Polymer-coated glass substrate

Abstract

To provide a polymer-coated glass substrate having a surface with controlled asperity and low elasticity.SOLUTION: A polymer-coated glass substrate is provided, comprising a hydrophilic polymer layer made up of a blend of hydrophilic polymers with different molecular weights formed on a surface of a glass substrate.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a polymer-coated glass substrate.

Background Art

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

[0003] However, among special polymers, there are some that are difficult to produce a smooth surface by coating, and the surface unevenness affects the adsorption performance of specific cells. Therefore, it is desired to provide a substrate having a surface with excellent adsorption performance for specific cells such as cancer cells, stem cells, and T cells, in which the unevenness is controlled (see Patent Document 1, etc.). Furthermore, when the surface unevenness is large, clouding of the coating surface easily occurs, and it is desired to suppress the clouding of the coating surface by controlling the surface unevenness. In addition, by suppressing the clouding 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

Summary of the Invention

Problems to be Solved by the Invention

[0005] An object of the present invention is to solve the above problems and provide a polymer-coated glass substrate in which the surface unevenness is controlled and the surface elastic modulus is low.

Means for Solving the Problems

[0006] The present invention relates to a polymer-coated glass substrate having a hydrophilic polymer layer formed of a blend of hydrophilic polymers having different molecular weights on the surface of a glass substrate.

Effects of the Invention

[0007] According to the present invention, since it is a polymer-coated glass substrate having a hydrophilic polymer layer formed of a blend of hydrophilic polymers having different molecular weights on the surface of a glass substrate, the surface unevenness is controlled, and a polymer-coated glass substrate having a low surface elastic modulus can be provided. Therefore, the above polymer-coated glass substrate can improve the adsorption performance of specific cells such as cancer cells, stem cells, and T cells.

Modes for Carrying Out the Invention

[0008] The above polymer-coated glass substrate has a hydrophilic polymer layer formed of a blend of hydrophilic polymers having different molecular weights on the surface of a glass substrate. By forming a hydrophilic polymer layer on the surface of a glass substrate with a blend of hydrophilic polymers having different molecular weights, it becomes possible to control the unevenness of the substrate surface, and a polymer-coated glass substrate having a small surface roughness, high smoothness, and a low surface elastic modulus and coated with a hydrophilic polymer layer can be provided.

[0009] Tumor cells (such as cancer cells) that appear in body fluids such as circulating tumor cells in the blood (several to several hundred cells / mL of blood) are extremely few in number. For testing, it is considered important to adsorb as many tumor cells present in the collected body fluid as possible. The polymer-coated glass substrate forms a hydrophilic polymer layer composed of a blend of hydrophilic polymers with different molecular weights on the glass substrate, thereby controlling the surface unevenness, resulting in a hydrophilic polymer layer with a small surface roughness and high smoothness. In addition, the formed hydrophilic polymer layer has a low surface elastic modulus. The unevenness of the hydrophilic polymer layer affects the adsorption properties of specific cells such as cancer cells, stem cells, and T cells. By controlling the surface unevenness and increasing the smoothness, excellent adsorption performance of specific cells can be obtained. Also, the surface elastic modulus affects the adsorption properties of specific cells, and the lower the elastic modulus, the better the adsorption performance. On the other hand, if the surface elastic modulus is too low, the adsorption property will conversely decrease. That is, there is an optimal value for the surface elastic modulus. For example, by adsorbing tumor cells onto a hydrophilic polymer layer composed of a blend of hydrophilic polymers with different molecular weights formed on the surface of the glass substrate and measuring the number of adsorbed cells, the number of tumor cells in the body fluid can be determined and used for confirming the cancer treatment effect. Also, by culturing the adsorbed tumor cells and checking the effectiveness of anticancer agents and the like with the cultured cells, the effectiveness of anticancer agents and the like can be confirmed outside the body before administration, and at the same time, it can also be used for the selection of anticancer agents and the like. In addition, gene analysis can be performed on the adsorbed or cultured tumor cells to examine the mutation state and expression state of the tumor cells, which can be used for the selection of anticancer agents and the like and can also be helpful for elucidating the mechanism of cancer.

[0010] The type of glass constituting the polymer-coated glass substrate 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.), potassium glass, crystal glass (glass containing PbO, such as SiO2 - PbO-based glass, SiO2 - PbO - B2O3-based glass, SiO2 - B2O3 - PbO-based glass, etc.), titanium crystal glass, barium glass, boron glass (such as B2O3 - ZnO - PbO-based glass, B2O3 - ZnO - Bi2O3-based glass, B2O3 - Bi2O3-based glass, B2O3 - ZnO-based glass, etc.), strontium glass, alumina silicate glass, soda zinc glass, soda barium glass (such as BaO - SiO2-based glass, etc.) can be mentioned. These glasses may be used alone or two or more types may be mixed.

[0011] The glass substrate preferably has a contact angle of bubbles in water of 100 degrees or more and 120 degrees or less. The contact angle is more preferably 103 degrees or more and more preferably 117 degrees or less. In this specification, the "contact angle of bubbles in water" is measured by attaching a bubble (air) to the substrate surface in water and measuring the contact angle between the bubble and the substrate surface.

[0012] The thickness of the glass substrate is not particularly limited, but as an 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. The average thickness is a value obtained by measuring the thickness at any 10 locations using a micrometer and averaging the measured values.

[0013] The above polymer-coated glass substrate has a hydrophilic polymer layer (hereinafter also referred to as a "blended hydrophilic polymer layer") formed of a blend of hydrophilic polymers with different molecular weights on the surface of the above glass substrate. The hydrophilic polymers with different molecular weights include those containing two polymers of the same hydrophilic polymer with different molecular weights (for example, a blend 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 those containing polymers of different hydrophilic polymers (hydrophilic polymers with different structures) with different molecular weights (for example, a blend of hydrophilic polymers containing poly(2-methoxyethyl acrylate) with a number average molecular weight of 16,000 and polyacryloylmorpholine with a number average molecular weight of 80,000). Among them, from the viewpoint of obtaining more effects, it is desirable that the above blended hydrophilic polymer layer is formed of polymers of the same hydrophilic polymer with different molecular weights.

[0014] The hydrophilic polymers constituting the above blended hydrophilic polymer layer are not particularly limited. For example, known hydrophilic polymers can be appropriately selected. The above hydrophilic polymers may be used alone or in combination of two or more.

[0015] The above hydrophilic polymers can be produced by known methods. For example, they can be synthesized by polymerizing hydrophilic monomers by known methods using a solution of hydrophilic monomers constituting the hydrophilic polymers. The solvent of the hydrophilic monomer solution is not particularly limited. For example, the solvents described below can be used. Among them, toluene and methanol are preferred.

[0016] Examples of the above hydrophilic polymers include homopolymers and copolymers of one or more hydrophilic monomers, and copolymers of one or more hydrophilic monomers and one or more other monomers. In the above polymer-coated glass substrate, a blended hydrophilic polymer layer containing a blend of such hydrophilic polymers with different molecular weights is formed.

[0017] The above hydrophilic monomer is not particularly limited, and for example, various monomers having a hydrophilic group can be used. Examples of the hydrophilic group include known hydrophilic groups such as an amide group, a sulfate group, a sulfonic acid group, a carboxylic acid group, a hydroxyl group, an amino group, an amide group, and an oxyethylene group.

[0018] Specific examples of the above hydrophilic monomer include (meth)acrylic acid; (meth)acrylate; alkoxyalkyl (meth)acrylate such as methoxyethyl (meth)acrylate; hydroxyalkyl (meth)acrylate such as hydroxyethyl (meth)acrylate; (meth)acrylamide; (meth)acrylamide derivatives having a cyclic group such as (meth)acryloylmorpholine; and the like. Among them, (meth)acrylic acid, (meth)acrylate, alkoxyalkyl (meth)acrylate, and (meth)acrylamide derivatives having a cyclic group are preferable, alkoxyalkyl (meth)acrylate and (meth)acryloylmorpholine are more preferable, alkoxyalkyl (meth)acrylate is still more preferable, and 2-methoxyethyl acrylate is particularly preferable. These may be used alone or in combination of two or more.

[0019] The above other monomers may be appropriately selected within a range that does not inhibit the action and effect of the hydrophilic polymer. Specific examples of the above other monomers include, for example, aromatic monomers such as styrene, vinyl acetate, N-isopropylacrylamide that can impart temperature responsiveness, and the like. These may be used alone or in combination of two or more.

[0020] As the above homopolymer and copolymer, specifically, homopolymers composed of one kind of hydrophilic monomer such as polyacrylic acid, polyacrylate, polymethacrylic acid, polymethacrylate, polyacryloylmorpholine, polymethacryloylmorpholine, polyacrylamide, polymethacrylamide, polyalkoxyalkyl acrylate, polyalkoxyalkyl methacrylate, etc.; copolymers composed of two or more kinds of the above-exemplified hydrophilic monomers; copolymers composed of one or more kinds of the above-exemplified hydrophilic monomers and one or more kinds of the above-exemplified other monomers; and the like can be mentioned.

[0021] Among them, the hydrophilic polymer constituting the blend hydrophilic polymer layer preferably contains a hydrophilic polymer represented by the following formula (I) having different molecular weights. Here, the hydrophilic polymer represented by the following formula (I) having different molecular weights may contain two or more hydrophilic polymers having the same structural unit and different molecular weights (for example, poly(2-methoxyethyl acrylate) having a number average molecular weight of 16,000 and poly(2-methoxyethyl acrylate) having a number average molecular weight of 80,000), or may contain two or more hydrophilic polymers having different structural units and different molecular weights (for example, poly(2-methoxyethyl acrylate) having a number average molecular weight of 16,000 and poly(2-ethoxyethyl methacrylate) having a number average molecular weight of 80,000). The hydrophilic polymer represented by the following formula (I) may be either one kind or two or more kinds. [Chemical formula] (In the formula, R 51 represents a hydrogen atom or a methyl group, and R 52 represents an alkyl group. p represents 1 to 8, m represents 1 to 5, and n represents the number of repetitions.)

[0022] As the hydrophilic polymer represented by the above formula (I), for example, a hydrophilic polymer represented by the following formula (I-1) is preferable. The hydrophilic polymer represented by the following formula (I-1) may be either one kind or two or more kinds. [Chemical formula] (In the formula, R51 is a hydrogen atom or a methyl group, R 52 represents an alkyl group. m represents 1 to 5, and n represents the number of repetitions.)

[0023] As the above hydrophilic polymer, a copolymer of a hydrophilic monomer represented by the following formula (II) having different molecular weights and another monomer can also be preferably used. Here, the copolymer of a hydrophilic monomer represented by the following formula (II) having different molecular weights and another monomer may include two or more hydrophilic polymers having the same constitutional units and different molecular weights (for example, a copolymer of 2-methoxyethyl acrylate and styrene having a number average molecular weight of 16,000 and a copolymer of 2-methoxyethyl acrylate and styrene having a number average molecular weight of 80,000), or may include two or more hydrophilic polymers having different constitutional units and different molecular weights (for example, a copolymer of 2-methoxyethyl acrylate and styrene having a number average molecular weight of 16,000 and a copolymer of 2-methoxyethyl acrylate and vinyl acetate having a number average molecular weight of 80,000). The hydrophilic polymer represented by the following formula (II) may be any one or two or more kinds. The other monomer may be any one or two or more kinds.

[0024]

Chemical formula

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

Chemical formula

[0026] In the above formulas (I), (I-1), (II), and (II-1), R 52 The alkyl group 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 (the number of repeating units) is preferably 15 to 1500, more preferably 40 to 1200.

[0027] Among the above-mentioned hydrophilic polymers, from the viewpoints of controlling the unevenness of the substrate surface, forming a hydrophilic polymer layer with a small surface roughness and high smoothness, etc., 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] In the hydrophilic polymers with different molecular weights as described above, the molecular weight is not particularly limited, and examples include the number average molecular weight (Mn), weight average molecular weight (Mw), etc. Among them, a blend of hydrophilic polymers with different Mn can be preferably used.

[0029] When using a blend of hydrophilic polymers with different Mn, from the viewpoint of obtaining better effects, it is preferable to include hydrophilic polymers with a difference in Mn of 30,000 or more, and more preferably to include hydrophilic polymers with a difference in Mn of 50,000 or more.

[0030] The blend of hydrophilic polymers with different molecular weights as described above preferably includes a low molecular weight hydrophilic polymer with a number average molecular weight (Mn) of 5000 or more and 25000 or less from the viewpoint of obtaining better effects. The Mn of the above low molecular weight hydrophilic polymer is preferably 10000 or more, more preferably 14000 or more, and preferably 23000 or less. In addition, from the viewpoint of obtaining better effects, the blend of hydrophilic polymers with different molecular weights preferably contains a high-molecular-weight hydrophilic polymer having a number-average molecular weight (Mn) of 35,000 or more and 200,000 or less. The Mn of the high-molecular-weight hydrophilic polymer is preferably 40,000 or more, more preferably 50,000 or more, and preferably 150,000 or less, more preferably 130,000 or less.

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

[0032] The thickness of the blend hydrophilic polymer layer is preferably 30 to 3000 nm, more preferably 30 to 1500 nm, and still more preferably 50 to 1000 nm. By adjusting within the above range, there is a tendency to obtain more effects, and low adsorption to good proteins and cells, and selective adsorption to specific cells such as cancer cells, stem cells, and T cells can be expected.

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

[0034] The above blend hydrophilic polymer layer can be formed by known methods such as (1) injecting a hydrophilic polymer solution or dispersion in which two or more hydrophilic polymers with different molecular weights are dissolved and dispersed in various solvents onto the surface of a glass substrate (such as a substrate recess), holding for a predetermined time and drying as necessary, or (2) coating (spraying) the hydrophilic polymer solution or dispersion onto the surface of a glass substrate (such as a substrate recess) and drying as necessary. By forming a blend hydrophilic polymer layer on the surface of a glass substrate using such known methods, a polymer-coated glass substrate can be manufactured. Further, by adding other components to the polymer-coated glass substrate as necessary, an apparatus capable of adsorbing, culturing, and inspecting 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. (1) and (2)'s holding and drying times may be appropriately set according to the size of the glass substrate, the type of liquid to be introduced, etc. The holding time is preferably 10 seconds to 10 hours, more preferably 1 minute to 5 hours, and even more preferably 5 minutes to 2 hours. Drying is preferably performed at room temperature (about 23°C) to 80°C, and more preferably at room temperature to 60°C. Also, drying may be performed under reduced pressure. Further, after holding for a certain time, the excess hydrophilic polymer solution or dispersion may be appropriately discharged and then dried.

[0036] The solvent is not particularly limited as long as it can dissolve the hydrophilic polymer, and may be appropriately selected according to the hydrophilic polymer to be used. For example, water, organic solvents, and mixed solvents thereof can be mentioned. Examples of organic solvents include alcohols such as methanol, ethanol, n-propanol, i-propanol, and methoxypropanol; ketones such as acetone and methyl ethyl ketone; tetrahydrofuran, acetonitrile, ethyl acetate, toluene, etc. Also, a heated solvent may be used. The solvent may be used alone or in combination of two or more.

[0037] The concentration of the above hydrophilic polymer solution / dispersion is not particularly limited and may be appropriately selected in consideration of injectability, coatability, sprayability, productivity, etc. However, 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 blend hydrophilic polymer layer may have a scaffold protein adsorbed on the surface of the layer. By further adsorbing a scaffold protein on the surface of the above blend hydrophilic polymer layer, the adsorption of specific cells such as cancer cells, stem cells, and T cells can be improved.

[0039] In the present specification, the "scaffold protein" means a protein having a function of promoting the selective adsorption of the specific cells, and examples thereof include proteins having a function of specifically binding to a protein present on the surface of the specific cells. It also means a protein having a function of interacting (adsorbing, binding, associating, etc.) with the above hydrophilic polymer. For example, a protein that mediates the function of promoting the selective adsorption of the specific cells on the surface of the above hydrophilic polymer by adsorbing to the above hydrophilic polymer can be mentioned.

[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 a scaffold protein to the above blend hydrophilic polymer layer is not particularly limited, and a known method can be applied. For example, a buffer solution of a scaffold protein (such as phosphate buffered saline PBS) is brought into contact with the above blend hydrophilic polymer layer by a known method, left at a predetermined temperature for a predetermined time, and washed as necessary to adsorb it. The temperature and time can be appropriately set, and for example, it can be carried out at 10 to 60 °C for about 0.1 to 24 hours.

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

[0043] In the above polymer-coated glass substrate, it is desirable that the surface of the blend hydrophilic polymer layer has a lower elastic modulus in water or an aqueous solution. Compared with normal cells such as blood cells, specific cells such as cancer cells, stem cells, and T cells are generally known to be soft. This is related to the fact that when specific cells such as cancer cells metastasize, they greatly deform their cell shape and move through gaps. For this reason, normal cells such as blood cells that are difficult to deform and hard are less likely to be adsorbed onto a polymer-coated glass substrate coated with a hydrophilic polymer with a soft surface, while specific cells such as cancer cells that have acquired deformability tend to be easily adsorbed onto a polymer-coated glass substrate with a soft surface. Note that if the polymer-coated glass substrate is too soft, the adsorbability will decrease. Therefore, by forming the blend hydrophilic polymer layer on the glass substrate, a hydrophilic polymer layer with high smoothness and a low elastic modulus (high flexibility) is formed, and excellent adsorption performance can be obtained for specific cells such as cancer cells, stem cells, and T cells.

[0044] From the viewpoint of suppressing the adsorption of normal cells such as blood cells and selectively adsorbing specific cells such as cancer cells, stem cells, and T cells, the above elastic modulus is preferably 1.20 MPa or less, more preferably 0.80 MPa or less, still more 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 water or an aqueous solution can be adjusted by changing the molecular weight of the hydrophilic polymer forming the blend hydrophilic polymer layer and the film thickness. Specifically, as the molecular weight of the hydrophilic polymer increases, the elastic modulus tends to increase, and as the film thickness of the hydrophilic polymer layer increases, the elastic modulus tends to decrease.

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

[0047] The atomic force microscope (AFM) is a type of scanning probe microscope that detects the force acting between the atoms of a sample and a probe. The probe is attached to the tip of a cantilever (a single - supported spring), and while changing the distance between the sample and the probe, the force (deflection amount) acting on the cantilever is measured to obtain a curve (force curve) plotting their relationship. By analyzing this force curve, the elastic modulus (hardness) of the sample surface can be obtained, and the elastic modulus can be measured at the nanolevel. The method of obtaining the elastic modulus from the force curve is a method known to those skilled in the art, and it is possible to obtain the elastic modulus by such a known method.

[0048] As a method for calculating the elastic modulus from the force curve, for example, there is a method of fitting the force curve according to the JKR (Johnson - Kendall - Roberts) theory to calculate the elastic modulus. In the JKR theory, the force F applied to the cantilever and the sample deformation amount δ are represented by the following formulas (1) and (2) with the adhesion energy as w.

Equation

[0049] The elastic modulus can be obtained by fitting the F - δ curve obtained by force curve measurement with formulas (1) and (2).

[0050] Here, the elastic modulus in water or an aqueous solution specifically means a measured value measured by the following method. The measured value in water or an aqueous solution can be measured by AFM by dropping water or an aqueous solution onto the surface of the sample so that a liquid droplet (convex meniscus) is formed. As the aqueous solution, for example, phosphate buffered saline (PBS) can be preferably used.

[0051] And the elastic modulus of the sample (the surface of the blend hydrophilic polymer layer) can be obtained, for example, by scanning within a predetermined range on the sample surface to obtain force curves at a large number of points within the predetermined range, obtaining the elastic modulus from each force curve, calculating the average value thereof.

[0052] The polymer-coated glass substrate can control the surface unevenness by forming a hydrophilic polymer layer composed of a blend of two or more hydrophilic polymers having different molecular weights on the surface of the glass substrate, and a hydrophilic polymer layer with small surface roughness and high smoothness is formed. Thereby, excellent adsorption performance for specific cells such as cancer cells, stem cells, and T cells can be obtained. In addition, the formed blend hydrophilic polymer layer has a low elastic modulus on the surface, thereby obtaining even more excellent adsorption performance for specific cells. Therefore, by measuring the number of adsorbed specific cells, the number of specific cells in the collected blood or body fluid can be known and can be used for confirming the cancer treatment effect, etc. Also, by culturing the adsorbed specific cells, it can be used for confirming the efficacy of anticancer agents, etc. and selecting anticancer agents.

Example

[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 prepare poly(2-methoxyethyl acrylate) (PMEA) (Mn 16,000).

[0055] (Preparation 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 prepare poly(2-methoxyethyl acrylate) (PMEA) (Mn 80,000).

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

[0057] (Example 2) The PMEA (Mn 16,000) prepared in the preparation of the above polymer 1 was dissolved in methanol so that the concentration became 0.33 wt% (the methanol solution was heated to 40 °C). To the obtained methanol solution, the PMEA (Mn 80,000) prepared in the preparation of the above polymer 2 was dissolved so that the concentration became 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) (non-coated, manufactured by Matsunami Glass Industry Co., Ltd., 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 prepare a polymer-coated glass substrate.

[0058] (Example 3) The PMEA (Mn 16,000) prepared in the preparation of the above polymer 1 was dissolved in methanol so that the concentration became 0.48 wt% (the methanol solution was heated to 40 °C). To the obtained methanol solution, the PMEA (Mn 80,000) prepared in the preparation of the above polymer 2 was dissolved so that the concentration became 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 slide chamber (2-well type) (non-coated, manufactured by Matsunami Glass Industry Co., Ltd., 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 prepare a polymer-coated glass substrate.

[0059] (Comparative Example 1) The PMEA (Mn 80,000) prepared in the preparation of the above polymer 2 was dissolved so that the concentration became 0.25 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) (non-coated, manufactured by Matsunami Glass Industry Co., Ltd., 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 prepare a polymer-coated glass substrate.

[0060] Regarding the polymer-coated glass substrates prepared in the above Examples and Comparative Examples, the presence or absence of cloudiness in the polymer layer, the elastic modulus of the surface by AFM in PBS (phosphate buffer 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) at an acceleration voltage of 200 kV using TEM (JEOL, JEM-2800).

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

[0063] 〔Elastic modulus of the surface〕 Phosphate buffered saline was dropped onto the surface of the hydrophilic polymer layer of the polymer-coated glass substrate so that a droplet (convex meniscus) was formed, and the elastic modulus of the surface was measured by the following method using the following apparatus (AFM). The obtained elastic modulus was taken as the elastic modulus measured in water or an aqueous solution. When measuring the elastic modulus, analysis based on the JKR contact theory was performed on the obtained force curve to obtain the elastic modulus. <Method for measuring elastic modulus> Apparatus (AFM): MFP-3D-SA manufactured by Oxford Instruments Measurement mode: AFM force curve mapping Cantilever: Material: Si, tip radius of curvature R = 150 nm, spring constant 0.67 N / m Measurement range: 20 μm × 20 μm scan, elastic modulus calculated by JKR two-point method Scan 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 polymer-coated glass substrate having a hydrophilic polymer layer formed of a blend of hydrophilic polymers having different molecular weights on the surface of a glass substrate.

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

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

[0068] The present invention (4) is a polymer-coated glass substrate in any combination of the present inventions (1) to (3), wherein the hydrophilic polymer contains a hydrophilic polymer represented by the following formula (I) having different number average molecular weights. [Chemical formula] (In the formula, R 51 represents a hydrogen atom or a methyl group, and R 52 represents an alkyl group. p represents 1 to 8, m represents 1 to 5, and n represents the number of repetitions.)

[0069] The present invention (5) is a polymer-coated glass substrate in any combination of the present inventions (1) to (3), wherein the hydrophilic polymer contains a copolymer of a hydrophilic monomer represented by the following formula (II) and another monomer having different number average molecular weights. [Chemical formula] (In the formula, R 51 represents a hydrogen atom or a methyl group, and R 52 represents an alkyl group. p represents 1 to 8, and m represents 1 to 5.)

[0070] The present invention (6) is a polymer-coated glass substrate in any combination of any of the present inventions (1) to (5) wherein the thickness of the hydrophilic polymer layer is 30 to 3000 nm.

[0071] The present invention (7) is a polymer-coated glass substrate in any combination of any of the present inventions (1) to (6) wherein a scaffold protein is adsorbed on the surface of the hydrophilic polymer layer.

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

Claims

1. A polymer-coated glass substrate having a hydrophilic polymer layer formed on the surface of a glass substrate, the hydrophilic polymer layer being composed of a blend of hydrophilic polymers having different molecular weights.

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

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

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

5. The polymer-coated glass substrate according to Claim 1, wherein the hydrophilic polymer contains a copolymer of a hydrophilic monomer represented by the following formula (II) having different number average molecular weights and another monomer. [Chemical Formula 2] (wherein, R ５１ represents a hydrogen atom or a methyl group, and R ５２ represents an alkyl group. p represents an integer of 1 to 8, and m represents an integer of 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 a 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.

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