CN109081935A - A kind of anisotropic microstructure becomes structural material and the preparation method and application thereof - Google Patents

A kind of anisotropic microstructure becomes structural material and the preparation method and application thereof Download PDF

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CN109081935A
CN109081935A CN201810609903.8A CN201810609903A CN109081935A CN 109081935 A CN109081935 A CN 109081935A CN 201810609903 A CN201810609903 A CN 201810609903A CN 109081935 A CN109081935 A CN 109081935A
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micro
substrate
layer
cellular adhesion
boundary layer
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聂壹峰
刘思迪
李想
韩东
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National Center for Nanosccience and Technology China
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National Center for Nanosccience and Technology China
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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
    • C08J7/00—Chemical treatment or coating of shaped articles made of macromolecular substances
    • C08J7/12—Chemical modification
    • C—CHEMISTRY; METALLURGY
    • C23—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; CHEMICAL SURFACE TREATMENT; DIFFUSION TREATMENT OF METALLIC MATERIAL; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL; INHIBITING CORROSION OF METALLIC MATERIAL OR INCRUSTATION IN GENERAL
    • C23C—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; SURFACE TREATMENT OF METALLIC MATERIAL BY DIFFUSION INTO THE SURFACE, BY CHEMICAL CONVERSION OR SUBSTITUTION; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL
    • C23C14/00—Coating by vacuum evaporation, by sputtering or by ion implantation of the coating forming material
    • C23C14/06—Coating by vacuum evaporation, by sputtering or by ion implantation of the coating forming material characterised by the coating material
    • C23C14/14—Metallic material, boron or silicon
    • C23C14/20—Metallic material, boron or silicon on organic substrates
    • C23C14/205—Metallic material, boron or silicon on organic substrates by cathodic sputtering
    • C—CHEMISTRY; METALLURGY
    • C23—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; CHEMICAL SURFACE TREATMENT; DIFFUSION TREATMENT OF METALLIC MATERIAL; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL; INHIBITING CORROSION OF METALLIC MATERIAL OR INCRUSTATION IN GENERAL
    • C23C—COATING METALLIC MATERIAL; COATING MATERIAL WITH METALLIC MATERIAL; SURFACE TREATMENT OF METALLIC MATERIAL BY DIFFUSION INTO THE SURFACE, BY CHEMICAL CONVERSION OR SUBSTITUTION; COATING BY VACUUM EVAPORATION, BY SPUTTERING, BY ION IMPLANTATION OR BY CHEMICAL VAPOUR DEPOSITION, IN GENERAL
    • C23C14/00—Coating by vacuum evaporation, by sputtering or by ion implantation of the coating forming material
    • C23C14/22—Coating by vacuum evaporation, by sputtering or by ion implantation of the coating forming material characterised by the process of coating
    • C23C14/34—Sputtering
    • C23C14/35—Sputtering by application of a magnetic field, e.g. magnetron sputtering
    • 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
    • C08J2383/00—Characterised by the use of macromolecular compounds obtained by reactions forming in the main chain of the macromolecule a linkage containing silicon with or without sulfur, nitrogen, oxygen, or carbon only; Derivatives of such polymers
    • C08J2383/04—Polysiloxanes
    • 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
    • C08J2471/00—Characterised by the use of polyethers obtained by reactions forming an ether link in the main chain; Derivatives of such polymers
    • C08J2471/02—Polyalkylene oxides
    • 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
    • C08J2489/00—Characterised by the use of proteins; Derivatives thereof

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  • Organic Chemistry (AREA)
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Abstract

Become structural material and the preparation method and application thereof the present invention relates to a kind of anisotropic microstructure.The anisotropic microstructure structural material that becomes includes substrate that at least one surface is equipped with raised micro-structure, anti-cellular adhesion boundary layer is additionally provided on the substrate surface, phase the same side on the anti-cellular adhesion boundary layer of each raised micro-structure is also successively arranged layers of chrome, gold nano layer, promotees cell adhesion layer.The PEG strand modification of anti-cellular adhesion is obtained anti-cellular adhesion interface on the surface PDMS by the present invention, gold nano layer is deposited in the side of microtrabeculae by way of inclination vapor deposition, by the rgd peptide modification for promoting cell adhesion to gold-plated surface, finally obtain cell anisotropic stick it is micro- become structure.The structure has the function of inducing and changing cell migration, influences cell behavior, the migratory behaviour mechanism of the various cells of research and modulate tumor cell for no medicine irritation and spread in vivo all with good research significance and application value.

Description

A kind of anisotropic microstructure becomes structural material and the preparation method and application thereof
Technical field
The invention belongs to technical field of biological materials, and in particular to a kind of anisotropic microstructure becomes structural material and its preparation side Method and application.
Background technique
It is numerous research shows that can be generated many peculiar when cell is in contact with the material surface with micro-nano structure Biobehavioral, meeting after being in contact this is mainly due to micro-nano structure with cell produce so that the albumen of cell membrane surface responds Raw polarization phenomena, and an important factor for cell migration it is exactly the polarization of cytoskeleton.Controllable inducing cell is constructed on the surface of the material Migration interface suffers from important meaning for research cell behavior and progress biomaterial implantation.
Anisotropic microstructure was got up in micro-nano structure field by extensive research in recent years, since its is not special right Title property, in optical field, magnetic fields suffer from good application, and this special structure biological field research still Seldom, it can be envisaged that the asymmetry of micro-nano structure will necessarily also cause the asymmetry of cell adhesion when contacting with cell, The cell behavior that this asymmetry is caused is worth studying.The design on previous cell migration surface is usually to construct gradient The micro structure array of variation or the soft or hard substrate of change of gradient, the cell migration behavior that this micro-structure is caused are moved Power is mainly derived from the change of gradient on surface, and variation is bigger, and active force is more obvious, and the range of this change of gradient is limited , therefore the range that micro-structure can construct is similarly limited, is greatly limited it in terms of practical application.
Summary of the invention
For problem of the prior art, the present invention provides the thinking and design that a kind of anisotropic microstructure becomes structure, i.e., it is micro- become The anisotropy of structure be based on each microtrabeculae rather than some region of anisotropy, and can be unlimited on the surface of the material It is micro- become extension go down.
The present invention goes out a kind of anisotropic microstructure in PDMS surface construction by way of pirate recordings and becomes structural material: by anti-cell The PEG strand modification sticked obtains anti-cellular adhesion interface on the surface PDMS, the one of microtrabeculae by way of inclination vapor deposition Side deposits gold nano layer, and the rgd peptide for promoting cell adhesion is modified to gold-plated surface, cell anisotropic is finally obtained and sticks It is micro- become structure.By SEM, AFM observation is proved: the present invention is prepared for microtrabeculae that diameter is 1-5 micron in a wide range of as tying The ordered anisotropic of structure unit it is micro- become array, this novel structure all shows on cyto-mechanics behavior, cell adhesion Interesting and significant anisotropy feature.Anisotropic microstructure constructed by the present invention becomes structure with induction and changes cell The function of migration, it is swollen that this influences cell behavior, the migratory behaviour mechanism of the various cells of research and regulation for no medicine irritation Oncocyte is spread in vivo all has good research significance and application value.
The anisotropy of micro-structure designed by the present invention is the anisotropy based on each microtrabeculae rather than a certain area Domain, and on the surface of the material can it is unlimited it is micro- become extension go down, it is very long that this constructs cell migration surface, it is sufficient to Length needed for reaching practical application has good practical application potential quality.
To achieve the above object, the invention adopts the following technical scheme:
A kind of anisotropic microstructure becomes structural material, the substrate of raised micro-structure is equipped with including at least one surface, in institute It states and is additionally provided with anti-cellular adhesion boundary layer on substrate surface, in the anti-cellular adhesion boundary layer of all raised micro-structures On phase the same side be also successively arranged layers of chrome and gold nano layer.
That is, only phase the same side on the anti-cellular adhesion boundary layer of each raised micro-structure also according to It is secondary to be equipped with layers of chrome, gold nano layer, the anti-cellular adhesion boundary layer is provided only on the surface of the protrusion micro-structure other side；From And form soft or hard different anisotropy.
Further, it is glutinous that rush cell is additionally provided on the gold nano layer and/or on the anti-cellular adhesion boundary layer Attached layer.And/or
Further, be successively arranged layers of chrome on the surface of each raised micro-structure, the region of gold nano layer account for it is each The 20-60% of the surface area of the protrusion micro-structure, more preferably 50%；And/or
Further, the surface of each raised micro-structure is equipped with the region for promoting cell adhesion layer and accounts for each institute State the 20-100% of the surface area of raised micro-structure, more preferably 50%.
Further, at least one surface of the substrate have the surface bulge micro-structure, such as can one side or Two sides all has surface bulge micro-structure.
Further, the base material can be selected from PDMS (dimethyl silicone polymer), polymethyl methacrylate (PMMA), one or more of polycarbonate (PC), glass, silicon wafer etc., preferably PDMS.
Further, the substrate with a thickness of 500nm-5mm.And/or
Further, the anti-cellular adhesion boundary layer with a thickness of 0.5-1nm.And/or
Further, the layers of chrome with a thickness of 2-10nm, preferably 5nm.And/or
Further, the gold nano layer with a thickness of 10-30nm, preferably 20nm.And/or
Further, it is described promote cell adhesion layer with a thickness of 0.1-1nm.
In the present invention, the surface bulge micro-structure is also referred to as microtrabeculae.
Further, the height of the surface bulge micro-structure is 1.5-2 microns；And/or the surface bulge micro-structure Height and maximum width ratio be 1:1；And/or the maximum width of the surface bulge micro-structure is 1.5-2 microns.
Further, the surface bulge micro-structure is cylindrical or irregular shape etc..
Further, the surface bulge micro-structure be cylinder, be highly 1.5-2 microns, highly with the ratio of diameter For 1:1 or so.
Further, spacing of the surface bulge micro-structure in substrate is 2-5 microns；Or base every square centimeter On bottom, the number of the surface bulge micro-structure is (4 × 106)-(4×107) a.
Further, the material for constituting the anti-cellular adhesion boundary layer is PEG (polyethylene glycol).
Specifically, the anti-cellular adhesion boundary layer is made by polyethylene glycol methacrylate-styrene polymer and isopropanol reaction 's；The mass ratio of polyethylene glycol methacrylate-styrene polymer and isopropanol is preferably (6:4)-(9:1), more preferably 8:2；It reacted Appropriate platinum can also be added in journey as catalyst.
In a specific embodiment of the invention, the substrate is put by polyethylene glycol methacrylate-styrene polymer 8g and different Propyl alcohol 2g is made in mixed solution, and the Karst platinum catalyst of 30 μ L is added, is stirred at room temperature 15 hours；Then using acetone, Distilled water washs for several times repeatedly, and vacuum drying can prepare anti-cellular adhesion boundary layer in substrate.
Further, it can be used magnetron sputtering vapour deposition method (magnetron sputtering vapor deposition instrument) first on anti-cellular adhesion boundary layer Layers of chrome is deposited in (surface), then layer gold is deposited in (surface) in layers of chrome.
Specifically, the substrate tilt certain angle of anti-cellular adhesion boundary layer is made into each micro- knot of protrusion when vapor deposition Layers of chrome, gold nano layer is successively deposited in phase the same side on the anti-cellular adhesion boundary layer of structure, and the other side can not be deposited, table Face is only anti-cellular adhesion boundary layer, to form soft or hard different anisotropy.General tilt angle is 20-80 degree angle, often Tilt angle is 45 degree of angles.
Further, the rush cell adhesion layer is by rgd peptide (Arg-Gly-Asp, i.e. arginine, glycine, lucid asparagus Propylhomoserin polypeptide) it constitutes, i.e., it is modified by rgd peptide to the layer gold surface, constitutes and promote cell adhesion layer.
In a preferable embodiment of the invention, the base material is PDMS；The surface bulge micro-structure is Cylinder, height is 5 microns, width is 5 microns, the spacing in substrate is 2-5 microns；The anti-cellular adhesion interface Layer, layers of chrome, gold nano layer, promote cell adhesion layer thickness be respectively 1nm, 5nm, 20nm, 1nm；The anti-cellular adhesion interface Layer is as made from polyethylene glycol methacrylate-styrene polymer and isopropanol reaction；The rush cell adhesion layer is modified by rgd peptide The gold nano layer surface is constituted.
Become the preparation method of structural material the present invention also provides a kind of anisotropic microstructure as described above, including walks as follows It is rapid:
1) substrate is put into silicate-containing oil (i.e. cured liquid PDMS) and the mixed solution of anhydrous methanol, is added suitable It measures concentrated sulfuric acid solution (such as concentrated sulfuric acid solution of silicone oil volume 2% or so), under room temperature magnetic agitation, handles certain time (30 minutes or so) then use n-hexane and methanol repeated flushing, are dried in vacuo (general 24 hours) after cleaning；It is sealed, It is spare；
Wherein, the volume ratio of silicone oil and anhydrous methanol is (1:2)-(3:4), preferably 3:5；
2) substrate by step 1) processing is put into the mixed solution containing polyethylene glycol methacrylate-styrene polymer and isopropanol, is added Enter platinum catalyst, (general 15 hours) are stirred at room temperature, anti-cellular adhesion boundary layer is prepared in substrate；Then third is used Ketone, distilled water wash for several times repeatedly, vacuum drying；It is sealed, it is spare；
Wherein, the mass ratio of polyethylene glycol methacrylate-styrene polymer and isopropanol is preferably (6:4)-(9:1), more preferably 8:2。
3) by step 2) processing gained substrate (being modified with peg molecule chain), using magnetron sputtering vapour deposition method, (magnetic control splashes Penetrate vapor deposition instrument) first layers of chrome is deposited in (surface) on anti-cellular adhesion boundary layer, then layer gold is deposited in (surface) in layers of chrome；
Specifically, keep all protrusions micro- the substrate tilt certain angle of the boundary layer containing anti-cellular adhesion when vapor deposition Layers of chrome, gold nano layer is successively deposited in phase the same side on the anti-cellular adhesion boundary layer of structure, and the other side can not be deposited, Surface is only anti-cellular adhesion boundary layer, to form soft or hard different anisotropy.General tilt angle is 20-80 degree angle, Common tilt angle is 45 degree of angles.
4) RGD is modified on the layer gold surface of step 3) processing gained substrate and/or on the anti-cellular adhesion boundary layer Polypeptide；
Conventional method in that art can be used in the step, such as step 3) processing gained substrate is immersed in and is connected to sulfydryl In rgd peptide aqueous solution (2% mass-volume concentration), by modification in 30 minutes, sample is taken out using deionized water repeatedly It rinses, removal is not connected to the rgd peptide of gold surface, is dried with nitrogen.
Substrate of the present invention can be prepared with conventional method in that art.
Specifically, the present invention provides a kind of preparation method of substrate, includes the following steps:
1) the silicon substrate end template containing microcellular structure is subjected to surface fluorination with dimethyldichlorosilane；
The clean silicon substrate end template containing microcellular structure can be specifically put into closed container, instill appropriate dimethyl two Chlorosilane carries out surface fluorination to the silicon substrate end template to reduce its apparent activation energy, and the fluorination time is generally 30 minutes left sides It is right；
2) PDMS performed polymer mixed liquor is filled on the silicon substrate end template containing microcellular structure of surface fluorination, is placed in (refrigerator cold-storage layer) is stood overnight under the conditions of 4 DEG C；Then it toasts, solidifies PDMS performed polymer；By cured PDMS from described It is separated on silicon substrate end template containing microcellular structure, die plate pattern pirate recordings to the surface PDMS can be obtained the micro- of ordered arrangement The substrate of column array.
Such as the size in hole is on the silicon substrate end template containing microcellular structure, and 5 microns of hole depth, 5 microns of the diameter in hole, hole 5 microns of spacing, constructing range is 2cm × 2cm, and dehydrated alcohol, acetone can be used respectively to be cleaned by ultrasonic under low power conditions twice, One time 5 minutes, using being dried with nitrogen surface after cleaning.
The PDMS performed polymer mixed liquor can be prepared by conventional method in that art, such as PDMS performed polymer is pressed with curing agent It is mixed according to the mass ratio of 10:1, is moved into mixed liquor in vacuum desiccator after stirring evenly, be evacuated 1 hour, it is mixed to drain performed polymer Close the bubble in liquid.
Baking temperature and time will affect the hardness of PDMS, can carry out selection adjustment as needed.Generally, the baking Temperature is 40-90 DEG C, and the time is -12 hours 40 minutes；Preferably, the baking temperature is 70 DEG C, and the time is 3 hours.
PDMS performed polymer curing reaction is as follows:
nSi(CH3)2Cl2+nH2O→[Si(CH3)2O]n+2nHCl
The invention also includes the anisotropic microstructures of above method preparation to become structural material.
The anisotropic microstructure of the present invention structural material that becomes can infinitely extend (in the in-plane direction).
The invention also includes above-mentioned anisotropic microstructure become structural material research in terms of application.
The raw materials used in the present invention is commercially available to buy, or prepares by conventional method in that art.
On the basis of common knowledge of the art, above-mentioned each optimum condition can be combined with each other each preferably to get the present invention Example.
Furthermore, it is to be understood that herein, also included in the meaning of terms such as "include", "comprise", " containing " " by ... Composition ", " by ... constitute ", " by ... it is made " etc..
The invention has the following advantages:
1) anisotropic microstructure that the present invention is prepared in uniform, a wide range of becomes structural material, this novel anisotropy Structure, with two different anisotropic properties, one: utilizing cell adhesion anisotropy caused by molecular modification；Two: benefit Generated soft or hard (Young's modulus) anisotropic properties are deposited with inclination.
2) present invention provides the thinking and design that a kind of anisotropic microstructure becomes structure, i.e., it is micro- become structure anisotropy be base In each microtrabeculae rather than some region of anisotropy, and can on the surface of the material it is unlimited it is micro- become extension go down.
3) anisotropic microstructure of the present invention becomes the migration for the promotion cell that structural material can be obvious.
Detailed description of the invention
Fig. 1: 1 anisotropic microstructure of embodiment become structural material preparation and flow diagram applied to cell experiment；
Fig. 2: the structure picture that the anisotropic microstructure prepared in embodiment 1 becomes under scanning of materials Electronic Speculum SEM；
Fig. 3: cell becomes the scanning electron microscopic picture migrated on material in anisotropic microstructure in experimental example 1.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..It is not specified in embodiment specific Technology or conditions person, described technology or conditions according to the literature in the art, or carried out according to product description.It is used Production firm person is not specified in reagent or instrument, is the conventional products that can be commercially available by regular distributor.
Embodiment 1
A kind of anisotropic microstructure becomes structural material, and preparation method is as shown in Figure 1, specifically include:
1. by the silicon substrate end template containing microcellular structure, (size in hole is in silicon base, and 5 microns of hole depth, the diameter 5 in hole is micro- Rice, 5 microns of the spacing in hole, constructing range is 2cm × 2cm), using dehydrated alcohol, acetone, each ultrasound is clear under low power conditions It washes twice, one time 5 minutes, using being dried with nitrogen surface after cleaning；
2. the surface after drying is put into a closed container, a few drop dimethyldichlorosilanes are instilled in a reservoir, into For row surface fluorination to reduce apparent activation energy, the fluorination time is 30 minutes；
3. PDMS performed polymer is mixed with curing agent according to the mass ratio of 10:1 in substrate fluorination process, after stirring evenly Mixed liquor is moved into vacuum desiccator, is evacuated 1 hour, drains the bubble in performed polymer mixed liquor；
4. being poured in performed polymer mixed liquor in the microcellular structure substrate that surface fluorination finishes after pumping, substrate is connected The same performed polymer mixed liquor for being covered on its surface (refrigerator cold-storage layer) under the conditions of 4 DEG C is stood overnight；
5. the sample after overnight is put into 70 DEG C of baking ovens 3 hours, solidify PDMS performed polymer；
6. the PDMS after hot setting is changed into harder solid-state by flowable state, using knife blade in template One jiao is carefully uncovered PDMS, die plate pattern pirate recordings to the surface PDMS can be obtained the micro-pillar array (pillar height of ordered arrangement 5 microns, 5 microns of the diameter of column, 5 microns of the spacing of column, constructing range is 2cm × 2cm).
7. the surface PDMS of above-mentioned acquisition is put into the mixed solution of silicone oil (3mL) and anhydrous methanol (5mL), it is added 60 The concentrated sulfuric acid solution of μ L, it is small to be dried in vacuo 24 with n-hexane and methanol repeated flushing for magnetic agitation 30 minutes under room temperature When, it is sealed；
8. the sample after dry is put into the mixed solution of polyethylene glycol methacrylate-styrene polymer (8g) and isopropanol (2g), add The Karst platinum catalyst for entering 30 μ L is stirred at room temperature 15 hours；
9. after the reaction was completed washing PDMS for several times repeatedly using acetone, distilled water, it is dried in vacuo 24 hours, sealing is protected It deposits；
10. condition of the PDMS of peg molecule chain using magnetron sputtering vapor deposition instrument at 45 degree of angles of inclination will be modified with Under, make its surface that the layers of chrome after one layer of 5nm be deposited, the layer gold after one layer of 20nm is then deposited again.
11. the PDMS that gained is coated with layer gold to be immersed in the rgd peptide aqueous solution (2% mass-volume concentration) for being connected to sulfydryl In, by modification in 30 minutes, sample to be taken out and uses deionized water repeated flushing, removal is not connected to the rgd peptide of gold surface, It is dried with nitrogen.
The structural material SEM figure that becomes of anisotropic microstructure made from the present embodiment is shown in Fig. 2.
Experimental example 1
Using anisotropic microstructure made from embodiment 1 become structural material research cell its surface migration:
1. the anisotropic material after modification is transferred to radiation sterilization 2 hours under ultraviolet lamp, cell is carried out as substrate Migration experiment:
2. getting off the melanoma cells of culture dish are covered with using trypsin digestion, it is configured to the every milli of 10000 cells The cell suspension risen；
3. the PDMS sample modified is put into one small culture dish (diameter 35mm, height 10mm), into culture dish The above-mentioned cell suspension of 3mL is added and is transferred in living cells work station；
4. living cells work station is connected to inverted fluorescence microscope, it is set as 37 DEG C, is passed through 5%CO2 gas, is continued Living cells culture for 24 hours；
5. being taken pictures during the cultivation process using the camera system of microscope institute band to observation area, by continuous during culture Migration situation of the photo evaluation and test cell of shooting in body structure surface.
Melanoma cells are shown in Fig. 3 in the anisotropic microstructure scanning electron microscope (SEM) photograph migrated on material that becomes.The result shows that melanin Oncocyte morphologic change, hence it is evident that it stretches, it is more apparent to find out that fusiformis shape, which occurs, to side migrates.
Although above the present invention is described in detail with a general description of the specific embodiments, On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.

Claims (10)

  1. The structural material 1. a kind of anisotropic microstructure becomes, which is characterized in that be equipped with raised micro-structure including at least one surface Substrate is additionally provided with anti-cellular adhesion boundary layer on the substrate surface；In the anti-cell of all raised micro-structures The phase the same side sticked on boundary layer is also successively arranged layers of chrome and gold nano layer；
    Preferably, it is additionally provided on the gold nano layer and/or on the anti-cellular adhesion boundary layer and promotees cell adhesion layer； And/or
    Preferably, be successively arranged layers of chrome on the surface of each raised micro-structure, the region of gold nano layer account for it is each described convex The 20-60% of the surface area of Ultrastructure, more preferably 50%；And/or
    Preferably, the surface of each raised micro-structure is equipped with the region for promoting cell adhesion layer and accounts for each protrusion The 20-100% of the surface area of micro-structure, more preferably 50%.
  2. The structural material 2. anisotropic microstructure according to claim 1 becomes, which is characterized in that the base material is selected from PDMS, one or more of polymethyl methacrylate, polycarbonate, glass, silicon wafer；
    And/or constitute the anti-cellular adhesion boundary layer material be PEG, preferably by polyethylene glycol methacrylate-styrene polymer with it is different Made from propyl alcohol reaction；
    And/or the rush cell adhesion layer is made of rgd peptide.
  3. The structural material 3. anisotropic microstructure according to claim 1 or 2 becomes, which is characterized in that
    The substrate with a thickness of 500nm-5mm；And/or
    The anti-cellular adhesion boundary layer with a thickness of 0.5-1nm；And/or
    The layers of chrome with a thickness of 2-10nm, preferably 5nm；And/or
    The gold nano layer with a thickness of 10-30nm, preferably 20nm；And/or
    It is described promote cell adhesion layer with a thickness of 0.1-1nm.
  4. The structural material 4. anisotropic microstructure according to claim 1-3 becomes, which is characterized in that the surface bulge The height of micro-structure is 1.5-2 microns；And/or the height of the surface bulge micro-structure and the ratio of maximum width are 1:1； And/or the maximum width of the surface bulge micro-structure is 1.5-2 microns；
    And/or the surface bulge micro-structure is cylindrical or irregular shape；
    And/or spacing of the surface bulge micro-structure in substrate is 2-5 microns；Or institute in substrate every square centimeter The number for stating surface bulge micro-structure is (4 × 106)-(4×107) a.
  5. The preparation method of structural material 5. a kind of anisotropic microstructure becomes, which comprises the steps of:
    1) substrate with surface bulge micro-structure is put into the mixed solution of silicate-containing oil and anhydrous methanol, appropriate dense sulphur is added Acid solution, magnetic agitation, processing after a certain period of time, with n-hexane and methanol repeated flushing, it is dry to clean rear vacuum under room temperature It is dry；It is sealed, it is spare；
    2) substrate by step 1) processing is put into the mixed solution containing polyethylene glycol methacrylate-styrene polymer and isopropanol, and platinum is added Au catalyst is stirred at room temperature, and anti-cellular adhesion boundary layer is prepared in substrate；Then it is washed repeatedly using acetone, distilled water For several times, it is dried in vacuo；It is sealed, it is spare；
    3) layers of chrome first is deposited on anti-cellular adhesion boundary layer using magnetron sputtering vapour deposition method in step 2) processing gained substrate, then Layer gold is deposited in layers of chrome；By the substrate tilt certain angle of the boundary layer containing anti-cellular adhesion when vapor deposition, make all protrusions Layers of chrome, gold nano layer is successively deposited in phase the same side on the anti-cellular adhesion boundary layer of micro-structure；
    4) rgd peptide will be modified on the layer gold surface of step 3) processing gained substrate and/or on the anti-cellular adhesion boundary layer.
  6. 6. preparation method according to claim 5, which is characterized in that the volume ratio of silicone oil and anhydrous methanol is in step 1) (1:2)-(3:4), preferably 3:5；And/or
    In step 2), the mass ratio of polyethylene glycol methacrylate-styrene polymer and isopropanol is preferably (6:4)-(9:1), and preferably 8: 2。
  7. 7. preparation method according to claim 5 or 6, which is characterized in that step 3) will contain anti-cellular adhesion circle when being deposited The substrate tilt 20-80 degree angle of surface layer, preferably 45 degree of angles.
  8. 8. according to the described in any item preparation methods of claim 5-7, which is characterized in that the preparation method of the substrate, including Following steps:
    1) the silicon substrate end template containing microcellular structure is subjected to surface fluorination with dimethyldichlorosilane；
    2) PDMS performed polymer mixed liquor is filled on the silicon substrate end template containing microcellular structure of surface fluorination, is placed at 4 It is stood overnight under the conditions of DEG C；Then it toasts, solidifies PDMS performed polymer；By cured PDMS from the silicon containing microcellular structure It is separated in substrate template, obtains the substrate of the micro-pillar array of ordered arrangement.
  9. The structural material 9. anisotropic microstructure of any one of claim 5-9 the method preparation becomes.
  10. Application of the structural material in terms of studying cell migration 10. any one of claim 1-4,9 anisotropic microstructure become.
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