molecular weights preferably contains a high molecular weight hydrophilic polymer with a number average molecular weight (Mn) of 35,000 or more and 200,000 or less. The Mn of the above high molecular weight hydrophilic polymer is preferably 40,000 or more, more preferably 50,000 or more, and in addition, preferably 150,000 or less, more preferably 130,000 or less.

[0031] In addition, in this specification, the number average molecular weight (Mn) and weight average molecular weight (Mw) can be calculated as standard polystyrene based on the measured values obtained by gel permeation chromatography (GPC) (GPC-8000 series manufactured by Tosoh Corporation, detector: differential refractometer, column: TSK GEL SUPERMALTPORE HZ-M manufactured by Tosoh Corporation).

[0032] The thickness of the above mixed hydrophilic polymer layer is preferably 30 to 3000 nm, more preferably 30 to 1500 nm, and further preferably 50 to 1000 nm. By adjusting within the above range, it tends to obtain better effects. In addition, good low adsorption properties for proteins and cells, as well as selective adsorption properties for specific cells such as cancer cells, stem cells, and T cells, can be expected.

[0033] The contact angle of water on at least a part (part or all) of the surface of the above mixed hydrophilic polymer layer is preferably 65 degrees or less, more preferably 60 degrees or less. The lower limit is not particularly limited, and the smaller the better.

[0034] The above-mentioned mixed hydrophilic polymer layer can be formed by known methods such as (1) injecting a hydrophilic polymer solution / dispersion prepared by dissolving / dispersing two or more hydrophilic polymers with different molecular weights in various solvents onto the surface of a glass substrate (such as a recess of the substrate), maintaining for a specified time as required and then drying; (2) coating (spraying) the hydrophilic polymer solution / dispersion onto the surface of the glass substrate (such as a recess of the substrate) and drying as required. Then, by forming a mixed hydrophilic polymer layer on the surface of the glass substrate using such a known method, a glass substrate coated with a polymer can be manufactured. In addition, by additionally attaching other components to the glass substrate coated with the polymer as required, a device capable of adsorbing, culturing, and examining specific cells such as cancer cells, stem cells, and T cells can be manufactured.

[0035] For the solvent, injection method, coating (spraying) method, etc., conventionally known materials and methods can be applied. The holding and drying times in (1) and (2) can be appropriately set according to the size of the glass substrate, the type of liquid introduced, etc. The holding time is preferably 10 seconds to 10 hours, more preferably 1 minute to 5 hours, and still more preferably 5 minutes to 2 hours. Drying is preferably carried out at room temperature (about 23 °C) to 80 °C, more preferably at room temperature to 60 °C. In addition, drying under reduced pressure is also possible. Further, after maintaining for a certain time, the remaining hydrophilic polymer solution / dispersion can be appropriately discharged and then dried.

[0036] The solvent is not particularly limited as long as it can dissolve the hydrophilic polymer, and it can be appropriately selected according to the hydrophilic polymer used. For example, water, organic solvents, and their mixed solvents can be cited. As the organic solvents, alcohols such as methanol, ethanol, n-propanol, isopropanol, and methoxypropanol; ketones such as acetone and methyl ethyl ketone; tetrahydrofuran, acetonitrile, vinyl acetate, toluene, etc. can be cited. In addition, the above solvents can also use the solvents after heating. The solvent can be used alone or in combination of two or more.

[0037] The concentration of the above-mentioned hydrophilic polymer solution / dispersion is not particularly limited and can be appropriately selected considering injectability, coatability, sprayability, productivity, etc. The concentration of the hydrophilic polymer in the hydrophilic polymer solution / dispersion (100% by mass) is preferably 0.01 to 10.0% by mass, more preferably 0.10 to 5.0% by mass.

[0038] The above-mentioned mixed hydrophilic polymer layer can also be a mixed hydrophilic polymer layer with a scaffold protein adsorbed on the surface of the layer. By further adsorbing a scaffold protein on the surface of the above-mentioned mixed hydrophilic polymer layer, the adsorption of specific cells such as cancer cells, stem cells, and T cells can be improved.

[0039] In addition, in this specification, "scaffold protein" means: a protein having a function of promoting the selective adsorption of a specific cell, for example, a protein having a function of specifically binding to a protein protruding from the surface of the specific cell, etc. In addition, it also refers to a protein having a function of interacting (adsorbing, binding, associating, etc.) with the above hydrophilic polymer, for example, a protein having a function of promoting the selective adsorption of the specific cell on the surface of the hydrophilic polymer by adsorbing to the above hydrophilic polymer, etc.

[0040] The above scaffold protein preferably has an RGD (arginine - glycine - aspartic acid) sequence. As the above scaffold protein, for example, fibronectin can be preferably used.

[0041] The method for adsorbing the above scaffold protein to the mixed hydrophilic polymer layer is not particularly limited, and known methods can be applied. For example, the mixed hydrophilic polymer layer can be brought into contact with a buffer solution (such as phosphate - buffered saline PBS) of the scaffold protein by a known method, left at a specified temperature for a specified time, and adsorbed by a method such as washing as needed. The temperature and time can be appropriately set. For example, it can be carried out at 10 - 60°C for about 0.1 - 24 hours.

[0042] From the aspect of adsorbing the above scaffold protein onto the mixed hydrophilic polymer layer, it is preferable to use a solution, dispersion, etc. in which the concentration of the scaffold protein is preferably adjusted to 0.5 - 500 μg / ml, more preferably adjusted to 1 - 250 μg / ml. By adjusting the concentration within the above range, excellent adsorption properties for specific cells can be obtained. In addition, the concentration of fibronectin is also preferably in the same range.

[0043] In the glass substrate of the coated polymer, it is preferable that the surface of the mixed hydrophilic polymer layer has a low elastic modulus in water or an aqueous solution. It is known that specific cells such as cancer cells, stem cells, and T cells are generally softer compared to normal cells such as blood cells. This is related to the fact that when specific cells such as cancer cells metastasize, the shape of the cells undergoes a large deformation and they move through gaps. Therefore, normal cells such as blood cells that are difficult to deform and are hard are difficult to be adsorbed onto the glass substrate of the coated polymer coated with a hydrophilic polymer with a soft surface. On the other hand, specific cells such as cancer cells with the ability to deform tend to be easily adsorbed onto the glass substrate of the coated polymer with a soft surface. In addition, when the glass substrate of the coated polymer is too soft, the adsorption property decreases. Therefore, by forming the above mixed hydrophilic polymer layer on the glass substrate, a hydrophilic polymer layer with high smoothness and low elastic modulus (high softness) can be formed, and excellent adsorption properties for specific cells such as cancer cells, stem cells, and T cells can be obtained.

[0044] From the viewpoint of being able to suppress the adsorption of normal cells such as blood cells and selectively adsorb specific cells such as cancer cells, stem cells, and T cells, the above-mentioned elastic modulus is preferably 1.20 MPa or less, more preferably 0.80 MPa or less, further preferably 0.50 MPa or less, and particularly preferably 0.30 MPa or less. The lower limit is preferably 0.01 MPa or more, more preferably 0.05 MPa or more.

[0045] The elastic modulus in the above-mentioned water or aqueous solution can be adjusted by changing the molecular weight and film thickness of the hydrophilic polymer forming the above-mentioned mixed hydrophilic polymer layer. Specifically, when the molecular weight of the hydrophilic polymer increases, the elastic modulus tends to increase. In addition, when the film thickness of the hydrophilic polymer layer increases, the elastic modulus tends to decrease.

[0046] In addition, in this specification, unless otherwise specified, the elastic modulus in water or aqueous solution refers to the elastic modulus in water or aqueous solution measured using an atomic force microscope (AFM).

[0047] An atomic force microscope (AFM) is one type of scanning probe microscope and is a microscope that detects the force between atoms acting on a specimen and a probe. The probe is installed at the tip of a cantilever (cantilever spring). While changing the distance between the specimen and the probe, the force (bending amount) acting on the cantilever is measured, and a curve (force curve) plotting the relationship between the two is obtained. By analyzing this force curve, the elastic modulus (hardness) of the specimen surface can be obtained, and the elastic modulus can be measured at the nanometer level. The method of obtaining the elastic modulus of the specimen surface by measuring the force curve is a method known to those skilled in the art, and the elastic modulus can be obtained by such a well-known method.

[0048] As a method for calculating the elastic modulus from the force curve, for example, a method of fitting the force curve according to the JKR (Johnson-Kendall-Roberts) theory and calculating the elastic modulus can be cited. In the JKR theory, when the adhesion energy is set to w, F applied to the cantilever and the specimen deformation amount δ are as shown in the following formulas (1) and (2).

Mathematical formula 1

[0049] By fitting the F-δ curve obtained by measuring the force curve and using formulas (1) and (2), the elastic modulus can be obtained.

[0050] Here, the elastic modulus in water or aqueous solution specifically represents the measured value obtained by the following method. The measured value in water or an aqueous solution can be measured by dropping water or an aqueous solution onto the surface of the sample to form a droplet (convex lens) and measuring it by AFM. As the aqueous solution, for example, phosphate buffered saline (PBS) can be preferably used.

[0051] Then, the elastic modulus of the sample (the surface of the above-mentioned mixed hydrophilic polymer layer), for example, can be obtained by scanning within a specified range on the surface of the sample, obtaining force curves at many points within the specified range, calculating the elastic modulus from each force curve, and calculating the average value thereof.

[0052] In the glass substrate coated with the polymer, by forming a hydrophilic polymer layer composed of a mixture of two or more hydrophilic polymers having different molecular weights on the surface of the glass substrate, the unevenness of the substrate surface can be controlled, and a hydrophilic polymer layer with a small surface roughness and high smoothness can be formed. Thus, excellent adsorption properties for specific cells such as cancer cells, stem cells, and T cells can be obtained. In addition, the elastic modulus of the surface of the formed mixed hydrophilic polymer layer is low, and thus, further excellent adsorption properties for specific cells can be obtained. Therefore, by measuring the number of adsorbed specific cells, the number of specific cells in the sampled blood or body fluid can be determined, which can be used to confirm the cancer treatment effect, etc. In addition, by culturing the adsorbed specific cells, it can be used to confirm the effects of anticancer agents, etc. and the selection of anticancer agents.

Examples

[0053] Hereinafter, the present invention will be specifically described based on examples, but the present invention is not limited thereto.

[0054] <Production of hydrophilic polymer> (Production of Polymer 1) Using an AIBN (azobisisobutyronitrile) 1.25 mg / ml toluene solution, 2-methoxyethyl acrylate (25 wt% toluene solution) was thermally polymerized at 60 °C for 7 hours to produce poly(2-methoxyethyl acrylate) (PMEA) (Mn 16,000).

[0055] (Production of Polymer 2) Using an AIBN (azobisisobutyronitrile) 1.25 mg / ml methanol solution, 2-methoxyethyl acrylate (50 wt% methanol solution) was thermally polymerized at 60 °C for 7 hours to produce poly(2-methoxyethyl acrylate) (PMEA) (Mn 80,000).

[0056] <Production of glass substrate coated with polymer> (Example 1) A solution prepared by mixing a 0.38 wt% methanol solution of PMEA (Mn 16,000) prepared in the production of the above polymer 1 (the methanol solution was heated to 40°C) and a 0.08 wt% methanol solution of PMEA (Mn 80,000) prepared in the production of the above polymer 2 (the methanol solution was heated to 40°C) at a mass ratio of 90:80 was injected with 85 μl (the methanol solution was heated to 40°C) into one well of a chamber slide (2-well type) (manufactured by Matsunami Glass Industry Co., Ltd., uncoated, bottom surface: made of soda-lime glass, contact angle of air bubbles in water: 109.9 degrees, average thickness 1.35 mm). Then, it was immediately vacuum-dried in an oven at 50°C for 20 minutes to produce a glass substrate coated with the polymer.

[0057] (Example 2) PMEA (Mn 16,000) prepared in the production of the above polymer 1 was dissolved in methanol at a concentration of 0.33 wt% (the methanol solution was heated to 40°C). Into the obtained methanol solution, PMEA (Mn 80,000) prepared in the production of the above polymer 2 was dissolved at a concentration of 0.04 wt% (the methanol solution was heated to 40°C). 85 μl of the prepared solution (the methanol solution was heated to 40°C) was injected into one well of a slide chamber (2-well type) (manufactured by Matsunami Glass Industry Co., Ltd., uncoated, bottom surface: made of soda-lime glass, contact angle of air bubbles in water: 109.9 degrees, average thickness 1.35 mm). Then, it was immediately vacuum-dried in an oven at 50°C for 20 minutes to produce a glass substrate coated with the polymer.

[0058] (Example 3) PMEA (Mn 16,000) prepared in the production of the above polymer 1 was dissolved in methanol at a concentration of 0.48 wt% (the methanol solution was heated to 40°C). Into the obtained methanol solution, PMEA (Mn 80,000) prepared in the production of the above polymer 2 was dissolved at a concentration of 0.13 wt% (the methanol solution was heated to 40°C). 85 μl of the prepared solution (the methanol solution was heated to 40°C) was injected into one well of a chamber slide (2-well type) (manufactured by Matsunami Glass Industry Co., Ltd., uncoated, bottom surface: made of soda-lime glass, contact angle of air bubbles in water: 109.9 degrees, average thickness 1.35 mm). Then, it was immediately vacuum-dried in an oven at 50°C for 20 minutes to produce a glass substrate coated with the polymer.

[0059] (Comparative Example 1) The PMEA (Mn 80,000) produced in the production of the above polymer 2 was dissolved at a concentration of 0.25 wt% (the methanol solution was heated to 40 °C). 85 μl of the prepared solution (the methanol solution heated to 40 °C) was injected into one well of a chamber slide (2-well type) (manufactured by Matsunami Glass Industry Co., Ltd., uncoated, bottom surface: made of soda-lime glass, contact angle of bubbles in water: 109.9 degrees, average thickness 1.35 mm). Then, it was immediately vacuum-dried in an oven at 50 °C for 20 minutes to obtain a glass substrate coated with the polymer.

[0060] For the glass substrates coated with the polymer produced in the above Examples and Comparative Examples, the presence or absence of cloudiness in the polymer layer, the elastic modulus of the surface based on AFM in PBS (phosphate-buffered aqueous solution), and the thickness of the hydrophilic polymer layer were measured by the following methods. The results are shown in Table 1.

[0061] [Thickness of hydrophilic polymer layer] The cross-section of the hydrophilic polymer layer was measured (photographed) using TEM (JEOL, JEM-2800) at an accelerating voltage of 200 kV.

[0062] [Presence or absence of cloudiness in polymer layer] Within 2 hours after coating, the surface was visually observed. Cloudiness indicates a surface with a large surface roughness. The less cloudiness, the higher the smoothness of the surface.

[0063] [Elastic modulus of surface] Phosphate-buffered saline was dropped onto the surface of the hydrophilic polymer layer of the glass substrate coated with the polymer to form a droplet (convex lens), and the elastic modulus of the surface was measured by the following method using the following device (AFM). The obtained elastic modulus was set as the elastic modulus measured in water or an aqueous solution. In addition, when measuring the elastic modulus, based on the obtained force curve, analysis was performed based on the JKR contact theory to obtain the elastic modulus. [Method for measuring elastic modulus] Device (AFM): MFP-3D-SA manufactured by Oxford Instruments Measurement mode: AFM force curve plotting Cantilever: Material: Si, tip curvature radius R = 150 nm, spring constant 0.67 N / m Measurement range: 20 μm × 20 μm scanning, elastic modulus calculated by the JKR two-point method Scanning speed: 1 Hz Measurement atmosphere: In PBS Measurement temperature: 23 °C The lower the elastic modulus, the better the adsorption performance of specific cells such as cancer cells.

[0064]

Table 1

[0065] The present invention (1) is: a glass substrate coated with a polymer, on the surface of which a hydrophilic polymer layer composed of a mixture of hydrophilic polymers with different molecular weights is formed.

[0066] The present invention (2) is: the glass substrate coated with a polymer according to the present invention (1), wherein the hydrophilic polymer contains hydrophilic polymers with a difference in number-average molecular weight of 30,000 or more.

[0067] The present invention (3) is: the glass substrate coated with a polymer according to the present invention (1), wherein the hydrophilic polymer contains hydrophilic polymers with a difference in number-average molecular weight of 50,000 or more.

[0068] The present invention (4) is: the glass substrate coated with a polymer according to any arbitrary combination of the present invention (1) to (3), wherein the hydrophilic polymer contains the hydrophilic polymer represented by the following formula (I) with different number-average molecular weights.

Chemical formula 5

[0069] The present invention (5) is: the glass substrate coated with a polymer according to any arbitrary combination of the present invention (1) to (3), wherein the hydrophilic polymer contains a copolymer of the hydrophilic monomer represented by the following formula (II) with different number-average molecular weights and other monomers.

Chemical formula 6

[0070] The present invention (6) is: the glass substrate coated with a polymer according to any arbitrary combination of the present invention (1) to (5), wherein the thickness of the hydrophilic polymer layer is 30 to 3000 nm.

[0071] The present invention (7) is: the glass substrate coated with a polymer according to any arbitrary combination of the present invention (1) to (6), wherein a scaffold protein is adsorbed on the surface of the hydrophilic polymer layer.

[0072] The present invention (8) is: a glass substrate coated with a polymer according to the present invention (7), wherein the scaffold protein is fibronectin.

Claims

1. A polymer-coated glass substrate, characterized in that: A hydrophilic polymer layer composed of a mixture of hydrophilic polymers having different molecular weights is formed on the surface of the glass substrate.

2. The polymer-coated glass substrate according to claim 1, wherein The hydrophilic polymer includes a hydrophilic polymer having a number average molecular weight difference of 30,000 or more.

3. The polymer-coated glass substrate according to claim 1, wherein The hydrophilic polymer includes a hydrophilic polymer having a number average molecular weight difference of 50,000 or more.

4. The polymer-coated glass substrate according to claim 1, wherein The hydrophilic polymer contains hydrophilic polymers represented by the following formula (I) having different number average molecular weights, In the formula, R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group, p represents 1 to 8, m represents 1 to 5, and n represents the number of repetitions.

5. The polymer-coated glass substrate according to claim 1, wherein The hydrophilic polymer comprises a copolymer of a hydrophilic monomer represented by the following formula (II) having different number average molecular weights and other monomers, In the formula, R 51 represents a hydrogen atom or a methyl group, R 52 represents an alkyl group, p represents 1 to 8, and m represents 1 to 5.

6. The polymer-coated glass substrate according to claim 1, wherein The thickness of the hydrophilic polymer layer is 30 to 3000 nm.

7. The polymer-coated glass substrate according to claim 1, wherein The scaffold protein is adsorbed on the surface of the hydrophilic polymer layer.

8. The polymer-coated glass substrate according to claim 7, wherein: The scaffold protein is fibronectin.

9. The polymer-coated glass substrate according to claim 1, wherein The elastic modulus of the surface of the hydrophilic polymer layer is 0.01 MPa to 1.20 MPa.

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