CN105311683A - Bionic tissue engineering scaffold containing inner channel network and oriented pore structure as well as preparation method and application of bionic tissue engineering scaffold - Google Patents
Bionic tissue engineering scaffold containing inner channel network and oriented pore structure as well as preparation method and application of bionic tissue engineering scaffold Download PDFInfo
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Abstract
The invention discloses a bionic tissue engineering scaffold containing an inner channel network and an oriented pore structure as well as a preparation method and application of the bionic tissue engineering scaffold. The bionic tissue engineering scaffold comprises a scaffold main body and a channel network, wherein the channel network is arranged inside the scaffold main body; the scaffold main body has the oriented micro-pore structure, and oriented big holes of the oriented micro-pore structure are mutually communicated by virtue of transverse small holes; the channel network has a hierarchical multi-branch structure, and the branches of the channel network are mutually communicated; and the scaffold main body is made from a degradable natural high polymer material. The bionic tissue engineering scaffold disclosed by the invention, which combines a 'core' manufacturing process with an 'oriented pore' directional crystallization and thermally induced phase separation process on the basis of thinking from 'presetting' to 're-dissolving', can break through the technical bottleneck of a complex micro-channel structure which is directly formed. By virtue of a rapid forming technology or a mold casting method, and in particular by virtue of the rapid forming technology, a core forming structure becomes flexible, so that a complex channel structure in any morphologies are formed; and the channel structure is good in passing-through characteristic.
Description
Technical field
The present invention relates to a kind of bionical tissue engineering bracket containing internal channel network and directional pore structure and preparation method thereof and application, belong to organizational project and Biotechnology field.
Background technology
Organizational project (TissueEngineering) is one in front of the door along interdisciplinary science, its ultimate principle is: by building the three-dimensional composite of cell and support in vitro, simulation organism environment, and cultivated and train, obtain the tissue possessing reparation or alternative functions.Current organizational project is developed rapidly in fields such as skin, bone/cartilage, bladder, blood vessel, liver, nerves, for most of difficult medical problem of facing mankind bring new hope, as the treatment of the diseases such as cardiovascular disease, alzheimer disease, parkinson disease, congenital hereditary defect and various injuries of tissues and organs.
Tissue engineering bracket is by bionic extracellular matrix, and for cell provides mechanical support, promoting cell adhesion, growth, propagation, migration and metabolism, is one of key factor of organizational project.For some natural tissues (as cardiac muscle, skeletal muscle, nerve), wherein parenchyma Orienting ordered arrangement, connection, as cardiac muscular tissue's cardiac myocyte along collagen fiber arrangement and connected by intercalated disc, this tissue characteristics plays an important role in the conduction of myocardial electrical signals.Now there are some researches show, there is the tissue engineering bracket of the directed micropore structure of microcosmic, to a certain extent can the aligning and be connected (HTHAu of inducing cardiomyocytes by the microstructure of material, BCui, etal.Cellculturechipsforsimultaneousapplicationoftopogra phicalandelectricalcuesenhancephenotypeofcardiomyocytes [J] .LabChip, 2009,9:564 – 575.); Electrical field stimulation can promote myocardial cell respectively along growth and the arrangement (RadisicM of direction of an electric field, ParkH, ShingH, etal.Functionalassemblyofengineeredmyocardiumbyelectrica lstimulationofcardiacmyocytesculturedonscaffolds.Proceed ingsoftheNationalAcademyofSciencesoftheUnitedStatesofAme rica, 2004,101 (52): 18129-18134.); External mechanical stretch stimulates can promote that cell is along stress direction growth and arrangement (Tandonetal.AlignmentandElongationofHumanAdipose-DerivedS temCellsinResponsetoDirect-CurrentElectricalStimulation. ConfProcIEEEEngMedBiolSoc.2009,1:6517 – 6521.), but corresponding action rule there is not yet deep research report.Traditional tissue engineering bracket manufacture method is as solution-cast/porogen deposition method, thermally induced phase separation, electrical spinning method, air pressure port-creating method etc., and the support of preparation mostly is isotropism, evenly random pore structure.Method at present for the directed micropore structure support that is shaped mainly contains crystallographic orientation-thermally induced phase separation and directed Electrospun two kinds.
On the other hand, vascularization problem is the common problem of Tissue Engineering Study, is also the bottleneck that restriction organizational project further develops.For the vigorous tissue of metabolism as cardiac muscle, liver, because cell has very high material and energy metabolism level, and oxygen and the nutrient substance transmission range in support is limited, therefore how sets up effective blood vessel network, the necrosis of solution internal layer cell hypoxia is the important problem that organizational project faces always.The modes such as existing perfusion cultures, raising surrounding oxygen content, the interpolation carrier of oxygen and the interior transplanting of body can make the artificial organ living cells layer thickness of structure reach 500 μm.There are some researches show, add the material such as endotheliocyte, angiogenic growth factor and can promote that transplanting rear blood capillary sample tissue in body generates.But said method does not form the through blood vessel network of similar classification, the oxygen supply problem of internal stent cell fundamentally can not be solved.Therefore, build containing the directed microscopic void support of internal channel network, significant for cardiac muscle tissue engineering and skeletal muscle, neural tissue engineering.
Summary of the invention
The object of this invention is to provide a kind of bionical tissue engineering bracket containing internal channel network and directional pore structure and preparation method thereof and application, the present invention fully simulates some natural tissues as cardiac muscle containing the bionical tissue engineering bracket of internal channel network and directional pore structure, skeletal muscle, neural feature-" cell directional ordered arrangement, containing multistage blood vessel network " structure, there is directed micropore structure and multiple-limb channel network, wherein there is mutually through horizontal aperture between directed macropore, and multiple-limb channel network configuration is directly through or connect through by directed micropore structure.The bionical tissue engineering bracket of the present invention can inducing cell oriented alignment, connection and growth, for internal stent cell provides oxygen and nutrient substance, promote angiogenesis, and improve support inner cell planting density and the degree of depth, form bulk tissue substitute, be expected to the efficiency greatly improving the reparation of disease damage histoorgan.
The bionical tissue engineering bracket containing internal channel network and directional pore structure provided by the present invention, comprises rack body and is arranged at the channel network in described rack body;
Described rack body has directed micropore structure, and realizes mutually through by horizontal aperture between the directed macropore of described directed micropore structure;
Described rack body is made up of degradable natural macromolecular material;
Described channel network has classification multiple branching construction;
Mutually through between the branch of described channel network.
In above-mentioned bionical tissue engineering bracket, be directly through between the branch of described channel network.
In above-mentioned bionical tissue engineering bracket, the hole by described rack body between the branch of described channel network is mutually through.
In above-mentioned bionical tissue engineering bracket, the diameter of section of described directed macropore can be 20 μm ~ 250 μm, specifically can be 20 ~ 100 μm, 50 ~ 200 μm or 100 ~ 250 μm, the degree of orientation (OrientationIndex, OI) can be 0.8 ~ 1, specifically can be 0.8 ~ 0.9,0.9 ~ 0.95 or 0.92 ~ 0.95;
The diameter of section of the branch of described channel network can be 50 μm ~ 3000 μm, specifically can be 50 ~ 1500 μm, 200 ~ 1000 μm or 1000 ~ 3000 μm;
Described channel network has 1 ~ 4 grade of branched structure, as 2 grades or 3 grades of hierarchies.
In above-mentioned bionical tissue engineering bracket, described degradable natural macromolecular material can be at least one in collagen, chitosan, sodium alginate, gelatin, fibroin albumen, hyaluronic acid, Fibrinogen and albumin.
Invention further provides the described preparation method containing the bionical tissue engineering bracket of internal channel network and directional pore structure, comprise the steps:
1) by speed forming method or injection molding, by inner core material molding, the core arrangement with described channel network configuration is obtained;
2) described core arrangement is fixed in directed bracket mould, then the solution of described degradable natural macromolecular material is filled the die cavity of described directed bracket mould; Described directed bracket mould is placed in the orientation temperature field lower than 0 DEG C, realize crystallographic orientation and the Thermal inactive of the solution of described degradable natural macromolecular material, then obtain containing core arrangement, the tissue engineering bracket with directive construction through lyophilization;
3) removal step 2) described core arrangement in the described tissue engineering bracket that obtains, namely obtain the bionical tissue engineering bracket containing internal channel network and directional pore structure.
In above-mentioned preparation method, step 1) in, described inner core material is the material with good solubility, biocompatibility and mechanical property, specifically can be at least one in saccharide compound, gelatin and alkali soluble light-sensitive polymer;
Described saccharide compound can be at least one in glucose, maltose and sucrose;
The molecular weight of described gelatin can be 15000 ~ 250000Da, can be A type gelatin, also can be Type B gelatin;
Described alkali soluble light-sensitive polymer can be the copolymer of dimethyl-acrylamide, methacrylic acid and polyvinyl pyrrolidone, and the mass ratio of three is 11:11:3.
In above-mentioned preparation method, step 1) in, described speed forming method can adopt the one in the techniques such as continuous expressing technique, stereolithography processes and laser micro-cladding sintering; Described injection molding is specially: by preparation containing the PDMS mould of described channel network configuration, liquid inner core material of casting wherein, by cooling or photopolymerization curing, and the solubility inner core of shaping multiple-limb form.
In above-mentioned preparation method, described inner core material be described saccharide compound and/or gelatin time, step 2) before, described method also comprises the step described core arrangement being placed in PLGA solution (if concentration is 0.1 ~ 1mg/mL), to form protective film on the surface of described core arrangement;
Step 2) in, the temperature of described orientation temperature field changes along the length direction of described directed bracket mould;
The temperature in the hot junction of described orientation temperature field can be 0 DEG C, and the temperature of cold junction can be-20 DEG C ~-196 DEG C, and the value of thermograde can be 2 ~ 20K/mm; The temperature of described cold junction specifically can be-20 DEG C ,-80 DEG C ,-196 DEG C ,-20 DEG C ~-80 DEG C or-80 DEG C ~-196 DEG C; The value of described thermograde specifically can be 2K/mm, 8K/mm, 20K/mm, 2 ~ 8K/mm or 8 ~ 20K/mm;
Step 3) in, before removing described core arrangement, described method also comprises the step utilizing the mode of physical crosslinking and/or chemical crosslinking to stablize described tissue engineering bracket;
When described inner core material be described saccharide compound and/or described alkali soluble light-sensitive polymer time, adopt and remove described core arrangement with the following method: be soaked in deionized water, distilled water, NaOH aqueous solution or PBS buffer;
When described core arrangement is described gelatin, adopts and remove described core arrangement with the following method: the temperature-sensing property utilizing its " gel-sol " to change, be placed in the stripping of gelling temp point.
Bionical tissue engineering bracket containing internal channel network and directional pore structure provided by the invention can be used for preparation tissue substituent, medicaments sifting model and pathological study model etc.;
Described tissue substituent can be myocardium substitute, skeletal muscle substitute or neural substitute etc.
Compared with prior art, the present invention has the following advantages and salience effect:
(1) tissue engineering bracket of the present invention possesses significant biomimetic features feature.Main part: the extra-cellular matrix structure of simulation natural tissues, constructs the support with directed micropore structure, can promote oriented alignment and the connection of parenchyma; Longitudinal through or non-fully through channel network, simulate the multi-branch complex vascular access structure in natural tissues, be conducive to the transmission of oxygen and nutrient substance, and promote the vascularization of endotheliocyte further, be conducive to improving cell seeding density and the degree of depth, thus provide possibility for building bulk engineering tissue.
(2) crystallographic orientation of " inner core " manufacturing process and " directed porosity ", thermal phase separation process combine by the present invention " preset-heavy molten " thinking, overcome the technical bottleneck of direct forming complexity MCA.Adopt rapid shaping technique or mold castings, especially the former, can make the flexibility of inner core shaped structure, has been shaped and had the complex passages structure of any pattern, and channel design has good through feature.
Accompanying drawing explanation
Fig. 1 is the structural representation containing the bionical tissue engineering bracket of internal channel network and directional pore structure provided by the invention, wherein Fig. 1 (a) is for comprising the structural representation of the bionical tissue engineering bracket of directly through multiple branching construction class blood vessel access, and Fig. 1 (b) is for comprising the structural representation of the bionical tissue engineering bracket by the indirectly through multiple branching construction class blood vessel access of directed porosity.
Fig. 2 is the mould of the solubility core arrangement for being shaped with multiple-limb passage.
Fig. 3 is for having multiple-limb shape core arrangement schematic diagram, and wherein Fig. 3 (a) is corresponding with the class blood vessel network form in Fig. 1 (a), and Fig. 3 (b) is corresponding with the class blood vessel network form in Fig. 1 (b).
Fig. 4 is the cross section electromicroscopic photograph that the present invention contains channel network in the bionical tissue engineering bracket of internal channel network and directional pore structure.
Fig. 5 is the electromicroscopic photograph containing directed macropore (Fig. 5 (a)) and horizontal aperture (Fig. 5 (b)) in the bionical tissue engineering bracket of internal channel network and directional pore structure provided by the invention.
Wherein, in accompanying drawing, each labelling is as follows:
1 brace aperture main structure body, 2 bionical class blood vessel access network structures, 3 core arrangement, 4 surface protective films, 5 horizontal apertures.
Detailed description of the invention
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Fig. 1 is the schematic diagram containing the bionical tissue engineering bracket of internal channel network and directional pore structure provided by the invention.The bionical class blood vessel access network structure 2 that this tissue engineering bracket comprises brace aperture main structure body 1 and is arranged in rack body; Wherein rack body 1 has directed micropore structure, there is mutually through horizontal aperture 5 between directed macropore wherein; Bionical class blood vessel access network 2 has classification multiple branching construction, and it is indirectly through or directly through with the form of Fig. 1 (b) by directed porosity with the form of Fig. 1 (a); Rack body 1 adopts degradable natural macromolecular material to make.
The diameter of section of the directed macropore in rack body 1 is 20 μm ~ 250 μm, and the OI value of the degree of orientation is 0.8 ~ 1; The diameter of section of bionical class blood vessel access network 2 is 50 μm ~ 3000 μm, has 1 ~ 4 grade of branched structure.
Degradable natural macromolecular material adopts the complex of one or more materials in collagen, chitosan, sodium alginate, gelatin, fibroin albumen, hyaluronic acid, Fibrinogen and albumin.
Following step is adopted to prepare the bionical tissue engineering bracket containing internal channel network and directional pore structure of the present invention:
1) solubility inner core material is dissolved, make the solution that mass percentage concentration is 0.1% ~ 5%;
2) inner core material is passed through quick molding method or die methods, the core arrangement 3 of shaping classification multiple branching construction; Core arrangement 3 is immersed in airtight preservation a period of time in PLGA solution (0.1 ~ 10mg/mL), takes out and air-dry more than 12 hours, form surface protective film 4.
3) core arrangement 3 is fixed on the longitudinal direction of directed bracket mould, again the solution of degradable natural polymer is filled the die cavity of directed bracket mould, mould is placed in the orientation temperature field lower than 0 DEG C, realize crystallographic orientation and the Thermal inactive of solution, then lyophilization, forms Embedded Web structure, has the tissue engineering bracket of directional pore structure;
4) support is full cross-linked, can be one or more in physical crosslinking, chemical crosslinking, then dissolve the network structure (i.e. core arrangement 3) in support, obtain the directed porosity support containing bionical class blood vessel access network structure 2.After sterilizing, can should use containing internal channel network, directive construction tissue engineering bracket.
Solubility inner core material used should have good solubility, biocompatibility and mechanical property, can be the complex of one or more materials of saccharide compound, gelatin and alkali soluble light-sensitive polymer.
Speed forming method is shaped, and can be the one in the techniques such as continuous expressing technique, stereolithography processes, laser micro-cladding sintering.
Injection molding is shaped, by preparation containing the PDMS mould of channel network configuration, and liquid inner core material of casting wherein, by cooling or photopolymerization curing, the solubility inner core of shaping multiple-limb form.
Embodiment 1, biomimetic features tissue engineering bracket are used for rat heart muscle defect repair
Gelatin (Gelatin, 15000 ~ 250000Da) is dissolved in PBS solution, is prepared into the gelatin solution that mass fraction is 0.5%; Sodium alginate is dissolved in PBS solution, is prepared into the sodium alginate soln that mass fraction is 3.0%; Above-mentioned two kinds of solution are prepared into evenly, bubble-free, without precipitation mixed solution, the mass ratio of gelatin and sodium alginate is 1:1; By PLGA, (viscosity is 0.93dl/g, CHCl for PLA:PGA=75:25, mol ratio
3/ 25 ° of C) be dissolved in chloroform, compound concentration is the PLGA solution of 2mg/mL.Sucrose, maltose, glucose are pressed the mass ratio Homogeneous phase mixing of 1:12:2, insert build melt extrude in the shower nozzle material chamber of equipment based on vapour-pressure type, and at 130 DEG C heat fused, extruded continuously by shower nozzle and partly solidify sugar material, according to the solubility inner core (as Suo Shi Fig. 3 (a)) of the structure designed in advance and path construction multiple-limb form.Inner core is immersed in airtight preservation 20min in PLGA solution (2mg/mL) after being shaped.Inner core is taken out from PLGA solution, air-dry 12 hours, makes surface protective film drying kinetics, obtain stable multiple-limb shape inner core.Inner core is fixed on the longitudinal direction of directed bracket mould, again " gelatin-sodium alginate " mixed solution is filled the die cavity of mould, mould is placed on the orientation temperature field of 0 ~-80 DEG C, and (length direction along mould changes, the temperature of mould upper end (hot junction) is 0 DEG C, the temperature of lower end (cold junction) is-80 DEG C, support longitudinal length is 10mm, thermograde is 8K/mm) in, realize crystallographic orientation and the Thermal inactive of solution, then form the tissue engineering bracket containing inner core with directive construction through lyophilization.Support is full cross-linked in calcium chloride solution, then in PBS solution, soak a few hours, dissolve the solubility inner core of multiple-limb shape, obtain bionical class blood vessel access network.
In the biomimetic features tissue engineering bracket of above-mentioned preparation, the diameter of section of directed macropore is 50 ~ 200 μm, and the degree of orientation (OI) value is 0.9 ~ 0.95; The diameter of section of bionical class blood vessel access network structure 2 is 200 ~ 1000 μm, has 3 grades of branched structures.
In the biomimetic features tissue engineering bracket of above-mentioned preparation, as shown in Figure 4, as can be seen from this figure, channel design form trait is better for the cross section electromicroscopic photograph of bionical class blood vessel access network 2.
The directed macropore of the micropore structure of the biomimetic features tissue engineering bracket of above-mentioned preparation and the electromicroscopic photograph of horizontal aperture are respectively Ru shown in (Fig. 5 (a)) and (Fig. 5 (b)), as can be seen from this figure, directional pore structure has the higher degree of orientation, simultaneously through by horizontal fine pore.
Extract neonatal rat myocyte and endotheliocyte as seed cell, put into 37 DEG C of cell culture incubators and hatch respectively, make it completely adherent, and in addition external static culture (37 DEG C, 5%CO
2).Then by external " perfusion-perfusion " combined type culture bioreactors, initial stage is at directed rack body position plantation myocardial cell and endotheliocyte mixture, modify endothelial cell growth factor (ECGF) in channel interior and pour into endotheliocyte, later stage realizes the dynamic cultivation of simulated body fluid circulation, and applies certain mechanics training (strain 5 ~ 10%) and electricity irritation (electric field intensity 5 ~ 8V/cm).After 7 days, obtain the bulk cardiac muscle substitute with class cardiac muscular tissue feature in vitro.
The organizational project of above-mentioned acquisition cardiac muscle is implanted rat heart infarcted region, detects visible defect after two months and occurred newborn cardiac muscular tissue, and have the feature of local vascular.
Embodiment 2, containing internal channel, directive construction biomimetic scaffolds be used for three-dimensional skeletal muscle tissue external structure
Preparation 1wt% acetum.Mus tail type i collagen (SIGMA company, production code member C7661) is dissolved in acetum, is prepared into the collagen solution that mass fraction is 1.0%; Chitosan is dissolved in acetum, is prepared into the chitosan solution that mass fraction is 1.0%; Above-mentioned two kinds of solution are prepared into evenly, bubble-free, without precipitation mixed solution, the mass ratio of collagen and chitosan is 1:5.PDLGA is dissolved in chloroform, the PDLGA solution of compound concentration 0.4mg/mL.Preparation is containing the PDMS mould of channel design, and as shown in Figure 2, gelatin of casting wherein, builds the gelatin network structure of multiple-limb form.After inner core is shaped, gelatin network structure is immersed in airtight preservation 10min in PDLGA solution.Inner core is taken out from PDLGA solution, air-dry 12 hours, makes surface protective film drying kinetics, obtain stable multiple-limb shape inner core.Inner core is fixed on the longitudinal direction of directed bracket mould, again " collagen-chitin " mixed solution is filled the die cavity of directed bracket mould, mould is placed on the orientation temperature field of 0 ~-20 DEG C, and (length direction along mould changes, the temperature of mould upper end (hot junction) is 0 DEG C, the temperature of lower end (cold junction) is-20 DEG C, support longitudinal length is 10mm, thermograde is 2K/mm) in, realize crystallographic orientation and the Thermal inactive of solution, then form the tissue engineering bracket containing inner core with directive construction through lyophilization.Support is full cross-linked in the PBS solution of 0.5wt% sodium polyphosphate, be then placed in 37 DEG C of baking oven a few hours, make gelatin inner core that " gel-sol " occur and change stripping, obtain bionical class blood vessel access network.
In the biomimetic features tissue engineering bracket of above-mentioned preparation, the diameter of section of directed macropore is 100 ~ 250 μm, and the degree of orientation (OI) value is 0.8 ~ 0.9; The diameter of section of bionical class blood vessel access network 2 is 1000 ~ 3000 μm, has 2 grades of branched structures.
After sterilizing, with Matrigel drug delivery medical device surface, support directive construction position plantation SD newborn rat sarcoplast, and apply cyclic tension stimulation (strain 10%), cultivate after 7 days, observing sarcoplast under scanning electron microscope is spindle shape, arrangement regulation, cell major axis is parallel to each other; Cultivate after 21 days, the expression of Myogenin, Desmin gene detected, prove it comparatively close to the biological characteristics at body muscular tissue, prove that the present invention contains internal channel, directive construction biomimetic scaffolds for building skeletal muscle tissue substitute, and will build vascularization from now on further, the muscular tissue of neuralization provides probability.
Embodiment 3, biomimetic features tissue engineering bracket are used for the reparation of peripheral nerve defect in rats
Preparation 2wt% acetum.Fibroin albumen (silkfiber) is dissolved in acetum, is prepared into the silk fibroin protein solution that mass fraction is 0.4%; Chitosan is dissolved in acetum, is prepared into the chitosan solution that mass fraction is 2.0%; Above-mentioned two kinds of solution are prepared into evenly, bubble-free, without precipitation mixed solution, the mass ratio of fibroin albumen and chitosan is 1:1.By the alkali soluble light-sensitive polymer (copolymer of dimethyl-acrylamide, methacrylic acid and polyvinyl pyrrolidone, the mass ratio of three is 11:11:3), according to the structure designed in advance and path (as shown in Fig. 3 (b), be shaped (Stereolithography) by stereolithography processes, build the alkali solubility network structure of multiple-limb form.Network structure is fixed on the longitudinal direction of directed bracket mould, again " fibroin-chitosan " mixed solution is filled the die cavity of directed bracket mould, mould is placed on the orientation temperature field of 0 ~-196 DEG C, and (length direction along mould changes, the temperature of mould upper end (hot junction) is 0 DEG C, the temperature of lower end (cold junction) is-196 DEG C, support longitudinal length is 10mm, thermograde is 20K/mm) in, realize crystallographic orientation and the Thermal inactive of solution, then form the tissue engineering bracket containing inner core with directive construction through lyophilization.Support is full cross-linked in sodium polyphosphate, then in NaOH water liquid, dissolve multiple-limb shape core arrangement, obtain bionical class blood vessel access network.
In the biomimetic features tissue engineering bracket of above-mentioned preparation, the diameter of section of directed macropore is 20 ~ 100 μm, and the degree of orientation (OI) value is 0.92 ~ 0.95; The diameter of section of bionical class blood vessel access network 2 is 50 ~ 1500 μm, has 4 grades of branched structures.
After sterilizing, with Matrigel drug delivery medical device surface, by its bridge joint SD rat sciatic nerve 10mm defect.12 weeks after operation observes lower limb exercise, and by transmission electron microscope and the visible a large amount of regenerated nervous fibers of immunofluorescence dyeing, electro physiology detection nerve conduction velocity and wave amplitude are close to nerve autograft.
Claims (10)
1. contain a bionical tissue engineering bracket for internal channel network and directional pore structure, it is characterized in that: described bionical tissue engineering bracket comprises rack body and is arranged at the channel network in described rack body;
Described rack body has directed micropore structure, and realizes mutually through by horizontal aperture between the directed macropore of described directed micropore structure;
Described channel network has classification multiple branching construction;
Mutually through between the branch of described channel network;
Described rack body is made up of degradable natural macromolecular material.
2. bionical tissue engineering bracket according to claim 1, is characterized in that: for directly through between the branch of described channel network.
3. bionical tissue engineering bracket according to claim 1, is characterized in that: the hole by described rack body between the branch of described channel network is mutually through.
4. the bionical tissue engineering bracket according to any one of claim 1-3, is characterized in that: the diameter of section of described directed macropore is 20 μm ~ 250 μm, and the degree of orientation is 0.8 ~ 1;
The diameter of section of the branch of described channel network is 50 μm ~ 3000 μm;
Described channel network has 1 ~ 4 grade of branched structure.
5. the bionical tissue engineering bracket according to any one of claim 1-4, is characterized in that: described degradable natural macromolecular material is at least one in collagen, chitosan, sodium alginate, gelatin, fibroin albumen, hyaluronic acid, Fibrinogen and albumin.
6. the preparation method of bionical tissue engineering bracket according to any one of claim 1-5, comprises the steps:
1) by speed forming method or injection molding, by inner core material molding, the core arrangement with described channel network configuration is obtained;
2) described core arrangement is fixed in directed bracket mould, then the solution of described degradable natural macromolecular material is filled the die cavity of described directed bracket mould; Described directed bracket mould is placed in the orientation temperature field lower than 0 DEG C, then obtains containing core arrangement, the tissue engineering bracket with directive construction through lyophilization;
3) removal step 2) described core arrangement in the described tissue engineering bracket that obtains, namely obtain the bionical tissue engineering bracket containing internal channel network and directional pore structure.
7. preparation method according to claim 6, is characterized in that: step 1) in, described inner core material is at least one in saccharide compound, gelatin and alkali soluble light-sensitive polymer.
8. preparation method according to claim 7, is characterized in that: described inner core material be described saccharide compound and/or gelatin time, step 2) before, described method also comprises the step described core arrangement being placed in PLGA solution;
Step 2) in, the temperature of described orientation temperature field changes along the length direction of described directed bracket mould;
The temperature in the hot junction of described orientation temperature field is 0 DEG C, and the temperature of cold junction is-20 DEG C ~-196 DEG C, and the value of thermograde is 2 ~ 20K/mm;
Step 3) in, before removing described core arrangement, described method also comprises the step utilizing the mode of physical crosslinking and/or chemical crosslinking to stablize described tissue engineering bracket.
9. the application of bionical tissue engineering bracket according to any one of claim 1-5 in preparation tissue substituent, drug screening and case mode research.
10. application according to claim 9, is characterized in that: described tissue substituent is myocardium substitute, skeletal muscle substitute or neural substitute.
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