JP2023036217A - Copolymer, Surface Modifier, Composition, Medical Device, Silicone Substrate, and Cell Culture Vessel - Google Patents

Abstract

To provide a surface modifier or the like that can give biosubstance deposition inhibitory capacity excellent in the surface of versatile polymer material.SOLUTION: A surface modifier contains a copolymer having, in a side chain, a first structure that is at least one of an oxyethylene structure and a betaine structure, and also having, in a side chain, a second structure that is a dimethylsiloxane structure.SELECTED DRAWING: Figure 6

Description

The present invention relates to copolymers, surface modifiers, compositions, medical devices, silicone substrates, and cell culture vessels.

In recent years, microfabrication technology such as MEMS (Microelectromechanical Systems) technology has been used to develop various microdevices in which circuits and holes with a predetermined shape on the order of μm are fabricated on a substrate (chip). It is used for minute amount experiments, isolation, purification, analysis, etc. in devices and biochemical engineering.
The market for these microchannels is expected to grow to a scale of 10 billion to 20 billion dollars in the 2020s.

A device using a biological sample has a problem that cells or proteins derived from the biological sample adhere to the surface of the device, leading to clogging and a decrease in analysis accuracy. In order to solve such problems, it is possible to coat the surface of device circuits and substrate materials (for example, glass, metal-containing compounds or semi-metal-containing compounds, resins, etc.) with a simple operation, and to provide excellent biological materials. The use of a copolymer containing a specific anion structure and a specific cation structure has been reported as a coating agent capable of imparting adhesion-inhibiting properties (see, for example, Patent Document 1).

Microdevices are usually manufactured by bonding a circuit made of polydimethylsiloxane (hereinafter referred to as PDMS) PDMS or glass to a substrate made of glass or resin with an adhesive. The cured product) layer can also come into contact with the biological sample as well as the surface of the circuit or substrate. Therefore, it is desirable that the adhesive (or its cured product) used for devices using biological samples also have high hydrophilicity and biocompatibility.

It has been reported that protein adsorption was suppressed by adding DBE-712 (manufactured by Gelest) as a surface modifier when forming PDMS microchannels. In this study, the PDMS membrane surface to which the surface modifier was added showed inhibition of protein adsorption, but inhibition of cell adhesion was not achieved (see, for example, Non-Patent Document 1).

It has been reported that addition of polystyrene/oligoethylene oxide methacrylate copolymer as a surface modifier to polymethyl methacrylate and polystyrene inhibited protein adsorption and cell adhesion. In this research, the expression of biocompatibility was achieved by adding a surface modifier to a high concentration of 20% by mass (see, for example, Non-Patent Document 2).

Japanese Patent Application Laid-Open No. 2021-008618

A. Asatekin, et al. Sci. Rep. , 2019, 9, 1-14. H. Yokoyama, et al. Adv. Mater. 2005, 17, 2329-2332.

An object of the present invention is to provide a surface modifier capable of imparting excellent adhesion inhibitory properties to the surface of general-purpose polymeric materials, and a composition capable of imparting excellent adhesion inhibitory properties to the surfaces of general-purpose polymeric materials. It is to provide things.

The present inventors have found that a copolymer having a specific structure can provide the surface of a general-purpose polymer material with an excellent ability to suppress adhesion of biological substances, and have completed the present invention.
The present invention is as follows.

（式（ａ１－１）中、Ｒ^１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^２は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表し、ｍは、１～１００を表す。＊は結合手を表す。
式（ａ１－２）中、Ｒ^１１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^１２は、炭素原子数１～６のアルキレン基を表し、Ｒ^１３及びＲ^１４は、それぞれ独立して、炭素原子数１～４のアルキル基を表し、Ｒ^１５は、炭素原子数１～６のアルキレン基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表す。＊は結合手を表す。
式（ａ１－３）中、Ｒ^２１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^２２は、炭素原子数１～６のアルキレン基を表し、Ｒ^２３及びＲ^２４は、それぞれ独立して、炭素原子数１～４のアルキル基を表し、Ｒ^２５は、炭素原子数１～６のアルキレン基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表す。＊は結合手を表す。
式（ａ１－４）中、Ｒ^３１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^３２は、炭素原子数１～６のアルキレン基を表し、Ｒ^３３は、炭素原子数１～６のアルキレン基を表し、Ｒ^３４～Ｒ^３６は、それぞれ独立して、炭素原子数１～４のアルキル基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表す。＊は結合手を表す。）

（式（ａ２－１）中、Ｒ^４１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^４２は、炭素原子数１～６のアルキレン基を表し、Ｒ^４３は、１価の有機基を表し、ｍは１～１００を表す。＊は結合手を表す。
式（ａ２－２）中、Ｒ^５１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^５２は、炭素原子数１～６のアルキレン基を表す。＊は結合手を表す。）
［３］ 前記共重合体における、前記繰り返し単位（ａ１）と前記繰り返し単位（ａ２）とのモル比（ａ１：ａ２）が、９９：１～１０：９０である、［２］に記載の表面改質剤。
［４］ 前記共重合体が、ブロック共重合体である、［１］～［３］のいずれかに記載の表面改質剤。
［５］ 生体物質の付着抑制に用いられる、［１］～［４］のいずれかに記載の表面改質剤。
［６］ 前記共重合体の数平均分子量（Ｍｎ）が、１，０００～１００，０００である、［１］～［５］のいずれかに記載の表面改質剤。
［７］ ［１］～［６］のいずれかに記載の表面改質剤を含有する、組成物。
［８］ 生体物質の付着抑制に用いられる、［７］に記載の組成物。
［９］ 更に樹脂を含有する、［７］又は［８］に記載の組成物。
［１０］ ［７］～［９］のいずれかに記載の組成物から形成される膜を有する、医療用デバイス。
［１１］ ［７］～［９］のいずれかに記載の組成物から形成される膜を有する、シリコーン基材。
［１２］ ［７］～［９］のいずれかに記載の組成物から形成される膜を有する、細胞培養容器。
［１３］ 繰り返し単位（ａ１）及び繰り返し単位（ａ２）を有し、
前記繰り返し単位（ａ１）が、下記式（ａ１－１）で表される繰り返し単位～（ａ１－４）で表される繰り返し単位の少なくともいずれかを有し、
前記繰り返し単位（ａ２）が、下記式（ａ２－１）で表される繰り返し単位及び（ａ２－２）で表される繰り返し単位の少なくともいずれかを有する、
共重合体。

（式（ａ１－１）中、Ｒ^１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^２は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表し、ｍは、１～１００を表す。＊は結合手を表す。
式（ａ１－２）中、Ｒ^１１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^１２は、炭素原子数１～６のアルキレン基を表し、Ｒ^１３及びＲ^１４は、それぞれ独立して、炭素原子数１～４のアルキル基を表し、Ｒ^１５は、炭素原子数１～６のアルキレン基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表す。＊は結合手を表す。
式（ａ１－３）中、Ｒ^２１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^２２は、炭素原子数１～６のアルキレン基を表し、Ｒ^２３及びＲ^２４は、それぞれ独立して、炭素原子数１～４のアルキル基を表し、Ｒ^２５は、炭素原子数１～６のアルキレン基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表す。＊は結合手を表す。
式（ａ１－４）中、Ｒ^３１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^３２は、炭素原子数１～６のアルキレン基を表し、Ｒ^３３は、炭素原子数１～６のアルキレン基を表し、Ｒ^３４～Ｒ^３６は、それぞれ独立して、炭素原子数１～４のアルキル基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表す。＊は結合手を表す。）

（式（ａ２－１）中、Ｒ^４１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^４２は、炭素原子数１～６のアルキレン基を表し、Ｒ^４３は、１価の有機基を表し、ｍは１～１００を表す。＊は結合手を表す。
式（ａ２－２）中、Ｒ^５１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^５２は、炭素原子数１～６のアルキレン基を表す。＊は結合手を表す。）
［１４］ ブロック共重合体である、［１３］に記載の共重合体。
［１５］ 生体物質の付着抑制に用いられる、［１３］又は［１４］に記載の共重合体。
［１６］ 数平均分子量（Ｍｎ）が、１，０００～１００，０００である、［１３］～［１５］のいずれかに記載の共重合体。
［１７］ 前記繰り返し単位（ａ１）と前記繰り返し単位（ａ２）とのモル比（ａ１：ａ２）が、９９：１～１０：９０である、［１３］～［１６］のいずれかに記載の共重合体。 [1] a first structure having at least one of an oxyethylene structure and a betaine structure in a side chain;
a second structure that is a dimethylsiloxane structure in the side chain;
A surface modifier containing a copolymer having
[2] The copolymer has repeating units (a1) having the first structure and repeating units (a2) having the second structure,
The repeating unit (a1) has at least one of repeating units represented by the following formulas (a1-1) to (a1-4),
The repeating unit (a2) has at least one of a repeating unit represented by the following formula (a2-1) and a repeating unit represented by (a2-2).
The surface modifier according to [1].

(In formula (a1-1), R ¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ² represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and X is -O- or -NR- (R represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms), m represents 1 to 100, * represents a bond.
In formula (a1-2), R ¹¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ¹² represents an alkylene group having 1 to 6 carbon atoms, and R ¹³ and R ¹⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ¹⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-3), R ²¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ²² represents an alkylene group having 1 to 6 carbon atoms, and R ²³ and R ²⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ²⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-4), R ³¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ³² represents an alkylene group having 1 to 6 carbon atoms, and R ³³ represents the number of carbon atoms. represents an alkylene group of 1 to 6, each of R ³⁴ to R ³⁶ independently represents an alkyl group having 1 to 4 carbon atoms, X is —O— or —NR— (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond. )

(In formula (a2-1), R ⁴¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ⁴² represents an alkylene group having 1 to 6 carbon atoms, and R ⁴³ represents a monovalent represents an organic group, m represents 1 to 100. * represents a bond.
In formula (a2-2), R ⁵¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and R ⁵² represents an alkylene group having 1 to 6 carbon atoms. * represents a bond. )
[3] The surface according to [2], wherein the molar ratio (a1:a2) between the repeating unit (a1) and the repeating unit (a2) in the copolymer is 99:1 to 10:90. Modifier.
[4] The surface modifier according to any one of [1] to [3], wherein the copolymer is a block copolymer.
[5] The surface modifier according to any one of [1] to [4], which is used for suppressing adhesion of biological substances.
[6] The surface modifier according to any one of [1] to [5], wherein the copolymer has a number average molecular weight (Mn) of 1,000 to 100,000.
[7] A composition containing the surface modifier according to any one of [1] to [6].
[8] The composition according to [7], which is used for suppressing adhesion of biological substances.
[9] The composition according to [7] or [8], which further contains a resin.
[10] A medical device having a film formed from the composition according to any one of [7] to [9].
[11] A silicone substrate having a film formed from the composition according to any one of [7] to [9].
[12] A cell culture vessel having a membrane formed from the composition according to any one of [7] to [9].
[13] having a repeating unit (a1) and a repeating unit (a2),
The repeating unit (a1) has at least one of repeating units represented by the following formulas (a1-1) to (a1-4),
The repeating unit (a2) has at least one of a repeating unit represented by the following formula (a2-1) and a repeating unit represented by (a2-2).
copolymer.

(In formula (a1-1), R ¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ² represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and X is -O- or -NR- (R represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms), m represents 1 to 100, * represents a bond.
In formula (a1-2), R ¹¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ¹² represents an alkylene group having 1 to 6 carbon atoms, and R ¹³ and R ¹⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ¹⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-3), R ²¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ²² represents an alkylene group having 1 to 6 carbon atoms, and R ²³ and R ²⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ²⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-4), R ³¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ³² represents an alkylene group having 1 to 6 carbon atoms, and R ³³ represents the number of carbon atoms. represents an alkylene group of 1 to 6, each of R ³⁴ to R ³⁶ independently represents an alkyl group having 1 to 4 carbon atoms, X is —O— or —NR— (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond. )

(In formula (a2-1), R ⁴¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ⁴² represents an alkylene group having 1 to 6 carbon atoms, and R ⁴³ represents a monovalent represents an organic group, m represents 1 to 100. * represents a bond.
In formula (a2-2), R ⁵¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and R ⁵² represents an alkylene group having 1 to 6 carbon atoms. * represents a bond. )
[14] The copolymer according to [13], which is a block copolymer.
[15] The copolymer according to [13] or [14], which is used for suppressing adhesion of biological substances.
[16] The copolymer according to any one of [13] to [15], which has a number average molecular weight (Mn) of 1,000 to 100,000.
[17] According to any one of [13] to [16], wherein the molar ratio (a1:a2) between the repeating unit (a1) and the repeating unit (a2) is 99:1 to 10:90. copolymer.

INDUSTRIAL APPLICABILITY According to the present invention, a surface modifier capable of imparting an excellent ability to suppress the adhesion of biological substances to the surface of a general-purpose polymer material, and an agent capable of imparting an excellent ability to suppress the adhesion of a biological substance to the surface of a general-purpose polymer material. A composition can be provided.

Atomic force microscope (AFM) images of EbS31-containing resin films obtained from each EbS31-containing resin composition 1-1 to 1-4 of Example 1 (height image and phase contrast image) Results of X-ray photoelectron spectroscopy (XPS) measurement of the surface of PMMA film containing EbS31 and PMMA film not containing EbS31 Results of X-ray photoelectron spectroscopy (XPS) measurement of the surface of PS film containing EbS31 and PS film not containing EbS31 Results of X-ray photoelectron spectroscopy (XPS) measurement of the surface of EbS31-containing PDMS film and EbS31-free PDMS film Results of X-ray photoelectron spectroscopy (XPS) measurement of the surface of the EbS31-containing PP film and the EbS31-free PP film Cell adhesion test results for EbS31-containing PMMA membrane, EbS31-containing PS membrane, EbS31-free PMMA membrane, and EbS31-free PS membrane Cell adhesion test results for EbS31-containing PDMS membrane, EbS31-containing PP membrane, EbS31-free PDMS membrane, and EbS31-free PP membrane AFM images of PMMA films with different EbS31 contents (height images (a) to (d) and phase contrast images (e) to (h)) AFM images of PS films with different EbS31 contents (height images (a) to (d) and phase contrast images (e) to (h)) Results of X-ray photoelectron spectroscopy (XPS) measurement of the surface of PMMA films with different EbS31 contents Results of X-ray photoelectron spectroscopy (XPS) measurement of the surface of PS films with different EbS31 contents Results of cell adhesion experiments on PMMA membranes with different EbS31 contents Cell adhesion experiment results of PEOMA-containing PMMA membrane, EbS31-containing PMMA membrane, PSiOMA-containing PMMA membrane, DBE-712-containing PMMA membrane, and PMMA membrane Cell adhesion test results for EbS31-containing PMMA membrane, EbS80-containing PMMA membrane, ErS31-containing PMMA membrane, PMMA membrane, and PS membrane

(Surface modifier)
The surface modifier of the present invention contains a copolymer and, if necessary, other components.
Surface modifiers are used, for example, to suppress adhesion of biological substances.
Copolymers are also subject of the present invention.
The surface modifier may be the copolymer itself.

<Copolymer>
A copolymer has a 1st structure and a 2nd structure in a side chain.
The first structure is at least one of an oxyethylene structure and a betaine structure.
The second structure is a dimethylsiloxane structure.
An oxyethylene structure and a betaine structure have common properties in that a polymer having that structure functions as an antifouling polymer (see, for example, Polymer Journal (2014) 46, 436-443).

Copolymers are, for example, addition polymers. Examples of addition polymers include polymers of monomers having polymerizable unsaturated groups.

An oxyethylene structure refers to a —OCH ₂ CH ₂ — structure.
A dimethylsiloxane structure refers to a —Si(CH ₃ )(CH ₃ )—O— structure.

A betaine structure refers to a structure having a positive charge and a negative charge in the same molecule and having neutralized charges. The betaine structure has positive and negative charges, preferably at non-adjacent positions, and preferably at positions across one or more atoms.
The betaine structure includes, for example, a sulfobetaine structure, a carboxybetaine structure, a phosphobetaine structure, and the like.
The sulfobetaine structure is due to the dissociated sulfo group of the negative charge of the betaine structure.
The carboxybetaine structure is due to the dissociated carboxy group of the negative charge of the betaine structure.
The phosphobetaine structure is due to the dissociated phosphate group of the negative charge of the betaine structure.

The copolymer preferably has repeating units (a1) having the first structure and repeating units (a2) having the second structure.
The repeating unit (a1) has at least one of repeating units represented by the following formulas (a1-1) to (a1-4).
The repeating unit (a2) has at least one of a repeating unit represented by the following formula (a2-1) and a repeating unit represented by (a2-2).
The copolymer may have other repeating units.

（式（ａ１－１）中、Ｒ^１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^２は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表し、ｍは、１～１００を表す。＊は結合手を表す。
式（ａ１－２）中、Ｒ^１１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^１２は、炭素原子数１～６のアルキレン基を表し、Ｒ^１３及びＲ^１４は、それぞれ独立して、炭素原子数１～４のアルキル基を表し、Ｒ^１５は、炭素原子数１～６のアルキレン基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表す。＊は結合手を表す。
式（ａ１－３）中、Ｒ^２１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^２２は、炭素原子数１～６のアルキレン基を表し、Ｒ^２３及びＲ^２４は、それぞれ独立して、炭素原子数１～４のアルキル基を表し、Ｒ^２５は、炭素原子数１～６のアルキレン基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表す。＊は結合手を表す。
式（ａ１－４）中、Ｒ^３１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^３２は、炭素原子数１～６のアルキレン基を表し、Ｒ^３３は、炭素原子数１～６のアルキレン基を表し、Ｒ^３４～Ｒ^３６は、それぞれ独立して、炭素原子数１～４のアルキル基を表し、Ｘは、－Ｏ－又は－ＮＲ－（Ｒは、水素原子又は炭素原子数１～４のアルキル基を表す。）を表す。＊は結合手を表す。）

(In formula (a1-1), R ¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ² represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and X is -O- or -NR- (R represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms), m represents 1 to 100, * represents a bond.
In formula (a1-2), R ¹¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ¹² represents an alkylene group having 1 to 6 carbon atoms, and R ¹³ and R ¹⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ¹⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-3), R ²¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ²² represents an alkylene group having 1 to 6 carbon atoms, and R ²³ and R ²⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ²⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-4), R ³¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ³² represents an alkylene group having 1 to 6 carbon atoms, and R ³³ represents the number of carbon atoms. represents an alkylene group of 1 to 6, each of R ³⁴ to R ³⁶ independently represents an alkyl group having 1 to 4 carbon atoms, X is —O— or —NR— (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond. )

（式（ａ２－１）中、Ｒ^４１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^４２は、炭素原子数１～６のアルキレン基を表し、Ｒ^４３は、１価の有機基を表し、ｍは１～１００を表す。＊は結合手を表す。
式（ａ２－２）中、Ｒ^５１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^５２は、炭素原子数１～６のアルキレン基を表す。＊は結合手を表す。）

(In formula (a2-1), R ⁴¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ⁴² represents an alkylene group having 1 to 6 carbon atoms, and R ⁴³ represents a monovalent represents an organic group, m represents 1 to 100. * represents a bond.
In formula (a2-2), R ⁵¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and R ⁵² represents an alkylene group having 1 to 6 carbon atoms. * represents a bond. )

Examples of alkyl groups having 1 to 5 carbon atoms in R ¹ , R ² , R ¹¹ , R ²¹ , R ³¹ , R ⁴¹ and R ⁵¹ include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t-butyl group, n-pentyl group, 1-methylbutyl group, 2-methylbutyl group, 3-methylbutyl group, 1,1-dimethylpropyl group, 1,2- dimethylpropyl group, 2,2-dimethylpropyl group, 1-ethylpropyl group and the like.
R ¹ , R ¹¹ , R ²¹ , R ³¹ , R ⁴¹ and R ⁵¹ are preferably a hydrogen atom, a methyl group or an ethyl group, more preferably a hydrogen atom or a methyl group.
R ² is preferably a hydrogen atom or a methyl group.

Examples of the alkylene group having 1 to 6 carbon atoms in R ¹² , R ¹⁵ , R ²² , R ²⁵ , R ³² , R ³³ , R ⁴² and R ⁵² include methylene group, ethylene group, propylene group and trimethylene group. , tetramethylene group, 1-methylpropylene group, 2-methylpropylene group, dimethylethylene group, ethylethylene group, pentamethylene group, 1-methyl-tetramethylene group, 2-methyl-tetramethylene group, 1,1-dimethyl -trimethylene group, 1,2-dimethyl-trimethylene group, 2,2-dimethyl-trimethylene group, 1-ethyl-trimethylene group, hexamethylene group and the like.
The alkylene group having 1 to 6 carbon atoms may be a linear alkylene group or a branched alkylene group.
Among these, the alkylene group having 1 to 6 carbon atoms is preferably a methylene group, an ethylene group, a triethylene group, or a tetramethylene group.

Examples of alkyl groups having 1 to 4 carbon atoms in R ¹³ , R ¹⁴ , R ²³ , R ²⁴ and R ³⁴ to R ³⁶ include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t-butyl group and the like.
Among these, the alkyl group having 1 to 4 carbon atoms is preferably a methyl group or an ethyl group, more preferably a methyl group.

The monovalent organic group for R ⁴³ may be linear, branched, or cyclic.
Examples of monovalent organic groups include alkyl groups, alkenyl groups, alkynyl groups, and aryl groups, with alkyl groups being preferred.
The alkyl group when R ⁴³ represents an alkyl group is preferably an alkyl group having 1 to 12 carbon atoms, more preferably an alkyl group having 1 to 6 carbon atoms, and particularly an alkyl group having 1 to 4 carbon atoms. preferable. Examples of alkyl groups having 1 to 4 carbon atoms include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group and t-butyl group. Among these, a methyl group is preferred.
When R ⁴³ represents an alkenyl group, the alkenyl group includes, for example, a vinyl group, an allyl group and a 2-propenyl group, with the vinyl group being preferred.
When R ⁴³ represents an alkynyl group, the alkynyl group includes, for example, an ethynyl group, a propargyl group and a prop-2-yn-1-yl group, with the ethynyl group being preferred.
When R ⁴³ represents an aryl group, the aryl group includes, for example, a phenyl group, a tolyl group, a pyridyl group and the like, with the phenyl group being preferred.

When X in formulas (a1-1) to (a1-4) is —NR—, R represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms.
Examples of the alkyl group having 1 to 4 carbon atoms in R include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t-butyl group and the like. be done.
R is preferably a hydrogen atom.

Monomers from which the repeating unit represented by formula (a1-1) can be derived include, for example, 2-hydroxyethyl (meth)acrylate, methoxyethyl (meth)acrylate, ethylene oxide (meth)acrylate, and ethylene glycol (meth)acrylate. etc.
In addition, in this specification, "(meth)acrylate" means an acrylate or a methacrylate.

Examples of monomers from which the repeating unit represented by formula (a1-2) can be derived include the following monomers.

Examples of commercially available monomers from which repeating units represented by formula (a1-2) can be derived include M1971, M3295, A3367, B6130, U0119, M3296, A3361 manufactured by Tokyo Kasei Kogyo.

Examples of monomers from which the repeating unit represented by formula (a1-3) can be derived include the following monomers.

Commercially available monomers from which the repeating unit represented by formula (a1-3) can be derived include, for example, M3185, M2359 and A3279 manufactured by Tokyo Kasei Kogyo.

Examples of monomers from which the repeating unit represented by formula (a1-4) can be derived include the following monomers.

Commercially available monomers from which the repeating unit represented by formula (a1-4) can be derived include, for example, M2005 manufactured by Tokyo Kasei Kogyo.

Examples of commercially available monomers capable of deriving the repeating unit represented by formula (a2-1) include X-22-2404, X-22-174ASX, X-22-174BX and KF manufactured by Shin-Etsu Chemical Co., Ltd. -2012, X-22-2426; Silaplane FM-711 and FM-0721 manufactured by JNC.

Commercially available monomers from which the repeating unit represented by formula (a2-2) can be derived include, for example, Silaplane TM-0701T manufactured by JNC.

The mol% of the repeating unit (a1) with respect to the total repeating units of the copolymer is not particularly limited as long as it exhibits the effect of the present invention, but is preferably 10 to 99 mol%, more preferably 20 to 95 mol%. Preferably, 30 to 90 mol % is particularly preferred.

The mol% of the repeating unit (a2) with respect to the total repeating units of the copolymer is not particularly limited as long as the effect of the present invention is exhibited, but is preferably 1 to 90 mol%, more preferably 5 to 80 mol%. 10 to 70 mol % is particularly preferred.

The molar ratio (a1:a2) between the repeating unit (a1) and the repeating unit (a2) in the copolymer is not particularly limited as long as the effect of the present invention is exhibited, but is 99:1 to 10:90. is preferred, 95:5 to 80:20 is more preferred, and 90:10 to 30:70 is particularly preferred.

When the copolymer has repeating units (a1) and repeating units (a2), the copolymer may have repeating units other than the repeating units (a1) and (a2).
When the copolymer has the repeating unit (a1) and the repeating unit (a2), the total mol% of the repeating unit (a1) and the repeating unit (a2) with respect to the total repeating units of the copolymer is the effect of the present invention. Although it is not particularly limited as long as it exhibits the above, it is preferably 50 mol% or more, more preferably 70 mol% or more, even more preferably 90 mol% or more, and particularly preferably 99 mol% or more.

The copolymer may be a random copolymer or a block copolymer, but a block copolymer is preferable because the effects of the present invention are more excellent.
The block copolymer may be a diblock copolymer, a triblock copolymer, or a copolymer having 4 or more blocks.
When the copolymer is a block copolymer, it has a block (A1) having repeating units (a1) and a block (A2) having repeating units (a2).
The molar ratio % of the repeating unit (a1) to all repeating units in the block (A1) is not particularly limited as long as the effect of the present invention is exhibited, but is preferably 50 mol % or more, and 70 mol % or more. More preferably, 90 mol % or more is even more preferable, and 99 mol % or more is particularly preferable.
The molar ratio % of repeating units (a2) to all repeating units in block (A2) is not particularly limited as long as the effect of the present invention is exhibited, but is preferably 50 mol % or more, and 70 mol % or more. More preferably, 90 mol % or more is even more preferable, and 99 mol % or more is particularly preferable.

The molecular weight of the copolymer is not particularly limited as long as it exhibits the effects of the present invention, but the number average molecular weight (Mn) is preferably 1,000 to 100,000, more preferably 5,000 to 50,000. More preferably, 10,000 to 30,000 is particularly preferred.
The polydispersity [weight average molecular weight (Mw)/number average molecular weight (Mn)] of the copolymer is not particularly limited as long as the effect of the present invention is exhibited, but is preferably 2.00 or less. 50 or less is more preferable.
The molecular weight of the copolymer can be determined by size exclusion chromatography (SEC).

<<Method for producing copolymer>>
The method for producing the copolymer is not particularly limited.
A copolymer can be produced, for example, by polymerization using a monomer having a first structure and a monomer having a second structure.
When the monomer is a (meth)acrylate monomer, the copolymer can be produced by subjecting it to radical polymerization, anionic polymerization, cationic polymerization, etc., which are general methods for synthesizing acrylic polymers or methacrylic polymers. Various polymerization methods such as solution polymerization, suspension polymerization, emulsion polymerization and bulk polymerization are possible. Among them, it is preferable to synthesize by a living radical polymerization method, but it is not limited to this method.

A living radical polymerization method is a catalytic reaction using a metal complex or a non-metallic organic catalyst. When a metal complex is used, the metal complex typically reacts with a polymerization initiator such as an organic halide compound to generate a radical species that initiates polymerization with the monomer. Examples of metal salts that form metal complexes here include copper (I) chloride, (II), copper (I) bromide, (II), iron chloride (II), (III), iron bromide (II ), (III), cobalt (II) chloride, cobalt (II) bromide, titanium (IV) chloride, ruthenium complex compounds, etc. Examples of ligands that form metal complexes include tris[2- (dimethylamino)ethyl]amine, tris(2-pyridylmethyl)amine (TPMA), 1,1,4,7,10,10-hexamethyltriethylenetetramine, N,N,N',N'',N Nitrogen-containing ligands such as ''-pentamethyldiethylenetriamine (PMDETA), n-butylamine and the like can be mentioned. Examples of polymerization initiators include organic halogen compounds such as 2-bromoisobutyric anhydride, methyl 2-bromoisobutyrate, ethyl 2-bromoisobutyrate, and propargyl 2-bromoisobutyrate. A copolymer having a predetermined molecular weight and a narrow molecular weight distribution can be obtained by carrying out living radical polymerization using these in accordance with a conventional method. One specific example of such a method is described in the Examples below.

<Biomaterial>
In the present invention, biological substances include proteins, sugars, viruses, nucleic acids, cells, and combinations of two or more thereof.

Examples of proteins include fibrinogen, bovine serum albumin (BSA), human albumin, various globulins, β-lipoproteins, various antibodies (IgG, IgA, IgM), peroxidase, various complements, various lectins, fibronectin, lysozyme, phone • Willebrand factor (vWF), serum γ-globulins, pepsin, ovalbumin, insulin, histones, ribonucleases, collagen, cytochrome c.

Sugars include, for example, glucose, galactose, mannose, fructose, heparin, and hyaluronic acid.

Examples of nucleic acids include deoxyribonucleic acid (DNA) and ribonucleic acid (RNA).

Cells include, for example, fibroblasts, bone marrow cells, B lymphocytes, T lymphocytes, neutrophils, erythrocytes, platelets, macrophages, monocytes, osteocytes, pericytes, dendritic cells, keratinocytes, adipocytes, Mesenchymal cells, epithelial cells, epidermal cells, endothelial cells, vascular endothelial cells, liver parenchymal cells, chondrocytes, cumulus cells, nervous system cells, glial cells, neurons, oligodendrocytes, microglia, astrocytes, heart cells, esophageal cells, muscle cells (e.g., smooth muscle cells or skeletal muscle cells), pancreatic beta cells, melanocytes, hematopoietic progenitor cells, mononuclear cells, embryonic stem cells (ES cells), embryonic tumor cells, embryonic germ Stem cells, induced pluripotent stem cells (iPS cells), neural stem cells, hematopoietic stem cells, mesenchymal stem cells, liver stem cells, pancreatic stem cells, muscle stem cells, germ stem cells, intestinal stem cells, cancer stem cells, hair follicle stem cells, and various cell lines ( For example, HCT116, Huh7, HEK293 (human embryonic kidney cells), HeLa (human cervical cancer cell line), HepG2 (human liver cancer cell line), UT7/TPO (human leukemia cell line), CHO (Chinese hamster ovary cell line). , MDCK, MDBK, BHK, C-33A, HT-29, AE-1, 3D9, Ns0/1, Jurkat, NIH3T3, PC12, S2, Sf9, Sf21, High Five, Vero) and the like.

The surface modifier of the present invention may be the copolymer itself, or may contain components other than the copolymer.

The method of using the surface modifier is not particularly limited. A method of transferring the surface modifier to the molded resin surface by pouring the resin by forming or the like can be mentioned.
Moreover, the method of using the surface modifier includes, for example, the method of using it by including it in the following composition.

(Composition)
The composition of the present invention contains a surface modifier and, if necessary, may contain other components such as a resin (excluding the above copolymer) and a solvent.

Hereinafter, as a method of using the composition of the present invention, there are, for example, the following two methods of use.
(1) A method of applying the composition of the present invention containing a resin and a surface modifier to form a resin film in which a copolymer is segregated on the surface.
(2) A method of applying the composition of the present invention containing a surface modifier to the surface of a resin to form a thin film of the copolymer on the surface of the resin.

The method of use (1) above is used, for example, in fabricating a device that is in contact with a biological material.
The method of use (2) above is used, for example, for producing a cell culture vessel.
The method of use (2) above may be used to fabricate a device in contact with a biological material, and the method of use (1) above may be used to fabricate a cell culture vessel.

The content of the copolymer in the composition when the composition does not contain a resin is not particularly limited, but is preferably 0.1 to 20% by mass, more preferably 0.5 to 10% by mass, and 1 to 5% by mass. % by weight is particularly preferred.

<Resin>
The resin contained in the composition is not particularly limited, and may be a synthetic resin, a natural resin, or a derivative of a natural resin.
Examples of synthetic resins include polymethyl methacrylate (PMMA), polydimethylsiloxane (PDMS), polystyrene (PS), polypropylene (PP), polyacrylonitrile (PAN), polyester polymer alloy (PEPA), polysulfone (PSF), polyethylene terephthalate (PET), polyvinyl alcohol (PVA), polyurethane (PU), ethylene vinyl alcohol (EVAL), polyethylene (PE), polyester, polyvinylidene fluoride (PVDF), polyethersulfone (PES), polycarbonate (PC), polyvinyl chloride (PVC), polytetrafluoroethylene (PTFE), ultra-high molecular weight polyethylene (UHPE), acrylonitrile-butadiene-styrene resin (ABS), Teflon (registered trademark), and the like.
Examples of natural resins or derivatives thereof include cellulose, cellulose triacetate (CTA), nitrocellulose (NC), cellulose with immobilized dextran sulfate, and the like.

The resin content in the composition is not particularly limited, but is preferably 0.1 to 20% by mass, more preferably 0.5 to 15% by mass, and particularly preferably 1 to 10% by mass.

The mass ratio of the copolymer and the resin in the composition is not particularly limited, but the content of the copolymer is preferably 0.1 to 20 parts by mass, preferably 0.2 to 0.2 parts by mass, with respect to 100 parts by mass of the resin. 10 parts by weight is more preferred, and 0.5 to 5 parts by weight is particularly preferred.

<Solvent>
Solvents when included in the composition include, for example, water, phosphate buffered saline (PBS), alcohols, glycol ethers, ketones, aromatic compounds, sulfoxides, amides, and the like.

Alcohols, ketones, aromatic compounds, sulfoxides, and sulfoxides are preferred as solvents when the composition contains a resin and the composition is used to fabricate a device in contact with a biological material.
When the composition is used to prepare a cell culture vessel, preferred solvents are water, phosphate buffered saline (PBS), alcohol, and glycol ether.

Alcohols include alcohols having 1 to 6 carbon atoms.
Examples of alcohols include methanol, ethanol, propanol, isopropanol, 1-butanol, 2-butanol, isobutanol, t-butanol, 1-pentanol, 2-pentanol, 3-pentanol, 1-heptanol, 2- Heptanol, 2,2-dimethyl-1-propanol (neopentyl alcohol), 2-methyl-1-propanol, 2-methyl-1-butanol, 2-methyl-2-butanol (t-amyl alcohol), 3-methyl -1-butanol, 3-methyl-3-pentanol, cyclopentanol, 1-hexanol, 2-hexanol, 3-hexanol, 2,3-dimethyl-2-butanol, 3,3-dimethyl-1-butanol, 3,3-dimethyl-2-butanol, 2-ethyl-1-butanol, 2-methyl-1-pentanol, 2-methyl-2-pentanol, 2-methyl-3-pentanol, 3-methyl-1 -pentanol, 3-methyl-2-pentanol, 3-methyl-3-pentanol, 4-methyl-1-pentanol, 4-methyl-2-pentanol, 4-methyl-3-pentanol and cyclo Hexanol is mentioned. These can be used individually by 1 type or in combination of 2 or more types.
Alcohols having 2 to 6 carbon atoms are preferable as the alcohol when the composition is used for producing a cell culture vessel.

Examples of glycol ethers include ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, methyl cellosolve acetate, ethyl cellosolve acetate, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, propylene glycol, propylene glycol monomethyl ether, propylene glycol monoethyl ether, and propylene. Glycol monomethyl ether acetate and propylene glycol propyl ether acetate can be mentioned. These can be used individually by 1 type or in combination of 2 or more types.

When the composition is used to prepare a cell culture vessel, the composition may use water, phosphate-buffered saline (PBS), alcohol, or a water-soluble organic solvent alone as a solvent. In addition, two or more of water, phosphate-buffered saline (PBS), alcohol, and water-soluble organic solvents may be used in combination as the solvent for the composition.
When the composition is used for producing a cell culture vessel, the following combinations of solvents are preferred.
- Water and alcohol - Water, alcohol and water-soluble organic solvent - Alcohol and water-soluble organic solvent As a combination of solvents, the following combinations are more preferable.
・Water and ethanol ・Water, ethanol and propylene glycol monomethyl ether ・Ethanol and propylene glycol monomethyl ether

Ketones include, for example, acetone, methyl ethyl ketone, and the like.
Examples of aromatic compounds include toluene and xylene.
Examples of sulfoxides include dimethylsulfoxide and the like.
Amides include, for example, N,N-dimethylformamide.

The solvent content in the composition is not particularly limited, but is preferably 0.1 to 50% by mass, more preferably 0.5 to 20% by mass, and particularly preferably 1 to 20% by mass.

<Other ingredients>
Other ingredients include pH adjusters, preservatives, surfactants, antifungal agents, sugars, inorganic fillers, reinforcing agents, coloring agents, stabilizers, extenders, viscosity modifiers, tackifiers, flame retardants, UV absorbers, antioxidants, discoloration inhibitors, antibacterial agents, anti-aging agents, antistatic agents, plasticizers, lubricants, smoothing agents, foaming agents, releasing agents and the like.

When the composition contains a resin and the composition is used for the production of a device in contact with a biological material, the other components include, among the above other components, inorganic fillers, reinforcing materials, coloring agents, stabilizing agents, and the like. agents, bulking agents, viscosity modifiers, tackifiers, flame retardants, UV absorbers, antioxidants, anti-tarnishing agents, antibacterial agents, anti-mold agents, anti-aging agents, anti-static agents, plasticizers, lubricants, smoothing agents agents, foaming agents, mold release agents, antifungal agents, and the like.
When the composition is used for producing a cell culture vessel, other ingredients include, for example, pH adjusters, preservatives, surfactants, antifungal agents, sugars, and the like.

<Method for producing a film using the composition>
From the composition of the present invention, a film having the ability to suppress adhesion of biological substances can be obtained.
A method for producing a membrane using the composition includes, for example, at least a supply step and a drying step, and optionally includes other steps such as a washing step and a sterilization step.
In addition, the membrane may be manufactured without performing the drying process after the supplying process is performed.

When the composition contains a resin, the obtained film is, for example, a resin film in which a copolymer is segregated on the surface.
When the composition does not contain a resin, the resulting film is, for example, a copolymer film formed on a substrate.
As described above, in one embodiment of the present invention, when a resin film is formed from a composition containing a copolymer and a resin, the copolymer segregates on the surface of the resin film. Anti-adhesion ability of biological substances is expressed.
In another embodiment of the present invention, when a copolymer film is formed on a substrate from a composition containing a copolymer and not containing a resin, The film itself exhibits the ability to suppress the adhesion of biological substances to the surface of the base material.

<<Supply Process>>
The supply step is not particularly limited as long as it is a step in which the composition of the present invention is supplied onto the substrate.

<<<base material>>>
The material, size, shape and structure of the substrate are not particularly limited.

Examples of the material of the substrate include glass, metal, metal-containing compound or metalloid-containing compound, activated carbon, and resin.
As metals, typical metals: (alkali metals: Li, Na, K, Rb, Cs; alkaline earth metals: Ca, Sr, Ba, Ra), magnesium group elements: Be, Mg, Zn, Cd, Hg; aluminum Group elements: Al, Ga, In; Rare earth elements: Y, La, Ce, Pr, Nd, Sm, Eu; Tin group elements: Ti, Zr, Sn, Hf, Pb, Th; Iron group elements: Fe, Co, Ni; earth-acid elements: V, Nb, Ta; chromium group elements: Cr, Mo, W, U; manganese group elements: Mn, Re; noble metals: Cu, Ag, Au; platinum group elements: Ru, Rh, Pd, Os, Ir, Pt, and the like.
Examples of metal-containing compounds or semi-metal-containing compounds include ceramics, which are sintered bodies whose basic component is a metal oxide and is sintered by heat treatment at high temperature, semiconductors such as silicon, metal oxides or semi-metal oxides ( inorganic solid materials such as moldings of inorganic compounds such as silicon oxides, alumina, etc.), metal carbides or semi-metal carbides, metal nitrides or semi-metal nitrides (silicon nitrides, etc.), metal borides or semi-metal borides, etc. , aluminum, nickel titanium, and stainless steel (SUS304, SUS316, SUS316L, etc.).
The resin may be a natural resin or a derivative thereof, or a synthetic resin. Examples of the natural resin or a derivative thereof include cellulose, cellulose triacetate (CTA), nitrocellulose (NC), cellulose with immobilized dextran sulfate, and synthetic resins. Resins include polyacrylonitrile (PAN), polyester polymer alloy (PEPA), polystyrene (PS), polysulfone (PSF), polyethylene terephthalate (PET), polymethyl methacrylate (PMMA), polyvinyl alcohol (PVA), and polyurethane (PU). , ethylene vinyl alcohol (EVAL), polyethylene (PE), polyester, polypropylene (PP), polyvinylidene fluoride (PVDF), polyethersulfone (PES), polycarbonate (PC), polyvinyl chloride (PVC), polytetrafluoroethylene (PTFE), ultra-high molecular weight polyethylene (UHPE), polydimethylsiloxane (PDMS), acrylonitrile-butadiene-styrene resin (ABS) or Teflon (registered trademark) is preferably used.
The material of the substrate may be of one type or a combination of two or more types.

Among these materials, glass, silicon, silicon oxide, polystyrene (PS), polypropylene (PP), polyethersulfone (PES), polyethylene terephthalate (PET), polycarbonate (PC), polyvinyl chloride (PVC), Teflon (registered trademark), cycloolefin polymer (COP), polydimethylsiloxane (PDMS) or stainless steel (SUS304, SUS316, SUS316L, etc.) alone or preferably a combination selected from these, glass, polystyrene (PS), Particularly preferred are polypropylene (PP), stainless steel (SUS304, SUS316, SUS316L, etc.), and polydimethylsiloxane (PDMS).

The shape of the substrate is not particularly limited, and the substrate may be a flat flat plate, or a vessel-shaped structure having depressions such as wells and dishes (so-called cell culture vessel). may

Substrates include, for example, petri dishes commonly used for cell culture, tissue culture dishes, dishes (petri dishes) such as multi-dishes, cell culture flasks, flasks such as spinner flasks, plastic bags, and Teflon (registered trademark). ) bags such as bags and culture bags, plates such as microplates, microwell plates, multiplates and multiwell plates, chamber slides, tubes, trays, bottles such as roller bottles, and the like. Dishes, plates, and trays are preferred.

<<<supply method>>>
A method for supplying the composition of the present invention onto a substrate is not particularly limited, and examples thereof include general coating methods.
Examples of coating methods include spin coating, inkjet, screen printing, slit coating, roll-to-roll, dip coating, and solvent casting. A printing technique such as an inkjet method or screen printing is preferable.
More specifically, the coating method includes, for example, a method of immersing a base material in the composition of the present invention, a method of adding the composition of the present invention to a base material, and a method of standing still for a predetermined time. be done.
When the base material is a container, the supply method can be carried out, for example, by adding the composition of the present invention to the container and allowing it to stand for a predetermined period of time. The addition can be carried out, for example, by adding the composition of the present invention in an amount of 0.5 to 1 times the total volume of the container using a syringe or the like.

<<Drying process>>
The drying step is not particularly limited as long as it is a step in which the composition of the present invention supplied onto the substrate is dried.

Drying can be carried out, for example, in the air or under vacuum at a temperature in the range of -200°C to 200°C.
The film can be formed, for example, by drying at room temperature (10° C. to 35° C., for example, 25° C.), but may be dried at, for example, 40° C. to 100° C. for faster film formation. .
A preferred drying temperature is 10°C to 180°C, and a more preferred drying temperature is 20°C to 150°C.
The drying time is not particularly limited, but is, for example, 1 minute to 24 hours.

<<Washing process>>
The washing step is not particularly limited as long as it is a step in which the film obtained by the drying step is washed. The cleaning step is performed, for example, to remove impurities remaining in the film, unreacted monomers, and the like from the film.

Washing can be performed by a known method, but washing with running water or ultrasonic washing is preferred.
Solvents used for washing include water, an aqueous solution containing an electrolyte, and the like. The solvent is usually used at room temperature (eg, 10 to 35°C), but may be heated to a range of, for example, 40 to 95°C. Aqueous solutions containing electrolytes are preferably PBS, physiological saline (containing only sodium chloride), Dulbecco's phosphate-buffered saline, Tris-buffered physiological saline, HEPES-buffered physiological saline, and Veronal-buffered physiological saline, and PBS is preferred. Especially preferred.

In addition, when the obtained membrane is used for cell culture, washing may be performed using a sample medium (for example, water, buffer solution, medium, etc.) used for cell culture. A preferred medium is a culture medium, and more preferred mediums are BME medium (Eagle's basal medium) and DMEM medium (Dulbecco's Modified Eagle's Medium).

<< Sterilization process >>
When the obtained membrane is used for cell culture, the obtained membrane needs to undergo a sterilization step by irradiation.
The sterilization step is typically carried out at ambient temperature (eg, about 0°C to about 40°C, preferably about 10°C to about 30°C, more preferably about 25°C). The radiation to be irradiated is not limited as long as it can sterilize, but γ-rays, X-rays or electron beams are preferable. γ-rays or X-rays are more preferred, and γ-rays are even more preferred. The irradiation dose of γ-rays may be, for example, the dose adopted in a normal sterilization process. For example, irradiation of about 5 to 40 kGy is sufficient, preferably 10 to 25 kGy.

(medical device)
The medical device of the invention has a membrane formed from the composition of the invention.
Medical devices are, for example, devices in contact with biological material. In the medical device, the film formed from the composition of the present invention is arranged, for example, at least part of the part that comes into contact with the biological material.

The medical device of the present invention is not particularly limited as long as it is a medical instrument. Examples include blood bags, urine collection bags, blood transfusion sets, sutures, drain tubes, various catheters, blood accesses, blood circuits, artificial blood vessels, artificial Examples include kidneys, heart-lung machines, artificial valves, plasma exchange membranes, various adsorbents, CAPDs, IABPs, pacemakers, artificial joints, artificial femoral heads, dental materials, and various shunts.

A medical device is a medical structure that is used in vivo, and such structures are divided into those that are used by being implanted in the body and those that are used inside the body (without being implanted). classified roughly. The size and length of the medical device are not particularly limited, and devices with minute circuits and devices for detecting minute amounts of samples are also included.
Structures to be implanted in the body include, for example, functional assisting devices such as cardiac pacemakers for supplementing the functions of diseased living organisms; devices for detecting abnormalities in living organisms such as implantable biochips; Examples include medical instruments such as implants, bone fixation materials, sutures, artificial blood vessels, and the like.
In addition, structures used inside the body (without being implanted) include catheters, gastroscopes, and the like.

(Silicone base material)
The silicone substrate of the present invention has a film formed from the composition of the present invention.
A silicone substrate is, for example, a silicone substrate that is in contact with a biological material. In the silicone base material, the film formed from the composition of the present invention is disposed, for example, on at least part of the portion that is in contact with the biological material.

The silicone base material is not particularly limited as long as it contains a silicone-based resin. Specific examples of such silicone substrates include silicone-containing medical devices and silicone substrates having microchannels, preferably silicone-containing microchannel devices.

Examples of microfluidic devices include microanalysis devices such as microreaction devices (specifically, microreactors, microplants, etc.), integrated nucleic acid analysis devices, microelectrophoresis devices, and microchromatography devices; microdevices for analysis sample preparation such as chromatography; physicochemical processing devices used for extraction, membrane separation, dialysis, etc.; environmental analysis chip, clinical analysis chip, gene analysis chip (DNA chip), protein analysis chip (proteome chip), Examples include microfluidic chips such as sugar chain chips, chromatography chips, cell analysis chips, and pharmaceutical screening chips. Among these, a microchannel chip is preferable.

The microchannel is a portion through which a small amount of sample (preferably a liquid sample) flows, and the channel width and depth are not particularly limited, but both are usually about 0.1 μm to 1 mm, preferably is between 10 μm and 800 μm.
The channel width and depth of the microchannel may be the same over the entire length of the channel, or may be partially different in size and shape.

(cell culture vessel)
The cell culture vessel of the present invention has a membrane formed from the composition of the present invention.
A cell culture vessel can be used, for example, for producing cell aggregates.

（式（Ｉ）中、Ｕ^ａ１及びＵ^ａ２は、それぞれ独立して、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^a１は、水素原子又は炭素原子数１～５のアルキル基を表し、Ｒ^a２は、炭素原子数１～５のアルキレン基を表す。）

（式（ＩＩ）中、Ｒ^ｂは、水素原子又は炭素原子数１～５のアルキル基を表す。）
炭素原子数１～５のアルキレン基の具体例としては、式（ａ１－２）のＲ^１２等の説明で挙げたアルキレン基の具体例が挙げられる。 The cell culture vessel may have a cell culture substrate membrane having cell adhesiveness on the membrane formed from the composition of the present invention.
Examples of the cell culture base film include the base film described in International Publication No. 2020/040247. It should be noted that the contents of WO2020/040247 are incorporated herein in full to the same extent as if explicitly set forth.
An example of the base film described in WO2020/040247 is a base film containing a polymer containing at least a repeating unit derived from a monomer represented by formula (I) below. The polymer may contain repeating units derived from a monomer represented by formula (II) below.

(In formula (I), U ^a1 and U ^a2 each independently represent a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and R ^a1 is a hydrogen atom or an alkyl group having 1 to 5 carbon atoms. and R ^a2 represents an alkylene group having 1 to 5 carbon atoms.)

(In formula (II), R ^b represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms.)
Specific examples of the alkylene group having 1 to 5 carbon atoms include the specific examples of the alkylene group mentioned in the description of R ¹² of the formula (a1-2).

The cell culture substrate membrane is, for example, arranged in a pattern rather than uniformly on the membrane formed from the composition of the present invention.
Examples of patterns include dot patterns.
In other words, an example of a cell culture vessel is a membrane formed from the composition of the present invention, which is capable of suppressing cell adhesion, and a cell culture base membrane having cell adhesiveness, which is arranged in a dot pattern on the membrane. and a cell seeding surface.

The thickness of the cell culture base membrane is, for example, 1 to 1,000 nm, preferably 5 to 500 nm, preferably 5 to 300 nm, preferably 5 to 200 nm, preferably 5 to 150 nm, preferably 10 to 150 nm. is between 20 and 150 nm.

Hereinafter, the present invention will be described in more detail based on Synthesis Examples, Examples, Test Examples, etc., but the present invention is not limited thereto.

In the examples, the equipment and conditions used for sample preparation and physical property analysis are as follows.
(1) Size exclusion chromatography (SEC)
Apparatus 1: High performance liquid chromatograph (HLC-8120GPC) manufactured by Tosoh Corporation
SEC column: TSKgel guard column Super AW-H (4.6 mm ID x 3.5 cm) x 1, TSKgel Super AW5000 (6.0 mm ID x 15 cm) x 1, Super AW2500 (6.0 mm ID x 15 cm) x 1 are connected .
Flow rate: 0.5mL/min
Eluent: N,N-dimethylformamide (DMF) (containing 10 wt% lithium bromide)
Column temperature: 40°C
Detection: Differential Refractive Index Detector (RI)
Injection concentration: polymer solid content 0.5% by mass
Injection volume: 100 μL
Calibration curve: Cubic approximation curve Standard sample: PMMA (manufactured by Sigma-Aldrich) x 6 types

(2) ¹ H NMR spectrum Apparatus: JNM-ECP400 manufactured by JEOL Ltd.
Solvent: deuterated chloroform (CDCl ₃ ), deuterated dimethylsulfoxide ((CD ₃ ) ₂ C(=O))
Standard: _CHCl3 (7.26 ppm)
Tetramethylsilane (0.00 ppm)
( _CH3 ) _2C (=O) (2.10 ppm)

(3) Spin coater device: 1H-D7 manufactured by Mikasa Co., Ltd.
(4) X-ray photoelectron spectroscopy (XPS)
Apparatus: PHI5000 VersaProbe III manufactured by ULVAC-Phi, Inc.
Measurement conditions: 14.0 kV, 14 mA
(5) Contact angle Device: DropMaster 500 manufactured by Kyowa Interface Science Co., Ltd.
(6) Optical microscope device: All-in-one fluorescence microscope (Biozero BZ-8100) manufactured by Keyence Corporation
(7) Atomic force microscope (AFM)
Apparatus: Asylum Research Cypher ES manufactured by Oxford Instruments Co., Ltd.

[Measurement of Substituent Composition Ratio by NMR (Nuclear Magnetic Resonance)]
The ratio of introduction of substituents shown in Synthesis Examples below is the result of measurement by NMR. All of Synthesis Examples 1 to 3 were measured using JNM-ECP400 (manufactured by JEOL Ltd.).

[Molecular weight and molecular weight distribution]
The number average molecular weight and molecular weight dispersity shown in Synthesis Examples below are the results of measurement by SEC.

[Synthesis Example 1]
Poly[oligo(ethylene oxide methacrylate)] (hereinafter referred to as PEOMA) was synthesized as a macroinitiator for EbS (hereinafter referred to as macroinitiator).
PEOMA was synthesized with reference to reference 1 (H. Matsuno, K. Tanaka. J. Mater. Chem. B, 2019, 7, 1045).
As a macromonomer, 30 g of oligo(ethylene oxide methacrylate) (referred to as EOMA) (manufactured by Sigma-Aldrich) was dissolved in 50.1 mL of anisole (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.). Furthermore, copper (I) bromide (manufactured by Kishida Chemical Co., Ltd.) 172.1 mg, copper (II) bromide (manufactured by Kishida Chemical Co., Ltd.) 67 mg, N, N, N', N''', 0.627 mL of N''-pentamethyldiethylenetriamine (PMDETA) (manufactured by Sigma-Aldrich) and 0.44 mL of 2-ethyl-2-bromoisobutyrate (EBiB) (manufactured by Tokyo Kasei Co., Ltd.) were added. Dissolved oxygen was removed by bubbling argon gas (manufactured by Japan Air Liquid Co., Ltd.) for 2 minutes. After that, it was immersed in an oil bath at 45° C. to initiate polymerization. Two hours after the initiation of the reaction, the reaction solution was cooled with ice water to terminate the polymerization reaction.
From the results of ¹ H NMR measurement, a peak derived from PEOMA was confirmed, suggesting that PEOMA was obtained by the polymerization method described above. SEC measurement revealed a number average molecular weight of 7,200 and a molecular weight dispersity of 1.40.
From the above results, it became clear that a polymer of PEOMA was obtained.

[Synthesis Example 2]
Poly[oligo(ethylene oxide methacrylate)-block-oligo(dimethylsiloxy methacrylate)] was synthesized. Conditions were set so that the polymer obtained here had a composition ratio of 31 mol % PEOMA and 69 mol % PSiOMA. The polymer obtained is hereinafter referred to as EbS31.
1.48 g of PEOMA [the compound obtained in Synthesis Example 1] and 2.94 g of oligo(dimethylsiloxy methacrylate) (manufactured by Shin-Etsu Chemical Co., Ltd., X-22-2404) were added to N,N-dimethylformamide (DMF) (Fuji Film Wada It was dissolved in 5.01 mL (manufactured by Kojunyaku Co., Ltd.). Furthermore, copper (I) bromide (manufactured by Kishida Chemical Co., Ltd.) 11.5 mg, copper (II) bromide (manufactured by Kishida Chemical Co., Ltd.) 4.47 mg, and N, N, N', N'' , N″-pentamethyldiethylenetriamine (PMDETA) (manufactured by Sigma-Aldrich) and 0.4 mL of DMF solution were added. Dissolved oxygen was removed by bubbling argon gas (manufactured by Japan Air Liquid Co., Ltd.) for 2 minutes. After that, it was immersed in an oil bath at 45° C. to initiate polymerization. Three hours after the initiation of the reaction, the reaction solution was cooled with ice water to terminate the polymerization reaction.
From the results of ¹ H NMR measurement, a peak derived from EbS was confirmed, suggesting that PEOMA-block-PSiOMA (EbS31) was obtained by the polymerization method described above. SEC measurement gave a number average molecular weight of 12,600 and a molecular weight dispersity of 1.25.
From the above results, it became clear that a polymer of EbS31 was obtained.

[Synthesis Example 3]
Poly[oligo(ethylene oxide methacrylate)-block-oligo(dimethylsiloxy methacrylate)] was synthesized. The composition ratio of the polymer obtained here was 80 mol % of PEOMA and 20 mol % of PSiOMA. Therefore, it is called EbS80.
2.22 g of PEOMA [the compound obtained in Synthesis Example 1] and 2.21 g of oligo(dimethylsiloxy methacrylate) (manufactured by Shin-Etsu Chemical Co., Ltd., X-22-2404) were added to N,N-dimethylformamide (DMF) (Fuji Film Wa It was dissolved in 4.62 mL (manufactured by Kojunyaku Co., Ltd.). Furthermore, copper (I) bromide (manufactured by Kishida Chemical Co., Ltd.) 17.2 mg, copper (II) bromide (manufactured by Kishida Chemical Co., Ltd.) 6.7 mg, and N, N, N', N'' , N″-pentamethyldiethylenetriamine (PMDETA) (manufactured by Sigma-Aldrich) and 0.6 mL of DMF solution were added. Dissolved oxygen was removed by bubbling argon gas (manufactured by Japan Air Liquid Co., Ltd.) for 2 minutes. After that, it was immersed in an oil bath at 45° C. to initiate polymerization. Three hours after the initiation of the reaction, the reaction solution was cooled with ice water to terminate the polymerization reaction.
From the result of ¹ H NMR spectrum, a peak derived from EbS was confirmed, suggesting that EbS80 was obtained by the polymerization method described above. SEC measurement revealed a number average molecular weight of 8,500 and a molecular weight dispersity of 1.07.
From the above results, it became clear that a polymer of EbS80 was obtained.

[Synthesis Example 4]
Poly[oligo(dimethylsiloxy methacrylate)] (referred to as PSiOMA) was synthesized.
8.84 g of oligo(dimethylsiloxy methacrylate) (manufactured by Shin-Etsu Chemical Co., Ltd., X-22-2404) was dissolved in 2.73 mL of anisole (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). Furthermore, copper (I) bromide (manufactured by Kishida Chemical Co., Ltd.) 40.2 mg, copper (II) bromide (manufactured by Kishida Chemical Co., Ltd.) 15.6 mg, N, N, N', N'' , N″-Pentamethyldiethylenetriamine (PMDETA) (Sigma-Aldrich) DMF solution 1.4 mL and 2-ethyl-2-bromoisobutyrate (EBiB) (Tokyo Kasei Co., Ltd.) DMF solution 1 .4 mL was added. Dissolved oxygen was removed by bubbling argon gas (manufactured by Japan Air Liquid Co., Ltd.) for 2 minutes. After that, it was immersed in an oil bath at 70° C. to initiate polymerization. One hour after the start of the reaction, the reaction solution was cooled with ice water to terminate the polymerization reaction.
From the results of the ¹ H NMR spectrum, a peak derived from PSiOMA was confirmed, suggesting that PSiOMA was obtained by the polymerization method described above. SEC measurement gave a number average molecular weight of 9,100 and a molecular weight dispersity of 1.18.
From the above results, it became clear that a polymer of PSiOMA was obtained.

[Synthesis Example 5]
Poly[oligo(dimethylsiloxymethacrylate)-random-oligo(dimethylsiloxymethacrylate)] (termed ErS31) was synthesized.
Oligo (ethylene oxide methacrylate) (EOMA) (manufactured by Sigma-Aldrich) 3.33 g and oligo (dimethylsiloxy methacrylate) (manufactured by Shin-Etsu Chemical Co., Ltd., X-22-2404) 5.89 g of anisole (Fujifilm Wako Pure Chemical Industries, Ltd. ) was dissolved in 6.87 mL. Copper (I) bromide (manufactured by Kishida Chemical Co., Ltd.) 80.4 mg, copper (II) bromide (manufactured by Kishida Chemical Co., Ltd.) 31.2 mg, N, N, N', N'', N 3.08 mL of an anisole solution of ''-pentamethyldiethylenetriamine (PMDETA) (manufactured by Sigma-Aldrich) and 0.76 mL of an anisole solution of 2-ethyl-2-bromoisobutyrate (EBiB) (manufactured by Tokyo Kasei Co., Ltd.) and added. Dissolved oxygen was removed by bubbling argon gas (manufactured by Japan Air Liquid Co., Ltd.) for 2 minutes. After that, it was immersed in an oil bath at 70° C. to initiate polymerization. After 1.6 hours from the initiation of the reaction, the reaction solution was cooled with ice water to terminate the polymerization reaction.
From the result of ¹ H NMR spectrum, a peak derived from PSiOMA was confirmed, suggesting that ErS31 was obtained by the polymerization method described above. SEC measurements gave a number average molecular weight of 13,600 and a molecular weight dispersity of 1.28.
From the above results, it became clear that a polymer of ErS31 was obtained.

The results of Synthesis Examples 1 to 5 are summarized in Table 1.

<Example 1>
EbS31 obtained in Synthesis Example 2 was mixed with a 3% by mass toluene solution of PMMA so as to be 5% by mass with respect to polyethylene methacrylate (referred to as PMMA), and a PMMA coating solution containing EbS31 (EbS31-containing resin composition 1-1). The coating solution was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds) to obtain an EbS31-containing PMMA film.
The EbS31 obtained in Synthesis Example 2 was mixed with a 3% by mass toluene solution of PS so as to be 5% by mass with respect to polystyrene (referred to as PS), and a PS coating liquid containing EbS31 (EbS31-containing resin composition 1 -2). The coating solution was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds) to obtain an EbS31-containing PS film.
EbS31 obtained in Synthesis Example 2 was mixed with polydimethylsiloxane (hydrogenated-terminated polydimethylsiloxane (manufactured by Gelest) and vinyl-terminated polydimethylsiloxane (manufactured by Gelest) were mixed at a ratio of 9 to 1) (the following was PDMS ) was mixed with a 3% by mass tetrahydrofuran solution of PDMS to obtain a PDMS coating solution containing EbS31 (EbS31-containing resin composition 1-3). The coating solution was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds) to obtain an EbS31-containing PDMS film.
EbS31 obtained in Synthesis Example 2 was mixed with a 0.75% by mass p-xylene solution of PP so as to be 5% by mass with respect to polypropylene (hereinafter referred to as PP), and a PP coating solution containing EbS31 ( EbS31-containing resin composition 1-4). The coating solution was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds, 85° C.) to obtain an EbS31-containing PP film.

<Comparative Example 1>
A 3% by mass toluene solution of PMMA was prepared and used as a PMMA coating solution. The coating solution was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds) to obtain a PMMA film.
A 3 mass % toluene solution of PS was prepared and used as a PS coating liquid. The coating liquid was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds) to obtain a PS film.
A 3% by mass tetrahydrofuran solution of PDMS was prepared and used as a PDMS coating solution. The coating solution was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds) to obtain a PDMS film.
A 0.75% by mass p-xylene solution of PP was prepared and used as a PP coating solution. The coating solution was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds, 85° C.) to obtain a PP film.

The substrates used in the following test examples are as follows.
(glass substrate)
A commercially available slide glass for microscope observation (manufactured by Matsunami Glass Industry Co., Ltd.) was cut into 1 cm×1 cm and used.
(silicon substrate)
A silicon wafer (manufactured by SUMCO Co., Ltd.) was cut into 1.0 cm×1.0 cm and used.

<Test Example 1-1>
[Observation of coating surface by atomic force microscope]
Each EbS31-containing resin composition 1-1 to 1-4 obtained in Example 1 or the coating liquid obtained in Comparative Example 1 was spin-coated on a silicon substrate, and then dried under reduced pressure by heating to obtain a sample. . The structure of the prepared sample surface was observed in the atmosphere using an atomic force microscope (AFM) (manufactured by Oxford Instruments Co., Ltd.). A cantilever (manufactured by Nanoworld) having a length of 160 μm, a resonance frequency of 285 kHz, and a spring constant of 42 N/m was used. Observation conditions were a scan range of 10 μm×10 μm and a scan speed of 0.5 Hz. This was subjected to image analysis using data processing software built into the device.
FIG. 1 shows the following height image and phase contrast image obtained from AFM.
Height image ・(a) EbS31/PMMA (RMS: 1.56 nm)
・(b) PMMA (RMS: 0.23 nm)
・(c) EbS31/PS (RMS: 4.66 nm)
・(d) PS (RMS: 0.26 nm)
(i) EbS31/PDMS (RMS: 3.28 nm)
(j) PDMS (RMS: 0.86 nm)
・(k) EbS31/PP (RMS: 5.93 nm)
(l) PP (RMS: 4.73 nm)
Phase contrast image ・(e) EbS31/PMMA
・(f) PMMA
・(g) EbS31/PS
・(h) PS
(m) EbS31/PDMS
(n) PDMS
・(o) EbS31/PP
・(p) PP
It was found that the EbS31-containing PMMA film, the EbS31-containing PS film, and the EbS31-containing PDMS film had unevenness on the surface, and the surface root mean square roughness (RMS) was about 1.56 to 5.93 nm.
On the other hand, the EbS31-free PMMA film, the EbS31-free PS film, and the EbS31-free PDMS film have no unevenness on the surface and have a surface root mean square roughness of about 0.23 to 0.86 nm. was smoother than
Similarly, phase difference images reflecting height images on the surfaces of the EbS31-containing PMMA film, the EbS31-containing PS film, and the EbS31-containing PDMS film were observed.
No difference was observed between the EbS31-containing PP film and the EbS31-free PP film in the AFM analysis results.

<Test Example 1-2>
The chemical compositions of the films prepared in Example 1 and Comparative Example 1 were evaluated based on X-ray photoelectron spectroscopy (XPS) measurements. An X-ray photoelectron spectrometer (PHI5000 VersaProbe III, ULVACPHI, PHI., INC) was used for the XPS measurement. The X-ray source was a monochromatic Al Kα ray, the applied voltage was 15.0 kV, 25 W, and the measurement was performed at room temperature. Further, the photoelectron extraction angle was set to 15°. The analysis depth of carbon 1s electrons calculated from the mean free path is about 3 nm. During the measurement, the sample surface was charged up and the binding energy shifted over time, so the carbon 1s peak (285 eV) was used as a reference and corrected using a neutralization gun.
The results are shown in FIGS. 2-5.

FIG. 2 shows the results of XPS measurement of the EbS31-containing PMMA film and the EbS31-free PMMA film. FIG. 2(a) is the spectrum of carbon atoms, FIG. 2(b) is the spectrum of oxygen atoms, and FIG. 2(c) is the spectrum of silicon atoms.
From the results of C _1s in FIG. 2(a), the peak intensity of the peak (289.5 eV) derived from the ether bond (—C—O—) of the PEOMA chain increased on the surface of the EbS31-containing PMMA film.
In addition, from the result of O _1s in FIG. 2(b), on the surface of the EbS31-containing PMMA film, an increase in the peak (532.5 eV) derived from the ether bond (—C—O—) of the PEOMA chain of EbS31 was observed. was taken.
Furthermore, from the result of Si _2p in FIG. 2(c), a peak (102 eV) derived from siloxane (—Si—O—) of PSiOMA was confirmed on the surface of the EbS31-containing PMMA film.

FIG. 3 shows the results of XPS measurement of the EbS31-containing PS film and the EbS31-free PS film. FIG. 3(a) is the spectrum of carbon atoms, FIG. 3(b) is the spectrum of oxygen atoms, and FIG. 3(c) is the spectrum of silicon atoms.
From the results of C _1s in FIG. 3(a), on the surface of the EbS31-containing PS film, the peak (289.5 eV) derived from the ether bond (-C-O-) of the PEOMA chain and the ester (-C(=O)O −), the intensity of the peak (289 eV) was increased.
In addition, from the result of O _1s in FIG. An increase in the peak (534 eV) derived from C(=O)O-) was observed.
Furthermore, from the result of Si _2p in FIG. 3(c), a peak (102 eV) derived from siloxane (—Si—O—) of PSiOMA was confirmed on the surface of the EbS31-containing PS film.

FIG. 4 shows the results of XPS measurement of the EbS31-containing PS film and the EbS31-free PDMS film. FIG. 4(a) is the spectrum of carbon atoms, FIG. 4(b) is the spectrum of oxygen atoms, and FIG. 4(c) is the spectrum of silicon atoms.
4(a), (b), and (c) show the above O _1s , O _1s , and Si _2p spectra. Here, the peaks of EbS31 and PDMS alone overlap, making it difficult to separate the peaks. there were.

FIG. 5 shows the results of XPS measurement of the EbS31-containing PP film and the EbS31-free PP film. FIG. 5(a) is the spectrum of carbon atoms, FIG. 5(b) is the spectrum of oxygen atoms, and FIG. 5(c) is the spectrum of silicon atoms.
From the result of C1s in FIG. 5(a), it was difficult to confirm the peak derived from the ether bond (--C--O--) of the PEOMA chain on the surface of the EbS31-containing PP film.
On the other hand, from the result of O _1s in FIG. 5(b), on the surface of the EbS31-containing PP film, the peak (532.5 eV) derived from the ether bond (—C—O—) of the PEOMA chain of EbS31 increased. Admitted.
Furthermore, from the result of Si _2p in FIG. 5(c), a peak (102 eV) derived from siloxane (—Si—O—) of PSiOMA was confirmed on the surface of the EbS31-containing PMMA film.

<Test Example 1-3>
[Evaluation of Static Contact Angle]
Each of the EbS31-containing resin compositions 1-1 to 1-4 obtained in Example 1 was spin-coated on a silicon substrate, and then dried by heating under reduced pressure to obtain a sample. The contact angle of the obtained film surface was evaluated by a fully automatic contact angle (DM-500, manufactured by Kyowa Interface Science Co., Ltd.). In all the samples, the water contact angle was defined as the value 60 seconds after the water droplet had landed. As for the bubble contact angle, the value 1 second after contact with the bubble was taken as the bubble contact angle.
Also, the static contact angle of each film prepared in Comparative Example 1 was measured.
In addition to the water contact angle obtained by measuring the contact angle of water droplets in air, the static contact angle is evaluated by both the bubble contact angle obtained by placing various substrates upside down in water and measuring the contact angle of bubbles. gone.
The water contact angle of the EbS31-containing PMMA film surface on the silicon substrate was shown to be 68±1.9°. The bubble contact angle showed 115±1.8°. The EbS31-free PMMA film of Comparative Example 1 has a water contact angle of 97±0.9° and a bubble contact angle of 111±0.2°. It was shown to be more hydrophilic in water than on the membrane surface.
Similarly, the water contact angle of the EbS31-containing PS film surface on the silicon substrate was shown to be about 91±0.3°. The bubble contact angle was about 109±6.8°. The EbS31-free PS film of Comparative Example 1 has a water contact angle of 89±2.0° and a bubble contact angle of 68±1.9°. It was shown to be more hydrophilic in water than the membrane surface.
Similarly, the water contact angle of the EbS31-containing PDMS film surface on the silicon substrate was shown to be about 114±0.5°. The bubble contact angle was about 133±0.9°. The EbS31-free PDMS film of Comparative Example 1 has a water contact angle of 68±1.9° and a bubble contact angle of 119±1.1°. It was shown to be more hydrophilic than the membrane surface.
Similarly, the water contact angle of the EbS31-containing PP film surface on the silicon substrate was shown to be about 100±0.5°. The bubble contact angle was about 98±1.3°. The EbS31-free PP film of Comparative Example 1 has a water contact angle of 103 ± 0.2 ° and a bubble contact angle of 93 ± 2.6 °. It was shown to be more hydrophilic than the membrane surface.

Tables 2 and 3 show a summary of the results of Test Examples 1-3. From the above, it has been clarified that the EbS31-containing resin film exhibits a more pronounced hydrophilic modification effect by being immersed in water.

From the results of Test Examples 1-1, 1-2, and 1-3, it was found that aggregate structures were formed on the surfaces of the PMMA film, PS film, and PDMS film containing EbS31. From the XPS spectrum, a peak derived from EbS31 was confirmed in this aggregate structure. On the other hand, regarding the PP resin, although there was no difference in the surface morphology between the PP film containing EbS31 and the PP film not containing EbS31, the XPS results confirmed a peak derived from EbS31 on the surface of the PP film containing EbS31. rice field. As for the air contact angle in water, it became clear that the EbS31-containing resin film of the example had improved hydrophilicity compared to the EbS31-free resin film of the comparative example. From the above results, it was clarified that when EbS31 was added to PMMA, PS, PDMS, or PP and applied, EbS31 was concentrated on the surface.

<Test Example 1-4>
[Cell adhesion experiment]
Using a glass substrate (hereinafter referred to as a coated substrate) on which an EbS31-containing resin film was formed in Example 1, a cell adhesion experiment was performed according to the following experimental method.
The coated substrate was placed on the bottom of a 24-well cell culture plate. Each EbS31-containing resin film was washed with sterilized water three times and then immersed in PBS at room temperature for 3 hours. Mouse fibroblasts (NIH3T3) were cultured in Dulbecco's modified Eagle's medium (FBS-containing DMEM) containing 10% bovine serum (FBS) in an environment of 5% CO ₂ and 37°C. Cultured NIH3T3 cells were washed once with sterile PBS and detached from cell culture plates by adding 3 mL of 0.25% trypsin/0.02% ethylenediaminetetraacetic acid (EDTA) mixture. The cell suspension was centrifuged to collect NIH3T3 cells, which were then resuspended in FCS-containing DMEM to obtain a solution of 7×10 ⁴ cells/mL.
A cell suspension was prepared in a medium so as to have 5×10 ⁴ cells/cm ² in each well of the cell culture plate on which the coated substrate was fixed, and seeded onto the coated substrate. After culturing for 12 hours in an environment of 5% CO ₂ and 37° C., the coated substrate was washed with PBS three times, and the cells adhered to the EbS31-containing resin membrane were fixed using a 2% glutaraldehyde PBS solution. After washing with ultrapure water three times, it was air-dried. The adhered cells were stained with a crystal violet-PBS staining solution having a concentration of 0.01% by mass, and observed using an optical microscope to observe the adhered cells.
FIG. 6 shows the results of cell adhesion experiments on the surfaces of the EbS31-containing PMMA film and EbS31-containing PS film prepared in Example 1, and the EbS31-free PMMA film and EbS31-free PS film prepared in Comparative Example 1 (optical microscope images). ).
6(a) to (d) are optical microscope images of the membrane before washing and staining. Figures 6(e)-(l) are optical microscope images of the membrane after washing and staining.
On the EbS31-containing PMMA membrane (Fig. 6(a)) and on the EbS31-containing PS membrane (Fig. 6(c)), the seeded cells floated without adhering to the membrane even after 12 hours of culture, and further aggregated. rice field. The seeded cells adhered on the EbS31-free PMMA membrane. On the EbS31-free PS membrane, suppression of spreading of adherent cells was observed, but a large number of cells adhered (FIGS. 6(b) and (d)). All coated substrates were washed with PBS and stained. Most of the cells were removed by the washing process on the EbS31-containing PMMA membrane and the EbS31-containing PS membrane (FIGS. 6(e) and (g)). After the staining step, although a few adhered cells were confirmed, it was found that the spreading of the cells was suppressed (FIGS. 6(i) and (k)). After washing and staining, the EbS31-free PMMA membrane and the EbS31-free PS membrane were found to have a large number of adherent cells remaining and spreading.
In addition, a similar cell adhesion test was performed on the EbS31-containing PDMS membrane and the EbS31-containing PP membrane prepared in Example 1. As comparative examples, an EbS31-free PDMS film and an EbS31-free PP film were used.
7(a) to (d) are optical microscope images of the membrane before washing and staining. Figures 7(e)-(l) are optical microscope images of the membrane after washing and staining.
FIG. 7 shows the results of cell adhesion experiments on the surface of the EbS31-containing PDMS film and EbS31-containing PP film prepared in Example 1, and the EbS31-free PDMS film and EbS31-free PP film prepared in Comparative Example 1 (optical microscope images). ). On the EbS31-containing PDMS membrane (Fig. 7(a)) and on the EbS31-containing PP membrane (Fig. 7(c)), the seeded cells remained floating without adhering to the membrane even after 12 hours, and further aggregated. . Some of the seeded cells adhered and spread on the EbS31-free PDMS membrane and the EbS31-free PP membrane (FIGS. 7(b) and (d)). All coated substrates were washed with PBS and stained. Most of the cells were removed by the washing process on the EbS31-containing PDMS membrane and the EbS31-containing PP membrane (FIGS. 7(e) and (g)). After the staining step, although a few adhered cells were confirmed, it was found that the spreading of the cells was suppressed (FIGS. 7(i) and (k)). It was found that the EbS31-free PDMS membrane and the EbS31-free PP membrane had adherent cells remaining and spreading after washing and staining.
From the above results, in the EbS31-containing resin film of Example 1, the EbS31 has a SiOMA composition, so that the surface is concentrated, and when the surface is in contact with water, the PEOMA chains are exposed and modified to be hydrophilic. It was shown that the interface has a high ability to suppress cell adhesion.

<Example 2>
In order to examine the effect of the content of EbS31, the experiment of Example 2 was conducted.
EbS31 obtained in Synthesis Example 2 was mixed with a 4.5% by mass toluene solution of PMMA so as to be 2% by mass, 1% by mass, or 0.5% by mass with respect to PMMA, and a PMMA coating solution containing EbS31 was obtained. (EbS31-containing resin compositions 2-1 to 2-3). The coating liquid was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds).
Similarly, the EbS31 obtained in Synthesis Example 2 was mixed with a 3.0% by mass toluene solution of PS so as to be 2% by mass, 1% by mass, or 0.5% by mass with respect to PS, and EbS31 was contained. PS coating liquids (EbS31-containing resin compositions 2-4 to 2-6) were used. The coating liquid was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds).

<Test Example 2-1>
[Observation of coating surface by atomic force microscope]
Each EbS31-containing resin composition obtained in Example 2 was spin-coated on a glass substrate, and then dried by heating under reduced pressure to obtain a sample. The structure of the prepared sample surface was observed in the atmosphere using an atomic force microscope (AFM) (manufactured by Oxford Instruments Co., Ltd.). A cantilever (manufactured by Nanoworld) having a length of 160 μm, a resonance frequency of 285 kHz, and a spring constant of 42 N/m was used. Observation conditions were a scan range of 10 μm×10 μm and a scan speed of 0.5 Hz. This was subjected to image analysis using data processing software built into the device.
FIG. 8 shows height images and phase contrast images obtained by AFM of PMMA films with different EbS31 contents. A PMMA film containing 2% by mass of EbS31 (FIG. 8A), a PMMA film containing 1% by mass of EbS31 (FIG. 8B), and a PMMA film containing 0.5% by mass of EbS31 (FIG. 8C )) and the EbS31-free PMMA film (FIG. 8(d)) had a surface root-mean-square roughness (PMS) of 0.46 nm or less, and a smooth film was formed. It was also revealed from the phase contrast image that the film surface was uniform.
Similarly, FIG. 9 shows height images and phase contrast images obtained by AFM of PS films with different EbS31 contents. A PS film containing 2% by mass of EbS31 (FIG. 9A), a PS film containing 1% by mass of EbS31 (FIG. 9B), and a PS film containing 0.5% by mass of EbS31 (FIG. 9C )) and the EbS31-free PS film (FIG. 9(d)) each had a surface root-mean-square roughness (PMS) of 0.59 nm or less, and a smooth film was formed. It was also revealed from the phase contrast image that the film surface was uniform.
From the above results, it was found that when the EbS31 content was 2% by mass or less, a smooth resin film having a surface root-mean-square roughness (PMS) of 0.59 nm or less was formed.

<Test Example 2-2>
The chemical compositions of PMMA films and PS films with different contents of EbS31 prepared in Example 2 were evaluated based on X-ray photoelectron spectroscopy (XPS) measurements. An X-ray photoelectron spectrometer (PHI5000 VersaProbe III, ULVACPHI, PHI., INC) was used for the XPS measurement. The X-ray source was a monochromatic Al Kα ray, the applied voltage was 15.0 kV, 25 W, and the measurement was performed at room temperature. Further, the photoelectron extraction angle was set to 15°. The analysis depth of carbon 1s electrons calculated from the mean free path is about 3 nm. During the measurement, the sample surface was charged up and the binding energy shifted over time, so the carbon 1s peak (285 eV) was used as a reference and corrected using a neutralization gun.
FIG. 10 shows the XPS spectra of PMMS films containing 2 wt % to 0.5 wt % EbS31. FIG. 10(a) is the spectrum of carbon atoms, FIG. 10(b) is the spectrum of oxygen atoms, and FIG. 10(c) is the spectrum of silicon atoms.
In the C _1s spectrum of FIG. 10(a), the proportion of carbon-carbon bond (C—C—C, 285 eV) peaks increased as the content of EbS31 increased. In the O _1s spectrum of FIG. 10(b), the percentage of siloxane bond (Si—O—Si, 532.5 eV) peaks increased as the content of EbS31 increased. In the Si _2p spectrum of Fig. 10(c), a siloxane bond (Si-O-Si, 102 eV) peak was obtained on the PMMA film containing EbS31.
Similarly, FIG. 11 shows XPS spectra of PS films containing 2% to 0.5% by weight of EbS31. FIG. 11(a) is the spectrum of carbon atoms, FIG. 11(b) is the spectrum of oxygen atoms, and FIG. 11(c) is the spectrum of silicon atoms.
In the C _1s spectrum of FIG. 11(a), as the content of EbS31 increases, the carbon-carbon bond (C-C-C, 285 eV), ether bond (C-O, 286.5 eV) and ester bond (C ( C═O) O, 288.2 eV) peak increased. In the O _1s spectrum of FIG. 11(b), as the content of EbS31 increases, the ratio of siloxane bond (Si—O—Si, 532.5 eV) peak and ester bond (C(=O)O, 534 eV) increases. bottom. In the Si _2p spectrum of Fig. 11(c), a siloxane bond (Si-O-Si, 102 eV) peak was obtained on the EbS31-containing PS film.
From the above results, chemical bonding of the silicone moiety, which is the hydrophobic unit of EbS31, was confirmed on the PMMA film and PS film containing EbS31. This suggests that EbS31 can concentrate on the resin surface even at low concentrations. Aggregation structures were observed on the PMMA film and PS film containing 5% by mass of EbS31, but surface concentration of EbS31 was also observed on sample surfaces without such aggregation structures.

<Test Example 2-3>
[Cell adhesion experiment]
Using the glass substrate (hereinafter referred to as the coated substrate) on which the EbS31-containing resin film was formed in Example 2, a cell adhesion experiment was performed according to the following experimental method.
The coated substrate was placed on the bottom of a 24-well cell culture plate. Each EbS31-containing resin film was washed with sterilized water three times and then immersed in PBS at room temperature for 3 hours. Mouse fibroblasts (NIH3T3) were cultured in Dulbecco's modified Eagle's medium (FBS-containing DMEM) containing 10% bovine serum (FBS) in an environment of 5% CO2 and 37°C. Cultured NIH3T3 cells were washed once with sterile PBS and detached from cell culture plates by adding 3 mL of 0.25% trypsin/0.02% ethylenediaminetetraacetic acid (EDTA) mixture. The cell suspension was centrifuged to collect NIH3T3 cells, which were then resuspended in FCS-containing DMEM to obtain a solution of 7×10 ⁴ cells/mL.
A cell suspension was prepared in a medium so as to have 5×10 ⁴ cells/cm ² in each well of the cell culture plate on which the coated substrate was fixed, and seeded onto the coated substrate. After culturing for 12 hours in an environment of 5% CO ₂ and 37° C., the coated substrate was washed with PBS three times, and the cells adhered to the EbS31-containing resin membrane were fixed using a 2% glutaraldehyde PBS solution. After washing with ultrapure water three times, it was air-dried. Adhered cells were stained with a crystal violet-PBS staining solution having a concentration of 0.01% by mass, and observed in 5 visual fields at will using an optical microscope to count the number of adhered cells.
12(a), (b), and (c) show the results (optical microscope images) of cell adhesion experiments on the surface of the EbS31-containing PMMA membrane prepared in Example 2. FIG. 12(a) to (d) are optical microscope images of the membrane after washing and staining.
The 2% by mass EbS31-containing PMMA membrane shown in FIG. 12(a) exhibited high inhibition of cell adhesion, similar to the 5% by mass EbS31-containing PMMA membrane. It was found that cell clusters adhered on the 1% by mass EbS31-containing PMMA membrane shown in FIG. 12(b). Cells clearly adhered and spread on the 0.5% by mass EbS31-containing PMMA membrane shown in FIG.
Similarly, FIGS. 12(d), (e), and (f) show the results (optical microscope images) of cell adhesion experiments on the surface of the EbS31-containing PS membrane prepared in Example 2. FIG. 12(d) to (f) are optical microscope images of the membrane after washing and staining.
The 2% by mass EbS31-containing PS membrane shown in FIG. 12(d) showed high inhibition of cell adhesion, similar to the 5% by mass EbS31-containing PS membrane. It was found that cell clusters adhered on the 1% by mass EbS31-containing PS membrane shown in FIG. 12(e). Cells clearly adhered and spread on the 0.5% by mass EbS31-containing PS membrane shown in FIG.
From the above results, PMMA membrane and PS membrane surfaces with an EbS31 content of 5% by mass or 2% by mass exhibited a high adhesion inhibitory effect, and cell adhesion was observed at 1% by mass or less. On the resin containing 2% by mass of EbS31, no aggregation structure was observed as on the resin containing 5% by mass of EbS31, but it was found that the cell adhesion inhibitory activity was not reduced.
Even when the content of EbS31 is 1% by mass or 0.5% by mass, the cell adhesion inhibitory ability is maintained as compared with the case where EbS31 is not contained.

<Example 3>
PEOMA, EbS31, and PSiOMA obtained in Synthesis Example 1, Synthesis Example 2, and Synthesis Example 4 were each mixed with a 4.5% by mass toluene solution of PMMA so as to be 2% by mass with respect to PMMA, and the surface was modified. A PMMA coating solution containing an agent was prepared. The coating liquid was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds) to form a PEOMA-containing PMMA film, an EbS31-containing PMMA film, and a PSiOMA-containing PMMA film (in Test Example 3-1, , collectively referred to as a “surface modifier-containing PMMA film”).

<Comparative Example 3-1>
A 4.5% by mass toluene solution of PMMA was prepared and used as a PMMA coating solution. The coating liquid was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds).

<Comparative Example 3-2>
For comparison with the surface modifier composition of the present invention, DBE-712 (manufactured by Gelest) having a composition of oligoethylene oxide and oligodimethylsiloxane, which is a commercial product, was added to PMMA at 10% by mass or 2% by mass. This was mixed with a 4.5% by mass toluene solution of PMMA to obtain a PMMA coating solution containing DBE-712. The coating solution was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds) to obtain a PMMA film containing DBE-712.

<Test Example 3-1>
[Cell adhesion experiment]
Using a glass substrate (hereinafter referred to as a coated substrate) on which a PMMA film containing a surface modifier was formed in Example 3, a cell adhesion experiment was performed according to the following experimental method.
The coated substrate was placed on the bottom of a 24-well cell culture plate. After washing the surface modifier-containing PMMA film with sterilized water three times, it was immersed in PBS at room temperature for 3 hours. Mouse fibroblasts (NIH3T3) were cultured in Dulbecco's modified Eagle's medium (FBS-containing DMEM) containing 10% bovine serum (FBS) in an environment of 5% CO ₂ and 37°C. Cultured NIH3T3 cells were washed once with sterile PBS and detached from cell culture plates by adding 3 mL of 0.25% trypsin/0.02% ethylenediaminetetraacetic acid (EDTA) mixture. The cell suspension was centrifuged to collect NIH3T3 cells, which were then resuspended in FCS-containing DMEM to obtain a solution of 7×10 ⁴ cells/mL.
A cell suspension was prepared in a medium so as to have 5×10 ⁴ cells/cm ² in each well of the cell culture plate on which the coated substrate was fixed, and seeded onto the coated substrate. After culturing for 12 hours in an environment of 5% CO ₂ and 37° C., the coated substrate was washed with PBS three times, and the cells adhered to the surface modifier-containing resin were fixed using a 2% glutaraldehyde PBS solution. bottom. After washing with ultrapure water three times, it was dried under reduced pressure with a rotary vacuum pump. Adhered cells were stained with a crystal violet-PBS staining solution having a concentration of 0.01% by mass, and observed in 5 visual fields at will using an optical microscope to count the number of adhered cells.
13(a), (b), and (c) show the results (optical microscope images) of cell adhesion experiments on the surface of the PMMA membrane containing the surface modifier prepared in Example 3. FIG. 13(a)-(d), (i) and (j) are optical microscope images of the membrane before washing and staining. Figures 13(e)-(h), (k) and (l) are optical microscope images of the membrane after washing and staining.
On the 2% by mass PEOMA-containing PMMA membranes shown in FIGS. 13(a) and (e), there was a tendency of cell adhesion suppression, but a slight tendency of cell adhesion was observed. The 2% by mass EbS31-containing PMMA membranes shown in FIGS. 13(b) and (f) exhibited high cell adhesion inhibition as before. 2% by mass PSiOMA-containing PMMA shown in FIGS. 13(c) and (g) did not inhibit cell adhesion.
Similarly, FIGS. 13(d) and (h) show the results of the cell adhesion test to the PMMA membrane prepared in Comparative Example 3-1.
Similarly, FIGS. 13(i), (j), (k), and (l) show the results of the cell adhesion test to the DBE-712-containing PMMA membrane prepared in Comparative Example 3-2. After 12 hours of culture, it was confirmed that the cells adhered well to the PMMA membrane containing 10% by mass or 2% by mass (FIGS. 13(i) and (j)). In addition, as a result of staining the cells adhered to the PMMA membrane containing 10% by mass or 2% by mass of DBE-712 (manufactured by Gelest) relative to the PMMA membrane, it was found that the cells were dispersed and spread well. It became clear. From these results, no inhibition of cell adhesion was observed on membranes containing DBE-712.

<Example 4>
EbS31, EbS80, and ErS31 obtained in Synthesis Example 2, Synthesis Example 3, and Synthesis Example 5 were mixed with a 4.5% by mass toluene solution of PMMA so as to be 2% by mass with respect to PMMA, and a surface modifier was added. A PMMA coating solution containing The coating solution was dropped onto a glass substrate or a silicon substrate and coated using a spin coater (2000 rpm, 60 seconds) to form an EbS31-containing PMMA film, an EbS80-containing PMMA film, and an ErS31-containing PMMA film (in Test Example 4-1, , collectively referred to as a “surface modifier-containing PMMA film”).

<Comparative Example 4>
A 24-well cell culture plate (Thermo Scientific ^™ Nunc ^™ cell culture multi-dish, cat# 142475) was used as the TCPS substrate for comparative samples.

<Test Example 4-1>
[Cell adhesion experiment]
Using a glass substrate (hereinafter referred to as a coated substrate) on which a PMMA film containing a surface modifier was formed in Example 4, a cell adhesion experiment was performed according to the following experimental method.
The coated substrate was placed on the bottom of a 24-well cell culture plate. After washing the coated substrate three times with sterilized water, it was immersed in PBS at room temperature for 3 hours. Mouse fibroblasts (NIH3T3) were cultured in Dulbecco's modified Eagle's medium (FBS-containing DMEM) containing 10% bovine serum (FBS) in an environment of 5% CO ₂ and 37°C. The cultured NIH3T3 cells were washed once with sterile PBS, added with 1 mL of 0.02% ethylenediaminetetraacetic acid (EDTA) solution and 1 mL of 0.25% trypsin solution, and detached from the cell culture plate. The cell suspension was centrifuged to collect NIH3T3 cells, which were then resuspended in FCS-containing DMEM to obtain a solution of 7×10 ⁴ cells/mL.
A cell suspension was prepared in a medium so as to have 5×10 ⁴ cells/cm ² in each well of the cell culture plate on which the coated substrate was fixed, and seeded onto the coated substrate. Observation was performed after culturing for 12, 24, 48 or 72 hours in a 5% CO ₂ environment at 37°C. After 72 hours, the coated substrate was washed with PBS three times, and the adherent cells on the surface modifier-containing PMMA membrane were immobilized using a 2% glutaraldehyde PBS solution. After washing three times with ultrapure water, the adhered cells were stained with a crystal violet-PBS staining solution having a concentration of 0.01% by mass, and air-dried. Adhered cells were observed using an optical microscope.
FIG. 14 shows the results (optical microscope image) of a cell adhesion experiment to the surface of the surface modifier-containing PMMA film produced in Example 4. FIG. 14(a) to (o) are optical microscope images of the membrane before washing and staining. Figures 14(p)-(y) are optical microscope images of the membrane after washing and staining.
On PMMA membranes containing EbS31 or EbS80, cells aggregated from 12 hours to 72 hours to form spherical aggregates (Fig. 14 (a), (b), (f), (g), ( k) and (l)). After 72 hours, the cell aggregates were slightly larger, suggesting the formation of spheroids (FIGS. 14(k) and (l)).
In Figures 14(p), (q), (u) and (v), after 72 hours of culture on PMMA membranes containing EbS31 or EbS80, the membrane surface was washed and fixed and stained as described above and observed again under a microscope. shows the results of Most cell clumps were removed from the surface by washing the membrane surface. It was found that although slight adhesion of cell aggregates remained, spreading of cells was suppressed.
FIGS. 14(c), (h) and (m) show micrographs of cells cultured on the ErS31-containing PMMA membrane prepared in Example 4 from 12 hours to 72 hours (the photographs are 24 hours and 48 hours). , or the photograph after 72 hours). The formation of cell aggregates was also observed on the ErS31-containing PMMA membrane after 72 hours of culturing. However, a state in which cell spreading did not progress was also observed. On the ErS31-containing PMMA membrane after 72 hours of washing and fixation staining, it was revealed that the cell clumps were strongly adhered without being removed so much.
Similarly, FIGS. 14(d), (i), (n), (s), and (x) show the results of the cell adhesion test to the PMMA membrane prepared in Comparative Example 3-1.
Similarly, FIGS. 14(e), (j), (o), (t), and (y) show the results of the cell adhesion test to the TCPS substrate prepared in Comparative Example 4.
It was also found that cells adhered well to the surface-modifying agent-free PMMA film used in the comparative example or on the TCPS substrate, and that cells proliferated after 72 hours (Fig. 14; PMMA film surface: (d ), (i), (n), (s), and (x), and the TCPS substrate surface: (e), (j), (o), (t), and (y)).
As a result, EbS31 had a high ability to suppress cell adhesion even when the composition ratio of EOMA and SiOMA was different. On the other hand, although ErS31 made of a random copolymer had the same composition ratio as EbS31, EbS31 had a higher ability to suppress cell adhesion. Since no cell adhesion inhibitory effect was observed on the PMMA film and TCPS substrate containing no surface modifier, the cell adhesion inhibitory ability exhibited by the resin surface containing EbS (PEOMA-block-PSiOMA) was is concentrated on the resin surface and the PEOMA chains are exposed at the water interface.

Claims (17)

a first structure that is at least one of an oxyethylene structure and a betaine structure in the side chain;
a second structure that is a dimethylsiloxane structure in the side chain;
A surface modifier containing a copolymer having The copolymer has repeating units (a1) having the first structure and repeating units (a2) having the second structure,
The repeating unit (a1) has at least one of repeating units represented by the following formulas (a1-1) to (a1-4),
The repeating unit (a2) has at least one of a repeating unit represented by the following formula (a2-1) and a repeating unit represented by (a2-2).
The surface modifier according to claim 1.

(In formula (a1-1), R ¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ² represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and X is -O- or -NR- (R represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms), m represents 1 to 100, * represents a bond.
In formula (a1-2), R ¹¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ¹² represents an alkylene group having 1 to 6 carbon atoms, and R ¹³ and R ¹⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ¹⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-3), R ²¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ²² represents an alkylene group having 1 to 6 carbon atoms, and R ²³ and R ²⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ²⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-4), R ³¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ³² represents an alkylene group having 1 to 6 carbon atoms, and R ³³ represents the number of carbon atoms. represents an alkylene group of 1 to 6, each of R ³⁴ to R ³⁶ independently represents an alkyl group having 1 to 4 carbon atoms, X is —O— or —NR— (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond. )

(In formula (a2-1), R ⁴¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ⁴² represents an alkylene group having 1 to 6 carbon atoms, and R ⁴³ represents a monovalent represents an organic group, m represents 1 to 100. * represents a bond.
In formula (a2-2), R ⁵¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and R ⁵² represents an alkylene group having 1 to 6 carbon atoms. * represents a bond. ) 3. The surface modifier according to claim 2, wherein the copolymer has a molar ratio (a1:a2) between the repeating unit (a1) and the repeating unit (a2) of 99:1 to 10:90. . The surface modifier according to any one of claims 1 to 3, wherein the copolymer is a block copolymer. 5. The surface modifier according to any one of claims 1 to 4, which is used for suppressing adhesion of biological substances. The surface modifier according to any one of claims 1 to 5, wherein the copolymer has a number average molecular weight (Mn) of 1,000 to 100,000. A composition comprising the surface modifier according to any one of claims 1-6. The composition according to claim 7, which is used for suppressing adhesion of biological substances. 9. A composition according to claim 7 or 8, further comprising a resin. A medical device having a membrane formed from the composition according to any one of claims 7-9. A silicone substrate having a film formed from the composition according to any one of claims 7-9. A cell culture vessel having a membrane formed from the composition according to any one of claims 7-9. Having a repeating unit (a1) and a repeating unit (a2),
The repeating unit (a1) has at least one of repeating units represented by the following formulas (a1-1) to (a1-4),
The repeating unit (a2) has at least one of a repeating unit represented by the following formula (a2-1) and a repeating unit represented by (a2-2).
copolymer.

(In formula (a1-1), R ¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ² represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and X is -O- or -NR- (R represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms), m represents 1 to 100, * represents a bond.
In formula (a1-2), R ¹¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ¹² represents an alkylene group having 1 to 6 carbon atoms, and R ¹³ and R ¹⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ¹⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-3), R ²¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ²² represents an alkylene group having 1 to 6 carbon atoms, and R ²³ and R ²⁴ are Each independently represents an alkyl group having 1 to 4 carbon atoms, R ²⁵ represents an alkylene group having 1 to 6 carbon atoms, X is -O- or -NR- (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond.
In formula (a1-4), R ³¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ³² represents an alkylene group having 1 to 6 carbon atoms, and R ³³ represents the number of carbon atoms. represents an alkylene group of 1 to 6, each of R ³⁴ to R ³⁶ independently represents an alkyl group having 1 to 4 carbon atoms, X is —O— or —NR— (R is a hydrogen atom or represents an alkyl group having 1 to 4 carbon atoms.). * represents a bond. )

(In formula (a2-1), R ⁴¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, R ⁴² represents an alkylene group having 1 to 6 carbon atoms, and R ⁴³ represents a monovalent represents an organic group, m represents 1 to 100. * represents a bond.
In formula (a2-2), R ⁵¹ represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, and R ⁵² represents an alkylene group having 1 to 6 carbon atoms. * represents a bond. ) 14. The copolymer of claim 13, which is a block copolymer. 15. The copolymer according to claim 13 or 14, which is used for suppressing adhesion of biological substances. The copolymer according to any one of claims 13 to 15, having a number average molecular weight (Mn) of 1,000 to 100,000. 17. The copolymer according to any one of claims 13 to 16, wherein the molar ratio (a1:a2) between the repeating unit (a1) and the repeating unit (a2) is from 99:1 to 10:90.

Images