CN110606963A - Preparation method of high-adhesion anti-oxidation self-repairable film for promoting cell adhesion - Google Patents

Preparation method of high-adhesion anti-oxidation self-repairable film for promoting cell adhesion Download PDF

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Publication number
CN110606963A
CN110606963A CN201910890912.3A CN201910890912A CN110606963A CN 110606963 A CN110606963 A CN 110606963A CN 201910890912 A CN201910890912 A CN 201910890912A CN 110606963 A CN110606963 A CN 110606963A
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film
self
grafted
cell adhesion
preparation
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葛丽芹
苑仁强
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Southeast University
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Southeast University
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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J5/00—Manufacture of articles or shaped materials containing macromolecular substances
    • C08J5/18—Manufacture of films or sheets
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0068—General culture methods using substrates
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
    • C08J2305/04—Alginic acid; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
    • C08J2305/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/10—Mineral substrates
    • C12N2533/12—Glass
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/20—Small organic molecules
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2533/00—Supports or coatings for cell culture, characterised by material
    • C12N2533/70—Polysaccharides
    • C12N2533/74—Alginate

Abstract

The invention discloses a preparation method of a high-adhesiveness antioxidant self-repairing film for promoting cell adhesion, which is characterized in that a layer-by-layer self-assembly technology is adopted to prepare the film, firstly, an RGD polypeptide grafted carboxymethyl chitosan (CCS-R) solution is dripped onto a substrate, a layer of CCS-R film is obtained through spin coating, dopamine grafted oxidized sodium alginate (OALG-D) is continuously spin coated after being washed by ultrapure water, the substrate is washed by the ultrapure water to remove the unbound OALG-D, and a (CCS-R/OALG-D) film is obtained after drying, wherein the grafted RGD polypeptide plays a role in promoting cell adhesion, the grafted dopamine provides a high-adhesiveness and an antioxidant effect for the film, and a Schiff base connecting bond is formed between an amino group of carboxymethyl chitosan and an aldehyde group of the oxidized sodium alginate, so that the film has excellent self-repairing performance. The preparation method provided by the invention is simple and convenient to operate, simple in process, low in cost and short in required time, and can be used for large-scale production.

Description

Preparation method of high-adhesion anti-oxidation self-repairable film for promoting cell adhesion
Technical Field
The invention relates to a preparation technology of a high-adhesion and anti-oxidation self-repairable film for promoting cell adhesion, and belongs to the technical field of biological materials.
Background
Regenerative medicine has recently attracted increasing attention because it brings great hope for patients suffering from tissue deterioration and organ failure caused by disease, aging or trauma. Rapid advances in regenerative medical technology are not open to the study of precisely designed intelligent and smart biomaterials that resemble physiological microenvironments. Therefore, the development of a platform that mimics the extracellular matrix that supports cell adhesion is of great interest. The effect of cell adhesion to the material determines cell migration, differentiation and proliferation. Studies have shown that cell adhesion is influenced by the physicochemical properties of the matrix, including surface charge, surface wettability and hardness, and the material assembly method.
The self-repairing film material plays a role in promoting cell adhesion at multiple angles: the grafted viscous Arg-Gly-Asp (RGD) peptide in the self-repairing film is used as an integrin binding sequence in extracellular matrix protein to play a role in promoting cell adhesion; the grafted dopamine provides high adhesion and an anti-oxidation effect for the film, a Schiff base connecting bond is formed between the amino group of the carboxymethyl chitosan and the aldehyde group of the oxidized sodium alginate, the self-repairing performance is excellent, and when the material is damaged by the outside, the integrity of the material can be recovered through the self-repairing function, so that the service life is prolonged.
Disclosure of Invention
The technical problem is as follows: the invention discloses a preparation method of a high-adhesion antioxidant self-repairable film for promoting cell adhesion, wherein a dynamic Schiff base bond is formed between an aldehyde group of oxidized sodium alginate and an amino group of carboxymethyl chitosan to endow the film with self-repairing performance, meanwhile, the RGD polypeptide grafted on the carboxymethyl chitosan can enhance the capacity of promoting cell adhesion, and dopamine endows the film with high adhesion stress and antioxidant performance; in addition, the film is assembled by polysaccharide with excellent biocompatibility and has no in-vitro cytotoxicity, so that the film prepared by the layer-by-layer self-assembly method can enhance the cell adhesion in multiple aspects.
The technical scheme is as follows: the preparation method of the high-adhesion antioxidant self-repairable film for promoting cell adhesion comprises the following steps: firstly, dropwise adding an RGD polypeptide grafted carboxymethyl chitosan CCS-R solution onto a substrate, carrying out spin coating to obtain a layer of CCS-R film, washing with ultrapure water, then continuously carrying out spin coating with dopamine grafted oxidized sodium alginate OALG-D, washing the substrate with ultrapure water to remove unbound OALG-D, and drying to obtain the CCS-R/OALG-D self-repairing film.
Wherein the RGD polypeptide comprises linear polypeptide and cyclic polypeptide, and the oxidized sodium alginate is polysaccharide containing aldehyde groups.
The RGD grafted carboxymethyl chitosan and the dopamine grafted oxidized sodium alginate are spin-coated on the surface of the substrate layer by a layer-by-layer self-assembly method, and the process can be repeated for multiple times to obtain the multilayer self-repairing film.
The spin coating mode is 1000rpm to 3000rpm, and the time is 2min to 10 min.
The concentration of the RGD grafted carboxymethyl chitosan is 1 mg/mL-10 mg/mL.
The substrate comprises glass, plastics, metal, silicon chips, epoxy plates or cell culture pore plates.
The concentration of the grafted dopamine grafted oxidized sodium alginate is 1 mg/mL-10 mg/mL.
Has the advantages that: the self-repairing film prepared by the RGD polypeptide grafted carboxymethyl chitosan and dopamine grafted oxidized sodium alginate solution through the layer-by-layer self-assembly method has the following advantages:
1. the method is simple and effective, the operation is simple and convenient, the required time is short, and the required equipment is simple.
2. The selected materials are polysaccharide materials with good biocompatibility and have no in vitro cytotoxicity.
3. The prepared film provides high adhesion, an anti-oxidation function and a self-repairing characteristic, and can realize a remarkable cell adhesion enhancing effect.
Drawings
FIG. 1a is a graph showing the effect of cell adhesion of a film to which RGD polypeptide and dopamine are not grafted, and FIG. 1b is a graph showing the effect of cell adhesion of a film to which RGD polypeptide and dopamine are grafted;
fig. 2a is a metallographic microscope image of a scratched film grafted with RGD polypeptide and dopamine, and fig. 2b is a metallographic microscope image of a self-repaired film grafted with RGD polypeptide and dopamine.
Detailed Description
The invention is explained in more detail below with reference to exemplary embodiments and the accompanying drawings. The following examples are provided only for illustrating the present invention and are not intended to limit the scope of the present invention.
The invention provides a preparation method of a self-repairing film, which comprises the following steps: the preparation method comprises the steps of preparing a film by adopting a layer-by-layer self-assembly technology, firstly, dropwise adding a RGD polypeptide grafted carboxymethyl chitosan (CCS-R) solution onto a substrate, carrying out spin coating to obtain a layer of CCS-R film, washing the CCS-R film by using ultrapure water, then continuously carrying out spin coating to dopamine grafted sodium alginate oxide (OALG-D), washing the substrate by using the ultrapure water to remove the unbound OALG-D, and drying to obtain the (CCS-R/OALG-D) film.
Wherein the content of the first and second substances,
the material prepared by the film is RGD grafted carboxymethyl chitosan and dopamine grafted oxidized sodium alginate, wherein the RGD polypeptide comprises linear polypeptide and cyclic polypeptide, and the oxidized sodium alginate is polysaccharide containing aldehyde groups.
The RGD grafted carboxymethyl chitosan and the dopamine grafted oxidized sodium alginate are spin-coated on the surface of the substrate layer by using a layer-by-layer self-assembly method, and the process can be repeated for multiple times to obtain the multilayer self-repairing film.
The spin coating mode is 1000rpm to 3000rpm, and the time is 2min to 10 min.
The concentration of the RGD grafted carboxymethyl chitosan is 1 mg/mL-10 mg/mL.
The substrate types include glass, plastic, metal, silicon wafer, epoxy plate, cell culture well plate, etc.
The concentration of the grafted dopamine grafted oxidized sodium alginate is 1 mg/mL-10 mg/mL.
The preparation method specifically comprises the following steps:
the self-repairing film is prepared by utilizing RGD polypeptide grafted carboxymethyl chitosan and dopamine grafted oxidized sodium alginate solution through a layer-by-layer self-assembly method.
Specifically, firstly, dropwise adding a RGD polypeptide grafted carboxymethyl chitosan (CCS-R) solution onto a substrate, spin-coating for a period of time at a certain speed to obtain a layer of CCS-R film, washing for 3 times with ultrapure water, then continuously spin-coating dopamine grafted sodium alginate oxide (OALG-D), washing the substrate with ultrapure water to remove unbound OALG-D, and drying to obtain the (CCS-R/OALG-D) film.
Example 1:
firstly, 1mg/mL RGD polypeptide grafted carboxymethyl chitosan (CCS-R) solution is dripped on a substrate, spin-coating is carried out for 2min at the speed of 1000rpm to obtain a layer of CCS-R film, 1mg/mL dopamine grafted sodium alginate oxide (OALG-D) is continuously spin-coated after the substrate is washed for 3 times by ultrapure water, the substrate is washed by the ultrapure water to remove the unbound OALG-D, the steps are repeated for 15 times, and a 15-layer (CCS-R/OALG-D) film is obtained after drying.
Example 2:
firstly, 2.5mg/mL RGD polypeptide grafted carboxymethyl chitosan (CCS-R) solution is dripped on a substrate, spin-coating is carried out for 5min at the speed of 1500rpm to obtain a layer of CCS-R film, 7.5mg/mL dopamine grafted sodium alginate oxide (OALG-D) is continuously spin-coated after the CCS-R film is washed for 3 times by ultrapure water, the substrate is washed by ultrapure water to remove the unbound OALG-D, the steps are repeated for 10 times, and 10 layers of (CCS-R/OALG-D) films are obtained after drying.
Example 3:
firstly, 10mg/mL RGD polypeptide grafted carboxymethyl chitosan (CCS-R) solution is dripped on a substrate, spin-coating is carried out for 10min at the speed of 3000rpm to obtain a layer of CCS-R film, after washing is carried out for 3 times by using ultrapure water, 10mg/mL dopamine grafted sodium alginate oxide (OALG-D) is continuously spin-coated, the substrate is washed by using the ultrapure water to remove unbound OALG-D, the steps are repeated for 5 times, and 5 layers of (CCS-R/OALG-D) films are obtained after drying.

Claims (7)

1. A preparation method of a high-adhesiveness antioxidant self-repairing film for promoting cell adhesion is characterized by comprising the following steps: firstly, dropwise adding an RGD polypeptide grafted carboxymethyl chitosan CCS-R solution onto a substrate, carrying out spin coating to obtain a layer of CCS-R film, washing with ultrapure water, then continuously carrying out spin coating with dopamine grafted oxidized sodium alginate OALG-D, washing the substrate with ultrapure water to remove unbound OALG-D, and drying to obtain the CCS-R/OALG-D self-repairing film.
2. The method for preparing the cell adhesion promoting high-adhesion antioxidant self-healing film according to claim 1, wherein the RGD polypeptide comprises linear polypeptide and cyclic polypeptide, and the oxidized sodium alginate is polysaccharide containing aldehyde groups.
3. The preparation method of the cell adhesion promoting high-adhesiveness antioxidant self-repairing film according to claim 1, characterized in that the RGD grafted carboxymethyl chitosan and the dopamine grafted oxidized sodium alginate are spin-coated on the surface of the substrate layer by a layer-by-layer self-assembly method, and the above processes can be repeated many times to obtain the multilayer self-repairing film.
4. The preparation method of the cell adhesion promoting high-adhesion oxidation-resistant self-repairable film according to claim 1, wherein the spin coating mode is 1000-3000 rpm, and the time is 2-10 min.
5. The preparation method of the cell adhesion promoting high-adhesion oxidation-resistant self-repairable film according to claim 1, wherein the concentration of the RGD grafted carboxymethyl chitosan is 1 mg/mL-10 mg/mL.
6. The method for preparing the cell adhesion promoting high-adhesion oxidation resistant self-healing film according to claim 1, wherein the substrate comprises glass, plastic, metal, silicon wafer, epoxy plate or cell culture well plate.
7. The preparation method of the cell adhesion promoting high-adhesion antioxidant self-repairable film according to claim 1, wherein the concentration of the grafted dopamine grafted oxidized sodium alginate is 1mg/mL to 10 mg/mL.
CN201910890912.3A 2019-09-20 2019-09-20 Preparation method of high-adhesion anti-oxidation self-repairable film for promoting cell adhesion Pending CN110606963A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111138696A (en) * 2020-01-13 2020-05-12 北京科技大学 Alginic acid dialdehyde/chitosan antifogging antibacterial film and preparation method thereof
CN111317852A (en) * 2020-02-27 2020-06-23 吉林大学 Medical dressing compounded by chitosan and polypropylene carbonate and preparation method thereof
CN113058830A (en) * 2021-03-16 2021-07-02 安徽华能电缆集团有限公司 Preparation method of underwater adhesive coating capable of being adhered for long time based on alloy substrate
CN113802364A (en) * 2021-09-01 2021-12-17 昆明理工大学 Preparation method of self-repairing, acid-base-resistant and antibacterial multilayer film coating

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104623725A (en) * 2014-12-31 2015-05-20 深圳清华大学研究院 Bioadhesive and preparation method thereof
CN107375995A (en) * 2017-07-21 2017-11-24 临沂市人民医院 A kind of preparation method based on layer assembly function selfreparing aquogel type dressing materials

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104623725A (en) * 2014-12-31 2015-05-20 深圳清华大学研究院 Bioadhesive and preparation method thereof
CN107375995A (en) * 2017-07-21 2017-11-24 临沂市人民医院 A kind of preparation method based on layer assembly function selfreparing aquogel type dressing materials

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
LIU XUEFAN等: "Multilayer Edible Fresh-keeping Films" Characterization and Their Preservation Effect", 《CHEMISTRY SELECT》 *
葛丽芹、苑仁强: "仿生自修复涂膜材料促进珊瑚细胞粘附的研究", 《中国化学会第 17届胶体与界面化学学术会议论文（摘要）集》 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111138696A (en) * 2020-01-13 2020-05-12 北京科技大学 Alginic acid dialdehyde/chitosan antifogging antibacterial film and preparation method thereof
CN111317852A (en) * 2020-02-27 2020-06-23 吉林大学 Medical dressing compounded by chitosan and polypropylene carbonate and preparation method thereof
CN111317852B (en) * 2020-02-27 2021-10-22 吉林大学 Medical dressing compounded by chitosan and polypropylene carbonate and preparation method thereof
CN113058830A (en) * 2021-03-16 2021-07-02 安徽华能电缆集团有限公司 Preparation method of underwater adhesive coating capable of being adhered for long time based on alloy substrate
CN113058830B (en) * 2021-03-16 2022-05-13 安徽华能电缆集团有限公司 Preparation method of underwater adhesive coating capable of being adhered for long time based on alloy substrate
CN113802364A (en) * 2021-09-01 2021-12-17 昆明理工大学 Preparation method of self-repairing, acid-base-resistant and antibacterial multilayer film coating
CN113802364B (en) * 2021-09-01 2023-03-10 昆明理工大学 Preparation method of self-repairing, acid-base-resistant and antibacterial multilayer film coating

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