CN105935454A - Decellularized matrix-source tissue engineering scaffold and preparation method and application thereof - Google Patents
Decellularized matrix-source tissue engineering scaffold and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a decellularized matrix-source tissue engineering scaffold and a preparation method and an application thereof; the decellularized matrix-source tissue engineering scaffold takes a treated animal membrane material as a biological membrane base material, and the surface of the biological membrane base material is attached with a collagen loose layer. The preparation method of the decellularized matrix source tissue engineering scaffold comprises the process steps of degreasing, decellularizing, antigen removal, cross-linking fixation, collagen extraction, collagen compositing and the like. The flexible and smooth animal membrane material is used as the biological membrane base material, and the defects that a pure collagen scaffold has poor mechanical properties and too fast degradation rate are overcome; with cooperation of the collagen loose layer, a good microenvironment is provided for tissue regenerative repair and cell growth; after being implanted into a body, the decellularized matrix-source tissue engineering scaffold material is gradually degraded along with repairing of defect tissues, also has controllable degradation time, does not exist as a permanent foreign matter, and has no any residual toxicity.
Description
Technical field
The present invention relates to tissue engineering technique field, specifically, relate to a kind of acellular matrix source tissue
Engineering rack and its preparation method and application.
Background technology
Along with organizational project subject and the development of regenerative medicine, people are to the support required for tissue engineering technique
Material launches substantial amounts of research, including macromolecular material, metal material, bioceramic, foreign material
Deng, but in clinical practice, there is certain complication or untoward reaction in these materials.Therefore, in recent years,
The biomaterial in acellular matrix source is by substantial amounts of concern and research, the biomaterial conduct of acellular matrix source
The research of tissue engineering bracket is risen, and research contents is mainly acellular matrix source material soft group of human body
Knit the repairing effect after damage and occupation mode；As the treatment after skin burn, the reparation after cartilage injury,
Hernia repair etc..But, when acellular matrix material uses as support, it is the most single to there is structure in itself,
Biocompatibility is not good enough, it is difficult to be applicable to the problems such as multiple clinic is suitable for disease, and Clinical practice is limited.
The acellular matrix material in heteroplasm source is because having wide material sources, and structure is high with human body similarity
Advantage, has had multiple launch and has been successfully applied to clinic, but the de-cell of heterologous source
The problem that host material exists the congenital defects such as immunological rejection, mechanical strength is poor, structural controllability is inadequate.
Summary of the invention
The invention aims to solve the defect existing for above-mentioned existing tissue engineering bracket, it is provided that a kind of
Having excellent mechanical performances, good biocompatibility, relatively reduced immunogenicity, degradable absorbs, and can control fall
Solving speed, do not cause rejection after implantation, being suitable for again multiple clinic is suitable for the acellular matrix source of disease simultaneously
Tissue engineering bracket.
It is a further object of the present invention to provide the preparation side of a kind of above-mentioned acellular matrix source tissue engineering rack
Method, can obtain having excellent mechanical performances, good biocompatibility, relatively reduced immunogenicity, and degradable absorbs,
Controllable degradation speed, does not cause rejection after implantation, being suitable for again multiple clinic is suitable for the above-mentioned of disease simultaneously
Acellular matrix source tissue engineering rack.
The third object of the present invention is to provide the application of a kind of above-mentioned acellular matrix source tissue engineering rack.
For solving above-mentioned technical problem, the present invention adopts the following technical scheme that a kind of acellular matrix source group
Weaver's engineering support, using the animal membrane material after process as biomembrane base material, on the surface of described biomembrane base material
Adhere to collagen protein weaker zone.
Preferably, described collagen protein weaker zone is to be prepared by animal tendon tissue.
Preferably, described biomembrane base material has matte and bright finish, institute's collagen protein weaker zone to adhere on matte.
Preferably, the thickness of described collagen protein weaker zone is 0.1～3.0mm, and porosity is 65～86%,
Aperture is 50～300 μm.
A kind of preparation method of acellular matrix source tissue engineering rack, including step as follows:
(1) preparation of biomembrane base material；
(2) preparation of collagen protein；
(3) collagen composite: adhered to by collagen protein on the matte of biomembrane base material, is dried.
Concrete, the preparation method of described acellular matrix source tissue engineering rack, including step as follows:
(1) pretreatment: take fresh animal membrane material and tendon tissue, cleans, sterilization, and removes fat group
Knit, fiber and chorionic villi；Preferably, can be by pruning away except fatty tissue, fiber and chorionic villi；
(2) defat: use organic solvent extracting extraction animal membrane material and tendon tissue, remove fat and liposoluble
Property impurity；Preferably, organic solvent be mass concentration be the aqueous solution of 0.5～3.2%；
(3) de-cell: use surfactant solution associated proteins enzymatic solution, control surfactant
Quality final concentration of 0.1～0.5%, the quality final concentration 0.1～0.3% of protease, ultrasonic digestion animal membrane material
And tendon tissue, with the cell contained by removal or cell debris；
(4) antigen is removed: use activating agent, with animal membrane material and tendon under the conditions of pH value is 7～8
Tissue reaction 24～48h, then uses strong hydrogen bonding reagent, under the conditions of pH value is 7～8 with animal membrane material with
Tendon tissue reaction 24～48h, to replace in animal membrane material and tendon tissue in collagen molecules coiled strand
Special hydrogen bond, thoroughly removed the biomembrane base material after antigen and tendon tissue；
(5) crosslinking is fixing: the no-aldehyde chemical crosslinking agent solution using mass concentration to be 0.5～5%, at pH
Value is under 7～8 stirring conditions, and reacting with the biomembrane base material after removing antigen and processing 2～8 days (can
Corresponding crosslinking time is selected according to the required degree of cross linking), the collagen molecules in biomembrane base material is carried out
Crosslinking；
(6) collagen protein extracts and purification: the acid solution dissolving warp using mass concentration to be 0.05～0.2%
Remove the tendon tissue after antigen processes, smash to pieces, under the acid condition that pH value is 3～5, then use matter
Amount concentration 0.1～the protein enzyme solution continuation dissolving of 0.4%, separate out collagen protein, filter, re-use quality
Concentration is the surfactant solution immersion of 0.1～0.5%, obtains collagen protein sterling after washing；
The present invention uses (mass concentration is 0.05～0.2%) acid solution of low concentration in order to allow
Tendon tissue loosens and preliminary acidolysis so that it is can mash in bruisher solids, conveniently uses enzymolysis
Carry out the decomposition of protein fragments rank；
(7) collagen composite: collagen protein sterling be vacuum dried, pulverizes, mills, sieve and make collagen egg
White lead end, is dissolved into collagen protein powder mass concentration 2～the collagen solution of 5%, obtains I type glue
Former, type III collagen or the blend of I/III Collagen Type VI, put down gained blend (collagen solution)
It is laid on the matte of crosslinked fixing biomembrane base material, vacuum drying, obtains that there is double-deck tissue
Engineering rack.
Preferably, during described animal membrane material is pericardium, goldbeater's skin or fat nethike embrane one or more.Preferably
, described animal membrane material can be chosen at butchers 4 hours interior pigs or the pericardium of cattle, goldbeater's skin or fat nethike embrane
Deng internal organs film.
Preferably, described activating agent is small molecular organic acid acid anhydride, acyl chlorides, amide, epoxide or halogen first
In alkane one or more；Described strong hydrogen bonding reagent is guanidine compound.
Preferably, guanidine compound is guanidine hydrochloride solution.
Preferably, described no-aldehyde chemical cross-linking agent is epoxide, two acid diamides, diisocyanate, gathers
One or more in ethylene glycol or carbodiimide reagent.
Preferably, described epoxide is monoepoxideDi-epoxide
Or one or more in low polyepoxide, wherein R=H or CnH2n+1-, n=0-10.Preferably, institute
Stating low polyepoxide is one or more in poly(propylene oxide), poly-epoxy prapanol or alkylene oxide.
Preferably, in preparation method, animal membrane material and tendon tissue quality, organic solvent, activating agent,
Strong hydrogen bonding reagent, no-aldehyde chemical crosslinking agent solution mass ratio be 1:(5～10): (10～20): (10～
20): (10～20).
Preferably, described acid solution is hydrochloric acid and/or acetic acid aqueous solution.
Preferably, described organic solvent is acetate esters, chloroform, carbon tetrachloride, ether, acetone or anhydrous
In ethanol one or more；Described surfactant is TritonX, dodecyl sodium sulfate, 3-[(3-
Gallbladder amidopropyl) dimethylammonio]-1-propane sulfonic acid salt, a kind of in NaTDC or carboxymethylamino methane
Or it is two or more；Described protease be in trypsin, pepsin or papain one or both with
On.
Preferably, in step (2), extracting extraction time is 24～48h.
Preferably, in step (3), ultrasonic digestion 24～48h under the conditions of 30～60kHz.
Preferably, step (6) uses protein enzyme solution to continue to dissolve, and temperature controls at 10～20 DEG C, analysis
Go out collagen protein.
Preferably, in step (7), vacuum drying conditional sampling is elected as: vacuum pressure is 0～10Pa, dry
The dry time 48～72h.Preferably, take particle size range collagen protein powder below 200 μm and be dissolved into glue
Former protein solution.
Preferably, in step (7), the mass concentration of the collagen solution after dissolving is 2～5%.
Preferably, in above-mentioned preparation method, described solution is aqueous solution or buffer solution.
Preferably, sterilization is to use peracetic acid, benzalkonium bromide, hibitane, bromo geramine, Hydrazoic acid,sodium salt
4～24h are soaked Deng broad-spectrum high efficacy disinfectant solution.
The blend of described type i collagen, type III collagen or I/III Collagen Type VI adheres to through going antigen to process
On the matsurface of the biomembrane base material fixing with the crosslinking of no-aldehyde fixative, after vacuum drying, form collagen protein
Weaker zone, and biomembrane base material and collagen protein weaker zone collectively form and have double-deck organizational project and prop up
Frame, this tissue engineering bracket through cutting out, is shaped, is packed, sterilizing, obtain finished product.
Described have double-deck tissue engineering bracket by biological fibrin glue or physical absorption by gained
The blend of type i collagen, type III collagen or I/III Collagen Type VI adheres on the matte of biomembrane base material.
Described sterilizing both can be passed through the gamma-ray irradiation sterilizing of doses and kill virus, it is possible to by other
Method is effectively removed or inactivates animal virus, and the gamma-ray effective irradiation dosage understanding doses at present exists
More than 15kGy, but those skilled in the art can also select suitable irradiation dose according to actual requirement.
The acellular matrix source tissue engineering rack of the present invention is for as soft tissue healing material.This
Bright support can be used for the repairing after the soft tissue injurys such as oral mucosa, cartilage, cerebral dura mater, plays repair deficiency
And recover the effect of function.This support also can carry cell reparation after above-mentioned soft tissue injury.Due to
It preferably attaches performance, and this support can exempt from seam in operation, directly against being attached to soft tissue injury district.
The present invention has following beneficial effect relative to prior art: process preparation by above-mentioned special process
Obtain that there is excellent mechanical performances, preferable biocompatibility, relatively reduced immunogenicity, can repair along with defective tissue
Multiple gradually degraded and absorbed, controllable degradation speed, do not cause rejection, not as permanent foreign body after implantation
Exist, without the acellular matrix source tissue engineering scaffold structures of any residual toxicity；And this acellular matrix source
Tissue engineering bracket can be as cell (stem cell), the carrying carrier of somatomedin, it is possible to be prepared as different rule
Lattice, for the repairing of soft tissue injury and rebuild that (cartilage defect repair, oral mucosa are repaired, cerebral dura mater repaiies
Mend).
Accompanying drawing explanation
Fig. 1 is the cross-sectional view of the acellular matrix source tissue engineering rack that the present invention prepares
(wherein, 1 is biomembrane base material, and 2 is collagen protein weaker zone)；
Fig. 2 is the collagen protein weaker zone of the acellular matrix source tissue engineering rack that the present invention prepares
Scanning electron microscope (SEM) photograph.
Detailed description of the invention
In order to make the purpose of the present invention, technical scheme and beneficial effect clearer, below in conjunction with accompanying drawing
And embodiment, one of the present invention acellular matrix source tissue engineering rack and preparation method thereof is carried out in detail
Explanation.Specific embodiment described herein is only used for explaining the present invention, is not intended to limit the present invention.
Principle explanation
1. about biomembrane base material and collagen protein weaker zone
The animal membrane material of the present invention through go antigen to process and crosslinking fixing after, available a kind of mechanical strength is good,
The biomembrane base material that immunogenicity is low, degradation time is controlled, but itself there is hole in this biomembrane base material
The problems such as rate is relatively low, aperture is less.And animal tendon tissue is fixing through past antigen, crosslinking, collagen protein
Extract and after the process of purification, can get a kind of size tunable and be mainly made up of I, type III collagen protein
Collagen protein powder, collagen protein powder forms collagen solution, after vacuum drying, shape after water-soluble
Become the collagen protein weaker zone of sponge structure.Collagen protein weaker zone short texture, have certain aperture and
Porosity, but to still suffer from degradation time fast, without deficiencies such as mechanics support forces for this collagen protein weaker zone.
The preparation method of the present invention a kind of acellular matrix source tissue engineering rack, successfully by biomembrane base material with
Collagen protein weaker zone combines, and makes collagen protein weaker zone be adhesively fixed at the matte of biomembrane base material
On, it is achieved biomembrane base material and the mutual supplement with each other's advantages of collagen protein weaker zone.
Pliable and tough and smooth biomembrane base material both can maintain pattern and the structure of collagen weaker zone, the most permissible
Prevent surrounding connective tissue from growing into, play barrier and reduce the effect of articular surface friction, and biomembrane base material
The preferable mechanical property having can overcome simple collagen support poor mechanical property and the too fast shortcoming of degradation rate.
By what I type, type III or I/III Collagen Type VI formed, there is three-dimensional open structure collagen protein weaker zone,
Aperture is suitable, connects, can be conducive to sticking and growing and the transport of nutritional labeling of host cell between hole
With the discharge of metabolite, create good microenvironment for tissue regeneration reparation；And be also beneficial to cell and glue
Attached, breed and break up, provide good microenvironment for cell growth, more preferably induction and promote tissue repair regeneration；
Collagen protein itself has distinctive physical adsorbability, also functions to certain hemostasis and closes the effect of wound.
2. about removing antigen
Theoretical according to Immunology Today, the antigenicity of animal tissue is mainly by some specific position in protein
Active group and particular conformation cause, these active groups mainly-OH ,-NH2,-SH etc., and special
Isomery causes as then some special hydrogen bond mainly due to collagen molecules coiled strand.Processing animal membrane
During tissue, with one or more activating agent easily reacted with these active groups (as anhydride, acyl chlorides,
Amide, epoxide, halomethane etc.) be combined with these active groups, by the active group in collagen protein
It is closed, can effectively remove antigen.The while of so, then with the use of strong hydrogen bonding reagent (such as guanidine
Compound), displacement causes the hydrogen bond of particular conformation, changes collagen protein conformation, it is possible to remove antigen further.
3. about no-aldehyde fixative
Owing to animal membrane tissue is easily degraded by microorganisms or decomposes, it is therefore desirable to fixing with fixative crosslinking.
Traditional method is to use to have the glutaraldehyde of residual toxicity to make fixative, and the present invention selects epoxide, two acyls
Diamidogen, diisocyanate, Polyethylene Glycol or carbodiimides etc. easily crosslink with protein molecule react and
The no-aldehyde fixative of non-residual toxicity.Epoxide is as currently preferred no-aldehyde fixative, easily
Open loop cross-linking reaction, control reaction condition is occurred so that the cross-linking products arrived is the most stable, to degrade the most easily.
4. fixing about crosslinking
The group (hydroxyl and carboxyl etc.) that enlivens of cross-linking agent can be anti-with the amido generation in collagen molecules
Should, the webbed intermolecular and intramolecular crosslinking structure of shape, and this rock-steady structure can be preferably to anti-human
The enzyme action etc. of vivo protein enzyme, such that it is able to the degradation speed of the collagen protein that slows down.The present invention is non-by controlling
The concentration of aldehyde fixative and response time, the material of different crosslinking degree can be obtained, select suitably with this
Degree of cross linking material meet the clinical application demand to acellular matrix source tissue engineering scaffold material.Additionally,
It is fixing that the present invention can also omit crosslinking, prepares noncrosslinking acellular matrix source tissue engineering scaffold material.
In following example, described solution if no special instructions, is aqueous solution.In embodiment, guanidine hydrochloride
Solution is the Tris solution of guanidine hydrochloride.
Embodiment 1
A kind of acellular matrix source tissue engineering rack, as it is shown in figure 1, with animal membrane material by pretreatment,
The biomembrane base material 1 obtained after the step process such as defat, de-cell, fixing, the removal antigen of crosslinking, in institute
The surface adhesion stating biomembrane base material 1 is passed through pretreatment, defat, de-cell, crosslinking by animal tendon tissue
Fixing, remove antigen and glue that collagen protein extracts and the step such as purification prepares collagen protein is dried to obtain
Former albumen weaker zone 2.Wherein, described biomembrane base material 1 has matte and bright finish, institute's collagen protein weaker zone
Adhere on matte.
Embodiment 2
A kind of cartilage frame in acellular matrix source, with animal membrane material by pretreatment, defat, de-cell,
The biomembrane base material 1 obtained after the step process such as crosslinking is fixing, removal antigen, at described biomembrane base material 1
Surface adhesion by animal tendon tissue by pretreatment, defat, de-cell, crosslinking is fixing, remove antigen
And the collagen protein weaker zone that the collagen protein that collagen protein extracts and the step such as purification prepares is dried to obtain
2.Wherein said collagen protein weaker zone has hole, and porosity is 65～86%, the aperture of hole be 50～
500μm.Described collagen protein weaker zone is fixed on the matte of biomembrane base material by physical absorption.Or,
Described collagen protein weaker zone is adhesively fixed on the matte of biomembrane base material by biological fibrin glue.Preferably,
The thickness of described collagen protein weaker zone is 0.1～2.9mm.Described cartilage support material is square membrane-like
Structure.A length of the 10 of described cartilage support material～80mm, width is 5.0～60mm, thickness be 0.2～
3mm.Support described in the present embodiment can be as Autologous Chondrocyte, stem cell, somatomedin or medicine
Delivery vehicles, have simultaneously closely attach with cartilaginous tissue, histocompatibility is good, cell bearing capacity is strong and
Post operation can the progressively advantage such as degraded.
Embodiment 3
The preparation method of a kind of acellular matrix source tissue engineering rack of the present invention, comprises the following steps:
(1) pretreatment: take the equal animal pericardium of fresh quality and tendon tissue, cleans, and uses new
Geramine broad-spectrum high efficacy disinfectant solution soaks 15h, and removes fatty tissue, fiber and chorionic villi；
(2) defat: the acetone soln extracting extraction animal pericardium using mass concentration to be 3.0% and tendon
Tissue, to remove fat and oil-soluble impurities, extraction time is 24h, acetone soln and animal pericardium and
The mass ratio of tendon tissue is 10:1；
(3) de-cell: TritonX (Triton-100) the bond quality using mass concentration to be 0.1%
The pepsin (described concentration is final concentration) of concentration 0.1%, the ultrasonic digestion animal heart under the conditions of 30kHz
Peplos and tendon tissue 48h, to remove cell contained in tissue and fragment thereof completely；
(4) antigen is removed: use and the activating agent acyl of animal pericardium with tendon tissue mass ratio 20:1
Amine, reacts 48h under conditions of pH value is 7, and with being 20 with animal pericardium and tendon tissue mass ratio:
The 8mol/L guanidine hydrochloride solution of 1, under conditions of pH value is 8 react 24h, with replace animal pericardium and
In tendon tissue, the special hydrogen bond in collagen molecules coiled strand, is thoroughly removed the biomembrane after antigen
Base material and tendon tissue；
(5) crosslinking is fixing: use and animal pericardium and the polycyclic oxygen third that tendon tissue mass ratio is 20:1
Alkane solution, the mass concentration of poly(propylene oxide) solution is 0.5%, is 7 and under conditions of stirring at pH value,
React 2 days with the biomembrane base material after removing antigen and processing, the collagen protein in biomembrane base material is divided
Son cross-links；
(6) collagen protein extracts and purification: the acetic acid aqueous solution using mass concentration to be 0.2% dissolves and passes through
De-cell and remove the tendon tissue after antigen processes, smashs to pieces through tissue mashing machine, is then 3 at pH value
Acid condition under use mass concentration be 0.4% pepsin solution continue dissolve 2 days, temperature controls
10 DEG C, separate out collagen protein, filter, re-use the dodecyl sodium sulfate (SDS) that mass concentration is 0.5%
Solution soaking 24h, the lipid that may contain with removal, obtain collagen protein sterling after washing；
(7) collagen composite: collagen protein sterling is put in freezer dryer, vacuum condition is 10Pa,
Being dried 72h, use pulverizer to pulverize, ball mill is milled, and sieves, and takes particle size range below 200 μm
Collagen protein powder；Collagen protein powder is dissolved into the collagen solution of mass concentration 5%, obtains I type
Collagen, type III collagen or the blend of I/III Collagen Type VI, by the type i collagen obtained, type III collagen or
I/III Collagen Type VI mixture is laid on the matte of biomembrane base material, puts in freezer dryer, vacuum bar
Part is 10Pa, is dried 72h, obtains having double-deck tissue engineering bracket, through cutting out, and sizing, bag
Dress, is that 30kGy gamma-rays carries out irradiation sterilization with irradiation dose, obtains finished product.
Through the acellular matrix source tissue engineering scaffold material that above-mentioned preparation method obtains, there is degradation speed
Hurry up, the advantage such as many, the good biocompatibility of cell bearing capacity, it is adaptable to the tissue of reconstruction speed lacks
Damaging position, degradation time in vivo is postoperative 2 months to 6 months.
Embodiment 4
The preparation method of a kind of acellular matrix source tissue engineering rack of the present invention, comprises the following steps:
(1) pretreatment: take the equal animal pericardium of fresh quality and tendon tissue, cleans, and use is washed
4h must be soaked by safe broad-spectrum high efficacy disinfectant solution, and remove fatty tissue, fiber and chorionic villi；
(2) defat: use mass concentration be 3.5% carbon tetrachloride solution extracting extraction animal pericardium and
Tendon tissue, to remove fat and oil-soluble impurities, extraction time is 30h, carbon tetrachloride solution, animal
The mass ratio of pericardium and tendon tissue is 5:1；
(3) de-cell: carboxymethylamino methane (Tris) the connexus using mass concentration to be 0.5%
The papain (described concentration is final concentration) of amount concentration 0.3%, under the conditions of 50kHz, ultrasonic digestion is moved
Thing pericardium and tendon tissue 24h, to remove cell contained in tissue and fragment thereof completely；
(4) antigen is removed: use and the activating agent fourth of animal pericardium with tendon tissue mass ratio 10:1
Anhydride, reacts 24h under conditions of pH value is 8, and with animal pericardium and tendon tissue mass ratio is
The 6mol/L guanidine hydrochloride solution of 1:10, reacts 48h, to replace animal pericardium under conditions of pH value is 7
In film and tendon tissue, the special hydrogen bond in collagen molecules coiled strand, is thoroughly removed the life after antigen
Thing film base material and tendon tissue；
(5) crosslinking is fixing: use and animal pericardium and the polycyclic oxygen third that tendon tissue mass ratio is 20:1
Alcoholic solution, the mass concentration of polycyclic oxygen propanol solution is 3%, is 7 and under conditions of stirring at pH value, with
Biomembrane base material after removing antigen and processing reacts 5 days, to the collagen molecules in biomembrane base material
Cross-link；
(6) collagen protein extracts and purification: the aqueous hydrochloric acid solution using mass concentration to be 0.05% dissolves and passes through
De-cell and remove the tendon tissue after antigen processes, smashs to pieces through tissue mashing machine, is then 4 at pH value
Acid condition under use mass concentration be 0.2% papain solution continue dissolve 5 days, temperature control
At 15 DEG C, separate out collagen protein, filter, re-use the sodium deoxycholate solution that mass concentration is 0.1% and soak
48h, the lipid that may contain with removal, obtain collagen protein sterling after washing；
(7) collagen composite: put in freezer dryer by collagen protein sterling, vacuum condition is 0Pa, dry
Dry 48h, uses pulverizer to pulverize, and ball mill is milled, and sieves, and takes particle size range glue below 200 μm
Former protein powder；Collagen protein powder is dissolved into the collagen solution of mass concentration 2%, obtains I type glue
Former, type III collagen or the blend of I/III Collagen Type VI, by the type i collagen obtained, type III collagen or
I/III Collagen Type VI mixture is laid on the matte of biomembrane base material, puts in freezer dryer, vacuum bar
Part is 0Pa, is dried 48h, obtains having double-deck tissue engineering bracket, through cutting out, and sizing, packaging,
It is that 25kGy gamma-rays carries out irradiation sterilization with irradiation dose, obtains finished product.
Through the acellular matrix source tissue engineering scaffold material that above-mentioned preparation method obtains, there is bio-compatible
Property is good, the advantages such as immunogenicity is low, and degradation time is controlled, and cell bearing capacity is big, it is adaptable to reconstruction is inclined
Slow tissue defect site, can realize the using effect that tissue reconstruction is Tong Bu with timbering material degraded, internal fall
The solution time can be controlled in postoperative 6 months to 2 years.
Embodiment 5
The preparation method of a kind of acellular matrix source tissue engineering rack of the present invention, comprises the following steps:
(1) pretreatment: take the equal animal pericardium of fresh quality and tendon tissue, cleans, and uses new
Geramine broad-spectrum high efficacy disinfectant solution soaks 10h, and removes fatty tissue, fiber and chorionic villi；
(2) defat: using mass concentration is 2.5%1:1 chloroform alcohol solution extracting extraction animal pericardium
With the fat contained in tendon tissue and oil-soluble impurities, extraction time is 24h, and chloroform alcohol solution is with dynamic
The mass ratio of thing pericardium and tendon tissue is 5:1；
(3) de-cell: dodecyl sodium sulfate (SDS) the connexus using mass concentration to be 0.25%
Concentration described in the trypsin of amount concentration 0.25% is final concentration), ultrasonic digestion animal under the conditions of 40kHz
Pericardium and tendon tissue 48h, to remove cell contained in tissue and fragment thereof completely；
(4) antigen is removed: use and the activating agent halogen of animal pericardium with tendon tissue mass ratio 10:1
Methane, under conditions of pH value is 7.2 react 48h, and with animal pericardium and tendon tissue mass ratio
For the 3mol/L guanidine hydrochloride solution of 10:1, under conditions of pH value is 7.2, react 48h, dynamic with displacement
In thing pericardium and tendon tissue, the special hydrogen bond in collagen molecules coiled strand, is thoroughly removed antigen
After biomembrane base material and tendon tissue；
(5) crosslinking is fixing: use molten with animal pericardium and the alkylene oxide that tendon tissue mass ratio is 10:1
Liquid, the mass concentration of alkylene oxide solution is 3%, is 7 and under conditions of stirring at pH value, removes with passing through
Biomembrane base material after antigen processes reacts 8 days, cross-links the collagen molecules in biomembrane base material；
(6) collagen protein extracts and purification: the acetic acid aqueous solution using mass concentration to be 0.2% dissolves and passes through
De-cell and remove the tendon tissue after antigen processes, smashs to pieces through tissue mashing machine, is then 3 at pH value
Acid condition under use mass concentration be 0.3% pepsin solution continue dissolve 3 days, temperature controls
10 DEG C, separate out collagen protein, filter, re-use the dodecyl sodium sulfate (SDS) that mass concentration is 0.5%
Solution soaking 24h, the lipid that may contain with removal, obtain collagen protein sterling after washing；
(7) collagen composite: put in freezer dryer by collagen protein sterling, vacuum condition is 5Pa, dry
Dry 24h, uses pulverizer to pulverize, and ball mill is milled, and sieves, and takes particle size range glue below 200 μm
Former protein powder；Collagen protein powder is dissolved into the collagen solution of mass concentration 5%, obtains I type glue
Former, type III collagen or the blend of I/III Collagen Type VI, by the type i collagen obtained, type III collagen or
I/III Collagen Type VI mixture is laid on the matte of biomembrane base material, puts in freezer dryer, vacuum bar
Part is 5Pa, is dried 48h, obtains having double-deck tissue engineering bracket, through cutting out, and sizing, packaging,
It is that 35kGy gamma-rays carries out irradiation sterilization with irradiation dose, obtains finished product.
Embodiment 6
In the crosslinking technique for fixing of (5th) step of the inventive method, poly(propylene oxide), polycyclic can be selected
At least two in oxygen propanol or alkylene oxide；Non-epoxides cross-linking agent can also be selected, can be two acyls
One or more in diamidogen, diisocyanate, Polyethylene Glycol, carbodiimides etc., crosslinker concentration is
0.5%～5%, pH value be 7～8 and stirring under conditions of, react 2～8 days with biomembrane base material, can obtain
The biomembrane base material of different crosslinking degrees.
Embodiment 7
The acellular matrix source tissue engineering of the low crosslinking degree (0～10%) that the embodiment of the present invention 3 prepares
Timbering material, the heat shrink temperature of conventional method detection material is 56～69 DEG C, and the thickness of timbering material is
1.5～3.0mm.Wherein, the porosity of collagen protein weaker zone is 65～86%, and aperture is 50～300 μm,
Described acellular matrix source tissue engineering scaffold material has preferable cell bearing capacity.
Embodiment 8
The acellular matrix source tissue engineering of the low crosslinking degree (0～10%) that the embodiment of the present invention 3 prepares
Timbering material, the heat shrink temperature of conventional method detection material is 56～69 DEG C, and the thickness of timbering material is
0.5～1.0mm.Wherein, the porosity of collagen protein weaker zone is 65～86%, and aperture is 50～300 μm,
Described collagen protein is possible not only to preferably fit with the soft tissue of patient, but also can preferably be attached to
Tissue surface.
Embodiment 9
The acellular matrix source tissue engineering scaffold material of Example 3, for repairing of dog reconstruction of oral defect
Mend experiment, collagen protein weaker zone side is attached at dog reconstruction of oral defect district, simultaneously with silk suture auxiliary
Carrying out wrapping with gauze sheet fixing, after within postoperative 7 days, removing wrapping bag, timbering material is as Oral Repair film energy
Enough preferably being attached to defective region, 3 months after operation visual inspection defective region has been repaired complete, histological observation
Visible timbering material has begun to degraded, and repairing district has autologous epithelial and fibroblastic grow into, and
The collagenous tissue of visible new life.
As can be seen here, the acellular matrix source tissue engineering scaffold material that the present invention prepares has preferably
Biocompatibility and physics attaching property, its potential applicability in clinical practice is preferable.
Being shown by test, the acellular matrix source tissue engineering scaffold material that other embodiments obtain also has
Similar effect.
Embodiment 10
The acellular matrix source tissue engineering scaffold material of Example 4, by 1.0～4.0 × 106Individual/cm2's
Animal Autologous Chondrocyte is inoculated in the collagen egg of described acellular matrix source tissue engineering scaffold material by density
White weaker zone, for the repairing treatment of goat defects of knee, 3 months after operation after inoculating 2～4 hours
Visible defective region is repaired the most completely, the existence of chondrocyte seen from histological observation patch area, and implant
The existing Partial digestion of frame material, and generated by newborn collagen fiber.
As can be seen here, the acellular matrix source tissue engineering scaffold material that the present invention prepares has preferably
Biocompatibility and degradability, have certain potential applicability in clinical practice.
Being shown by test, the acellular matrix source tissue engineering scaffold material that other embodiments obtain also has
Similar effect.
Embodiment 11
The acellular matrix source tissue of the middle high-crosslinking-degree (11～35%) that the embodiment of the present invention 4 prepares
Engineering scaffold material, the heat shrink temperature of conventional method detection material is 70～90 DEG C, the thickness of timbering material
It is 0.5～1.5mm.Wherein, the porosity of collagen protein weaker zone is 65～86%, and aperture is 50～300
μm, described timbering material collagen layer is made up of collagen protein, itself has preferably absorption and bonding effect,
And have certain hemostasis and close the clinical effectiveness of wound.This timbering material basal layer has the friendship of middle high level
Connection degree, has degradation speed the most slowly in vivo, and about 2 years the most after surgery the most degradable.
Embodiment 12
The acellular matrix source tissue engineering scaffold material of Example 4, by 1.0～4.0 × 106Individual/cm2's
People source chondrocyte is seeded in the collagen egg of unit are acellular matrix source tissue engineering scaffold material by density
White weaker zone, inoculates latter 30 minutes, and unit are acellular matrix source tissue engineering scaffold material is learnt in detection
Collagen protein weaker zone adhere to cell number account for more than the 90% of total inoculating cell number.
As can be seen here, the acellular matrix source tissue engineering scaffold material that embodiment 4 prepares has preferably
Cell bearing capacity.
Being shown by test, the acellular matrix source tissue engineering scaffold material that other embodiments obtain also has
Similar effect.
Embodiment 13
The acellular matrix source tissue engineering scaffold material of Example 4, as exempting to stitch meninges sticking patch for dog
The reparative experiment of dura defect, is attached at dog cerebral dura mater face, biomembrane base material by collagen protein weaker zone side
Layer is towards skull face, and the area of timbering material is greater than defect of meninges district, i.e. with defect meninges at normal structure
Imbricate about 3～5mm, does not do the stitching of repair materials and meningeal tissue, successively closes scalp.Postoperative 6
Week opens field of operation, it is seen that timbering material with the bonding of dog autologous meningeal tissue firmly, leaks outside without cerebrospinal fluid etc.；
Meanwhile, timbering material substrate side and skull, muscle are all without any adhesion, smooth surface, collagen layer side and dog
Cerebral tissue is without any adhesion, smooth surface.Timbering material seen from histological observation and dog cerebral tissue, flesh
Meat etc. are without any adhesion point, and collagen layer is the most degradable, have a little fibroblast to grow into, and NIP reacts.
As can be seen here, the acellular matrix source tissue engineering scaffold material that the present invention prepares has preferably
Bonding effect, has preferable biocompatibility and structural stability, may be used in meninges repairing operation.
Being shown by test, the acellular matrix source tissue engineering scaffold material that other embodiments obtain also has
Similar effect.
In sum, animal membrane material and tendon tissue are after the present invention processes, and structural stability is higher, exempts from
Epidemic focus is extremely low, is applicable to embedded material and uses.Through repeatedly experiment in vitro and et al. Ke experimental verification,
The acellular matrix source tissue engineering rack of the present invention has stronger cell bearing capacity, can be by substantial amounts of
Cell or somatomedin are transported to soft tissue injury region, are greatly facilitated reparation and the tissue weight of soft tissue injury
Building, this using method is equally applicable to the clinical practice of other tissue engineering products.As in figure 2 it is shown, this
Bright acellular matrix source tissue engineering rack also has preferable physical absorption effect, can be with autologous patient
Mucosal tissue, cerebral dura mater tissue adhesion together, as Oral Repair film, are exempted from seam type dural patch etc. and are faced
Bed operation application.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by above-mentioned reality
Execute the restriction of example, the change made under other any spirit without departing from the present invention and principle, modification,
Substitute, combine, simplify, all should be the substitute mode of equivalence, within being included in protection scope of the present invention.
Claims (16)
1. an acellular matrix source tissue engineering rack, it is characterised in that: after processing
Animal membrane material is dredged as biomembrane base material, the surface adhesion collagen protein at described biomembrane base material
Pine layer.
A kind of acellular matrix source tissue the most according to claim 1 engineering rack, its
It is characterised by: described collagen protein weaker zone is to be prepared by animal tendon tissue.
A kind of acellular matrix source tissue the most according to claim 2 engineering rack, its
It is characterised by: described biomembrane base material has matte and bright finish, institute's collagen protein weaker zone to adhere to
On matte.
A kind of acellular matrix source tissue the most according to claim 1 engineering rack, its
Be characterised by: the thickness of described collagen protein weaker zone is 0.1～3.0mm, porosity be 65～
86%, aperture is 50～300 μm.
5. according to the arbitrary described acellular matrix source tissue engineering rack of claim 1-4
Preparation method, it is characterised in that: the step included is as follows:
(1) preparation of biomembrane base material；
(2) preparation of collagen protein；
(3) collagen composite: adhered to by collagen protein on the matte of biomembrane base material, is dried.
The system of a kind of acellular matrix source tissue the most according to claim 5 engineering rack
Preparation Method, it is characterised in that: the step included is as follows:
(1) pretreatment: take fresh animal membrane material and tendon tissue, cleans, sterilization, and
Remove fatty tissue, fiber and chorionic villi；
(2) defat: use organic solvent extracting extraction animal membrane material and tendon tissue, remove
Fat and oil-soluble impurities；
(3) de-cell: use surfactant solution associated proteins enzymatic solution, control table
The quality final concentration of 0.1～0.5% of face activating agent, the quality final concentration 0.1 of protease～
0.3%, ultrasonic digestion animal membrane material and tendon tissue；
(4) antigen is removed: use activating agent, with animal under the conditions of pH value is 7～8
Film material and tendon tissue react 24～48h, then use strong hydrogen bonding reagent, are 7～8 at pH value
Under the conditions of react 24～48h, after thoroughly being removed antigen with animal membrane material and tendon tissue
Biomembrane base material and tendon tissue；
(5) crosslinking is fixing: the no-aldehyde chemical cross-linking agent using mass concentration to be 0.5～5%
Solution, under pH value is 7～8 stirring conditions, with the biomembrane after removing antigen and processing
Base material reacts 2～8 days；
(6) collagen protein extract and purification: use mass concentration be 0.05～0.2% acid molten
Liquid dissolves the tendon tissue after removing antigen and processing, and smashs to pieces, is then 3～5 at pH value
Use mass concentration 0.1～the protein enzyme solution continuation dissolving of 0.4% under acid condition, separate out glue
Former albumen, filters, and re-uses the surfactant solution immersion that mass concentration is 0.1～0.5%,
Collagen protein sterling is obtained after washing；
(7) collagen composite: make collagen protein powder after collagen protein sterling being vacuum dried,
Crosslinked fixing biomembrane it is laid in after collagen protein powder is dissolved into collagen solution
On the matte of base material, vacuum drying.
The system of a kind of acellular matrix source tissue the most according to claim 6 engineering rack
Preparation Method, it is characterised in that: described animal membrane material is a kind of in pericardium, goldbeater's skin or fat nethike embrane
Or it is two or more.
The system of a kind of acellular matrix source tissue the most according to claim 6 engineering rack
Preparation Method, it is characterised in that: described activating agent be small molecular organic acid acid anhydride, acyl chlorides, amide,
In epoxide or halomethane one or more；Described strong hydrogen bonding reagent is guanidine chemical combination
Thing.
The system of a kind of acellular matrix source tissue the most according to claim 8 engineering rack
Preparation Method, it is characterised in that: guanidine compound is guanidine hydrochloride solution, described guanidine hydrochloride solution
Concentration is 3～8mol/L.
The system of a kind of acellular matrix source tissue the most according to claim 6 engineering rack
Preparation Method, it is characterised in that: described no-aldehyde chemical cross-linking agent be epoxide, two acid diamides,
One or more in diisocyanate, Polyethylene Glycol or carbodiimide reagent.
11. a kind of acellular matrix source tissue according to claim 10 engineering racks
Preparation method, it is characterised in that: described epoxide is monoepoxideDicyclo
OxideOr one or more in low polyepoxide, wherein
R=H or CnH2n+1-, n=0-10.
12. a kind of acellular matrix source tissue according to claim 11 engineering racks
Preparation method, it is characterised in that: described low polyepoxide is poly(propylene oxide), polycyclic oxygen third
One or more in alcohol or alkylene oxide.
The system of 13. a kind of acellular matrix source tissue according to claim 6 engineering racks
Preparation Method, it is characterised in that: animal membrane material and tendon tissue quality, organic solvent, vivaciously try
Agent, strong hydrogen bonding reagent, no-aldehyde chemical crosslinking agent solution mass ratio be 1:(5～10): (10～
20): (10～20): (10～20).
The system of 14. a kind of acellular matrix source tissue according to claim 6 engineering racks
Preparation Method, it is characterised in that: described acid solution is hydrochloric acid and/or acetic acid aqueous solution.
The system of 15. a kind of acellular matrix source tissue according to claim 6 engineering racks
Preparation Method, it is characterised in that: described organic solvent be acetate esters, chloroform, carbon tetrachloride,
In ether, acetone or dehydrated alcohol one or more；Described surfactant be TritonX,
Dodecyl sodium sulfate, 3-[(3-gallbladder amidopropyl) dimethylammonio]-1-propane sulfonic acid salt,
In NaTDC or carboxymethylamino methane one or more；Described protease is pancreas egg
In white enzyme, pepsin or papain one or more.
16. prop up according to the arbitrary described a kind of acellular matrix source tissue engineering of claim 1-4
Frame is for as soft tissue healing material.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1986007A (en) * | 2005-12-20 | 2007-06-27 | 广东冠昊生物科技有限公司 | Biological surgical patch |
| CN101172166A (en) * | 2007-11-16 | 2008-05-07 | 广东冠昊生物科技有限公司 | Ocular surface biomembrane and preparation method thereof |
| EP2079490B1 (en) * | 2006-10-23 | 2012-08-29 | Cook Biotech Incorporated | Processed ecm materials with enhanced component profiles |
| CN104117096A (en) * | 2013-04-23 | 2014-10-29 | 王伟 | Novel composite biological dura mater |
| CN205073351U (en) * | 2015-09-18 | 2016-03-09 | 广州市美昊生物科技有限公司 | Take off oral cavity patch in cell matrix source |
-
2016
- 2016-03-29 CN CN201610188734.6A patent/CN105935454A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1986007A (en) * | 2005-12-20 | 2007-06-27 | 广东冠昊生物科技有限公司 | Biological surgical patch |
| EP2079490B1 (en) * | 2006-10-23 | 2012-08-29 | Cook Biotech Incorporated | Processed ecm materials with enhanced component profiles |
| CN101172166A (en) * | 2007-11-16 | 2008-05-07 | 广东冠昊生物科技有限公司 | Ocular surface biomembrane and preparation method thereof |
| CN104117096A (en) * | 2013-04-23 | 2014-10-29 | 王伟 | Novel composite biological dura mater |
| CN205073351U (en) * | 2015-09-18 | 2016-03-09 | 广州市美昊生物科技有限公司 | Take off oral cavity patch in cell matrix source |
Cited By (29)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN111359020A (en) * | 2018-12-26 | 2020-07-03 | 深圳兰度生物材料有限公司 | Soft tissue repair material and preparation method and application thereof |
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