CN116850344A - Preparation method of bioengineering implant - Google Patents
Preparation method of bioengineering implant Download PDFInfo
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- CN116850344A CN116850344A CN202310780935.5A CN202310780935A CN116850344A CN 116850344 A CN116850344 A CN 116850344A CN 202310780935 A CN202310780935 A CN 202310780935A CN 116850344 A CN116850344 A CN 116850344A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 10
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- 229920001610 polycaprolactone Polymers 0.000 description 3
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- 230000002107 myocardial effect Effects 0.000 description 2
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 2
- 229920001308 poly(aminoacid) Polymers 0.000 description 2
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- 229960004889 salicylic acid Drugs 0.000 description 2
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- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
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Classifications
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- G—PHYSICS
- G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
- G03F—PHOTOMECHANICAL PRODUCTION OF TEXTURED OR PATTERNED SURFACES, e.g. FOR PRINTING, FOR PROCESSING OF SEMICONDUCTOR DEVICES; MATERIALS THEREFOR; ORIGINALS THEREFOR; APPARATUS SPECIALLY ADAPTED THEREFOR
- G03F7/00—Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor
- G03F7/20—Exposure; Apparatus therefor
- G03F7/2002—Exposure; Apparatus therefor with visible light or UV light, through an original having an opaque pattern on a transparent support, e.g. film printing, projection printing; by reflection of visible or UV light from an original such as a printed image
- G03F7/2004—Exposure; Apparatus therefor with visible light or UV light, through an original having an opaque pattern on a transparent support, e.g. film printing, projection printing; by reflection of visible or UV light from an original such as a printed image characterised by the use of a particular light source, e.g. fluorescent lamps or deep UV light
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/20—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
-
- 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
- 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
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0657—Cardiomyocytes; Heart cells
-
- G—PHYSICS
- G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
- G03F—PHOTOMECHANICAL PRODUCTION OF TEXTURED OR PATTERNED SURFACES, e.g. FOR PRINTING, FOR PROCESSING OF SEMICONDUCTOR DEVICES; MATERIALS THEREFOR; ORIGINALS THEREFOR; APPARATUS SPECIALLY ADAPTED THEREFOR
- G03F7/00—Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor
- G03F7/20—Exposure; Apparatus therefor
-
- 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
- C12N2533/72—Chitin, chitosan
-
- 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
- C12N2535/00—Supports or coatings for cell culture characterised by topography
- C12N2535/10—Patterned coating
-
- 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
- C12N2537/00—Supports and/or coatings for cell culture characterised by physical or chemical treatment
- C12N2537/10—Cross-linking
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Veterinary Medicine (AREA)
- General Engineering & Computer Science (AREA)
- Cell Biology (AREA)
- Public Health (AREA)
- Dermatology (AREA)
- Microbiology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Cardiology (AREA)
- Rheumatology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the field of biomedical materials and tissue engineering, and in particular relates to a preparation method of a bioengineered implant. The invention adopts a one-step light direct writing method to prepare the surface micropatterned hydrogel or elastomer as a bioengineering implant. The one-step optical direct writing method is different from traditional complex process technologies such as photoetching casting film reproduction, soft photoetching, imprinting, 3D printing and the like, takes natural biomacromolecules such as collagen with good biocompatibility and low immunogenicity as a bracket material, prepares uniform gel or elastomer prepolymer by adding photosensitive micromolecules for blending, and spontaneously forms ordered or unordered fold microstructured surfaces by exposure and crosslinking under an ultraviolet or visible light source with a certain wavelength.
Description
Technical Field
The invention belongs to the field of biomedical materials and tissue engineering, and in particular relates to a preparation method of a bioengineered implant.
Background
In the cell culture environment constructed by the traditional bioengineering graft, the vast majority uses a two-dimensional environment as a support, under the environment, the cell culture mainly imitates the in-vivo biochemical environment, the graft mainly shows the functions of the support and the bracket in the body, and the environment has a great gap compared with the three-dimensional environment in which the cells live in situ in the body. With the progressive depth of research, extracellular matrix components and their unique environments of construction have had a tremendous role in regulating cellular behavior.
In the construction of the traditional three-dimensional culture environment, a plurality of different technologies are utilized to prepare micropattern structural materials with different dimensions and different morphologies, such as a laser etching technology, a soft lithography technology, a 3D printing technology and the like, the technologies are developed to be perfect and stable in the continuous development process, the precision of micron-level or even nano-level can be realized, the technologies generally need a plurality of steps to complete the preparation of the final micropattern, and the process is complex. Meanwhile, the precision of the template manufactured by the photoetching technology is reduced after the template is used for a plurality of times.
Disclosure of Invention
In order to solve the problems, the invention provides a preparation method of a bioengineered implant, wherein the preparation method is a one-step optical direct writing method, and the bioengineered implant is surface micropatterned gel or elastomer.
Further, the one-step optical direct writing method specifically comprises the following steps:
(1) Preparing a dynamic polymer crosslinked network structure;
(2) Dissolving the polymer, and adding photosensitive molecules into a prepolymer system;
(3) The mixed liquid is made into a required structure, a photomask plate is used for being paved on an exposure frame, the distance between the photomask plate and an ultraviolet light source as well as the distance between the photomask plate and a sample are fixed, and the mixture is exposed under the ultraviolet light source or the visible light source.
Further, the dynamic polymer cross-linked network structure scaffold material in the step (1) is one or more of chitin, chitosan and chitosan.
Further, the dissolving solution in the step (2) comprises deionized water, physiological saline, PBS solution or formic acid solution.
Further, the photosensitive material in the step (2) comprises anthranilate, coumarin, cinnamate and vinyl pyrene.
Further, the ratio of the base material to the photosensitive molecules in the step (2) is 0.1% -30%.
Further, the prepolymer in the step (2) is dissolved at a concentration of 1 to 95%.
Further, the pattern that can be produced in step (3) is in the size range of 0.01-2000 microns.
The invention has the following beneficial effects:
The method simplifies the preparation flow of the micropattern, has a plurality of choices in the pattern selectivity, and simultaneously realizes the dynamic polymerization and depolymerization of the micropattern.
The stent material used in the invention has good biocompatibility, has advantages for subsequent cell culture, has lower immunogenicity as a bioengineered graft, and can reduce immune response of an organism to the graft after implantation. Thereby being better applied to the biomedical field.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art. FIG. 1 is a schematic diagram of a one-step optical direct writing process for preparing a surface patterned cell culture scaffold according to the present invention;
FIG. 2 preparation of a dynamic polymer crosslinked network from an amino group-containing polymer (exemplified by chitosan CS and polyacrylate copolymer PDASBA);
FIG. 3 is a view of a PAN material mirror prepared by a one-step optical direct writing method and having a concentric circle structure and a 'back' shape structure;
FIG. 4 is a fluorescent image of a material (PAN, example 1) with a surface micropattern structure prepared from the prepared "Hui" polyacrylate and photosensitive molecular vinyl pyrene, after the material was seeded with primary cardiomyocytes from a suckling mouse.
Fig. 5 is a fluorescent image of myoblasts L6 grown on the prepared grating patterned example 3 material.
FIG. 6 is a fluorescent photograph of a prepared grating patterned example 10 material after primary cardiomyocytes were seeded thereon.
Fig. 7 is a fluorescent photograph of myoblasts C2C12 after being planted on the prepared grating patterned example 18 material.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, with reference to the examples using conventional methods, unless otherwise indicated, and with reference to reagents, either conventional commercial reagents or reagents configured using conventional methods. The detailed description is not to be taken as limiting, but is to be understood as a more detailed description of certain aspects, features, and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
According to a first aspect of the present invention, there is provided a method for preparing a one-step optical direct-writing gel surface micro-nano structure, the method comprising (a) uniformly mixing an aqueous solution of a polymer system in a mold or tube having a certain depth to obtain a gel prepolymer having a certain thickness; and (B) exposing the above gel prepolymer layer to an ultraviolet light source or a visible light source having a wavelength of 300nm or more, thereby spontaneously forming a maze-like disordered pattern having a recognizable size in the range of 0.01 to 1000 μm, wherein the polymer system comprises a natural biomacromolecule or an artificially synthesized polymer having a group that can chemically react under the ultraviolet light exposure condition.
Example 1
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
the chitosan or chitosan derivative is used as a bracket material to prepare a dynamic polymer crosslinked network structure. After the materials are dissolved in deionized water, normal saline, PBS solution or formic acid solution (including but not limited to these solutions) and the like according to the concentration of the prepolymer of 3%, photosensitive molecules (including but not limited to these molecules) such as anthranilate, coumarin, cinnamate, vinyl pyrene and the like are added into the prepolymer system, and the proportion of the base material and the photosensitive molecules is 0.1% -30%. For example: chitosan, chitosan or its derivative, chitosan and anthranilate are polymerized to form supermolecule hydrogen bond photocrosslinking network with light and acid-base gas sensitivity. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. As shown in fig. 2. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 2
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
the chitosan or chitosan derivative is used as a bracket material to prepare a dynamic polymer crosslinked network structure. After the materials are dissolved in an acidic solution such as hydrochloric acid, formic acid, acetic acid, lactic acid, malic acid, ascorbic acid or some common organic acid (including but not limited to these solutions) according to the concentration of the prepolymer of 30%, photosensitive molecules such as anthranilate, coumarin, cinnamate, vinyl pyrene or the like (including but not limited to these molecules) are added into the prepolymer system, and the ratio of the base material to the photosensitive molecules is 0.1% -30%. For example: chitosan, chitosan or its derivative is polymerized with anthranilate through hydrogen bond to form supermolecule hydrogen bond photocrosslinking network with light and acid-base gas sensitivity. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 3
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
the chitosan or chitosan derivative is used as a bracket material to prepare a dynamic polymer crosslinked network structure. These materials are dissolved in an acidic solution such as hydrochloric acid, formic acid, acetic acid, lactic acid, malic acid, ascorbic acid or some common organic acid (including but not limited to these solutions) at a prepolymer concentration of 80%, and then a photosensitive molecule such as anthranilate, coumarin, cinnamate, vinyl pyrene or the like (including but not limited to these molecules) is added to the prepolymer system. The proportion of the substrate material and the photosensitive molecules is in the range of 0.1% to 30%. For example: chitosan, chitosan or its derivative is polymerized with anthranilate through hydrogen bond to form supermolecule hydrogen bond photocrosslinking network with light and acid-base gas sensitivity. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 4
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
dynamic polymer cross-linked network structures were prepared using amino group containing polyacrylate copolymers (PDASBA) as scaffold materials. After the material is dissolved in an organic solvent such as acetone, ethyl acetate and the like (including but not limited to the solutions) according to the concentration of the prepolymer of 3 percent, photosensitive molecules such as anthranilate, coumarin, cinnamate, vinyl pyrene and the like (including but not limited to the molecules) are added into the polyacrylate solution, and the proportion of the substrate material and the photosensitive molecules is 0.1 percent to 30 percent. For example: the polyacrylate forms a supramolecular ionomer network with vinyl pyrene (such polymers are also sensitive to carbon dioxide in addition to uv light). The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micro-nano pattern is prepared by a one-step illumination method under the ultraviolet light source or visible light exposure for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 5
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
dynamic polymer cross-linked network structures were prepared using amino group containing polyacrylate copolymers (PDASBA) as scaffold materials. After the material is dissolved in an organic solvent such as acetone, ethyl acetate and the like (including but not limited to the solutions) according to the concentration of the prepolymer of 90 percent, photosensitive molecules such as anthranilate, coumarin, cinnamate, vinyl pyrene and the like (including but not limited to the molecules) are added into the polyacrylate solution, and the proportion of the substrate material and the photosensitive molecules is 0.1 percent to 30 percent. For example: the polyacrylate forms a supramolecular ionomer network with vinyl pyrene (such polymers are also sensitive to carbon dioxide in addition to uv light). The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micro-nano pattern is prepared by a one-step illumination method under the ultraviolet light source or visible light exposure for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 6
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
a dynamic polymer cross-linked network structure is prepared by using cellulose as a scaffold material. Cellulose is dissolved in a solution (including but not limited to such solutions) such as cuprammonium or cuprammonium diamine according to the concentration of the prepolymer of 3%, and photosensitive molecules (including but not limited to such molecules) such as anthranilate, coumarin, cinnamate and vinyl pyrene are added into the prepolymer system, wherein the proportion of the substrate material and the photosensitive molecules is 0.1% -30%. For example: a hydrogen bonding polymer network is formed between the cellulose and the anthranilate. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 7
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
a dynamic polymer cross-linked network structure is prepared by using cellulose as a scaffold material. Cellulose is dissolved in a solution (including but not limited to such solutions) such as cuprammonium or cuprammonium diamine according to the concentration of the prepolymer of 90%, and photosensitive molecules (including but not limited to such molecules) such as anthranilate, coumarin, cinnamate and vinyl pyrene are added into the prepolymer system, wherein the proportion of the substrate material and the photosensitive molecules is 0.1% -30%. For example: a hydrogen bonding polymer network is formed between the cellulose and the anthranilate. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 8
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
the collagen or its derivative gelatin is used as the scaffold material to prepare the dynamic polymer cross-linked network structure. After these materials are dissolved in deionized water, physiological saline, PBS solution or solution of glycerin, propylene glycol, acetic acid, salicylic acid, phthalic acid (including but not limited to these solutions) and the like according to the concentration of the prepolymer of 5%, photosensitive molecules (including but not limited to these molecules) such as anthracene, anthranilate, coumarin, cinnamate, vinyl pyrene and the like are added into the prepolymer system, and the proportion of the base material and the photosensitive molecules is 0.1% -30%. For example: gelatin can form a hydrogen bond photo-crosslinking network with anthracene, and the system can generate stress in the system through anthracene group gradient photo-dimerization reaction so as to form a regular and ordered surface pattern, and the gelatin patterned scaffold material is prepared by one-step illumination. And the anthracene group can be reversibly photodimerized under the irradiation of ultraviolet light with different wavelengths, so that the fold pattern can be reversibly wiped off and regenerated, and thus has photoresponsivity. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 9
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
the collagen or its derivative gelatin is used as the scaffold material to prepare the dynamic polymer cross-linked network structure. After these materials are dissolved in deionized water, physiological saline, PBS solution or solution of glycerin, propylene glycol, acetic acid, salicylic acid, phthalic acid (including but not limited to these solutions) and the like according to the concentration of the prepolymer of 85%, photosensitive molecules (including but not limited to these molecules) such as anthracene, anthranilate, coumarin, cinnamate, vinyl pyrene and the like are added into the prepolymer system, and the proportion of the base material and the photosensitive molecules is 0.1% -30%. For example: gelatin can form a hydrogen bond photo-crosslinking network with anthracene, and the system can generate stress in the system through anthracene group gradient photo-dimerization reaction so as to form a regular and ordered surface pattern, and the gelatin patterned scaffold material is prepared by one-step illumination. And the anthracene group can be reversibly photodimerized under the irradiation of ultraviolet light with different wavelengths, so that the fold pattern can be reversibly wiped off and regenerated, and thus has photoresponsivity. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 10 (seemed to repeat with example 8, collagen concentration was close)
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
the collagen or its derivative gelatin is used as the scaffold material to prepare the dynamic polymer cross-linked network structure. After the materials are dissolved in deionized water, normal saline, PBS solution (including but not limited to the solutions) and the like according to the concentration of the prepolymer of 3 percent, photosensitive molecules (including but not limited to the molecules) such as anthracene, anthranilate, coumarin, cinnamate, vinyl pyrene and the like are added into the prepolymer system, and the proportion of the substrate material and the photosensitive molecules is 0.1 percent to 30 percent. For example: the alginate can form a hydrogen bond photo-crosslinking network with anthracene, the system can generate stress in the system through anthracene group gradient photo-dimerization reaction so as to form a regular and ordered surface pattern, and anthracene groups can generate reversible photo-dimerization under the irradiation of ultraviolet light with different wavelengths, so that the fold pattern can be reversibly wiped off and regenerated, and thus the system has photo-responsiveness. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 11
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
and preparing a dynamic polymer crosslinked network structure by using alginate as a scaffold material. After the alginate is dissolved in deionized water, normal saline, PBS (including but not limited to these solutions) and other solutions according to the concentration of the prepolymer of 85%, anthracene, anthranilate, coumarin, cinnamate, vinyl pyrene and other photosensitive molecules (including but not limited to these molecules) are added into the prepolymer system, and the proportion of the substrate material and the photosensitive molecules is 0.1% -30%. For example: the alginate can form a hydrogen bond photo-crosslinking network with anthracene, the system can generate stress in the system through anthracene group gradient photo-dimerization reaction so as to form a regular and ordered surface pattern, and anthracene groups can generate reversible photo-dimerization under the irradiation of ultraviolet light with different wavelengths, so that the fold pattern can be reversibly wiped off and regenerated, and thus the system has photo-responsiveness. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 12
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
the dynamic polymer cross-linked network structure is prepared by using hyaluronic acid as a scaffold material. Hyaluronic acid and alginate are similar, and are composed of uronic acid. After hyaluronic acid is dissolved in deionized water, normal saline, PBS (including but not limited to these solutions) and other solutions according to the concentration of the prepolymer of 5%, anthracene, anthranilate, coumarin, cinnamate, vinyl pyrene and other photosensitive molecules (including but not limited to these molecules) are added into the prepolymer system, and the proportion of the substrate material and the photosensitive molecules is 0.1% -30%. For example: hyaluronic acid can form a hydrogen bond photo-crosslinking network with cinnamate, and the system can generate photopolymerization reaction through the cinnamate to generate stress in the system so as to form a regular and ordered surface pattern. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 13
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
the dynamic polymer cross-linked network structure is prepared by using hyaluronic acid as a scaffold material. Hyaluronic acid and alginate are similar, and are composed of uronic acid. After hyaluronic acid is dissolved in deionized water, normal saline, PBS (including but not limited to these solutions) and other solutions according to the concentration of the prepolymer of 85%, anthracene, anthranilate, coumarin, cinnamate, vinyl pyrene and other photosensitive molecules (including but not limited to these molecules) are added into the prepolymer system, and the proportion of the substrate material and the photosensitive molecules is 0.1% -30%. For example: hyaluronic acid can form a hydrogen bond photo-crosslinking network with cinnamate, and the system can generate photopolymerization reaction through the cinnamate to generate stress in the system so as to form a regular and ordered surface pattern. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 14
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
the dynamic polymer cross-linked network structure is prepared by using human keratin or silk fibroin as a scaffold material. Human hair keratin and silk fibroin are fibrous proteins, the former is extracted from human hair or nails, the latter is extracted from silk, after the human hair keratin and silk fibroin are hydrolyzed and dissolved in deionized water, physiological saline, PBS (including but not limited to these solutions) and other solutions according to the concentration of the prepolymer, anthracene, anthranilate, coumarin, cinnamate, vinyl pyrene and other photosensitive molecules (including but not limited to these molecules) are added into the prepolymer system, and the proportion of the base material and the photosensitive molecules is 0.1% -30%. For example: human hair keratin and silk fibroin can form a hydrogen bond photo-crosslinking network with coumarin, and the system can generate a photopolymerization reaction through coumarin to generate stress in the system so as to form a regular and ordered surface pattern. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 15
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
two or more natural biological materials are used as the scaffold material to prepare the dynamic polymer cross-linked network structure. In physiological environments, most tissues and extracellular matrices contain multiple components at the same time, not single components, and different components play different roles and together support in vivo environments. For example: after gelatin or collagen and alginate or hyaluronic acid and the like are used and dissolved in deionized water, physiological saline, PBS (including but not limited to these solutions) and the like (the ratio of the two materials can be from 2:98 to 98:2) at a prepolymer concentration of 50%, photosensitive molecules (including but not limited to these molecules) such as anthracene, anthranilate, coumarin, cinnamate, vinyl pyrene and the like are added into the prepolymer system, and the ratio of the base material and the photosensitive molecules can be 0.1% -30%. For example: the collagen and alginate mixture can form a hydrogen bond photo-crosslinking network with the anthracene formate, and the system can generate photopolymerization reaction through the anthracene formate, so that stress in the system is generated, and a regular and ordered surface pattern is formed. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 16
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
two or more natural biological materials are used as the scaffold material to prepare the dynamic polymer cross-linked network structure. For example: cellulose and chitosan are used, after being dissolved in a solution of dilute inorganic acid such as hydrochloric acid, formic acid, acetic acid, lactic acid, malic acid, ascorbic acid and the like or a certain common organic acid (including but not limited to the solutions) and the like (the ratio of the two materials can be from 2:98 to 98:2), photosensitive molecules such as anthracene, anthranilate, coumarin, cinnamate, vinyl pyrene and the like (including but not limited to the molecules) are added into a prepolymer system, and the ratio of the base material to the photosensitive molecules can be 0.1% -30%. For example: cellulose and chitosan can form a hydrogen bond photo-crosslinking network with anthranilic acid, and the system can generate photopolymerization reaction through the anthranilic acid ester to generate stress in the system so as to form a regular and ordered surface pattern. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 17
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
dynamic polymer cross-linked network structures are prepared using synthetic materials such as polyvinyl alcohol, polysiloxanes, polyethylene glycol, polyamino acids, polycaprolactone, polyurethane, polyphosphazene, polyetheretherketone, polyphosphoester, polyanhydrides, polyesters, polyesteramides, poly (meth) acrylates, polylactic acid and derivatives thereof as scaffold materials. These materials are dissolved in deionized water, physiological saline, PBS solution or organic solvents such as dimethyl sulfoxide, acetone, tetrahydrofuran (including but not limited to these solutions) respectively according to the concentration of the prepolymer of 5%, and then photosensitive molecules such as anthranilate, coumarin, cinnamate, vinyl pyrene (including but not limited to these molecules) are added into the prepolymer system, wherein the proportion of the base material and the photosensitive molecules is 0.1% -30%. For example: polylactic acid or its derivative and anthracene formate are polymerized through hydrogen bond to form supermolecule hydrogen bond photo-crosslinking network with light and acid-base gas sensitivity. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 18
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
dynamic polymer cross-linked network structures are prepared using synthetic materials such as polyvinyl alcohol, polysiloxanes, polyethylene glycol, polyamino acids, polycaprolactone, polyurethane, polyphosphazene, polyetheretherketone, polyphosphoester, polyanhydrides, polyesters, polyesteramides, poly (meth) acrylates, polylactic acid and derivatives thereof as scaffold materials. After the materials are dissolved according to the prepolymer concentration of 90% (part of synthetic materials, such as polysiloxane, the prepolymer concentration can be 100%), and respectively dissolved in deionized water, physiological saline, PBS solution or organic solvents (including but not limited to dimethyl sulfoxide, acetone, tetrahydrofuran and the like) according to different solubilities, photosensitive molecules (including but not limited to molecules) such as anthranilate, coumarin, cinnamate, vinyl pyrene and the like are added into a prepolymer system, and the proportion of the substrate material and the photosensitive molecules can be 0.1% -30%. For example: polylactic acid or its derivative and anthracene formate are polymerized through hydrogen bond to form supermolecule hydrogen bond photo-crosslinking network with light and acid-base gas sensitivity. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 19
A surface patterned bioengineered graft or cell culture scaffold prepared using one-step optical direct writing method, prepared using the following method:
dynamic polymer cross-linked network structures are prepared using synthetic materials such as polyvinyl alcohol, polysiloxane, polyethylene glycol, polyamino acid, polycaprolactone, polyurethane, polyphosphazene, polyether ether ketone, polyphosphonate, polyanhydride, polyester, polyesteramide, poly (meth) acrylate, polylactic acid and derivatives thereof, a mixture of two or more of which is used as a scaffold material. For example: after polyacrylate and polyethylene glycol are mixed (the ratio of the two materials can be from 2:98 to 98:2)), the mixture is dissolved in an organic solvent (including but not limited to dimethyl sulfoxide, acetone, tetrahydrofuran and the like according to the concentration of the prepolymer to be 50%, and then photosensitive molecules (including but not limited to molecules) such as anthranilate, coumarin, cinnamate, vinyl pyrene and the like are added into the prepolymer system, wherein the ratio of the base material to the photosensitive molecules can be 0.1% -30%. For example: polyethylene glycol, polyacrylic acid, anthranilate and the like are polymerized through hydrogen bonds to form a supermolecule hydrogen bond photo-crosslinking network sensitive to light and acid-base gases. The mixed liquid is made into a film-like, columnar, cube-like and other (including but not limited to the above shape) structure, a photomask is used to be tiled on a self-made exposure frame (micropatterns with different shapes and resolutions can be obtained by using photomasks with different shapes and dimensions), the distance between the photomask and an ultraviolet light source as well as between the photomask and a sample is fixed, and the surface micropattern is prepared by a one-step illumination method under the exposure of the ultraviolet light source or visible light source for a proper time. Patterns can be produced having identifiable dimensions in the size range of 0.01 to 2000 microns, preferably 0.1 to 1000 microns, more preferably 10 to 500 microns. The prepared micropatterned gel or elastomer can be used as a cell culture bracket or as a bioengineering implant after being cleaned and sterilized in deionized water.
Example 20
Material with surface micro-pattern structure prepared by one-step light direct writing method for culturing myocardial cells
And (3) preparing materials: the method comprises the steps of preparing a material (PAN) with a surface micropattern structure by using artificial synthetic material polyacrylate and photosensitive molecular vinyl pyrene, preparing the mixed material into a film-like structure with the size of 1cm x 1cm, tiling the film-like structure on a self-made exposure frame by using a 'back' type photomask, fixing the distance between the photomask and an ultraviolet light source and between the photomask and a sample, and exposing the film-like structure under the ultraviolet light source for a proper time, so as to prepare the 'back' type micropattern by a one-step illumination method. And (3) cleaning the prepared material with deionized water, and sterilizing under high pressure for later use. Mice myocardial primary cells were seeded on PAN material at 100000 per square centimeter, cultured for seven days, and the cultured cells were single stained with DAPI dye. The results are shown in FIG. 4.
The microstructure feature morphology dimensions and cell culture results for the different examples described above are shown in table 1.
TABLE 1 comparison of microstructural feature morphology size and cell culture results for different examples
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The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (9)
1. A method for preparing a bioengineered implant, which is characterized in that the preparation method is a one-step optical direct writing method, and the bioengineered implant is surface micropatterned gel or elastomer.
2. The method according to claim 1, wherein the one-step optical direct writing method specifically comprises the steps of:
(1) Preparing a dynamic polymer crosslinked network structure;
(2) Dissolving the polymer, and adding photosensitive molecules into a prepolymer system;
(3) The mixed liquid is made into a required structure, a photomask plate is used for being paved on an exposure frame, the distance between the photomask plate and an ultraviolet light source as well as the distance between the photomask plate and a sample are fixed, and the mixture is exposed under the ultraviolet light source or the visible light source.
3. The method according to claim 2, wherein the dynamic polymer cross-linked network scaffold material in the step (1) is one or more of chitin, chitosan and chitosan.
4. The method according to claim 2, wherein the solvent in step (2) comprises deionized water, physiological saline, PBS solution or formic acid solution.
5. The method according to claim 2, wherein the photosensitive material in the step (2) comprises anthracene, anthracene formate, coumarin, cinnamate, vinyl pyrene.
6. The method according to claim 2, wherein the ratio of the base material to the photosensitive molecule in the step (2) is 0.1 to 30%.
7. The process according to claim 2, wherein the prepolymer in step (2) is dissolved at a concentration of 1 to 99%.
8. The method of producing according to claim 2, wherein the pattern that can be produced in the step (3) is in the size range of 0.01 to 2000 μm.
9. A bioengineered graft and method of in vitro and cell culture, characterised in that a surface micropatterned hydrogel or elastomer according to claim 1 is used.
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