CN107411844A - Lumen organization's construct and preparation method thereof, preparation facilities - Google Patents

Abstract

The present invention relates to a kind of lumen organization's construct and preparation method thereof, preparation facilities, the preparation method of wherein lumen organization's construct includes：Inwall (5) and outer wall (10) for preparing lumen organization's construct is provided, wherein, the inwall (5) is the biological construct comprising bioactive substance, and the outer wall (10) is tubulose；The outer wall (10) is fixed together with the inwall (5) by suit mode.Inner and outer wall is provided separately in the present invention, and the equal tubular structure of outer wall, inwall is the biological construct comprising bioactive substance, inwall and outer wall can be fixed together by way of suit, so as to form lumen organization's construct, this assemble method step is simple, and packaging efficiency is high, suitable for batch production.

Description

Lumen organization's construct and preparation method thereof, preparation facilities

Technical field

The present invention relates to field of tissue engineering technology, more particularly to a kind of lumen organization's construct and preparation method thereof, preparation dress Put.

Background technology

Blood vessel grafting can be used for the blood vessel of narrow, inaccessible, expansion, damage or deformity is rebuild or repaired.It is common Blood vessel graft source be autologous patient artery or vein, still, autologous patient vascular supply deficiency situation Under (such as patient with vascular disease or previously had been carried out blood vessel grafting), it is necessary to be made using artificial blood vessel or heterologous blood vessel For substitute.

Existing artificial blood vessel is made up of polymer fiber (such as nylon, terylene), silk or expanded PTFE. When carrying out vasotransplantation, blood vessel of problems is rebuild using complete artificial blood vessel, or also utilize sheet Or the artificial blood vessel such as bulk.Problematic blood vessel is repaired.Although using this artificial blood vessel to lesion or impaired blood Pipe, which is replaced or repaired, has clinically obtained huge effect, but it still faces insoluble problem, when being included in long Between be implanted into after thrombus again occur and tube chamber ISR appearance.The basic reason of these problems is caused to be, it is this artificial The inwall of blood vessel lacks complete endothelial layer.

Have lot of experiments at present to attempt to solve the above problems, correlation technique includes：Adhere in artificial blood vessel's inwall Inducible factor, attract adhesion, differentiation and the growth of the stem cell (such as endothelial progenitor cells) in blood；Applied in artificial blood vessel's inwall Biomaterial is smeared, promotes to be planted in the differentiation of stem cell or the adhesion of adult cell and growth thereon.But to being at present Only, these technologies can not be realized all the time forms complete endothelial layer in artificial blood vessel's inwall, is attached to artificial blood vessel's inwall Cell it is easy to fall off, it is difficult to normal differentiation and survival, do not possess preferably biological function, may influence vasotransplantation into Using effect after power and transplanting, thus be difficult to meet clinical demand.

Therefore, it is necessary to prepare a kind of new artificial blood vessel, to reduce thrombus, doped calcium, narrow or infection and biology Learn the problems such as function is not ideal.

It should be noted that the information for being disclosed in background of invention part is merely intended to totality of the increase to the present invention The understanding of background, and be not construed as recognizing or imply the information structure in any form by those skilled in the art public affairs The prior art known.

The content of the invention

The purpose of the present invention is to propose to a kind of lumen organization's construct and preparation method thereof, preparation facilities, to form one kind New lumen organization's construct.Further, additionally it is possible to improve preparation efficiency.

To achieve the above object, first aspect present invention provides a kind of preparation method of lumen organization's construct, including：

Inner and outer wall for preparing lumen organization's construct is provided, wherein, the inwall is to include biological active matter The biological construct of matter, the outer wall are tubulose；

The outer wall is fixed together with the inwall by suit mode.

Further, the outer wall and the inwall are fixed together by suit mode and operated including suit, The suit operation includes：

First the inwall is fixed, the outer wall is then moved and the outer wall is sleeved on the inner wall outside；Or

First the outer wall is fixed, the inwall is then moved and the inner wall sleeve is mounted in the inside outer wall；Or

The outer wall and the inwall move simultaneously, so that the outer wall is set with the inner wall outside.

Further, the outer wall and the inwall are fixed together by suit mode be additionally included in it is described Adhesive coating operation is carried out before suit operation：It is viscous in the outer surface coating of the inner surface and/or the inwall of the outer wall Mixture, to make the outer wall and the inwall inter-adhesive by described adhesive after the suit operation.

Further, described adhesive coating operation also includes：It is big according to the gap between the outer wall and the inwall The coating thickness of small adjustment described adhesive.

Further, also include before the suit operation：

Adjust the axial direction between the outer wall and the inwall and/or circumferential distance.

Further, the outer wall and/or the inwall are formed by the printing of 3D biometric prints machine.

Further, the biological construct is alimentary canal lumen organization construct, respiratory tract lumen organization construct, leaching Hand shaft lumen organization construct or vessel lumen tissue construct.

Further, in addition to：There is provided for before preparing the inner and outer wall of lumen organization's construct, using biofacies Capacitive material prepares the outer wall.

Further, the tubular structure that the inwall and/or the outer wall are sidewall opening or side wall is closed.

To achieve the above object, second aspect of the present invention provides a kind of preparation facilities of lumen organization's construct, including First supporting construction, the second supporting construction and bushing device, first supporting construction and second supporting construction are used respectively It is used for the outer wall and inwall for preparing lumen organization's construct in carrying, the inwall is the biology structure comprising bioactive substance Body, the outer wall are tubulose, and the bushing device is used to the outer wall is set with and is fixed together with the inwall.

Further, first supporting construction includes support column, and the outer wall is sheathed on the periphery of the support column；Or Person, first supporting construction include support set, and the support set is sheathed on the periphery of the outer wall.

Further, second supporting construction includes fixture, and the fixture is used for the support of inwall described in clamp standoff Part, the inwall are arranged at the periphery of the support member；Or the supporting construction includes support bar, the inwall is set It is placed in the periphery of the support bar.

Further, the support member is the rotation for supporting inwall when 3D biometric prints machine prints the inwall Bull stick.

Further, the bushing device includes drive mechanism, and the drive mechanism is used to drive the outer wall and/or institute Inwall is stated, so that with the inwall relative motion occurs for the outer wall, realizes the suit of the outer wall and the inwall.

Further, the bushing device also includes resetting-mechanism, and the resetting-mechanism is connected with the drive mechanism, and For resetting the drive mechanism.

Further, the bushing device also includes coating unit, and the coating unit is used for the interior table in the outer wall Adhesive is smeared in face and/or the outer surface of the inwall, so that the outer wall after inwall suit with passing through described adhesive It is inter-adhesive.

Further, the coating thickness of described adhesive is adjustable.

Further, the coating unit includes container, transfer passage and adhesive outlet, and the container is used to hold institute State adhesive, the transfer passage is connected with the container and is used to convey described adhesive, described adhesive outlet with it is described Transfer passage is connected so that described adhesive is coated on the outer wall or the inwall.

Further, the transfer passage is arranged on inside the first supporting construction；And/or described adhesive outlet is to set The aperture connected with the transfer passage being placed in the first supporting construction.

Further, the transfer passage includes delivery pipe and shower nozzle, and the delivery pipe is connected with the shower nozzle, the spray Head spout as described adhesive outlet, with by the shower nozzle by described adhesive be sprayed on the outer wall inner surface or On the outer surface of the inwall.

Further, the preparation facilities also includes positioner, and the positioner is used for by the outer wall and institute The outer wall and/or the inwall are positioned before stating inwall suit.

Further, the positioner can adjust at least one relative to another in the outer wall and the inwall Individual position.

Further, the outer wall and the inwall are coaxial, the positioner can adjust the outer wall with it is described interior Axial distance between wall.

Further, the positioner includes support base and the support being connected with first supporting construction, the branch Frame can move relative to the support base, and the support is provided with rack, and the support base is provided with mutual with the rack Meshed gears, to drive the rack to move by the rotation of the gear, the first supporting construction described in the rack drives Motion, so that the outer wall relatively moves relative to the inwall.

Further, the positioner also includes guiding mechanism, and the guiding mechanism is arranged on the support base, institute The first supporting construction is stated on the guiding mechanism, so that first supporting construction being capable of drawing in the guiding mechanism Lead lower motion.

Further, the guiding mechanism includes slide rail and sliding block, and the sliding block is slidably matched with the slide rail, the cunning Rail is arranged on the support base, and first supporting construction is connected with the sliding block.

Further, material is biocompatible materials used by the supporting part of first supporting construction.

To achieve the above object, third aspect present invention provides a kind of lumen organization's construct, lumen organization's structure Body is built to be prepared using the preparation method of above-mentioned lumen organization's construct.

Further, the outer wall is biocompatible materials.

Further, the outer wall is Biodegradable material or biological non-degradable material.

Further, the material of the outer wall is nylon, terylene, silk, polytetrafluoroethylene (PTFE) or the lumen organization of animal.

Further, the inwall includes cell.

Further, lumen organization's construct is alimentary canal lumen organization construct, respiratory tract lumen organization structure Body, lymphatic vessel lumen organization construct or vessel lumen tissue construct.

Based on above-mentioned technical proposal, inner and outer wall is provided separately in the present invention, and outer wall is tubular structure, and inwall is bag Inwall and outer wall, are then fixed together by the biological construct of matters of containing biological activities by way of suit, so as to Lumen organization's construct is formed, this assemble method step is simple, and packaging efficiency is high, suitable for batch production.

Brief description of the drawings

Accompanying drawing described herein is used for providing a further understanding of the present invention, forms the part of the application, this hair Bright schematic description and description is used to explain the present invention, does not form inappropriate limitation of the present invention.In the accompanying drawings：

Fig. 1 is the structural representation of preparation facilities one embodiment of lumen organization's construct of the present invention.

Fig. 2 is the left view of Fig. 1 embodiments.

Fig. 3 be lumen organization's construct of the present invention preparation facilities one embodiment in transfer passage structural representation.

Fig. 4 is the enlarged drawing of part shown in label P in Fig. 3.

Fig. 5 is structural representation of Fig. 1 embodiments in the first motion state.

Fig. 6 is structural representation of Fig. 1 embodiments in the second motion state.

Fig. 7 is structural representation of Fig. 1 embodiments in the 3rd motion state.

Fig. 8 is structural representation of Fig. 1 embodiments in the 4th motion state.

Fig. 9 is the structural representation of another embodiment of the preparation facilities of lumen organization's construct of the present invention.

In figure：1- bases, 2- installing plates, 3- fixtures, 4- swingles, 5- inwalls, 6- slide rails, 7- sliding blocks, 8- supports, 9- branch Dagger, 10- outer walls, 11- push rods, 12- springs, 13- racks, 14- gears, 15- handwheels, 16- containers, 17- transfer passages, 18- Aperture.

Embodiment

Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in embodiment is clearly and completely retouched State.Obviously, described embodiment is only the part of the embodiment of the present invention, rather than whole embodiments.Based on this hair Bright embodiment, the every other implementation that those of ordinary skill in the art are obtained under the premise of creative work is not made Example, belongs to the scope of protection of the invention.

In the description of the invention, it is to be understood that term " " center ", " transverse direction ", " longitudinal direction ", "front", "rear", The orientation or position relationship of the instruction such as "left", "right", " on ", " under ", " vertical ", " level ", " top ", " bottom ", " interior ", " outer " are Based on orientation shown in the drawings or position relationship, it is for only for ease of the description present invention and simplifies description, rather than instruction or dark Show that the device of meaning or element there must be specific orientation, with specific azimuth configuration and operation, thus it is it is not intended that right The limitation of the scope of the present invention.

In the present invention, unless otherwise stated, Science and Technology noun used herein has art technology The implication that personnel are generally understood that.For a better understanding of the present invention, the definition and explanation of relational language is provided below.

As used in this specification and the appended claims, singulative "one", " one kind " and " should/described " bag The indicant of plural number is included, unless the context clearly determines otherwise.In addition, herein any "or" referred to be intended to include " and/ Or ", unless otherwise indicated.

As used in this article, term " tissue " refers to be made up of homomorphosis or similar, function identical cell mass Cell aggregate, and the material (being referred to as cytoplasm, such as matrix, fiber etc.) generally also comprising acellular form.Tissue It may include one or more cells.

As used herein, term " micro-capsule " refers to, the micro-structural (example containing cell and biocompatible materials Such as, micron order is to millimetre-sized structure), wherein, cell is wrapped in the biocompatible materials.The micro-capsule of the present invention exists (such as 4-37 DEG C, such as pH is between 6-8, such as under the hydrodynamic shear of physiological environment) has stable under physiological environment Structure.Preferably, micro-capsule has the mechanical strength that will not cause micro-capsule broken in absorption or extruding.

In the present invention, term " biological construct " refers to the object built using the micro-capsule of the present invention, and it can have Two dimension or three-dimensional structure, can be used for preparing artificial organ precursor.

As used in this article, term " tube chamber " refers to be shaped as tubulose, has the organ of hollow cavity, such as circulation pipe Chamber, digestive tract cavity, breathing tube chamber, uropoiesis tube chamber or reproduction tube chamber, such as blood vessel, oesophagus, tracheae, stomach, bile duct, enteron aisle (including Small intestine and large intestine, such as duodenum, jejunum, ileum, caecum (including appendix), the colon ascendens, flexura coil dextra, transverse colon, colon Zuo Qu, colon descendens, sigmoid colon, rectum), fallopian tubal, vas deferens, ureter, bladder or lymphatic vessel).

As used herein, term " biocompatible materials " refers to such material, its (and its degraded production Thing) be avirulent for cell, and in implantation host (such as human body) afterwards and host compatibility, will not cause it is significant or Serious side effect, for example, toxic action will not be caused to host (such as tissue), the immunological rejection of host will not be caused Reaction, allergic reaction or inflammatory reaction etc..

As used herein, term " Biodegradable material " refers to such material, and it can be by cell or life Object is degraded and absorbed, and its catabolite is biocompatibility.Such material can be natural origin (such as source In animals and plants) or it is artificial synthesized.

As used in this article, term " biometric print " refers to：(included but is not limited to, biomolecule using biomaterial Such as protein, lipid, nucleic acid and metabolite；Cell such as cell solution, celliferous gel, cell suspending liquid, cell Concentrate, many cells aggregation and multicell；Subcellular structure such as organelle and cell membrane；Point related to biomolecule The sub biomolecule of such as synthesis or the analog of biomolecule) printing.As used in this article, term " printing " refers to, The process of deposition materials in predetermined patterns.In the present invention, biometric print preferably by with it is automatic or automanual, Method that computer assisted three-dimensional prototype device (such as biometric print machine) matches is realized.However, in the present invention, " printing " (such as biometric print) can be carried out by various methods, be included but is not limited to, and use printer (such as 3D printer Or biometric print machine) printed；Printed using automation or non-automated mechanical process (rather than printer)；Pass through hand Work is placed or manual deposition (such as using pipettor) is printed.

In order to improve efficiency prepared by lumen organization's construct, present invention firstly provides a kind of system of lumen organization's construct Preparation Method.

In a preferred embodiment, the preparation method of lumen organization's construct includes：

Inwall 5 and outer wall 10 for preparing lumen organization's construct is provided, wherein, inwall 5 is to include biological active matter The biological construct of matter, outer wall 10 are tubulose；

Outer wall 10 is fixed together with inwall 5 by suit mode.

Wherein, inwall 5 is the biological construct comprising bioactive substance, i.e. inwall 5 possesses bioactivity, can be with Formed, can also made manually by the printing of 3D biometric prints machine.

Biological construct can be alimentary canal lumen organization construct, respiratory tract lumen organization construct, lymphatic vessel tube chamber Tissue construct or vessel lumen tissue construct.

Inwall 5 can be passed through arrangement form, micro-capsule shape by cell encapsulation bioactive substance by one or more micro-capsules Into.The generation type of inwall 5 can have a variety of, for example can be shaped as tubulose or column (such as the circle that side wall is not open Tubulose, sidewall opening round tubular, cylindric or the column set along part-circular periphery) interim holder predeterminable area on It is bonded, can also be formed on the print platform with curved surface by the printing of 3D biometric prints machine, can also be in the pre- of plane If print and then roll on region as tubular structure, etc..

Outer wall 10 is tubulose, and outer wall 10 can be biological tissue wall, such as lumen organization's wall of animal etc.；Outer wall 10 also may be used Think abiotic tissue wall, i.e. outer wall 10 is that do not possess bioactivity, usually, can use polymer fiber (such as Buddhist nun Dragon, terylene), the material such as silk or expanded PTFE is made.

Outer wall 10 can be as the holder of inwall 5, and outer wall 10 can be the nondegradable material of biology or life The material of Biodegradable.For nondegradable material, after inwall 5 grows up to target tubular tissue, outer wall 10 can be still So exist；For degradation material, after inwall 5 grows up to target tubular tissue, its outer wall 10 eventually gradually decomposes, Disappear, that is, obtain complete biological tubular tissue.

Preferably, tubulose biological construct is prepared to carry out by the method comprised the steps of：

(1) one or more micro-capsules are provided, its all or part of surface attachment has the first component；Preferably, described One component is contained in the first reagent；

(2) second examination of the coating containing the second component on the predeterminable area of the support member surfaces for supporting inwall 5 Agent, wherein, when first component contacts with the second component, adhesion effect can be produced, realizes adhesive effect；It is described to be used for The support member of support inwall 5 for tubulose or column (such as side wall be not open it is round tubular, sidewall opening it is round tubular, Column that is cylindric or being set along part-circular periphery) thing, song of the predeterminable area positioned at the support member for being used to support inwall 5 Face；Optionally, before the second reagent is coated with, backing material is coated on to the pre- of support member surfaces for supporting inwall 5 If on region；

(3) micro-capsule that all or part of surface attachment in step (1) has the first component is positioned over and is coated with second The predeterminable area of reagent, the first component on the micro-capsule surface is contacted with the second component on predeterminable area, produce adhesion Effect, so as to which into the first Rotating fields, first Rotating fields are tubular structure by micro-capsule assembling (bonding)；

Optionally, methods described is further comprising the steps of：

(4) the second reagent is being coated with caused by previous step in structure；

(5) micro-capsule that all or part of surface attachment in step (1) has the first component is positioned over previous step production In raw structure, the first component on the micro-capsule surface is contacted with the second component caused by previous step in structure, produce Raw adhesion effect, so as to which in structure, the micro-capsule is being assembled into (bonding) into another Rotating fields caused by previous step；

(6) optionally, repeat step (4) and (5) are one or many；For example, at least 1 time, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 10 times, at least 15 times, at least 20 times, at least 30 times, at least 40 times, at least 50 times, at least 100 times, At least 200 times, at least 500 times, or more time；

So as to obtain tubulose biological construct.

Optionally, methods described also includes：The round tubular biological construct of sidewall opening is bonded, obtains side wall The round tubular biological construct not being open.

Optionally, methods described also includes：Tubulose biological construct is divided from the support member for supporting inwall 5 From.

Preferably, the support member for being used to support inwall 5 is the print platform with curved surface, such as 3D printer Swingle.

Preferably, the backing material is Thermo-sensitive material, such as gelatin, poly-N-isopropyl acrylamide, poly- N- isopropyls Base acrylamide-polyethyleneglycol block copolymer, ethylene glycol copolymer (such as Kollicoat IR), gather Hydroxy ethyl methacrylate, agarose, Matrigel, chitosan/sodium glycero-phosphate system or Pluronic F127.

Preferably, it is described to be used to supporting the support member of inwall 5 for Thermo-sensitive material (such as gelatin, poly-N-isopropyl third Acrylamide, poly-N-isopropyl acrylamide-polyethyleneglycol block copolymer, ethylene glycol copolymer, poly-hydroxyethyl acrylic acid Ester, agarose, Matrigel, chitosan/sodium glycero-phosphate system or Pluronic F127) made of cylinder or pipe Shape thing.

In certain preferred aspects, the support member for supporting inwall 5 is cylinder.

In certain preferred aspects, the support member for supporting inwall 5 is cylinder, described default Region is the whole side of cylinder, so as to which the first Rotating fields that step (3) obtains are the round tubular knot that side wall is not open Structure.

In certain preferred aspects, the support member for supporting inwall 5 is cylinder, described default Region is a rectangle on the side of the cylinder of expansion, and the predeterminable area along cylinder axially through cylinder The side of shape thing, so as to which the first Rotating fields that step (3) obtains are the round tubular structure that side wall is not open.

In certain preferred aspects, the support member for supporting inwall 5 is cylinder, described default Region is a rectangle on the side of the cylinder of expansion, and radial direction of the predeterminable area along cylinder penetrates column Thing side, so as to which the first Rotating fields that step (3) obtains are the round tubular structure that side wall is not open.

In certain preferred aspects, the support member for supporting inwall 5 is cylinder, described default Region is a rectangle on the side of the cylinder of expansion, and not radially or axially through cylinder side, so that, The first Rotating fields that step (3) obtains are the round tubular structure of sidewall opening.

Preferably, in step (3), step (2) is positioned in the micro-capsule that all or part of surface attachment is had into the first component Be coated with after the predeterminable area of the second reagent, stand 0.1-60s；(such as 0.1-1s, 1-5s, 5-10s, 10-15s, 15- 20s, 20-25s, 25-30s, 30-35s, 35-40s, 40-45s, 45-50s, 50-55s, or 55-60s).The standing step is favourable Fully contact, and interact with the second component on predeterminable area in the first component on the micro-capsule surface, so as to The micro-capsule is assembled into (bonding) into the first Rotating fields.

Preferably, the method for preparing tubulose biological construct is implemented by biometric print method.

Preferably, biometric print method is carried out using printer (such as 3D biometric prints machine)；Or using automation or Non-automated mechanical process carries out biometric print method；Or by using placing by hand or manual deposition method (such as uses shifting Liquid device) carry out biometric print method.

The method of artificial organ precursor is prepared in the present invention, it is preferable that first component and/or the second component are Biocompatible materials, and/or to be Biodegradable material from the material of biology.

In certain preferred aspects, first component contacts with the second component and caused adhesion effect can use It is bonded together in by two micro-capsules, forms construct；And the stretch modulus of the construct thus obtained is not less than 10Pa, Such as not less than 20Pa, not less than 30Pa, not less than 40Pa, not less than 50Pa, not less than 60Pa, not less than 70Pa, it is not less than 80Pa, not less than 90Pa, not less than 100Pa, not less than 200Pa, not less than 300Pa, not less than 400Pa, not less than 500Pa, Not less than 600Pa, not less than 700Pa, not less than 800Pa, not less than 900Pa, not less than 1000Pa.

Preferably, first component and the second component are to be selected from following combination：

(1) fibrinogen and fibrin ferment；

(2) alginate (such as sodium alginate) or oxidation alginate (such as sodium alginate of oxidation) and contain Ca2+, Mg2+, Ba2+, Sr2+ or Fe3+ material (such as solution containing Ca2+, Mg2+, Ba2+, Sr2+ or Fe3+ or half Solid (such as gel))；

(3) molecule containing maleimide base group (such as polyethylene glycol (MAL-PEG) containing maleimide base group) With the molecule (such as polyethylene glycol (PEG-SH) containing free sulfhydryl group) containing free sulfhydryl group；

(4) material containing anion (such as solution or semisolid (such as gel) containing anion) and alpha-cyano third Olefin(e) acid ester (such as Mecrilate, α-cyanoacrylate, isobutyl alpha-cyanoacrylate, alpha-cyanoacrylate Dissident's ester, α-n-octylcyanoacrylate)；

(5) fibrinogen and a-cyanoacrylate (such as Mecrilate, α-cyanoacrylate, Isobutyl alpha-cyanoacrylate, alpha-cyanoacrylate dissident's ester, α-n-octylcyanoacrylate)；

(6) seralbumin (for example, bovine serum albumin(BSA)) and glutaraldehyde；

(7) molecule of acid esters containing urethane groups (- NHCOO-) or isocyanate groups (- NCO) is (such as containing carbamic acid The polyethylene glycol of ester group or the polyethylene glycol containing isocyanate groups) and molecule (such as carboxylic poly- second containing active hydrogen Glycol)；

(8) gelatin-resorcinol and glutaraldehyde；

(9) carbodiimides cross-linked gelatin and L-glutamic acid (PLGA)；With

(10) aminated gelatin and aldehyde radical polysaccharide.

The size of the micro-capsule of the present invention can be selected according to being actually needed, and be not particularly limited.Spherical vesicles Size can generally be explicitly defined by its diameter.In the case of strict difinition, term " diameter " cannot be used for retouching State aspherical structure.However, in the present invention, the size of aspherical micro-capsule also is described using term " diameter ".Herein In the case of, term " diameter " represents, has the diameter of the spherical vesicles of same volume with aspherical micro-capsule.In other words, at this In invention, using the diameter of spherical vesicles come the size of aspherical micro-capsule that describes there is same volume.Therefore, some excellent In the embodiment of choosing, the size (that is, diameter defined herein) of micro-capsule of the present invention can be 20-2000 μm, such as 30- 1900 μm, 40-1800 μm, 50-1700 μm, 60-1600 μm, 70-1500 μm, 80-1400 μm, 90-1300 μm, 100-1200 μ M, 200-1000 μm, 300-800 μm, 400-600 μm, 100-500 μm.In certain preferred aspects, micro-capsule of the present invention Size (that is, diameter defined herein) can be 20-30,30-50,50-100,100-150,150-200,200-250, 250-300、300-350、350-400、400-450、450-500、500-600、600-700、700-800、800-900、900- 1000、1000-1500、1500-2000、20-50、20-100、100-200、200-400、500-600、600-800、800- 1000 or 1000-2000 μm.In certain preferred aspects, (that is, defined herein is straight for the size of micro-capsule of the present invention Footpath) be at least 20,30,50,100,120,150,200,250,300,350,400,450,500,600,700,800,900, 1000th, 1500 or 2000 μm.

The shape of the micro-capsule of the present invention can be selected according to being actually needed, and be not particularly limited.It is for example, of the invention Micro-capsule can be spherical or any desired shape (such as cube, rectangular prism, six prisms, cylinder, or irregular Shape).For example, some shapes (such as spherical, cube, rectangular prism, six prisms) it can be used for realizing micro-capsule in construct It is tightly packed.

In certain preferred aspects, micro-capsule of the invention is solid or semisolid.In some preferable embodiment party In case, micro-capsule of the invention is gel state.For example, the stratum nucleare and/or shell of the micro-capsule of the present invention can be gel state.Some In preferred embodiment, micro-capsule of the invention includes hydrogel.In certain preferred aspects, the hydrogel includes Alginate, agarose, gelatin, chitosan, or other water-soluble or hydrophilic polymers.

In certain preferred aspects, micro-capsule of the invention exists as a mixture.In such embodiment In, micro-capsule can be contacted or merged with another micro-capsule in mixture.In certain preferred aspects, micro-capsule of the invention It is the micro-capsule of separation.For example, in certain embodiments, micro-capsule does not contact directly with other micro-capsules.Some preferable real Apply in scheme, the micro-capsule of separation of the invention is provided in container.

Various methods can be used to prepare for the micro-capsule of the present invention.For example, in certain preferred aspects, it can be used and use The micro-capsule of the present invention is prepared in the method for manufacturing microsphere, such as is prepared using instrument is granulated.Some preferable real Apply in scheme, micro-capsule of the invention is aseptically prepared.In some preferred embodiments, micro-capsule of the invention is Prepared in GMP workplaces.In certain preferred aspects, micro-capsule of the invention is produced immediately before use. In some preferred embodiments, micro-capsule of the invention is stored in 4 DEG C after preparation, for example, storage 3 hours, 6 hours, it is 12 small When, 1 day, 2 days or 3 days.

The species for the cell that micro-capsule of the present invention includes can be selected according to being actually needed, and be not particularly limited.It is excellent Selection of land, endothelial cell (such as vascular endothelial cell), smooth muscle cell (such as vascular smooth muscle cells) are included in the micro-capsule And/or undifferentiated cell.

Preferably, the cell in the micro-capsule is undifferentiated cell, such as stem cell (such as fat mesenchymal is dry thin Born of the same parents, mesenchymal stem cells MSCs, induced multi-potent stem cell and embryonic stem cell).

Preferably, the undifferentiated cell can be divided into endothelial cell and/or smooth muscle cell.

Preferably, the undifferentiated cell is selected from stem cell (such as fat mesenchymal stem cell, medulla mesenchyma is dry thin Born of the same parents, induced multi-potent stem cell and embryonic stem cell) and progenitor cells (such as endothelial progenitor cells) in one or more.

The source for the cell that micro-capsule of the present invention includes can be selected according to being actually needed, and be not particularly limited.It is excellent Selection of land, the cell are obtained from animal, such as mammal, such as people, ape, monkey, gorilla, ox, pig, dog, sheep and goat.

Preferably, the cell derived is in selected from following tissues：Connective tissue is (for example, loose connective tissue, fine and close knot Form tissue, elastic fibrous tissue, reticular connective tissue and adipose tissue), musculature (for example, skeletal muscle, smooth muscle and cardiac muscle), secrete Germinal tissue, gastrointestinal tissue, lung tissue, bone tissue, nerve fiber and epithelial tissue are urinated (for example, on simple epithelium and cladding Skin), the tissue of endoderm origin, the tissue of the tissue of mesoderma origin and ectodermal origin.

The quantity for the cell that micro-capsule of the present invention includes can be selected according to being actually needed, and be not particularly limited.Example Such as, the stratum nucleare of micro-capsule of the present invention can include 1-10 independently of one another⁶Individual cell, such as 10-900,20-800,30-700,40- 600th, 50-500,60-400,70-300,80-200,10-100,10-10³Individual, 10-10⁴Individual, 10-10⁵Individual, 10-10⁶It is individual thin Born of the same parents.In certain preferred aspects, micro-capsule of the present invention include at least 1,2,4,6,8,10,15,20,25,30,40,50, 60、70、80、90、100、150、200、300、400、500、600、700、800、900、1000、2000、3000、4000、5000、 6000、7000、8000、9000、10⁴、2x10⁴、3x10⁴、4x10⁴、5x10⁴、6x10⁴、7x10⁴、8x10⁴、9x10⁴、10⁵、 2x10⁵、3x10⁵、4x10⁵、5x10⁵、6x10⁵、7x10⁵、8x10⁵、9x10⁵Or 10⁶Individual cell.In some preferred embodiments In, micro-capsule of the present invention includes 1-2,2-4,4-6,6-8,8-10,10-15,15-20,20-25,25-30,30-40,40-50,50- 60、60-70、70-80、80-90、90-100、100-150、150-200、200-300、300-400、400-500、500-1000、 1000-2000、2000-3000、3000-4000、4000-5000、5000-10⁴、10⁴-2x10⁴、2x10^4-3x10⁴、3x10⁴- 4x10⁴、4x10⁴-5x10⁴、5x10⁴-10⁵、10⁵-2x10⁵、2x10⁵-3x10⁵、3x10⁵-4x10⁵、4x10⁵-5x10⁵、5x10⁵- 10⁶、1-10、2-10、2-5、5-10、10-20、20-30、30-50、2-25、25-50、2-50、50-100、100-200、50- 250th, 250-500,500-2000,2-100,2-500 or 2-2000 cells.

In certain preferred aspects, except endothelial cell as described above, smooth muscle cell and/or undifferentiated Outside cell, the cell of the micro-capsule parcel also includes additional cell.In certain preferred aspects, the additional cell From selected from following tissues：Connective tissue is (for example, loose connective tissue, dense connective tissue, elastic fibrous tissue, netted knot Form tissue and adipose tissue), musculature (for example, skeletal muscle, smooth muscle and cardiac muscle), urogenital tissue, gastrointestinal tissue, lung Tissue, bone tissue, nerve fiber and epithelial tissue (for example, simple epithelium and stratified epithelium), the tissue of endoderm origin, middle embryo The layer tissue in source and the tissue of ectodermal origin.In certain preferred aspects, it is thin to be selected from muscle for the additional cell Born of the same parents (for example, Skeletal Muscle Cell, cardiac muscle cell, smooth muscle cell and sarcoblast), phoirocyte are (for example, osteocyte, soft Osteocyte, fibroblast and the cell for being divided into Gegenbaur's cell, cartilage cell or lymphoid tissue), bone marrow cell, skin it is thin Born of the same parents, epithelial cell, mammary glandular cell, vascular cell, haemocyte, lymphocyte, nerve cell, schwann cell, gastrointestinal cell, liver are thin Born of the same parents, pancreatic cell, pneumonocyte, tracheal cell, keratocyte, urogenital cell, nephrocyte, adipocyte, parenchyma, week are thin Born of the same parents, mesothelial cell, stroma cell, the cell of endoderm origin, the cell of mesoderma origin, the cell of ectodermal origin, cancer are come Cell, cell line or its any combinations in source.

Preferably, micro-capsule of the invention includes cell and wraps up the stratum nucleare of the cell.Preferably, the stratum nucleare can be The vital movement of cell provides microenvironment.In certain preferred aspects, micro-capsule provides suitable cell adherence and stretching, extension Space structure and microenvironment, so as to which cell can be normally carried out breeding in the structure, break up, migrate, secreting or new old generation Thank.The microenvironment refers to the environment that cell is grown, and its key element included includes physical factor, for example space structure, mechanics are strong Degree, temperature, humidity, osmotic pressure etc.；Chemical factor, such as acid-base value, ion concentration etc.；Biological factor, including cell, cell because Son etc..These key elements collectively form the environment of cell activities, and the propagation of the cell to growing in this environment, point Change, migrate, secretion and metabolism carry out dynamic regulation.Preferably, the stratum nucleare can provide battalion for the vital movement of cell Support material.

Preferably, it is made up of biocompatible materials the stratum nucleare.

In certain preferred aspects, the micro-capsule is also comprising the shell for encapsulating the stratum nucleare.

In certain preferred aspects, the shell of micro-capsule provides mechanics protection for the cell of parcel.Some excellent In the embodiment of choosing, the shell of the micro-capsule or micro-capsule has certain mechanical strength, so as to realize stereo stocking. In the present invention, particularly preferably, micro-capsule and its shell with appropriate mechanics protective value (for example, with suitable hardness and/ Or modulus of elasticity).On the one hand, (for example, during 3D printing) is easy to because the external world presses the cell in micro-capsule in operation The injury of power or shearing force and it is impaired or dead.Therefore, if the hardness and/or modulus of elasticity of micro-capsule and its shell are too low, that The cell survival rate in micro-capsule will be caused to be remarkably decreased after manual operation, and then cause the application of micro-capsule to be restricted, or Person needs to use substantial amounts of cell.On the other hand, if the hardness and/or modulus of elasticity of micro-capsule and its shell are too high, then will Cause the stretching, extension of the cell in micro-capsule, migration to be restricted, and hinder to establish cell connection between the cell of different micro-capsules, no Beneficial to structure organic whole (for example, artificial organ).Therefore, appropriate mechanics protective value not only makes it possible to the present invention Micro-capsule carry out various operations (such as carry out 3D biometric prints, carry out exact placement of micro-capsule etc.), and be advantageous in micro-capsule Cytochrome oxidase isozymes, migrate, establish cell connection, and form organic construct (such as artificial organ), be particularly preferred therefore 's.

In certain preferred aspects, the stratum nucleare of micro-capsule of the present invention and/or shell are each optionally past processing (example Such as handled using stratum nucleare fixer or shell fixer, for example, to improve the mechanical property of stratum nucleare or shell)

In certain preferred aspects, the shell of the micro-capsule, the stratum nucleare of micro-capsule or micro-capsule has independently of one another About 0.01,0.02,0.03,0.04,0.05,0.06,0.07,0.08,0.09,0.1,0.15,0.2,0.3 or 0.4GPa's is hard Degree.In certain preferred aspects, the shell of the shell micro-capsule of the micro-capsule or micro-capsule, the stratum nucleare of micro-capsule or micro-capsule is each Independently there is 0.01-0.02,0.02-0.03,0.03-0.04,0.04-0.05,0.05-0.06,0.06-0.07,0.07- 0.08、0.08-0.09、0.09-0.1、0.1-0.15、0.15-0.2、0.2-0.3、0.3-0.4、0.01-0.4、0.01-0.05、 0.05-0.1,0.1-0.2,0.2-0.4,0.05-0.15 or 0.06-0.1GPa hardness.In some preferred embodiments In, the shell of the micro-capsule, the stratum nucleare of micro-capsule or micro-capsule has about 0.083GPa hardness.In some preferred embodiments In, the shell of the micro-capsule, the stratum nucleare of micro-capsule or micro-capsule has about 0.01 independently of one another, 0.05,0.1,0.5,0.8,1, 1.2nd, 1.4,1.6,1.8,2,2.4,2.8,3.2,4,10,20,30,40,50,80 or 100MPa modulus of elasticity.Some excellent In the embodiment of choosing, the shell of the micro-capsule, the stratum nucleare of micro-capsule or micro-capsule has 0.01-0.05,0.05- independently of one another 0.1、0.1-0.5、0.5-0.8、0.8-1、1-1.2、1.2-1.4、1.4-1.6、1.6-1.8、1.8-2、2-2.4、2.4-2.8、 2.8-3.2、3.2-4、4-10、10-20、20-30、30-40、40-50、50-80、80-100、0.5-4、0.5-1、1-1.5、 1.5-2,2-3,0.8-1.6,1.4-2.4,0.8-3.2,0.01-100,1-100,10-100 or 0.5-50MPa springform Amount.The mechanics protective effect (for example, consistency and elasticity modulus) of stratum nucleare or shell can by the component to stratum nucleare or shell and/or The configuration of content controls.

In certain preferred aspects, the shell also can provide microenvironment for the vital movement of cell, such as Nutriment.In certain preferred aspects, the shell is made up of biocompatible materials.

In certain preferred aspects, the biocompatible materials for preparing stratum nucleare and shell can be identical It is or different.It is particularly preferred, however, that ground, according to its expected purpose, stratum nucleare and shell have different compositions.It is not limited to theory Limitation, it is generally accepted that shell provides main mechanics protective effect, and stratum nucleare then provides the master needed for cell activities The nutritional ingredient and microenvironment wanted.Therefore, in certain preferred aspects, compared with shell, stratum nucleare has more battalion Support material.In certain preferred aspects, compared with stratum nucleare, shell has relatively low degradation rate, but with higher Hardness and/or modulus of elasticity.In certain preferred aspects, cell is not included in shell.

In certain preferred aspects, stratum nucleare and shell include identical bio-compatible with different weight ratios respectively Property material.In other words, stratum nucleare and shell can be made up of identical biocompatible materials, but include life with different weight ratios Biodegradable material.

In certain preferred aspects, the shell is permeability independently of one another.For example, the shell for Water, oxygen, and nutriment (carbohydrate such as glucose, fat, protein, amino acid, small peptide, mineral matter, vitamin, cell The factor, nucleotides etc.) it is permeability.

It is generally believed that the use of semipermeable (that is, selecting penetrating) shell is probably favourable, because it is enabled to The nutriments such as water, oxygen, glucose, mineral matter, and amino acid pass through shell, into stratum nucleare, and are supplied to cell, and energy It is enough to prevent to enter stratum nucleare to the material (such as antibody protein from host immune system) that cell is harmful to.However, in the present invention Micro-capsule in, the use of permeability shell is preferable and favourable.Especially, the shell of permeability causes various nutriments (including macromolecular and small molecule nutriment, such as glucose, fat, protein, amino acid, small peptide, mineral matter, vitamin, Cell factor, nucleotides etc.) it can be more prone to, swimmingly swap, abundance can not be obtained by avoiding the cell of regional area Nutriment.For example, when building large-sized artificial organ using the micro-capsule of the present invention, the shell of permeability can promote Enter the exchange of various nutriments, promote inside artificial organ/micro-capsule of nucleus in cell obtain sufficient nutrients Matter.Connected in addition, the cell that the shell of permeability is advantageous between different micro-capsules carries out signal transmission with cell is established.Especially Ground, cell can secrete many kinds of substance (some components and multi-signal molecule that include extracellular matrix) in growth course, with Neighbouring, even distal end cell carries out signal transmission and/or material exchange, and thus the vital movement to cell itself and Neighbouring, the even cell of distal end vital movement has an impact or regulated and controled.Therefore, if shell using permselectivity If layer, then signal transmission and/or material exchange between cell would be possible to be affected/hinder, such as cell secretion Some macromolecular semiochemicalses (such as Cytokine protein) possibly can not pass through shell, so as to hinder different micro-capsules it Between cell signal transmission and cell establishment of connection, be unfavorable for build organic whole (for example, artificial organ).Therefore, The use of permeability shell is preferable for the micro-capsule of the present invention.In the present invention, state " permeability shell " it is meant that Various small molecules and macromolecular substances (such as protein) can pass freely through shell.For example, in some preferred embodiments In, the shell is penetrating in below 5000kDa molecule for molecular weight.For example, in certain embodiments, the shell Layer for molecular weight below 200kDa or molecular weight 200kDa-300kDa, 300kDa-400kDa, 400kDa-500kDa, 500kDa-800kDa、800kDa-1000kDa、1000kDa-1500kDa、1500kDa-2000kDa、2000kDa-3000kDa、 Molecule in the range of 3000kDa-4000kDa or 4000kDa-5000kDa is penetrating.In certain embodiments, the shell Layer is penetrating for immunoglobulin (such as IgG, IgM, IgA, IgD, IgE).

In certain preferred aspects, the shell has independently of one another is used for the logical of mass exchange inside and outside micro-capsule Road or hole.In certain preferred aspects, (carbohydrate such as glucose, fat, protein, amino acid are short for nutriment Peptide, mineral matter, vitamin, cell factor, nucleotides etc.) diffused into by the passage or hole in the micro-capsule.Some In preferred embodiment, the passage a diameter of at least 10,20,50,100,150,200,250,300,350,400 or 500nm.In certain preferred aspects, a diameter of such as 1nm-5 μm of the passage；10nm-2μm；100nm-1μm； 200-800nm etc..In certain preferred aspects, the hole a diameter of at least 100,200,400,600,800, 1000th, 1500,2000,4000 or 5000nm.

The thickness of the shell of the micro-capsule of the present invention can be selected according to being actually needed, and be not particularly limited.For example, The thickness of the shell of micro-capsule of the present invention can be 1-20 μm independently of one another, such as 5-15 μm, such as 8-12 μm.Some preferred Embodiment in, the thickness of the shell of micro-capsule of the invention can be about 0.1 independently of one another, 0.5,1,2,5,10,15, 20th, 25,30 or 50 μm.In certain preferred aspects, the thickness of the shell of micro-capsule of the invention independently of one another can be with For 0.1-0.5,0.5-1,1-2,2-5,5-10,10-15,15-20,20-25,25-30,30-50,50-100,100-200, 200-300,300-400,400-500,0.1-1,1-5,1-10,5-10,10-20,10-30,5-20 or 1-20 μm.

In certain preferred aspects, the shell of micro-capsule of the invention does not include cell.

Preferably, biocompatible materials of the present invention includes Biodegradable material.

In the present invention, using Biodegradable material come to prepare micro-capsule be particularly preferred.Especially, exist for micro-capsule For preparing the purposes in artificial organ precursor, the use for the material that can not be degraded is unfavorable.Because, on the one hand, this The material that can not be degraded a bit will be retained in obtained artificial organ, so as to limit the application of artificial organ；On the other hand, Cell connection is established between the cell that these materials that can not be degraded will hinder different micro-capsules, is unfavorable for building organic whole (for example, artificial organ).Therefore, before use of the Biodegradable material in shell using micro-capsule for preparing artificial organ Body is particularly advantageous and preferable.

In embodiments of the invention, the Biodegradable material for preparing micro-capsule can be naturally occurring (example Such as derive from the naturally occurring Biodegradable material of animals and plants, such as collagen, fibrin, chitosan, alginic acid Salt, starch, hyaluronic acid, laminin, agarose, gelatin, glucan, and its any combination), artificial synthesized, weight Caused by group, by modified, or its any combinations.

In certain preferred aspects, the Biodegradable material for preparing micro-capsule be it is naturally occurring can Degradation biological material.Preferably, the naturally occurring degradable biomaterial, selected from collagen, fibrin, shell gathers Sugar, alginate (such as sodium alginate or calcium alginate), starch, hyaluronic acid, laminin, agarose, gelatin, Portugal gathers Sugar, chitin, cellulose (such as carboxymethyl cellulose, oxidized regenerated cellulose, bacteria cellulose), fibroin, chondroitin sulfate Element, heparin, fibrinogen, fibronectin, mucopolysaccharide, mucoitin, and its any combination.In some preferred embodiments In, the Biodegradable material for preparing micro-capsule be by modified degradable biomaterial, such as by modification Alginate, such as oxidation alginate (such as oxidized sodium alginate), (such as dialdehyde starch DAS is cross-linking modified bright for modified gelatin Glue), and its any combination.

In certain preferred aspects, the Biodegradable material for preparing micro-capsule is the degradable of synthesis Biomaterial, such as polyphosphazene, polyacrylic acid and its derivative be (such as polymethylacrylic acid, acrylic acid and methacrylic acid Copolymer), PLA (PLA), polyglycolic acid (PGA), polylactic-co-glycolic acid (PLGA), poe (POE), Polycaprolactone (PCL), poly butyric ester (PHB), polyaminoacid (such as polylysine), degradability polyurethane (such as starch Modified polyurethane), PHA (PHAs), poly- hydroxyl valerate (PHV), poly butylene succinate (PBS), polyethylene Alcohol, PPDO, poly-p-dioxanone, poly- dioxane ketone, polytetramethylene carbonate diol, and its is any Combination.In certain preferred aspects, can be by enzyme (such as cell for preparing the Biodegradable material of micro-capsule The enzyme of secretion) degraded.The degradation rate of different Biodegradable materials is widely different, and it may range from one month to number Year.But in the present invention, particularly preferably, for preparing the Biodegradable material of shell in the time no more than 1 month Interior degraded, for example, no more than 30 days, no more than 25 days, no more than 20 days, no more than 15 days, no more than 10 days, no more than 5 My god, no more than 4 days, no more than being degraded in 3 days, the time no more than 2 days or no more than 1 day.For example, for preparing micro-capsule Biodegradable material can at 1-2 days, 2-3 days, 3-4 days, 4-5 days, 5-10 days, 10-15 days, 15-20 days, 20-25 days, or Degraded in the time of 25-30 days.It is particularly preferred that for preparing the Biodegradable material of micro-capsule in the time no more than 10 days Interior degraded.Molecular composition, molecular size range and the molecules align of degradation rate and Biodegradable material are (for example, straight chain or branch Chain) it is closely related.Generally, molecular weight is higher, molecules align is closer, and degradation time is longer.Therefore, the degraded of micro-capsule Speed can be controlled by the configuration of component and/or content to shell.For example, in order to obtain faster degradation rate, can make It is (such as low with the Biodegradable material of low content (such as less than 0.5%, 1%, 2%, 3%, 4% or 5%), low molecule amount In 500Da, 1kDa, 2kDa, 3kDa, 5kDa or 10kDa) Biodegradable material, and/or with loose molecular arrangement Biodegradable material.In order to obtain slower degradation rate, can be used high content (such as higher than 0.5%, 1%, 2%, 3%, 4% or Biodegradable material 5%), HMW (such as higher than 500Da, 1kDa, 2kDa, 3kDa, 5kDa or Biodegradable material 10kDa), and/or the Biodegradable material with close molecular arrangement.In addition, can also be by changing Become the structure of micro-capsule (such as：Multilayer parcel, porous surface, porosity size, specific surface area etc.) adjust Biodegradable material Degradation rate.In addition, the degradation rate of Biodegradable material can also by change synthesize the material polymerization methodses and Copolymer ratio is adjusted；Or it can be adjusted by the crosslinking to the material.In addition, for preparing micro-capsule The degradation rate of Biodegradable material can also be influenceed by cell activities.

In the present invention, it is therefore particularly preferred that the cell in micro-capsule can grow, stretches, breeds, migrate, and and other Cell in micro-capsule establishes cell connection, forms organic construct (such as artificial organ).Therefore, in some preferable implementations In scheme, the micro-capsule is in relatively short time (such as in the time no more than 30 days, such as in time no more than 10 days) Degraded, to promote the cell establishment of connection between different micro-capsules, the cell for avoiding hindering or influenceing between different micro-capsules is established Mutual cell connection.In certain preferred aspects, the micro-capsule no more than 30 days, no more than 25 days, be no more than 20 days, no more than 15 days, no more than 10 days, no more than 5 days, no more than 4 days, no more than 3 days, no more than 2 days or no more than 1 Degraded in it time.For example, the micro-capsule can be at 1-2 days, 2-3 days, 3-4 days, 4-5 days, 5-10 days, 10-15 days, 15- 20 days, 20-25 days, or the time interior degraded of 25-30 days.

Various Biodegradable materials are well known by persons skilled in the art, and its degradation property has been carried out extensively Research.See, for example, Alexander D.Augst, Hyun Joon Kong, David J.Mooney, Alginate Hydrogels as Biomaterials, Macromol.Biosci.2006,6,623-633, it is incorporated herein by reference.

In certain preferred aspects, the degraded of the micro-capsule can provide the life for maintaining or promoting the cell The microenvironment of activity, such as nutriment.In certain preferred aspects, the catabolite of shell is small molecule chemical combination Thing, such as organic acid, monose (such as glucose), oligosaccharides, amino acid, lipid etc..Such catabolite may participate in cell In metabolic activity, for synthetic cell epimatrix or the required energy of activity is converted into.

In certain preferred aspects, for prepare micro-capsule Biodegradable material and its catabolite for thin Born of the same parents are nontoxic, and/or are non-immunogenics for host.

In certain preferred aspects, for prepare the Biodegradable material of micro-capsule contain extracellular matrix or its Analog (such as elastin laminin).The use of extracellular matrix or its analog (such as elastin laminin) can be thin in micro-capsule The vital movement (particularly growth, adhesion, the stretching, extension of cell, and the foundation of Cell tracking) of born of the same parents, which provides, to be similar in vivo Favourable microenvironment, so as to be preferable.

In certain preferred aspects, it is selected from collagen (such as I for preparing the Biodegradable material of micro-capsule Type, II types, type III collagen), fibrin, chitosan, alginate (such as sodium alginate or calcium alginate), oxidation Alginate (such as oxidized sodium alginate), starch, hyaluronic acid, laminin, elastin laminin, gelatin, glucan, poly- ammonia Base acid (such as polylysine), agarose, or its any combinations.

In certain preferred aspects, the micro-capsule includes alginate (such as sodium alginate or calcium alginate), Such as comprising calcium alginate and gelatin, optionally also include elastin laminin.

In certain preferred aspects, the micro-capsule include alginate (such as sodium alginate or calcium alginate) and Gelatin.

In certain preferred aspects, the micro-capsule includes alginate (such as sodium alginate or calcium alginate), Such as comprising calcium alginate and gelatin, optionally also include elastin laminin.In certain preferred aspects, the micro-capsule bag The alginate containing oxidation (such as oxidized sodium alginate).In certain preferred aspects, the micro-capsule includes alginate (such as sodium alginate or calcium alginate) and agarose.

In certain preferred aspects, alginate (such as the sodium alginate of oxidation and the oxidation of oxidation can be used Calcium alginate) prepare micro-capsule, and its degradation speed can be adjusted by controlling the oxidizability of alginate, so that micro- The degradation speed of capsule matches with being wrapped in vitro growth rates therein.

In certain preferred aspects, the micro-capsule also includes extra reagent, for example, nutriment, extracellular Matrix, cell factor and/or active constituents of medicine.Preferably, the extra reagent can regulate and control (such as promotion) cell Propagation, differentiation, migration, secretion and/or metabolism.In certain preferred aspects, the micro-capsule includes at least one (such as 1,2,3,4,5 or more kinds) can regulate and control propagation, differentiation, migration, secretion and/or the new old generation of (such as promotion) cell The extra reagent thanked.In certain preferred aspects, the micro-capsule can discharge described extra in a controlled manner Reagent.

In certain preferred aspects, the nutriment includes but is not limited to, nucleotides, amino acid, polypeptide, carbon Hydrate (such as monose, oligosaccharides, polysaccharide), lipid, vitamin etc..

In certain preferred aspects, extracellular matrix is selected from polysaccharide, such as glycosaminoglycan, proteoglycans；Structure Albumen, such as collagen and elastin laminin；Adhesion protein, such as FTN and laminin.

In certain preferred aspects, the cell factor can be for regulating cell propagation, break up, move The cell factor of shifting, secretion and/or metabolism, include but is not limited to：

- the cell factor related to cell growth, such as insulin, insulin-like growth factor (such as IGF- I, IGF- II), TGF (such as TGF-α and TGF β), VEGF, EGF, fibroblastic growth because It is son, PDGF, osteosarcoma derived growth factor, growth hormone-release inhibiting factor, nerve growth factor, white Cytokine (such as IL-1, IL-11, IL-3), erythropoietin, colony stimulating factor, cortisol, thyroxine, or its What is combined；

- the cell factor related to cell differentiation, such as Oct3/4, Sox2, Klf4, c-Myc, GATA4, TSP1, β-sweet Oleophosphoric acid sodium, dexamethasone, vitamin C, insulin, IBMX, indomethacin, PDGF-BB (PDGF-BB), 5-azacitidine, or its any combinations；

- the cell factor related to cell migration, such as CAMP, triphosphoric acid phosphatidylinositols, stroma cell spread out The raw factor -1, N- cadherins, Nuclear factor kappa B, osteonectin, thromboxane A2, Ras, or its any combinations；And/or

- the cell factor related to cell metabolism, for example, insulin-like growth factor 1, TRIP-Br2, DKK-1, SRANKL, OPG, TRACP-5b, ALP, SIRT1 (2-7), PGC-1 α, PGC-1 β, OPG, IL-3, IL-4, IL-6, TGF-β, PGE2, G-CSF, TNF-α, or its any combinations.

In certain preferred aspects, the active constituents of medicine is the increasing that can regulate and control (such as promotion) cell Grow, break up, migrating, secreting and/or metabolism reagent.In certain preferred aspects, the active constituents of medicine Selected from rhIL-2, rhIL-11, rhEPO, IFN-α, IFN-β, IFN-γ, G-CSF, GM-CSF, rHuEPO, sTNF-R1 and rhTNF-α。

Preferably, the micro-capsule, which includes, can induce undifferentiated cell to the thin of smooth muscle cell or endothelial cell differentiation Intracellular cytokine, such as TGF-a1, PDGF-BB, VEGF or b-FGF.

In certain preferred aspects, the micro-capsule includes：Fat stem cell and the parcel fat stem cell are thin The stratum nucleare of born of the same parents, it is preferable that the stratum nucleare is made up of Biodegradable material；Preferably, it is dry thin to provide induced lipolysis for the stratum nucleare Born of the same parents are to the microenvironment of endothelial cell cell or SMC differentiation (for example, the stratum nucleare is inside comprising induced lipolysis stem cell The inducible factor of chrotoplast or SMC differentiation).In certain preferred embodiments, the induced lipolysis stem cell is to flat The inducible factor of sliding myocyte's differentiation is selected from TGF-a1 and PDGF-BB.In certain preferred embodiments, the induced lipolysis is done Cell is selected from VEGF and b-FGF to the inducible factor of endothelial cell differentiation.

In certain preferred aspects, the micro-capsule includes：Fat stem cell, wrap up the fat stem cell cell Stratum nucleare, and, encapsulate the shell of the stratum nucleare；Preferably, the stratum nucleare and shell are independently of one another by Biodegradable material It is made；Preferably, the stratum nucleare provides microenvironment of the induced lipolysis stem cell to endothelial cell cell or SMC differentiation (for example, the stratum nucleare includes induced lipolysis stem cell to endothelial cell or the inducible factor of SMC differentiation).Some In preferred embodiment, the shell of such micro-capsule also provides induced lipolysis stem cell to endothelial cell or SMC differentiation Microenvironment (for example, the shell includes induced lipolysis stem cell to endothelial cell or the inducible factor of mixed with smooth muscle).At certain In a little preferred embodiments, the induced lipolysis stem cell to the inducible factor of SMC differentiation be selected from TGF-a1 and PDGF-BB.In certain preferred embodiments, the induced lipolysis stem cell is selected to the inducible factor of endothelial cell differentiation VEGF and b-FGF.

Preferably, the biocompatible materials includes Biodegradable material.In the present invention, biodegradable material is used Material prepares outer wall 10, can cause artificial organ precursor implantation subject's body in after continuous growth course in, outer wall 10 by Step degraded, finally causes artificial organ and the autologous tissue for the person of being implanted to be fused into one completely.

Preferably, the Biodegradable material is selected from synthesized degradable material (such as aliphatic polyester (such as PLA (PLA), polycaprolactone (PCL), PHA (PHAs), poly- hydroxyl valerate (PHV), poly butyric ester (PHB), gather Succinic acid-butanediol ester (PBS)), polyglycolic acid (PGA), polylactic-co-glycolic acid (PLGA), poe (POE), Degradability polyurethane (such as starch conversion polyurethane), polyvinyl alcohol, PPDO, poly-p-dioxanone, Poly- dioxane ketone, polytetramethylene carbonate diol, polyphosphazene, and its any combinations).

Preferably, the biocompatible materials also comprising biological non-degradable material (such as nylon, terylene, polypropylene, Polyethylene, polytetrafluoroethylene (PTFE), silicon rubber, fluorosioloxane rubber, natural rubber, polyacrylate, aromatic polyester (such as it is poly- to benzene two Formic acid glycol ester (PET)), nondegradation polyurethane, polyether-ether-ketone, polyacrylonitrile, polysiloxanes, polyformaldehyde, polyvinyl chloride, And its any combinations).

Preferably, the biological non-degradable material is biologically inert.

Preferably, the outer wall 10 is outer tubular wall 10 or sheet outer wall 10.

Preferably, the outer wall 10 impregnated by mould, electrostatic spinning, extrusion forging, 3D printing or spraying be made.

In certain preferred aspects, the method that the outer wall 10 is impregnated by mould obtains.Preferably, the mould Tool infusion process comprises the steps of：

(1) will to be dissolved in suitable solvent for the material (such as Biodegradable material) for preparing outer wall 10 (such as organic Solvent, such as chloroform, tetrahydrofuran or DMA) in, it is configured to prepare solution；

(2) mould is immersed in the preparation solution, takes out mould, the solvent on mould is volatilized；

(3) repeat step (2) repeatedly, obtains outer wall 10；

Optionally, methods described is further comprising the steps of：

Outer wall 10 is dried, sheared and/or sterilized.

Compared in the prior art only with by polymer fiber (such as nylon, terylene), silk or e-PTFE second Mode of the outer tube as artificial blood vessel made of the materials such as alkene, in lumen organization's construct that the above embodiment of the present invention is provided Preparation method in, using made of the materials such as polymer fiber (such as nylon, terylene), silk or expanded PTFE The structure or structure made of biological tissue as outer wall, holder of the outer wall as inwall, and uses as inwall Inwall of biological tissue's wall as artificial blood vessel with bioactivity, it is this by outer wall as support structure it is more complete, Firm, particularly in the case where there is external force effect, inwall is not allowed easy to fall off, is advantageously implemented the biology work(of destination organization Can so that the inwall of manufactured lumen organization's construct can reduce thrombus, calcium sinks closer to the native blood vessel activity of human body The generation of the problems such as product, infection.By checking, the lumen organization prepared by preparation method provided according to embodiments of the present invention After construct implants, the layer of smooth muscle cells similar to normal vascular tissues and endothelial layer can be formed, and with just Normal vascular tissue is fused into one.

In the above-described embodiments, be provided separately inwall 5 and outer wall 10, and outer wall 10 is tubular structure, inwall be comprising Inwall and outer wall, are then fixed together by the biological construct of bioactive substance by way of suit, so as to shape Into lumen organization's construct, this assemble method step is simple, and packaging efficiency is high, suitable for batch production.

In the above-described embodiments, outer wall 10 is tubular structure, and inwall 5 is also tubular structure, if as inwall 5 and outer wall 10 Material be not originally tubular structure, the preparation method of lumen organization's construct can also include outer wall 10 and inwall 5 being made The step of tubulose.

Specifically, inwall 5 and outer wall 10 can be laminated structure before tubulose is formed, and be assembled by outer wall 10 and inwall 5 Also include before：Inwall 5 and outer wall 10 are rolled as tubulose.

Inwall 5 and outer wall 10 can each have multiple arcs structure before tubulose is formed, by 5 groups of outer wall 10 and inwall Also include before dress：By the multiple arcs structures to form for forming inwall 5 into tubulose, by multiple arcs for forming outer wall 10 Shape structures to form is into tubulose.

Certainly, in inwall 5 and outer wall 10 can also one be laminated structure, to form tubulose by rolling, another includes Multiple arcs structure, it is tubulose by the split of multiple arcs structure.

In the above-described embodiments, outer wall 10 can be had with the specific implementation that inwall 5 is set with it is a variety of, such as, can be with First inwall 5 is fixed, outer wall 10 is then sleeved on the outside of inwall 5；First outer wall 10 can also be fixed, then by 5 sets of inwall Mounted in the inner side of outer wall 10；It is, of course, also possible to make outer wall 10 and inwall 5 while move, so that outer wall 10 is set with the outside of inwall 5.

In order to avoid the problem of outer wall 10 and inwall 5 come off occurs in long-term blood flow, by outer wall 10 with it is interior Wall 5 is fixed together progress adhesive coating operation before being additionally included in the suit operation by suit mode.

Specifically, adhesive coating operation includes：Smeared in the inner surface of outer wall 10 and/or the outer surface of inwall 5 viscous Mixture, to make outer wall 10 and inwall 5 inter-adhesive by adhesive.

Wherein, adhesive can be coated in the inner surface of outer wall 10, can also be coated in the outer surface of inwall 5, can also be outside Smear on the inner surface of wall 10 and the outer surface of inwall 5, can specifically be determined according to the property of adhesive.

For example adhesive is applied in any one in the inner surface of outer wall 10 and the outer surface of inwall 5 when electing biogum as Smearing can be with, but needs anion on another in the outer surface of the inner surface of outer wall 10 and inwall 5.

Certainly, most of inwall 5 can all carry anion after medium culture, therefore can be only in outer wall 10 Surface coats biogum, and the outer surface of inwall 5 need not especially be handled.

If smearing biogum in the inner surface of outer wall 10, glue the anion that the outer surface of itself and inwall 5 inherently carries During conjunction, biogum first can also be smeared in the outer surface of inwall 5 as needed, advantage of this is that reducing the outer surface of inwall 5 The anion being exposed, the anion of the outer surface of inwall 5 it is remaining it is few if, the biogum of itself and the inner surface of outer wall 10 occurs The process of reaction will be relatively gentleer, can so be more beneficial for being set with.And the number of plies for specifically smearing biogum can be according to reality Depending on the situation of border.Solidified after the biogum and anionic reactive that the outer surface of inwall 5 is applied, inwall 5 can be made to form one more Stable entirety, convenient further suit, external force is on 5 integrally-built influence of inwall during overcoming suit.

Certainly, adhesive is not limited to biogum, can also be suitable for clinical AB glue, only need to be in the interior table of outer wall 10 One in the outer surface of face and inwall 5 upper to smear A glue, and the inner surface of outer wall 10 with it is another in the outer surface of inwall 5 Individual upper smearing B glue, when A glue and B glue meet, the two occurs chemical reaction and outer wall 10 is bonded together with inwall 5.Its In, A glue or B glue are likely to be a kind of component natively having on the inner surface of outer wall 10 or the outer surface of inwall 5, as long as can Bonded with corresponding surface.

In one embodiment of the preparation method of lumen organization's construct, adhesive coating operation also includes：According to outer The coating thickness of gap length adjustment adhesive between wall 10 and inwall 5.

Specifically, when the gap between outer wall 10 and inwall 5 is larger, the thickness of adhesive can be made larger, so not But be advantageous to the Stability Analysis of Structures of inwall 5, can also avoid in some cases due between outer wall 10 and inwall 5 gap it is too big and It can not cement；When the gap between outer wall 10 and inwall 5 is smaller, it can correspondingly reduce the coating thickness of adhesive.

To further facilitate suit, can also include adjusting outer wall 10 and/or interior before outer wall 10 is set with inwall 5 The step of position of wall 5.

Specifically, set-up procedure includes：Adjust axial direction and/or the circumferential distance of outer wall 10 and inwall 5.

Before being adjusted to outer wall 10 and inwall 5, outer wall 10 and the in the vertical direction of inwall 5 can be made coaxial, also may be used , can also be coaxial on other incline directions with coaxial in the horizontal direction, then by adjusting between outer wall 10 and inwall 5 Axial distance makes outer wall 10 and inwall 5 close to each other, to facilitate suit, improves suit efficiency.

Can have a variety of for the means of circumferential distance regulation, for example can be supportted the internal diameter of outer wall 10 by Stress control Greatly, to facilitate suit of inwall 5 etc..

In above-mentioned each embodiment, inwall 5 can be printed by 3D biometric prints machine and formed, and outer wall 10 can also pass through 3D Printer printing forms.

The preparation method of lumen organization's construct can also include：Inwall for preparing lumen organization's construct is being provided 5 and outer wall 10 before, prepare the outer wall 10 with biocompatible materials.

On the shape of inwall 5 and outer wall 10, inwall 5 and/or the pipe that outer wall 10 can be sidewall opening or side wall is closed Shape structure, to facilitate suit.

To improve efficiency prepared by lumen organization's construct, the present invention also proposes a kind of preparation dress of lumen organization's construct Put.

As shown in figure 1, the preparation facilities of lumen organization's construct includes the first supporting construction, the second supporting construction and set Assembling device, the first supporting construction and the second supporting construction are respectively used to the He of outer wall 10 that carrying is used to prepare lumen organization's construct Inwall 5, inwall 5 are the biological construct comprising bioactive substance, and outer wall 10 is tubulose, bushing device be used for by outer wall 10 with Inwall 5 is set with and is fixed together.

Relevant explanation wherein on inwall 5 and outer wall 10 refers to the related content in aforementioned preparation process, here no longer Repeat.

Outer wall 10 and inwall 5 are fixed together by way of suit, assembly program can be simplified as much as possible, It is time-consuming to shorten assembling, improves packaging efficiency.

In the above-described embodiments, the specific constructive form of the first supporting construction and the second supporting construction can be selected flexibly, As long as can be respectively that outer wall 10 and inwall 5 provide support.

As the preferred of the first supporting construction, the first supporting construction includes support column 9, and outer wall 10 is sheathed on support column 9 Periphery；Or first supporting construction include support set, support set is sheathed on the periphery of outer wall.

As the preferred of the second supporting construction, the second supporting construction includes fixture 3, and fixture 3 is used in described in clamp standoff The support member of wall 5, inwall 5 are arranged at the periphery of support member；Or supporting construction includes support bar, inwall is arranged at branch The periphery of strut.

Wherein, it is used to support inwall when the support member clamped by fixture 3 is preferably 3D biometric prints machine printing inwall 5 5 swingle 4, swingle 4 is clamped using fixture 3, operation can be made more convenient, and eliminated inwall 5 from swingle 4 On take off, the step being then placed on again in other supporting constructions, improve packaging efficiency.

In the above-described embodiments, the suit of outer wall 10 and inwall 5 can use manual type to carry out, and can also use machinery Automation equipment is carried out.For convenience of operation, bushing device includes drive mechanism, and drive mechanism is used to driving outer wall 10 and/or interior Wall 5, so that with inwall 5 relative motion occurs for outer wall 10, the suit of outer wall 10 and inwall 5 is realized, drive mechanism can be further Improve packaging efficiency.

The specific constructive form of drive mechanism has a variety of, for example can use stepper motor, be precisely controlled outer wall 10 and/or The amount of exercise of inwall 5.

As shown in Fig. 2 drive mechanism includes push rod 11, for convenience of exerting a force, push rod 11 could be arranged to L-type arm and the collar Combining structure, when pushing the L-type arm of push rod 11, the collar for the periphery for being sheathed on support column 9 is driven to move down, so as to promote set Periphery loaded on support column 9 simultaneously moves downward positioned at the outer wall 10 of collar bottom relative to inwall 5.

Further, bushing device also includes resetting-mechanism, and resetting-mechanism is connected with drive mechanism, and for making driving machine Structure resets.

Resetting-mechanism is preferably but not limited to be arranged on push rod 11 for spring 12, spring 12, with push rod 11 Compressed during pressure, after the distress resolves of push rod 11, spring 12 recovers, and drives push rod 11 to reset, and is operated for suit next time Prepare.

To enable outer wall 10 to be fitted together more securely with inwall 5, the preparation facilities of lumen organization's construct is also Including coating unit, coating unit is used to smear adhesive in the inner surface of outer wall 10 and/or the outer surface of inwall 5, so that outside Wall 10 is inter-adhesive by described adhesive after being set with inwall 5.

On the selection of adhesive, the associated description in above-mentioned preparation method can also be referred to, is repeated no more here.

The coating thickness of adhesive can be adjusted according to the gap length between inwall 5 and outer wall 10.Specifically, may be used By by adjust adhesive flow velocity or inwall 5, outer wall 10 translational speed etc. in a manner of be adjusted.

As a preferred embodiment of coating unit, as shown in Figure 2, Figure 3 and Figure 4, the coating unit includes container 16th, transfer passage 17 and adhesive outlet, the container 16 are used to hold described adhesive, the transfer passage 17 and the appearance Device 16 is connected and for conveying described adhesive, and described adhesive outlet is connected with the transfer passage 17 with by described adhesive Coated on the outer wall 10 or the inwall 5.

Wherein, adhesive is exported by adhesive and exported, for example adhesive infiltrates from adhesive outlet.

Specifically, transfer passage 17 can be the special delivery pipe being connected between container 16 and aperture 18, be used for Adhesive is delivered to aperture 18, or the pipeline being arranged on inside associated components.As shown in figure 4, transfer passage 17 is set Put inside the first supporting construction, adhesive outlet is the aperture connected with transfer passage 17 being arranged in the first supporting construction 18。

In this embodiment, the smearing of adhesive occurs during outer wall 10 or inwall 5 are with the relative motion of aperture 18, For example outer wall 10 can be made motionless, inner surface movement of the aperture 18 along outer wall 10, in motion process, adhesive is made in pressure Aperture 18 constantly is extruded with lower, so that adhesive is applied on the inner surface of outer wall 10；Aperture 18 can also be made motionless, outside Wall 10 moves relative to aperture 18, and in motion process, adhesive is constantly extruded aperture 18 under pressure, so that viscous Mixture is applied on the inner surface of outer wall 10.For inwall 5, situation is similar, repeats no more here.

In this embodiment, the coating thickness of adhesive can control the outflow of adhesive by adjusting the size of pressure Speed, and then adjust the bond thickness of adhesive.

In addition, transfer passage can also be the structure including delivery pipe and shower nozzle, delivery pipe is connected with shower nozzle, the spray of shower nozzle Mouth exports as adhesive, to be sprayed on adhesive by shower nozzle on the inner surface of outer wall 10 or the outer surface of inwall 5.

In this embodiment, adhesive is applied on outer wall 10 or inwall 5 by way of spraying, the reality of this mode Now more flexibly, outer wall 10 or inwall 5 can be fixed, and shower nozzle moves, and complete spraying process, the forms of motion of shower nozzle Also it is relatively more flexible, it be able to can also be rotated with translation, which smearing is more uniform, and controllability is also more preferable.

In this embodiment, the coating thickness of adhesive can be by controlling the time sprayed, the spraying number of plies, adhesive defeated Outflow etc. adjusts.

In a preferred embodiment of the preparation facilities of lumen organization's construct of the present invention, in addition to positioner, it is fixed Position device is used to position outer wall 10 and/or inwall 5 before outer wall 10 is set with inwall 5.

Outer wall 10 or inwall 5 are positioned by positioner, can to suit when moving line be adjusted, make Suit is more convenient, reduces inaccuracy and uncertainty during suit, improves suit efficiency.

The concrete form of positioner can have it is a variety of, such as positioner can be provided in groove on certain part or Person's clip, as long as outer wall 10 or inwall 5 are put into the groove or clamped with clip, you can realize to outer wall 10 or inwall 5 It is fixed.

In other embodiments, positioner can also be can adjust it is at least one relative in outer wall 10 and inwall 5 Device in another position, to cause outer wall 10 and inwall 5 coaxial, easily facilitate suit.Here in outer wall 10 and inwall 5 At least one position relative to another, can be horizontal, vertical or the relative position on a variety of different directions such as tilt Put, the adjustment of relative position on multiple different directions can also be included simultaneously.

It is further preferred that outer wall 10 and inwall 5 are coaxial, positioner can adjust the axle between outer wall 10 and inwall 5 To distance.First cause that outer wall 10 and inwall 5 are coaxial when mounted, then using positioner adjustment outer wall 10 and inwall 5 it Between axial distance.

As a specific embodiment of positioner, positioner includes support base and is connected with the first supporting construction Support 8, support 8 can move relative to support base, and support 8 is provided with rack 13, and support base is provided with and mutually nibbled with rack 13 The gear 14 of conjunction, to drive rack 13 to move by the rotation of gear 14, rack 13 drives the motion of the first supporting construction, so that Outer wall 10 relatively moves relative to inwall 5.

Wherein, support base can include the base 1 and installing plate 2 shown in Fig. 2, and installing plate 2 is arranged on base 1, support 8 It can be moved relative to base 1 and installing plate 2.

Certainly, the structure that rack-and-pinion cooperates, which can also replace with other, can drive what the first supporting construction was moved Mechanism, such as stepper motor etc..

Further, positioner also includes guiding mechanism, and guiding mechanism is arranged on support base, the first supporting construction peace On guiding mechanism, so that the first supporting construction can move under the guiding of guiding mechanism.

The concrete form of support base have it is a variety of, as shown in Fig. 2 support base includes base 1 and installing plate 2, guiding mechanism peace On installing plate 2, the first supporting construction is arranged on the support 8 being connected with installing plate 2, and the second supporting construction passes through fixture 3 On base 1.

As a preferred embodiment of guiding mechanism, guiding mechanism includes slide rail 6 and sliding block 7, and sliding block 7 is slided with slide rail 6 Dynamic to coordinate, slide rail 6 is arranged on support base, and the first supporting construction is connected with sliding block 7.

In an embodiment as illustrated in figure 2, base 1 is horizontal positioned, and installing plate 2 is placed vertically, correspondingly, slide rail 6 and tooth Bar 13 is vertically movable, to cause outer wall 10 to be moved straight down relative to inwall 5, completes the suit of outer wall 10 and inwall 5. In other embodiments, base 1, installing plate 2, slide rail 6 and rack 13 can also have other modes of emplacements, as long as enabling to outer With inwall 5 relative motion occurs for wall 10.Such as in embodiment as shown in Figure 9,1 vertical mode of base, installing plate 2 Horizontal positioned, slide rail 6 and rack 13 are horizontal movements, to cause outer wall 10 to be moved right relative to the level of inwall 5, are completed outer The suit of wall 10 and inwall 5.

In above-mentioned each embodiment, the supporting part of the first supporting construction and/or the second supporting construction can use biology Compatibility material, to be supported to possessing the outer wall 10 of bioactivity or inwall 5.

Based on the preparation method of above lumen organization construct, the present invention also proposes a kind of biological tissue's construct, the life Thing tissue construct uses the preparation method of the lumen organization's construct proposed in each embodiment above to be prepared.It is manufactured Lumen organization's construct can be alimentary canal lumen organization construct, respiratory tract lumen organization construct, lymphatic vessel lumen organization Construct or vessel lumen tissue construct.

Wherein, the outer wall 10 is biocompatible materials.Outer wall 10 can be that Biodegradable material or biology can not Degradable material.For example the material of outer wall 10 can be nylon, terylene, silk, polytetrafluoroethylene (PTFE) or the lumen organization of animal.It is interior Wall 5 can include cell.

Below to the concrete structure of lumen organization's construct of the present invention and preparation method thereof, one embodiment of preparation facilities It is described in detail with the course of work：

The structure of preparation facilities one embodiment of lumen organization's construct is introduced with reference to Fig. 1~Fig. 4 first.

With reference to figure 1 and Fig. 2, the device includes assembly platform and pusher, wherein, assembly platform includes base 1, installation Plate 2 and fixture 3, installing plate 2 and fixture 3 are respectively arranged in above base 1, and fixture 3 is used for the rotation for clamping 3D biometric print machines Bar 4, inwall 5 can be the biological tissue's wall being directly printed upon on swingle 4.

Pusher includes slide rail 6, sliding block 7, support 8, support column 9, push rod 11, spring 12, rack 13, gear 14, hand Wheel 15 and container 16, wherein slide rail 6 are arranged on installing plate 2, and sliding block 7 is arranged on slide rail 6, and support 8 is arranged on sliding block 7, Rack 13 is arranged on support 8, and gear 14 is arranged on installing plate 2, and gear 14 is connected with handwheel 15, by rotation hand wheel 15, Band moving gear 14 rotates, and makes rack 13 and support 8 while moves up and down, and support 8 can move up and down along slide rail 6, to realize The up and down motion of support column 9, complete the positioning of outer wall 10 before assembling.Support column 9 can be sleeve structure, and outer wall 10 is arranged on set The outside of pipe.

Push rod 11 is located at the top of outer wall 10, and push rod 11 may be mounted on the outer wall of support column 9, and push rod 11 can be along Support column 9 pushes downwards outer wall 10, and after push action is completed, push rod 11 can be resetted by the spring 12 at end mounted thereto. The smearing that adhesive is completed while outer wall 10 moves downward is pushed in push rod 11, and outer wall 10 is pushed to the periphery of inwall 5, Complete the assembling of inwall 5 and outer wall 10.

With reference to figure 3 and Fig. 4, adhesive is stored in container 16, and container 16 is arranged on the upper end of support column 9, support column 9 Inside be provided with transfer passage 17, and the front end of support column 9 is provided with aperture 18, and adhesive can export through aperture 18, and glue The inner surface of outer wall 10 is attached to, while being moved down with outer wall 10, controls the flow of adhesive, adhesive is according to a constant speed Degree continues to infiltrate from aperture 18, to complete the smearing of the whole adhesive of outer wall 10.By adjusting the size of pressure, can control The flow and smearing thickness of adhesive.

Below with reference to Fig. 5~Fig. 8, the specific implementation step of the preparation method based on device made above is illustrated：

(1) inwall 5 and outer wall 10 are installed

As shown in figure 5, inwall 5 is by the printed biological tissue's inwall of 3D biometric print machines, swingle 4 is inwall 5 carrier.First, the swingle 4 of printed inwall 5 is arranged in fixture 3, outer wall 10 is then arranged on support column 9 Outer surface.

(2) outer wall 10 is positioned

Rotate handwheel 15, by the coupled movements of gear 14 and rack 13, make support 8 drive support column 9 along slide rail 6 to Lower movement, outer wall 10 is set to complete the positioning of outer wall 10 close to inwall 5 (for example distance is preferred in 2mm or so).

(3) outer wall 10 is pushed

Push rod 11 is pushed down on, push rod 11 is moved down with outer wall 10, while by pressure control vessel 16 Adhesive exports from the aperture 18 of the front end of support column 9, and is attached to the inner surface of outer wall 10.When the lower top of outer wall 10 and inwall During 5 upper hanging, stop push, push rod 11 is automatic to reset upwards under the active force of spring 12.Now complete inwall 5 With the assembling of outer wall 10.

(4) device resets and taken out the lumen organization's construct assembled

Rotate counterclockwise handwheel 15, by the coupled movements of gear 14 and rack 13, make support 8 drive support column 9 along Slide rail 6 moves up, support column 9 is then taken out swingle 4 from fixture 3 away from swingle 4, will finally assemble Lumen organization's construct is removed above swingle 4.

, can by the explanation to lumen organization's construct of the present invention and preparation method thereof, multiple embodiments of preparation facilities To see that lumen organization's construct of the present invention and preparation method thereof, preparation facilities embodiment at least have one or more of excellent Point：

1st, by making inner and outer wall respectively, then the two is set in together, assembling speed is fast, takes short, efficiency It is high；

2nd, preparation facilities is provided with positioner and guiding mechanism, easy to operate, quick；

3rd, the coating thickness of adhesive is controllable, to adapt to gap length different between outer wall and inwall.

Finally it should be noted that：The above embodiments are merely illustrative of the technical scheme of the present invention and are not intended to be limiting thereof；To the greatest extent The present invention is described in detail with reference to preferred embodiments for pipe, those of ordinary skills in the art should understand that：Still The embodiment of the present invention can be modified or equivalent substitution is carried out to some technical characteristics；Without departing from this hair The spirit of bright technical scheme, it all should cover among the claimed technical scheme scope of the present invention.

Claims (33)

1. A kind of 1. preparation method of lumen organization's construct, it is characterised in that including：

   Inwall (5) and outer wall (10) for preparing lumen organization's construct is provided, wherein, the inwall (5) is to include biology The biological construct of active material, the outer wall (10) are tubulose；

   The outer wall (10) is fixed together with the inwall (5) by suit mode.
2. 2. the preparation method of lumen organization's construct according to claim 1, it is characterised in that by the outer wall (10) with The inwall (5) is fixed together by suit mode and operated including suit, and the suit operation includes：

   It is first that the inwall (5) is fixed, then move the outer wall (10) and the outer wall (10) is sleeved on the inwall (5) outside Side；Or

   It is first that the outer wall (10) is fixed, then move the inwall (5) and the inwall (5) is sleeved in the outer wall (10) Side；Or

   The outer wall (10) and the inwall (5) move simultaneously, so that the outer wall (10) is set with the outside of the inwall (5).
3. 3. the preparation method of lumen organization's construct according to claim 1, it is characterised in that by the outer wall (10) Progress adhesive coating before being additionally included in the suit operation is fixed together by suit mode with the inwall (5) Operation：In the inner surface of the outer wall (10) and/or the outer surface application of adhesive of the inwall (5), to be grasped in the suit Make the outer wall (10) and the inwall (5) inter-adhesive by described adhesive after work.
4. 4. the preparation method of lumen organization's construct according to claim 3, it is characterised in that described adhesive coating behaviour Also include：According to the coating thickness of the gap length adjustment described adhesive between the outer wall (10) and the inwall (5).
5. 5. the preparation method of lumen organization's construct according to claim 1, it is characterised in that in the suit operation It is preceding also to include：

   Adjust the axial direction between the outer wall (10) and the inwall (5) and/or circumferential distance.
6. 6. the preparation method of lumen organization's construct according to claim 1, it is characterised in that the outer wall (10) and/ Or the inwall (5) is formed by the printing of 3D biometric prints machine.
7. 7. the preparation method of lumen organization's construct according to claim 1, it is characterised in that the biological construct is Alimentary canal lumen organization construct, respiratory tract lumen organization construct, lymphatic vessel lumen organization construct or vessel lumen tissue Construct.
8. 8. the preparation method of lumen organization's construct according to claim 1, it is characterised in that also include：Use is being provided Before the inwall (5) and outer wall (10) of lumen organization's construct is prepared, the outer wall (10) is prepared with biocompatible materials.
9. 9. the preparation method of lumen organization's construct according to claim 1, it is characterised in that the inwall (5) and/or The tubular structure that the outer wall (10) is sidewall opening or side wall is closed.
10. 10. a kind of preparation facilities of lumen organization's construct, it is characterised in that including the first supporting construction, the second supporting construction And bushing device, first supporting construction and second supporting construction are respectively used to carrying and are used to prepare lumen organization's structure The outer wall (10) and inwall (5) of body, the inwall (5) are the biological construct comprising bioactive substance, the outer wall (10) For tubulose, the bushing device is used to the outer wall (10) is set with and is fixed together with the inwall (5).
11. 11. the preparation facilities of lumen organization's construct according to claim 10, it is characterised in that the first support knot Structure includes support column (9), and the outer wall (10) is sheathed on the periphery of the support column (9)；Or the first supporting construction bag Support set is included, the support set is sheathed on the periphery of the outer wall.
12. 12. the preparation facilities of lumen organization's construct according to claim 10, it is characterised in that the second support knot Structure includes fixture (3), and the fixture (3) is used for the support member of inwall (5) described in clamp standoff, and the inwall (5) is arranged at The periphery of the support member；Or the supporting construction includes support bar, the inwall (5) is arranged at the support bar Periphery.
13. 13. the preparation facilities of lumen organization's construct according to claim 12, it is characterised in that the support member is It is used for the swingle (4) for supporting the inwall (5) when 3D biometric prints machine prints inwall (5).
14. 14. the preparation facilities of lumen organization's construct according to claim 10, it is characterised in that the bushing device bag Drive mechanism is included, the drive mechanism is used to drive the outer wall (10) and/or the inwall (5), so that the outer wall (10) Relative motion occurs with the inwall (5), realizes the suit of the outer wall (10) and the inwall (5).
15. 15. the preparation facilities of lumen organization's construct according to claim 14, it is characterised in that the bushing device is also Including resetting-mechanism, the resetting-mechanism is connected with the drive mechanism, and for resetting the drive mechanism.
16. 16. the preparation facilities of lumen organization's construct according to claim 10, it is characterised in that the bushing device is also Including coating unit, the coating unit is used to apply in the inner surface of the outer wall (10) and/or the outer surface of the inwall (5) Adhesive is smeared, so that the outer wall (10) is inter-adhesive by described adhesive after being set with the inwall (5).
17. 17. the preparation facilities of lumen organization's construct according to claim 16, it is characterised in that the painting of described adhesive It is adjustable to cover thickness.
18. 18. the preparation facilities of lumen organization's construct according to claim 16, it is characterised in that the coating unit bag Container (16), transfer passage (17) and adhesive outlet are included, the container (16) is used to hold described adhesive, and the conveying is logical Road (17) is connected with the container (16) and for conveying described adhesive, described adhesive outlet and the transfer passage (17) Connect so that described adhesive is coated on the outer wall (10) or the inwall (5).
19. 19. the preparation facilities of lumen organization's construct according to claim 18, it is characterised in that the transfer passage (17) it is arranged on inside the first supporting construction；And/or described adhesive outlet for be arranged in the first supporting construction with it is described The aperture (18) of transfer passage (17) connection.
20. 20. the preparation facilities of lumen organization's construct according to claim 18, it is characterised in that the transfer passage (17) delivery pipe and shower nozzle are included, the delivery pipe is connected with the shower nozzle, and the spout of the shower nozzle goes out as described adhesive Mouthful, so that described adhesive to be sprayed on to the outer surface of the inner surface of the outer wall (10) or the inwall (5) by the shower nozzle On.
21. 21. the preparation facilities of lumen organization's construct according to claim 10, it is characterised in that the preparation facilities is also Including positioner, the positioner is used for before by the outer wall (10) and the inwall (5) suit to the outer wall (10) and/or the inwall (5) is positioned.
22. 22. the preparation facilities of lumen organization's construct according to claim 21, it is characterised in that the positioner energy Enough adjust at least one position relative to another in the outer wall (10) and the inwall (5).
23. 23. the preparation facilities of lumen organization's construct according to claim 22, it is characterised in that the outer wall (10) with Coaxially, the positioner can adjust the axial distance between the outer wall (10) and the inwall (5) to the inwall (5).
24. 24. the preparation facilities of lumen organization's construct according to claim 23, it is characterised in that the positioner bag Support base and the support (8) being connected with first supporting construction are included, the support (8) can move relative to the support base Dynamic, the support (8) is provided with rack (13), and the support base is provided with and the rack (13) pitch wheel (14), to drive the rack (13) to move by the rotation of the gear (14), the rack (13) drives described first Support structure moves, so that the outer wall (10) relatively moves relative to the inwall (5).
25. 25. the preparation facilities of lumen organization's construct according to claim 24, it is characterised in that the positioner is also Including guiding mechanism, the guiding mechanism is arranged on the support base, and first supporting construction is arranged on the Guiding machine On structure, so that first supporting construction can move under the guiding of the guiding mechanism.
26. 26. the preparation facilities of lumen organization's construct according to claim 25, it is characterised in that the guiding mechanism bag Slide rail (6) and sliding block (7) are included, the sliding block (7) is slidably matched with the slide rail (6), and the slide rail (6) is arranged on the support On seat, first supporting construction is connected with the sliding block (7).
27. 27. the preparation facilities of lumen organization's construct according to claim 10, it is characterised in that the first support knot Material is biocompatible materials used by the supporting part of structure.
28. 28. a kind of lumen organization's construct, it is characterised in that lumen organization's construct is used as claim 1~9 is any The preparation method of lumen organization's construct described in is prepared.
29. 29. according to lumen organization construct of the right described in require 28, it is characterised in that the outer wall (10) is bio-compatible Property material.
30. 30. according to lumen organization construct of the right described in require 28, it is characterised in that the outer wall (10) is that biology can drop Solve material or biological non-degradable material.
31. 31. according to lumen organization construct of the right described in require 28, it is characterised in that the material of the outer wall (10) is Buddhist nun Dragon, terylene, silk, the lumen organization of polytetrafluoroethylene (PTFE) or animal.
32. 32. according to lumen organization construct of the right described in require 28, it is characterised in that the inwall (5) includes cell.
33. 33. according to lumen organization construct of the right described in require 28, it is characterised in that lumen organization's construct is disappears Hua Dao lumen organizations construct, respiratory tract lumen organization construct, lymphatic vessel lumen organization construct or vessel lumen tissue structure Build body.