CN104841020B - A kind of three-dimensional order tissue engineering bracket of macroscopical Supramolecular Assembling and preparation - Google Patents
A kind of three-dimensional order tissue engineering bracket of macroscopical Supramolecular Assembling and preparation Download PDFInfo
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Abstract
The invention discloses a kind of macroscopical Supramolecular Assembling prepares ordered 3 D structure, in terms of prepared ordered structure is expected to be applied to cell induction differentiation masterplate and three-dimensional tissue's engineering scaffold material.What the method can be introduced constructs primitive includes the various biocompatible material such as polydimethylsiloxane, hydrogel, shitosan, above-mentioned material and magnetic nano-particle is pressed into prefabricated masterplate by minute-pressure impression method and can realize the modification constructed a large amount of preparations of primitive and specificity supermolecule functional group is carried out to its surface.Further, realize that constructing primitive is accurately positioned three-dimensional using external magnetic field, and construct primitive using supermolecule interaction fixation in specified location, so as to realize the controllable standby of ordered 3 D structure.In addition, by constructing primitive surface modification specific proteinses recognition site specified, somatomedin required for being introduced using intermolecular specific recognition in the method implementation process, so as to realize specifying orientation of the somatomedin inside three-dimensional stent material to introduce.
Description
Technical field
The present invention relates to Supramolecular Assembling and ordered 3 D structure field of material preparation.More particularly, to a kind of macroscopic view
The three-dimensional order tissue engineering bracket of Supramolecular Assembling and preparation.
Background technology
The controllable standby of ordered 3 D structure is obtained in multiple fields such as photonic crystal, reinforcing material and photoelectric devices
Extensive concern and further investigation.At present, the research work in the field pays close attention to the primitive of constructing of nanoscale mostly, and micro- to more than ten
Rice studies less with the primitive of constructing of up to mm-scale, and its reason is that the interval ordered 3 D structure of the size is prepared relatively
It is difficult：For traditional processing, the size is too little is difficult to accurate operation, for emerging micro-nano technology technology, should
Size then causes greatly very much technique complicated, time-consuming very long, not with actual operability.However, ten microns to one centimetre of three-dimensional
Ordered structure has important research meaning for tissue engineering bracket, longer-wave photons field of crystals.Especially, most cells are all
Within this size range, current cell culture technology has also been advanced to three-dimensional cell training from traditional two dimension culture
Support, construct the three dimensional scaffold structure comparable with cell size can carry for cell growth be more nearly biological internal milieu
Frame material.Therefore, develop ten microns of preparation methoies to a cm size ordered 3 D structure, conjunction can be provided for cell growth
Suitable support simultaneously produces significance to promotion organization engineering research.So far, due to the difficulty in preparation, ten microns to one
The relevant report of cm size ordered 3 D structure is less.For example, the three dimensional biological based on 3D printing technique prints (3D
Bioprinting) method can be by methods such as inkjet printing, micro- extrusion and laser assisted printings, will be containing cell, biofacies
The liquid mixture of culture environment needed for the cell such as capacitive material and somatomedin as " ink ", using computer aided technique
The threedimensional model of structure, organized or organ printing and reproduce.However, the problem of the method is its limited resolution,
It is difficult to build fine timbering material；Due to the particularity of printing equipment, the method to equipment, cell, biomaterial and growth because
Son has certain restriction and requires, so as to constrain its extensive application in field of tissue engineering technology.Therefore, develop one kind to have
Universality, gentle, biocompatible ordered 3 D structure preparation method remains a major challenge.
Supramolecular Assembling, as method that is a kind of gentle, biocompatible and not relying on special installation, can be compared with
Solve the above problems well.Since proposition, the research of traditional Supramolecular Assembling typically looks at molecular level or nanoscale
Construct primitive.In order to self-assembly system is extended to micron and scale above, Whitesides et al. proposition from molecular level
The concept of " full size self assembly ", and application interface minimization of free energy principle realizes millimeter in oil-water interfaces and constructs primitive
Assembling, form oldered array, the structure that the array can be obtained not relying on interface by follow-up curing operation and be existed.
In order to remove the dependence to interface, professor's Harada group introduces cyclodextrin/diphenyl diimide supermolecule on grade gel mass surface
By Host-guest Recognition effect, functional group, can realize that gel constructs the Supramolecular Assembling of primitive simultaneously under the conditions of simple concussion
Obtain self-existent assembly；They utilize illumination or competition molecule to realize the regulation and control to assembling process.The same year, applicant's
Research and development group reports 17 micrometer glass fiber of diameter and passes through controllable groups of the multiple tooth coordination in substrate surface between metal-carboxylic acid
Dress, constructs orderly pattern.For a further understanding of the Supramolecular Assembling mechanism of primitive more than micron is constructed, we introduce flexibility
The concept of wall, it is possible to achieve various rigidity construct the Supramolecular Assembling of primitive (PDMS, polymethyl methacrylate), and demonstrate,prove
Real its assembly mechanism is multiple interaction, former so as to propose a kind of design for macroscopical Supramolecular Assembling of non-aqueous gel system
Then.Additionally, the assembling such as DNA hybridization, electrostatic interaction motive force is used for the Supramolecular Assembling of micron above gel rubber system.This
A little results show Supramolecular Assembling have the advantages that from bottom to top, experiment condition it is gentle, and have in supramolecular chemistry and enrich
Biocompatible molecular recognition system can be used for constructing the specificity assembling of primitive.However, due to macroscopical Supramolecular Assembling
The forming process of body has certain randomness, constructs, so that the new method of development
To realize that macroscopic view constructs the rule of primitive, orientation arrangement, to form ordered 3 D structure.
The content of the invention
It is an object of the present invention to provide a kind of three-dimensional order tissue engineering bracket of macroscopical Supramolecular Assembling.
Further object is that providing a kind of macroscopical Supramolecular Assembling prepares three-dimensional order tissue engineering bracket
Method.
The present invention has for the difficulty in terms of current ten microns to a cm size ordered 3 D structure preparation, and three-dimensional
In sequence structure, orientation introduces the problem of various somatomedin, proposes and has developed a kind of Supramolecular Assembling method and realize ten microns extremely
The preparation of one centimetre of ordered 3 D structure, and particular growth factor is introduced in specific site.For constructing the biology of three dimensional structure
The compatibility is constructed primitive and is mainly obtained by methods such as frozen section, micro-embossing, micro Process, and its character shape, size can
With adjustable in the range of more than ten microns to cm size.In the preparation process for constructing primitive, the magnetic Nano of hydro-thermal method synthesis
Particle constructs primitive with magnetic responsiveness to prepare by mixing of adulterating.The prepared surface modification for constructing primitive has supermolecule
Effect or the functional group of bio-identification effect.The structure of ten microns to one centimetre of ordered 3 D structure is by combining external magnetic field
Construct to magnetic responsiveness that primitive is accurately positioned and macroscopical Supramolecular Assembling is carried out constructing primitive and fixes what is realized；This three
During dimension the constructing of structure, required somatomedin can be constructed the specific binding site on primitive surface draw by identification
Enter.
Size is obtained at ten microns to one centimetre by methods such as frozen section, micro-embossing, mechanical micro Process masterplate casting
Biocompatibility construct primitive.In the preparation for constructing primitive, magnetic nano-particle is introduced by way of mixing and doping with
Prepare ten microns of biocompatibility to one centimetre of magnetic response and construct primitive.Primitive surface is constructed by alternating layer in prepared
The modes such as shape package technique, doping mixing modify the complementation with supermolecular mechanism such as Host-guest Recognition, electrostatic, hydrogen bond, coordination
Functional group and bio-identification effect such as biotin/avidin (biotin/avidin), bacillus cereuss rna enzymes/bacillus cereuss
The complementary functional groups of rna enzyme inhibitors (barnase/barstar) so that above-mentioned complementation is constructed primitive and can pass through supermolecule phase
Macroscopical Supramolecular Assembling is realized in interaction or bio-identification effect.Further, with reference to external magnetic field to magnetic responsiveness biocompatibility structure
Build that primitive carries out being accurately positioned and macroscopical Supramolecular Assembling constructs the mode that primitive is fixed to magnetic responsiveness biocompatibility,
Ten micron ordered 3 D structures to a cm size scope are built from bottom to top progressively, and by constructing base during constructing
The supermolecule on first surface or bio-identification site introduce somatomedin, realize particular growth factor in a three-dimensional structure controllable fixed
Position, is finally expected to break up masterplate and tissue engineering bracket material for cell induction；Present invention introduces magnetic responsiveness biofacies
Capacitive is constructed primitive and is referred to：This is constructed primitive and has good biocompatibility, and in the presence of external magnetic field, can be with
Controlled by external magnetic field by magnetic response mode, moved and positioned.
The three-dimensional order tissue engineering bracket of macroscopical Supramolecular Assembling of the present invention be stacking shape ordered 3 D structure, institute
State ordered 3 D structure by it is multiple two-dimensional directional abreast, the primitive of constructing of periodically ordered arrangement constitute, two phase mutual connections
There is supermolecule interaction and realize the connection on two surfaces in tactile functional group of the primitive by modifying on respective surface of constructing；
The size of the support is 10-5～10-2m。
Primitive ordered arrangement of constructing in the present invention to refer to and construct primitive in conplane, be parallel to each other and adjacent,
Construct the arrangement that primitive spacing is at ten microns to a millimeter.
To reach above-mentioned purpose, the present invention adopts following technical proposals：
1. the preparation of magnetic nano-particle
Ferriferrous oxide nano-particle is prepared using hydro-thermal method or coprecipitation.
2. the preparation of primitive micron is constructed to grade magnetic responsiveness biocompatibility
For the biocompatibility of millimeter and scale above constructs primitive, can be poured in masterplate by processing to traditional mechanical
The casting liquid biological compatibility is constructed the molding of primitive solution and realizes that mass biocompatibility constructs primitive preparation, especially by such as
Lower step is realized：
1) computer aided technique is combined, using the laser-engraving technique in mechanical processing industry, institute is engraved on sheet material
The array of hollow pattern is needed, as the masterplate for preparing biocompatibility and constructing primitive.Shape on mm-scale to reticle pattern
Shape, size carry out accuracy controlling；
2) after cleaning-drying masterplate, smooth glass surface is fixed in, prepared magnetic nano-particle is scattered in
Crosslinkable biocompatibility is constructed in primitive liquid, and finally casting filling is to the pierced pattern of masterplate；
3) by way of mechanical grip, pressing plate-masterplate-base plate sandwich structure is formed, by modes such as heating, illumination
Solidification biocompatibility constructs primitive, and the demoulding obtains the biocompatibility of discrete magnetic response and constructs primitive.
Frozen sections prepare micro-meter scale magnetic responsiveness biocompatibility and construct primitive
1) choose the relatively large liquid biological compatibility of viscosity and construct primitive, dispersion is magnetic using " sandwich " mode
The cross-linking liquid biological compatibility of nanoparticle is constructed primitive and is closely suppressed between pressing plate and base plate, and biocompatibility is constructed
Primitive thickness passes through two ends clamping plate thickness adjusted.
2) base is constructed to the folded liquid biological compatibility for being dispersed with magnetic nano-particle by modes such as heating or illumination
Unit is thoroughly solidified, and forms the thin film of desired thickness, and the demoulding obtains the thin slice of desired thickness.
3) thin slice is cut into into length and width in grade under the microscope, and length-width ratio is 1：1 thin slice.
4) above-mentioned thin slice is coated with frost embedding medium OCT, set freezing microtome step distance, the thin slice for having embedded is put
Put perpendicular to the position of the freezing microtome edge of a knife, cut under the control of stepper motor, can obtain length in batches is
Grade, width and height are micron-sized strip magnetic responsiveness biocompatibility and construct primitive.
Micro-embossing method prepares micro-meter scale magnetic responsiveness biocompatibility and constructs primitive
1) the SU8 photoresist arrays of required figure are prepared with Ultraviolet lithography in silicon substrate surface.
2) on SU8 photoresist arrays spin coating PDMS performed polymers and its cross-linking agent mixture, above-mentioned array is carried out multiple
System, obtains the hole array of opposite pattern, as micro-embossing masterplate.
3) primitive is constructed in the cross-linking liquid biological compatibility for being dispersed with magnetic nano-particle and drops to above-mentioned PDMS masterplates
On, and polyethylene terephthalate (PET) thin slice is covered, compress.
4) PDMS- biocompatibility is constructed primitive liquid mixture-PET sandwiches to suppress by nanometer hot pressing instrument,
To be sufficient filling with hole array on PDMS masterplates, while remove and overflowing hole and remaining in the liquid between PDMS masterplates and PET
Layer.
5) the liquid biological compatibility is caused to construct the crosslinking of primitive mixture, solidification by heating or illumination.PET is opened, by
It is high more than the surface energy (～22mN/m) of PDMS masterplates in the surface energy (～42mN/m) of PET, so during opening PET, institute
The biocompatibility of solidification is constructed primitive and can be adhered on PET, can pass through to peel off and collect to obtain discrete magnetic responsiveness life
Primitive is constructed in the thing compatibility.
The functional group of primitive surface modification is constructed to distinguish magnetic responsiveness biocompatibility, can be mixed in mixing liquid
Enter a small amount of dyestuff to distinguish.
For without step 4) in nanometer hot pressing instrument suppress in the case of, due to liquid level between PDMS masterplates and PET
Exist, the magnetic responsiveness biocompatibility being fixed on thin film is can only obtain after the demoulding and constructs cell array, and be difficult to
Primitive is constructed to discrete magnetic responsiveness biocompatibility.
3. magnetic responsiveness biocompatibility is constructed primitive surface modification supermolecule or has the molecule of bio-identification effect, substrate
Surface modification supermolecule
1) using graft reaction or free-radical polymerized mode, supermolecule or the molecule with bio-identification effect are connect respectively
To polyelectrolyte；The supermolecule includes supermolecule donor or Supramolecular Receptors；The molecule with bio-identification effect
Including bio-identification donor or bio-identification receptor.
2) magnetic responsiveness biocompatibility is constructed primitive to clean by way of ethanol, deionized water ultrasound, prepares and hand over
For the polyelectrolyte solution needed for layered self-packaging (LbL) process, concentration is in 0.1-1.0mg/mL scopes.
3) in order to distinguish the primitive of constructing for being modified with different functional groups, before LbL modifications, primitive is constructed to dye differentiation,
Such as red and green.Viridescent magnetic responsiveness biocompatibility will be contaminated construct primitive alternate cycles and be soaked in and be grafted with oversubscription
In a polyelectrolyte solutions and b polyelectrolyte solutions of sub- donor, the b polyelectrolyte can be by quiet with a polyelectrolyte
Electricity, hydrogen bond, coordinate bond etc. interact carries out the polyelectrolyte of LbL, and cycle alternation immersion can construct primitive surface in green
The certain thickness polyelectrolyte multilayer film containing the supermolecule donor of modification.Meanwhile, will be the magnetic responsiveness for being infected with redness biological
The compatibility constructs primitive alternate immersion in the c polyelectrolyte solutions and d polyelectrolyte solutions of Supramolecular Receptors is grafted with, described
D polyelectrolyte can be to interact to carry out the polyelectrolyte of LbL with c polyelectrolyte by electrostatic, hydrogen bond, coordinate bond etc., follow
Ring alternate immersion can construct the certain thickness polyelectrolyte multilayer containing the Supramolecular Receptors of primitive surface modification in redness
Film.
Equally, the polyelectrolyte for being grafted with biological identification molecule can pass through step 3) modify magnetic responsiveness bio-compatible
Property constructs primitive surface.
Magnetic responsiveness biocompatibility is constructed into primitive and is changed to substrate, with above-mentioned steps 1) to step 3) it is identical, can be in base
Basal surface modifies supermolecule.
4. Magnetic guidance is modified with supramolecular magnetic responsiveness biocompatibility and constructs primitive and constructs ordered 3 D structure
1) place in having the culture dish of a certain amount of water and be modified with supramolecular substrate, supermolecule is modified with water surface dispersion
Magnetic responsiveness biocompatibility construct primitive；
2) apply local magnetic field with Magnet or three-dimensional magnetic actuating device, make magnetic responsiveness biocompatibility construct primitive sound
External magnetic field is answered, can be moved with the movement in magnetic field or change, magnetic responsiveness biocompatibility is constructed primitive by magnetic manipulation field
Position to specified location；
3) slow water suction to be reducing water level, makes to be modified with supramolecular magnetic responsiveness biocompatibility and constructs primitive with modification
There is supramolecular substrate surface to be fully contacted, reach intermolecular interaction distance, after a period of time, the magnetic responsiveness biocompatibility
Construct primitive and can be passed through super by the substrate of (to) body with supermolecule is modified with to (receiving) body by the supermolecule of its surface modification
Interaction of molecules is fixed, that is, construct the supermolecule that the supermolecule of primitive surface modification is modified to (receiving) body molecule and substrate surface
There is supermolecule by (giving) body molecule to interact, the magnetic responsiveness biocompatibility is constructed after primitive fixed, and no longer will be received
The impact of changes of magnetic field；
To receptor, heretofore described supermolecule refers to that a kind of supermolecule interaction (such as electrostatic interaction) is related to
Effect both sides.
4) add water again, constructed with magnetic responsiveness biocompatibility of the second modification supermolecule of induced by magnetic field to (receiving) body
Primitive, magnetic are carried to specified location, i.e., construct primitive strip with first fixed magnetic responsiveness biocompatibility and mutually put down
Row and the mutually concordant position of end points, the two spacing can be adjusted in the range of ten microns to one millimeter, according to above-mentioned steps 3)
Continue positioning and fixed, the magnetic responsiveness biocompatibility required for can constructing in substrate constructs primitive pattern, form the
One layer of magnetic responsiveness biocompatibility constructs primitive；
5) similar step is modified with supermolecule magnetic responsiveness biocompatibility structure to (receive) body with induced by magnetic field 3) with 4)
Build elementary motions, magnetic is carried to the magnetic responsiveness biocompatibility that ground floor assembled and constructs above primitive, is modified with this super
Molecule is constructed primitive and is fixed on ground floor and is assembled by supermolecule interaction to the magnetic responsiveness biocompatibility of (receiving) body
Magnetic responsiveness biocompatibility construct on primitive, formed second layer magnetic responsiveness biocompatibility construct primitive；
6) ordered 3 D structure of the required number of plies is constructed from bottom to up.
5. Magnetic guidance is modified with supermolecule and/or the magnetic responsiveness biocompatibility of molecule of bio-identification effect constructs base
The ordered 3 D structure for having particular growth factor in specific site is constructed by unit
1) for different somatomedin (belonging to protein matter), first, by 1- (3- dimethylamino-propyls) -3- second
Base carbodiimide hydrochloride/N- hydroxysuccinimides, i.e. EDC/NHS catalyst system and catalyzings, in aqueous respectively to somatomedin
Carry out specific recognition site labelling, such as biotin (biotin) labelling and bacillus cereuss rna enzyme inhibitors (barstar) mark
Note；
2) part magnetic responsiveness biocompatibility is constructed into primitive modification supermolecule, part magnetic responsiveness biocompatibility structure
Primitive is built while modifying supermolecule and the molecule (biotin or barstar) with bio-identification effect；
3) place in having the culture dish of a certain amount of water and be modified with supramolecular substrate, supermolecule is modified with water surface dispersion
Magnetic responsiveness biocompatibility construct primitive；
4) apply local magnetic field with Magnet or three-dimensional magnetic actuating device, make to be modified with supramolecular magnetic responsiveness biofacies
Capacitive constructs primitive response external magnetic field, can move with the movement in magnetic field or change, and any one magnetic is rung by magnetic manipulation field
Answering property biocompatibility is constructed primitive and is moved to and is modified with optional position above supramolecular substrate；
5) slow water suction to be reducing water level, makes to be modified with supramolecular biocompatibility and constructs primitive and be modified with supermolecule
Substrate surface be fully contacted, reach intermolecular interaction distance, after a period of time, it is biological that this is modified with supramolecular magnetic responsiveness
The compatibility construct primitive can by the supermolecule on its surface to (receiving) body be modified with substrate surface of the supermolecule by (to) body
Interacted by supermolecule and fixed, that is, construct what the supermolecule of primitive surface modification was modified to (receiving) body molecule and substrate surface
There is supermolecule by (giving) body molecule and interact in supermolecule, the magnetic responsiveness biocompatibility is constructed after primitive fixed, will
No longer affected by changes of magnetic field；
6) add water again, operate second to be modified with supramolecular magnetic responsiveness biocompatibility with magnetic field and construct primitive,
Magnetic is carried to specified location in substrate, i.e., construct primitive with first fixation and be parallel to each other and the mutually concordant position of end points,
The two spacing can be adjusted in the range of ten microns to one millimeter, 5) continued to position and fixed according to above-mentioned steps, formed first
The magnetic responsiveness biocompatibility of the parallel to each other, periodic arrangement of layer constructs primitive；
7) similar step is simultaneously modified with supermolecule and bio-identification effect molecule (biotin) with induced by magnetic field 5) with 6)
Magnetic responsiveness biocompatibility construct elementary motions, magnetic is carried to ground floor magnetic responsiveness biocompatibility and constructs on primitive
Side, its placement location have following characteristics：Primitive is constructed with ground floor intersection point, the folders constructed between primitive all with ground floor
Angle can be adjusted between 0-90 degree.Make the magnetic responsiveness biocompatibility construct primitive to be fixed on by supermolecule interaction
Ground floor magnetic responsiveness biocompatibility is constructed on primitive；
8) 7) similar step, is modified with supermolecule and another kind of bio-identification effect molecule simultaneously with induced by magnetic field
(barstar) magnetic responsiveness biocompatibility constructs primitive to same layer another location, or the desired location of other layers；
9) ordered 3 D structure of the required number of plies is constructed from bottom to up, and is introduced with specific biological in desired location
The biocompatibility of recognition site constructs primitive；
10) by step 9) prepared by ordered 3 D structure be soaked in can be with biotin, barstar specific recognition
In protein mixed solution (i.e. Avidin avidin, bacillus cereuss rna enzyme barnase), the biological knowledge in ordered 3 D structure is made
Other site can respectively by respective specific biological combination (biotin/avidin identifications, barstar/barnase
Identification), corresponding somatomedin is recognized, is realized that the orientation of different somatomedin in ordered 3 D structure is introduced, is further thin
Born of the same parents' differentiation provides complicated chemical, biological three-dimensional environment；
11) for prepared ordered 3 D structure, Cytotoxic evaluation is carried out by MTT standard methods, fibre is trained
Dimension cell, fat stem cell, endotheliocyte, mescenchymal stem cell etc. carry out cell adsorption experiment, do for mesenchyme is adsorbed with
The three-dimensional rack of cell carries out long-time culture, investigates differentiation situation of the stem cell under the effect of introduced somatomedin.
Described biocompatibility construct primitive include PDMS, hydrogel, polylactic acid, polylactic acid-glycol acid, gather oneself in
Ester, collagen, shitosan etc..
Described macroscopical Supramolecular Assembling yardstick is comprising ten microns to a centimeter scale scope.
The three-dimensional order tissue engineering bracket of macroscopical Supramolecular Assembling prepared by the present invention is stacking shape ordered 3 D structure,
The ordered 3 D structure by it is multiple two-dimensional directional abreast, the primitive of constructing of periodically ordered arrangement constitute, two are mutually
The reality to the generation supermolecule interaction of (receiving) body functional group for constructing complementation of the primitive by modifying on respective surface of contact
The connection on existing two surfaces；The size of the support is 10-5～10-2m。
Beneficial effects of the present invention are as follows：
Macroscopical Supramolecular Assembling method for preparing three-dimensional order tissue engineering bracket that the present invention is provided, micro- is added by various
Work method acquisition size constructs primitive in ten microns to one centimetre of biocompatibility, and the system of primitive is constructed in biocompatibility
Standby middle introducing magnetic nano-particle is so as to construct primitive response external magnetic field.Primitive is constructed to prepared biocompatibility carries out table
Face modification is obtained to be made with supermolecular mechanism such as Host-guest Recognition, electrostatic, hydrogen bond, the complementary functional groups of coordination and bio-identification
Complementary functional groups so that the above-mentioned complementary biological compatibility is constructed primitive and can realize that macroscopic view is super by supermolecule interaction
Molecule is assembled and is fixed up.Further, constructing primitive in three dimensions to biocompatibility using external magnetic field carries out accurately determining
Position, and the fixed life of supermolecule interaction constructed between primitive using operating platform substrate and biocompatibility in specified location
Primitive is constructed in the thing compatibility, so as to progressively build ten microns of ordered 3 D structures to a cm size scope from bottom to top；
During three dimensional structure is constructed, primitive surface biological recognition site is constructed by biocompatibility and can orient the specified growth of introducing
The factor, so as to realize that orientation of various somatomedin inside three-dimensional stent material is introduced.The method can provide a kind of new
Prepare with complicated chemical, the method for bioenvironmental ordered 3 D structure, be it is a kind of with universality, it is gentle, biological
Compatible preparation method, is expected to be used for preparing the masterplate of Cell differentiation inducing activity and three-dimensional tissue's engineering rack.
Description of the drawings
Below in conjunction with the accompanying drawings the specific embodiment of the present invention is described in further detail.
Fig. 1 a and Fig. 1 b illustrate the transmission electron microscope photo of synthesis ferriferrous oxide nano-particle；
Fig. 2 a illustrate the pure ferriferrous oxide nano-particle of synthesis and
Fig. 2 b illustrate that magnetic responsiveness constructs the hysteresis curve of primitive；
Fig. 3 illustrates square acrylic masterplate and mechanical grip optical photograph；
Fig. 4 illustrates that magnetic responsiveness PDMS cubes construct primitive；
Fig. 5 illustrates that magnetic responsiveness hydrogel constructs primitive；
Fig. 6 illustrates prepared by Frozen sections and constructs primitive schematic flow sheet；
Fig. 7 illustrates that PDMS strips prepared by Frozen sections construct primitive；
Fig. 8 illustrates that micro-embossing method prepares micron order and constructs primitive schematic diagram；
Fig. 9 a, Fig. 9 b, Fig. 9 c and Fig. 9 d illustrates SU8 photoresist arrays prepared by Ultraviolet lithography；
Figure 10 a and Figure 10 b illustrate that micron order hydrogel prepared by micro-embossing method constructs primitive fluorescence photo
Figure 10 c and Figure 10 d illustrate that micron order hydrogel prepared by micro-embossing method are constructed and peel off in primitive water ordinary optical photograph
Piece；
Figure 11 a illustrate sinteticses PAA-CD nuclear magnetic spectrograms；
Figure 11 b illustrate sinteticses PAA-Azo nuclear magnetic spectrograms；
Figure 12 illustrates that alternately primitive surface schematic flow sheet is constructed in the modification of stratiform self-assembling technique method；
Figure 13 illustrates that Magnetic guidance macroscopic view Supramolecular Assembling constructs ordered 3 D structure schematic flow sheet；
Figure 14 a, Figure 14 b, Figure 14 c and Figure 14 d illustrates the ordered 3 D structure constructed by Magnetic guidance macroscopic view Supramolecular Assembling；
Figure 15 a and Figure 15 b illustrate that specific site has the ordered 3 D structure of particular growth factor；
Figure 16 illustrates the Cytotoxic evaluation result of material；
Figure 17 a and Figure 17 b illustrate cell Adsorption experimental results.
Specific embodiment
In order to be illustrated more clearly that the present invention, the present invention is done further with reference to preferred embodiments and drawings
It is bright.In accompanying drawing, similar part is indicated with identical reference.It will be appreciated by those skilled in the art that institute is concrete below
The content of description is illustrative and be not restrictive, and should not be limited the scope of the invention with this.
1. hydro-thermal method prepares ferroferric oxide magnetic nano-particles
1), Iron(III) chloride hexahydrate is dissolved in ethylene glycol, stirring forms clear yellow solution.
2) sodium acetate and Polyethylene Glycol are added, continues to be stirred vigorously 30 minutes, formed colour of loess color cloud liquid, be added to poly- four
In fluorothene reactor, sealing is heated 10 hours at 200 degrees celsius.
3) room temperature, is cooled to, with the ferriferrous oxide nano-particle obtained by washing with alcohol, is washed three times, it is Celsius 60
Degree vacuum oven 6 hours, obtains black powder, and its size and hysteresis curve are respectively such as Fig. 1 a and Fig. 1 b, Fig. 2 a
With shown in Fig. 2 b.
2.1 mechanical micro Process masterplate casting methods prepare grade magnetic responsiveness biocompatibility and construct primitive
For the biocompatibility of millimeter and scale above constructs primitive, masterplate can be processed by traditional mechanical and be cast into
It is prepared by the mode of type.
1) using the laser-engraving technique in mechanical processing industry, engrave on polymethyl methacrylate (acrylic) sheet material
Hollow pattern needed for carving, as the masterplate for preparing biocompatibility and constructing primitive.By means of computer aided technique, the upper figure of masterplate
The shape of case, size are easily controlled and are adjusted in millimeter and scale above.By taking square pattern as an example, can conduct
Casting cube biocompatibility constructs the masterplate of primitive, as shown in Figure 3；
2) masterplate is cleaned successively with ethanol, deionized water, be fixed on smooth glass surface after being dried；
3) at room temperature, by the liquid biological compatibility construct primitive (by taking PDMS as an example, the mixing of its performed polymer and cross-linking agent
Thing) dispersion is sufficiently mixed with prepared ferriferrous oxide nano-particle, resulting mixture is filled into and is fixed on glass
In masterplate square hole on plate；
4) another glass plate is up withheld, and glass plate-masterplate-glass plate three is clamped, in 65 degrees Celsius of baking ovens
Middle heating a period of time, solidification, the demoulding obtain the cube PDMS fritters of discrete magnetic responsiveness and construct as biocompatibility
Primitive, as shown in Figure 4.
For hydrogel biocompatibility constructs the preparation of primitive, same step can be adopted, by hydrogel monomer, crosslinking
Liquid after agent, initiator mix homogeneously, is sufficiently mixed dispersion with magnetic nano-particle, is filled in masterplate, by thermal initiation
Or after light initiation polymerization forms gel, the demoulding, as shown in Figure 5.For polylactic acid, polylactic acid-glycol acid, gather in oneself
The low melting materials such as ester, collagen, shitosan, can become melt liquid at relatively high temperatures, mixed with magnetic nano-particle
Pour after conjunction and cast from metal matrix, be subsequently cooled to cold curing, the demoulding.
2.2 Frozen sections prepare micro-meter scale magnetic responsiveness biocompatibility and construct primitive
1), by microscope slide Piranha washing liquid (concentrated sulphuric acids：Hydrogen peroxide=3：1v/v) cleaned, deionized water punching
Wash, nitrogen is dried up.In hermetic container, a small amount of 1H of Deca, 1H, 2H, 2H- perfluoro capryl triethoxysilane, discrete placement are washed
Net microscope slide, at 60 c by the vapour deposition perfluor silane to microscope slide, makes microscope slide that hydrophobicity is presented.
2), as shown in fig. 6, by taking PDMS as an example, first by PDMS performed polymers and its cross-linking agent, magnetic nano-particle with certain
Ratio mixes, and drips on prepared hydrophobic microscope slide, and two ends mat thickness is 150 microns of coverslip, and top presses hydrophobic load
Slide, forms hydrophobic microscope slide-PDMS mixture-hydrophobic microscope slide sandwich, is heating and curing under 65 degrees Celsius, and the demoulding is obtained
To the PDMS thin slices that thickness is 150 microns.
3) above-mentioned 150 microns of PDMS thin slices are cut into into width for 2 millimeters × 2 millimeters × 150 microns, under the microscope,
Thin slice.
4), above-mentioned thin slice is coated with frost embedding medium OCT, set freezing microtome step distance, by the thin slice for having embedded
The position of the freezing microtome edge of a knife is disposed perpendicular to, is cut under the control of stepper motor, size can be obtained in batches
For 2 millimeters × 150 microns × 10～100 microns of strip, wherein 10～100 microns to set adjustable step distance,
As shown in Figure 7.
2.3 micro-embossing methods prepare micro-meter scale magnetic responsiveness biocompatibility and construct primitive
Micro-embossing method flow process is as shown in Figure 8：
1), by taking 200 microns × 25 microns × 20 microns hydrogel strips as an example, prepared in silicon chip surface with Ultraviolet lithography
The array of SU8 photoresists, as shown in figure 9, Fig. 9 a are strip SU8 photoresist arrays, Fig. 9 b construct base for one of strip
The enlarged drawing and corresponding size of unit, Fig. 9 c are the cross-sectional views that strip constructs primitive, and Fig. 9 d are strip cross-sectional views in Fig. 9 c
Size.
2), on SU8 photoresist arrays spin coating PDMS performed polymers and its cross-linking agent mixture, above-mentioned array is carried out multiple
System, obtains the strip hole of opposite pattern, as masterplate.
3), hydrogel monomer, cross-linking agent, initiator and viscosifier are mixed, and dispersed magnetic nanoparticle, this is mixed
Thing is dropped on above-mentioned PDMS masterplates, and covers polyethylene terephthalate (PET) thin slice, is compressed.
4), PDMS- hydrogel monomer mixture-PET sandwiches are compressed by nanometer hot pressing instrument, can be sufficient filling with
The upper long strip type holes of PDMS, while removing overflowed and remaining in the liquid between masterplate and PET.
5) hydrogel crosslinking, solidification are caused by 365nm ultraviolet lightings or heating,.PET is opened, due to the surface energy of PET
More match with the surface energy of gel strip, so during opening PET, the gel mass for being solidified can be adhered on PET, can
So that discrete hydrogel strip, such as Figure 10 a are come off and collected in water, shown in Figure 10 b, Figure 10 c and Figure 10 d.
The functional group of primitive surface modification is constructed to distinguish magnetic responsiveness biocompatibility, can be mixed in mixing liquid
Enter a small amount of dyestuff to distinguish.
3. magnetic responsiveness biocompatibility constructs functional group of the primitive surface modification with supermolecular mechanism
1) the supramolecular polyelectrolyte that is modified with for alternately stratiform package technique synthesizes：Using graft reaction or freedom
Base copolymerization mode, supermolecule cyclodextrin and diphenyl diimide are introduced in polyacrylic acid (PAA) this polyelectrolyte, obtain polypropylene
Acid-cyclodextrin (PAA-CD), polyacrylic acid-diphenyl diimide (PAA-Azo), nuclear-magnetism figure is as shown in figures 11a and 11b.
2) magnetic responsiveness biocompatibility is constructed into primitive (by taking PDMS cube fritters as an example) by ethanol, deionized water
The mode of ultrasound is cleaned.
3) prepare polyelectrolyte solution alternately needed for layered self-packaging (LbL) process：Polyacrylic acid-cyclodextrin (PAA-
CD), polyacrylic acid-diphenyl diimide (PAA-Azo)：0.1-1.0mg/mL.
4) a kind of biocompatibility is constructed into primitive (dying green) alternate cycles as shown in Figure 12, and is soaked in poly- (two
Allyl dimethyl ammonium chloride) (PDDA) and in PAA-CD solution, obtain certain thickness PDDA/PAA-CD multilayer films；Will be another
A kind of biocompatibility is constructed primitive (dying redness) alternate cycles and is soaked in PDDA and PAA-Azo solution, obtains certain thick
The PDDA/PAA-Azo multilayer films of degree.So, by biocompatibility constructs primitive surface, modification is with cyclodextrin or azo
The supermolecule of benzene.
Furthermore, it is possible to become PAH-biotin in PAH (PAH) side chain graft biotin (biotin),
Formed containing a kind of supramolecular PAH-biotin/PAA multilayer films of biotin by participating in above-mentioned alternately stratiform package technique, or
Person is simultaneous with cyclodextrin and the supramolecular PAH-biotin/PAA-CD multilayer films of two kinds of biotin.It is likewise possible to introduce contain
There is the polyelectrolyte PAH-barnase of bacillus cereuss rna enzymes, obtain the multilayer film of PAH-barnase/PAA-CD.
In pre-cast solution, mixing is with supramolecular compound, such as PAA-CD or PAA-Azo, it is also possible to realize institute
Prepare biocompatibility and construct supermolecule in the surface modification of primitive.
The surface modification of ordered 3 D structure operating platform substrate (piezoid, sheet glass or silicon chip)：First, use
Piranha washing liquid (concentrated sulphuric acids：Hydrogen peroxide=3：1v/v) piezoid or sheet glass or silicon chip are cleaned, deionized water punching
Wash, nitrogen is dried up；Then piezoid or sheet glass or silicon chip alternate cycles are soaked in PDDA and PAA-CD solution, obtain one
Determine the PDDA/PAA-CD multilayer films of thickness.
4. Magnetic guidance be modified with supramolecular magnetic responsiveness biocompatibility construct primitive assembling construct ordered 3 D structure
1) so that strip magnetic responsiveness PDMS biocompatibility constructs primitive as an example, put in having the culture dish of a certain amount of water
The piezoid or sheet glass or silicon chip substrate for being modified with PDDA/PAA-CD multilayer films is put, and diphenyl diimide is modified with water surface dispersion and is surpassed
The PDMS strips of molecule；
2) as shown in figure 13, apply local magnetic field with Magnet or three-dimensional magnetic actuating device, make to be modified with diphenyl diimide oversubscription
The long bar response external magnetic fields of PDMS of son, can move with the movement in magnetic field or change, and magnetic manipulation field will be modified with diphenyl diimide
Supramolecular PDMS strips position the optional position to substrate；
3) slow water suction makes to be modified with the supramolecular PDMS strips of diphenyl diimide and operating platform substrate surface to reduce water level
Be fully contacted, reach intermolecular interaction distance, after a period of time, the PDMS bars can pass through the diphenyl diimide supermolecule on its surface with
The cyclodextrin supermolecule of operating platform substrate surface is interacted by supermolecule and makes to be modified with the supramolecular PDMS bars of diphenyl diimide
The optional position being fixed in substrate, is no longer affected by changes of magnetic field after its fixation；
4) adding water again, second being operated with magnetic field and be modified with the supramolecular PDMS strips of diphenyl diimide, magnetic is carried to substrate
Upper to be parallel to each other with first PDMS strip and the mutually concordant position of end points, the two spacing can be in ten microns to one millimeter models
Interior regulation is enclosed, 3) continues to position and fixed according to above-mentioned steps, being modified with required for can constructing in operating platform substrate
The pattern of the supramolecular PDMS strips of diphenyl diimide, formation ground floor is parallel, periodic arrangement magnetic responsiveness biocompatibility structure
Build primitive；
5) adding water again, the supramolecular PDMS strips motion of cyclodextrin being modified with induced by magnetic field, magnetic is carried to ground floor
It is modified with above the supramolecular PDMS strips of diphenyl diimide, its placement location has feature：There is intersection point with ground floor PDMS bars,
Angle between PDMS bars all with ground floor can be adjusted between 0-90 degree.According to this arrangement mode, ring paste is modified with
The supramolecular PDMS strips of essence are fixed on by supermolecular mechanism and are modified with the supramolecular PDMS layer of diphenyl diimide, form the second layer
Magnetic responsiveness biocompatibility constructs primitive；
6) ordered 3 D structure of the required number of plies, for resulting ordered 3 D structure, its magnetic are constructed from bottom to up
Position, spacing and the number of plies that response biocompatibility constructs primitive is adjustable in three dimensions, such as Figure 14 a, Figure 14 b, figure
Shown in 14c and Figure 14 d.
5. Magnetic guidance is modified with supermolecule and/or the magnetic responsiveness biocompatibility of molecule of bio-identification effect constructs base
The ordered 3 D structure for having particular growth factor in specific site is constructed in unit's assembling
1) introducing two kinds of different somatomedin, VEGF (VEGF) and fibroblast growth factor
(FGF) as a example by, first, by 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides/N- hydroxysuccinimides,
I.e. two kinds of growth factor VEGFs and FGF are carried out biotin (biotin) labelling and bud by EDC/NHS catalyst system and catalyzings in aqueous
Spore bacillus rna enzyme inhibitors (barstar) labelling, obtains VEGF-biotin, FGF-barstar；
2) so that strip magnetic responsiveness PDMS biocompatibility constructs primitive as an example, by part PDMS strip modification cyclodextrins
Or diphenyl diimide supermolecule, part PDMS strips are while modification cyclodextrin supermolecule and Avidin (avidin) molecule, part PDMS
Strip is while modification cyclodextrin supermolecule and bacillus cereuss rna enzyme molecules, part PDMS strips are while avidin molecule and azo
Benzene supermolecule, part PDMS strips modify bacillus cereuss rna enzyme molecules and diphenyl diimide supermolecule simultaneously；
3) in having the culture dish of a certain amount of water place be modified with PDDA/PAA-CD multilayer films piezoid or sheet glass or
Silicon chip substrate, is modified with the supramolecular PDMS strips of diphenyl diimide in water surface dispersion；
4) apply local magnetic field with Magnet or three-dimensional magnetic actuating device, make to be modified with the supramolecular PDMS strips of diphenyl diimide
Response external magnetic field, can move with the movement in magnetic field or change, and magnetic manipulation field will be modified with the supramolecular PDMS of diphenyl diimide
Strip positions the specified location to substrate；
5) slow water suction makes to be modified with the supramolecular PDMS strips of diphenyl diimide and operating platform substrate surface to reduce water level
It is fully contacted, reaches intermolecular interaction distance, after a period of time, this is modified with the supramolecular PDMS bars of diphenyl diimide and can pass through which
The diphenyl diimide supermolecule on surface is interacted by supermolecule with the cyclodextrin supermolecule of operating platform substrate surface and is fixed, and which is solid
No longer affected by changes of magnetic field after fixed；
6) adding water again, second being operated with magnetic field and be modified with the supramolecular PDMS strips of diphenyl diimide, magnetic is carried to substrate
On specified location, 5) continue to position and fixed according to above-mentioned steps, repairing required for can constructing in operating platform substrate
The pattern of the supramolecular PDMS strips of diphenyl diimide is decorated with, ground floor magnetic responsiveness biocompatibility is formed and is constructed primitive；
7) add water again, the PDMS strips fortune of cyclodextrin supermolecule and avidin molecule is modified with simultaneously with induced by magnetic field
Dynamic, magnetic is carried to ground floor and is modified with above the supramolecular PDMS strips of diphenyl diimide, makes to be modified with cyclodextrin supermolecule and affine
The PDMS strips of plain molecule interact to be fixed on by supermolecule and are modified with the supramolecular PDMS layer of diphenyl diimide, form second
Layer magnetic responsiveness biocompatibility constructs primitive；
8) 7) similar step, contains bacillus cereuss rna enzyme molecules and the supramolecular PDMS of cyclodextrin simultaneously with induced by magnetic field
Strip, is fixed to same layer another location, or Magnetic guidance contains bacillus cereuss rna enzyme molecules simultaneously and diphenyl diimide is supramolecular
PDMS strips are fixed to the specified location, or Magnetic guidance of adjacent layer while containing avidin molecule and diphenyl diimide supermolecule
PDMS strips be fixed to the specified location of adjacent layer；
9) ordered 3 D structure of the required number of plies is constructed from bottom to up, and is introduced with specific recognition in desired location
The PDMS biocompatibility in site (such as avidin, barstar) constructs primitive, as shown in Figure 15 a and Figure 15 b；
10) by step 9) prepared by structure be soaked in the mixed solution of VEGF-biotin and FGF-barstar, make
Avidin sites and barnase sites in ordered 3 D structure can pass through biotin/avidin or barnase/ respectively
Barstar specific bindings are acted on, and are recognized corresponding VEGF-biotin and FGF-barstar somatomedin, are realized three-dimensional order
In structure, the orientation of different somatomedin is introduced, and provides chemical, the biological three-dimensional environment of complexity for further cell differentiation；
11) for step 9) prepared by structure, Cytotoxic evaluation is carried out by MTT standard methods, as a result shows institute
There is no cytotoxicity (as shown in figure 16) with material.Culture fibroblast, fat stem cell, endotheliocyte, mesenchyme are dry thin
Born of the same parents etc. carry out cell adsorption experiment, for the three-dimensional rack for being adsorbed with mescenchymal stem cell carries out long-time culture, investigate dry thin
Differentiation situation of the born of the same parents under the effect of introduced somatomedin.Cell Adsorption experimental results are embodied as shown in accompanying drawing 17a and Figure 17 b
Preferable cell absorption, migration, growth and differentiation effect.
Obviously, the above embodiment of the present invention is only intended to clearly illustrate example of the present invention, and is not right
The restriction of embodiments of the present invention, for those of ordinary skill in the field, may be used also on the basis of the above description
To make other changes in different forms, all of embodiment cannot be exhaustive here, it is every to belong to this
Bright technical scheme it is extended obvious change or change still in protection scope of the present invention row.
Claims (6)
1. a kind of method that macroscopical Supramolecular Assembling prepares three-dimensional order tissue engineering bracket, it is characterised in that：The support is
Stacking shape ordered 3 D structure, the ordered 3 D structure by it is multiple two-dimensional directional abreast, the structure of periodically ordered arrangement
Primitive composition is built, two functional groups of the primitive by modifying on respective surface of constructing for contacting with each other occur supermolecule interaction
And realize the connection on two surfaces；The size of the support is 10-5 ~10-2m；
Methods described includes：
Magnetic responsiveness biocompatibility constructs the preparation of primitive；
Magnetic responsiveness biocompatibility constructs primitive surface modification functional group；
The magnetic responsiveness biocompatibility for being modified with functional group is constructed primitive and constructs ordered 3 D structure；
The magnetic responsiveness biocompatibility constructs the preparation of primitive, is constructed to biocompatibility by way of mixing and doping
Magnetic nano-particle is introduced in primitive primitive is constructed to prepare magnetic responsiveness biocompatibility；
The magnetic responsiveness biocompatibility for being modified with functional group is constructed the step of primitive constructs ordered 3 D structure and is included：
Magnetic responsiveness biocompatibility constructs primitive surface modification functional group；
Magnetic guidance is modified with the magnetic responsiveness biocompatibility of functional group and constructs that primitive constructs ordered 3 D structure or Magnetic guidance is repaiied
Be decorated with functional group magnetic responsiveness biocompatibility construct primitive construct specific biological recognition site have particular growth because
The ordered 3 D structure of son；
The Magnetic guidance is modified with the magnetic responsiveness biocompatibility of functional group the step of construct primitive and construct ordered 3 D structure
Specifically include：
1) place in aqueous container and be modified with the substrate of functional group, primitive is constructed what water surface dispersion was modified with functional group；
The substrate is selected from sheet glass, piezoid or silicon chip；The substrate for being modified with functional group is to be modified with supramolecular substrate；Institute
State be modified with constructing primitive and supramolecular primitive being constructed to be modified with for functional group；
2) to step 1) in container apply magnetic field, the water surface is scattered be modified with functional group construct primitive in magnetic field
The lower positioning of effect is to the specified location being modified with the substrate of functional group；
3) to step 2) in container slowly absorb water, to reduce water level in container, make the scattered structure for being modified with functional group of the water surface
Build primitive and be fully contacted with the substrate surface for being modified with functional group, reach intermolecular interaction distance, realize after 1 minute to 5 minutes
It is fixed；
4) to step 3) in container add water again, construct primitive with what induced by magnetic field second was modified with functional group, in magnetic field
In the presence of position to the specified location being modified with the substrate of functional group；
3) and 4) 5) repeat step, constructs primitive pattern required for constructing in substrate；
6) primitive of constructing of functional group is modified with induced by magnetic field, is positioned to established ground floor under the influence of a magnetic field and is constructed
Above primitive, the ordered 3 D structure for constructing the required number of plies that this step then can be from bottom to top is repeated several times.
2. method according to claim 1, it is characterised in that the magnetic responsiveness that the Magnetic guidance is modified with functional group is biological
The compatibility is constructed primitive and constructed to be had the step of the ordered 3 D structure that specific biological recognition site has particular growth factor
Body includes：
1) carry out specific biological recognition site labelling in aqueous to somatomedin；
2) place in aqueous container and be modified with the substrate of functional group, primitive is constructed what water surface dispersion was modified with functional group；
The substrate is selected from sheet glass, piezoid or silicon chip；The substrate for being modified with functional group is to be modified with supramolecular substrate；Institute
State be modified with constructing primitive and supramolecular primitive being constructed to be modified with for functional group；
3) to step 2) in container apply magnetic field, the primitive of constructing for being modified with functional group is positioned to modification under the action of a magnetic field
The specified location having in the substrate of functional group；
4) to step 3) in container slowly absorb water, to reduce water level in container, make the scattered structure for being modified with functional group of the water surface
Build primitive and be fully contacted with the substrate surface for being modified with functional group, reach intermolecular interaction distance, realize after 1 minute to 5 minutes
It is fixed；
5) to step 4) in container add water again, construct primitive with what induced by magnetic field second was modified with functional group, in magnetic field
In the presence of position to the specified location being modified with the substrate of functional group；
4) and 5) 6) repeat step, constructs primitive pattern required for constructing in substrate；
7) primitive of constructing of functional group is modified with induced by magnetic field, is positioned to established ground floor under the influence of a magnetic field and is constructed
Above primitive, the ordered 3 D structure for constructing the required number of plies that this step then can be from bottom to top is repeated several times；Institute in this step
State and be modified with the primitive of constructing of functional group and include being modified with and supramolecular construct primitive or be modified with supermolecule and/or bio-identification
The molecule of effect constructs primitive；
8) by step 7) prepared by ordered 3 D structure be soaked in and step 1) in the complementary egg of specific biological recognition site
In white mixed solution, the various functional groups with bio-identification effect in ordered 3 D structure are allow respectively by respective
Specific biological combination, recognizes corresponding somatomedin, realizes that the orientation of different somatomedin in ordered 3 D structure is drawn
Enter.
3. method according to claim 1, it is characterised in that：The magnetic responsiveness biocompatibility constructs the size of primitive
For 10-5 ~10-2 m。
4. method according to claim 1, it is characterised in that：The magnetic responsiveness biocompatibility constructs primitive by machine
It is prepared by tool micro Process masterplate casting method, frozen section method or minute-pressure impression method.
5. method according to claim 1, it is characterised in that：The magnetic responsiveness biocompatibility is constructed primitive surface and is repaiied
Decorations functional group is modified by alternately stratiform package technique or surface grafting or monomer blend method；The biocompatibility structure
Building primitive includes PDMS, hydrogel, polylactic acid, polylactic acid-glycol acid, polycaprolactone, collagen or shitosan；The three-dimensional has
The size of sequence structure is 10 μm of -1 cm.
6. method according to claim 1, it is characterised in that：The specific biological recognition site, is with specificity
The bioactive sites of identification ability, the specific biological recognition site are biotin labeling or bacillus cereuss enzyme inhibitor mark
Note.
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| CN113968977B (en) * | 2021-10-29 | 2023-04-07 | 山东理工大学 | Precise assembly method and assembly for macro-functional construction elements in single-phase system |
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| Layer-by-Layer Self-Assembley under High Gravity Field;Lanxin Ma等;《Langmuir》;20120528;第28卷;9849-9856 * |
| Macroscopic Supramolecular Assembly of Rigid Building Blocks Through a Flexible Spacing Coating;Mengjiao Cheng等;《Advanced Materias》;20140122;第26卷;第3009页左栏第1段至第3010页左栏第1段、第3013页左栏第1段 * |
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