CN109078226A - A kind of de- cell porcine aorta matrix in micropore - Google Patents
A kind of de- cell porcine aorta matrix in micropore Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3641—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the site of application in the body
- A61L27/3679—Hollow organs, e.g. bladder, esophagus, urether, uterus, intestine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3691—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/22—Materials or treatment for tissue regeneration for reconstruction of hollow organs, e.g. bladder, esophagus, urether, uterus
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Abstract
The present invention provides a kind of micropores to take off cell porcine aorta matrix, have a feature in that micropore takes off cell porcine aorta matrix and has microcellular structure, the shape of the micropore of microcellular structure is round or oval, wherein, micropore take off cell porcine aorta matrix be by porcine aorta is placed under vacuum conditions in low concentration of detergent buffer carry out thawing after successively carry out concussion cleaning in low concentration of detergent buffer, aseptic deionized water and phosphate solution again after obtain.Micropore provided by the present invention takes off the three-dimensional structure and biomechanical property that cell porcine aorta matrix effectively remains porcine aorta tissue, and there is no residual detergent toxicity, it can promote fibroblast and vascular endothelial cell infiltrative growth, to promote blood vessel and collagen newborn, there is good internal remoldability.
Description
Technical field
The present invention relates to field of tissue engineering technology, and in particular to a kind of de- cell porcine aorta matrix in micropore.
Background technique
It is not only developed country with the vascular conditions that aortic aneurysm, vascular dissection, Atherosclerosis turn to representative, still
One of primary lethal lesion of developing country.Surgical replacement lesion vessels are the final hands for the treatment of vascular diseases
Section, thus need various vascular replacement materials.In addition, increasing year by year with Incidence of esophageal cancer, artificial esophagus displacement material
Demand it is also more and more.
Although homologue displacement is ideal displacement material, source is few, there are ethics limitations.And currently with life
The development of object science and technology has developed a variety of high-molecular biologic synthetic materials as artificial organ displacement material.For example, PET
(Polyethylene terephthalate, Dacron) and ePTFE (Expanded polytetra-
Fluoroethylene), polyurethane (Polyurethane) is because it is with preferable biomechanical property and due to internal compatibility
It is widely used in cardiovascular surgical procedure.But synthetic material also has limitation, such as synthesising biological material in clinical application
It is to increase mortality an important factor for leading to graft failure with natural tissues mismatch.In addition, lasting chronic inflammation is anti-
The risks such as caused transplanting tissue endometrial hyperplasia, chronic infection are answered to increase the incidence that decays of transplanting later period.
Therefore, a kind of similar with Normal aorta, oesophagus substitute in institutional framework and function is researched and developed to closing weight
It wants.Based on this reason, cell porcine aorta is taken off due to it substantially, under microscope and in physiological function is being similar to human aorta
It receives more and more attention.Also, there is research surface porcine aorta functionally also to can replace oesophagus.
De- cell is to reduce transplanting tissue immunogenicity in Tissue Engineering Study, retain graft engineering three-dimensional tissue structures, life
Object mechanical property, the important method of bioactivity, also, method for removing cells has been successfully applied to heart valve, prostate, angle
In the Tissue Engineering Studies such as film, heel string.It is cell free to realize that the prior art generally handles porcine aorta using detergent
Purpose, although this method can obtain good de- cell effect, the de- cell porcine aorta that obtains by this method
Matrix often remaining toxicity, and its biomechanical property is also destroyed.In addition, a large number of studies show that cell free pig master
The extra-cellular matrix structure of artery is fine and close, and therefore, cell is difficult endogenous growth when applied in host, the weight in host
It moulds ineffective.
Summary of the invention
The present invention is to carry out to solve the above-mentioned problems, and it is an object of the present invention to provide a kind of micropore takes off cell porcine aorta
Matrix.
The present invention provides a kind of micropores to take off cell porcine aorta matrix, has a feature in that micropore takes off cell
Porcine aorta matrix has microcellular structure, and the shape of the micropore of microcellular structure is round or oval, wherein micropore takes off cell
Porcine aorta matrix is to carry out thawing by the way that porcine aorta is placed under vacuum conditions in low concentration of detergent buffer
It is obtained after successively carrying out concussion cleaning in low concentration of detergent buffer, aseptic deionized water and phosphate solution again afterwards
's.
It takes off in cell porcine aorta matrix, is can also have the following features: wherein in micropore provided by the invention, it is micro-
Hole is not of uniform size, and the diameter maximum of micropore is up to 70 μm.
It takes off in cell porcine aorta matrix, is can also have the following features: wherein in micropore provided by the invention, it is low
Concentration detergent buffer includes Tris buffer, lauryl sodium sulfate and NaTDC, the concentration of Tris buffer
Range is 8~12mmol/L, and pH value is 7.4~7.8, and lauryl sodium sulfate is in the volume fraction range of Tris buffer
0.1%~0.5%W/V, NaTDC are 0~0.8%W/V in the volume fraction range of Tris buffer.
It takes off in cell porcine aorta matrix, is can also have the following features: wherein in micropore provided by the invention, it will
Porcine aorta, which is placed under vacuum conditions in low concentration of detergent buffer, carries out 2~10 freeze thawing, when the freezing of each freeze thawing
Between be 2~6h.
It takes off in cell porcine aorta matrix, is can also have the following features: wherein in micropore provided by the invention, warp
Porcine aortic valve after crossing freeze thawing carries out concussion processing in low concentration of detergent buffer and needs to shake in 30~50 DEG C of shaking tables
Swing 28~96h of processing.
It takes off in cell porcine aorta matrix, is can also have the following features: wherein in micropore provided by the invention, it is right
Porcine aorta aseptic deionized water shakes 12~48h of cleaning.
It takes off in cell porcine aorta matrix, is can also have the following features: wherein in micropore provided by the invention, it is right
Porcine aorta phosphate solution shakes 48~96h of cleaning.
The action and effect of invention
Related micropore takes off cell porcine aorta matrix according to the present invention, because having microcellular structure, microcellular structure
The shape of micropore be round or oval, micro-pore diameter is not of uniform size, and maximum gauge is up to 70 μm, such microcellular structure energy
Enough promote detergent intrusion organization internal to remove cell, and then the use concentration of detergent can be reduced, is effectively retained pig actively
The three-dimensional structure and biomechanical property of arteries and veins tissue, and significantly reduce the residual toxicity of detergent.In addition, such micropore knot
Structure can promote fibroblast and vascular endothelial cell infiltrative growth, to promote blood vessel and collagen newborn, so that micropore
Gap takes off cell porcine aorta matrix has good remoldability in host.Therefore, micropore takes off cell porcine aorta matrix
It is highly suitable for the transplanting of the clinical tissues engineering such as blood vessel, oesophagus.
In addition, due to this preparation method be using porcine aorta as research object, this preparation method it is low in cost,
The valuable medical resource for saving several hundred million members is engineering blood vessel, the clinical application of oesophagus and the optimization of acellular matrix
Research and development provide experimental basis and pointer.
Detailed description of the invention
Fig. 1 is the schematic diagram that porcine aorta and micropore take off cell porcine aorta matrix in the embodiment of the present invention；
Fig. 2 is the scanning electron microscope signal that porcine aorta and micropore take off cell porcine aorta matrix in the embodiment of the present invention
Figure；And
Fig. 3 is the histology signal that porcine aorta and micropore take off cell porcine aorta matrix in the embodiment of the present invention
Figure.
Specific embodiment
It is real below in order to be easy to understand the technical means, the creative features, the aims and the efficiencies achieved by the present invention
Example combination attached drawing is applied to be specifically addressed the de- cell porcine aorta matrix in micropore of the present invention.
Fig. 1 is the schematic diagram that porcine aorta and micropore take off cell porcine aorta matrix in the embodiment of the present invention.
Fig. 2 is the scanning electron microscope signal that porcine aorta and micropore take off cell porcine aorta matrix in the embodiment of the present invention
Figure.
Fig. 3 is the histology signal that porcine aorta and micropore take off cell porcine aorta matrix in the embodiment of the present invention
Figure.
As shown in Figures 1 to 3, Fig. 1 (a), Fig. 2 (a), Fig. 3 (a) indicate normal porcine aorta, Fig. 1 (b), Fig. 2 (b), Fig. 3
(b) indicate that micropore takes off cell porcine aorta matrix.As shown in Figures 1 to 3, the micropore in the present embodiment takes off cell porcine aorta
Matrix has continuous microcellular structure.It is not of uniform size that the shape of the micropore of microcellular structure is round or oval micro-pore diameter, most
Major diameter is up to 70 μm.Micropore takes off cell porcine aorta matrix and is free of cell and cell fragment.
The preparation process that micropore in the present embodiment takes off cell porcine aorta matrix is as follows:
Porcine aorta is placed in 2~6h of freezing in low concentration of detergent buffer under vacuum conditions by step 1, low dense
Spending detergent buffer includes: Tris buffer, lauryl sodium sulfate and NaTDC.The concentration model of Tris buffer
It encloses for 8~12mmol/L, pH value is 7.4~7.8, and lauryl sodium sulfate is in the volume fraction range of Tris buffer
0.1%~0.5%W/V, NaTDC are 0~0.8%W/V in the volume fraction range of Tris buffer.In the present embodiment
In, the proportion of buffer, lauryl sodium sulfate and NaTDC are as follows: the concentration of Tris buffer is 10mmol/L, PH
It is 7.6, the deoxycholic acid of 0.1~0.4g lauryl sodium sulfate and 0.1~0.5g is added in the Tris buffer of every 100mL
Sodium.Cooling time of the porcine aorta in low concentration of detergent buffer is 4~5h.
Step 2 after freezing, the porcine aorta after freezing is placed in the water-bath that temperature is 30~50 DEG C and is dissolved.
Step 3 repeats step 1 and step 2 1~10 time, porcine aorta is carried out thawing.
Porcine aorta after thawing is taken out from low concentration of detergent buffer, is then placed by step 4
In dirty agent buffer, and 28~96h of processing concussion in 30~50 DEG C of shaking tables.
Step 5, with aseptic deionized water to 12~48h of porcine aorta concussion cleaning after concussion.In the present embodiment,
Aseptic deionized water is 24~36h to the concussion scavenging period of porcine aorta, and the preferably concussion time is for 24 hours.
Step 6 obtains after 48~96h is cleaned in the porcine aorta concussion after being cleaned with phosphate solution to aseptic deionized water
To the acellular matrix of porcine aorta cell, i.e. micropore takes off cell porcine aorta matrix.In the present embodiment, phosphate solution
Concussion scavenging period to porcine aorta is 72h.
Cell porcine aorta matrix is taken off as experimental group, with normal porcine aorta (Tris using the micropore in the present embodiment
Buffer synchronization process) it is control group, it carries out structure and performance measurement, obtained result is as follows:
(1) micropore takes off cell porcine aorta matrix without cell and cell fragment
DNA quantitative detection result are as follows: the DNA content of control group is 1254.2 ± 263.9ng/mg wet tissue weight；Experimental group
DNA content be 497.5 ± 323.6ng/mg wet tissue weight.DNA content in experimental group is significantly lower than control group (P < 0.05),
Illustrate that micropore takes off cell porcine aorta matrix DNA content and significantly reduces, the result and result shown in Fig. 3 are completely the same, i.e.,
Contain many cells in normal porcine aorta, and micropore takes off cell porcine aorta matrix and does not contain cell and cell fragment.
(2) micropore, which takes off cell porcine aorta matrix, has continuous microcellular structure
Surface sweeping Electronic Speculum result is as shown in Fig. 2, the institutional framework of control group is very close, and experimental group is in loose spongy, tool
There is continuous microcellular structure.
(3) the de- cell porcine aorta matrix in micropore is able to maintain porcine aorta biomechanical property and three-dimensional structure
Biomechanical property testing result is as shown in table 1:
Table 1
As shown in table 1, long axis biomechanical analysis shows: compared with the control group, the maximum load of experimental group, maximum stress
And change (P > 0.05) of the elasticity modulus without obvious statistical significance.
Horizontal axis biomechanical analysis shows compared with the control group, maximum load, maximum stress and the springform of experimental group
Amount rises.
(4) micropore takes off cell porcine aorta matrix no cytotoxicity
Extracting cytotoxicity detection shows compared with the control group, experimental group can remarkably promote huve cell and
The growth of mesenchymal stem cells.The growth ability of experimental group huve cell is the 1.5 of normal incubation medium Cell growth ability
± 0.1 times, and the growth ability of cellular control unit is only 0.2 ± 0.1 times of normal incubation medium Cell growth ability.Between experimental group
The growth ability of mesenchymal stem cells is 1.1 ± 0.1 times of normal incubation medium Cell growth ability, and the growth energy of cellular control unit
Power is only 0.5 ± 0.1 times of normal incubation medium Cell growth ability.Experimental group is compared with control group has apparent statistics meaning
Adopted (P < 0.05).
Exposing cell toxicity detection shows compared with the cell that control group co-cultures, in the umbilical vein that experimental group co-cultures
The upgrowth situation of chrotoplast and mescenchymal stem cell is good.
(5) in vivo, the calcification and inflammatory reaction of the de- cell porcine aorta matrix in micropore are light
Control group and experimental group are subjected to subcutaneous rat embedding respectively, detection calcification is taken out in particular point in time and inflammation is anti-
It answers.
Calcification the result shows that: at the 28th day, the calcification of control group was the most serious, and calcium deposition amount is 71.2 ± 15.9/ single
Position；The calcification degree of experimental group is significantly lighter than control group, is 19.2 ± 5.4/ units, the two, which is compared, has apparent statistics meaning
Adopted (P < 0.05).
Inflammatory reaction the result shows that: control group mainly result in the T cell chronic lymphocytic based on CD4 and CD8 infiltration；Experimental group
Acute inflammatory reaction is light, it is early to disappear, and T lymphocyte infiltration degree is lower than control group.28th day, CD4 in control group and
The quantity of cd8 cell is respectively 106.0 ± 20.5/high power field of view, 109.3 ± 14.3/high power field of view；And experimental group
In CD4 and cd8 cell quantity then be respectively 61.2 ± 13.4/high power field of view, 57.8 ± 30.8/high power lens view
It is wild.Experimental group is compared with control group has apparent statistical significance (P < 0.05).
(6) in vivo, the blood vessel and tissue remodeling of the de- cell porcine aorta matrix in micropore are significant
Control group and experimental group are subjected to subcutaneous rat embedding respectively, carry out haematoxylin Yihong after particular point in time taking-up
(HE), immunohistochemical staining and collagen staining detection.
HE and immunohistochemical staining the result shows that: at 28 days, in the porcine aorta of control group myofibroblast only
In transplanted abdominal surrounding wetting, artery center is cell-free；It is female that the micropore of experimental group takes off muscle fibre in cell porcine aorta matrix
Cell is uniformly distributed in each layer of artery, and transplanted abdominal periphery and central area myofibroblast number are respectively 127.5 ± 20.3
A/high power field of view, 89.7 ± 15.0/high power field of view.
Myofibroblast is the important cells for secreting collagen, and consistent with myofibroblast distribution, collagen staining shows:
Control group only have around transplanted abdominal virgin rubber original shape at, artery center without virgin rubber original shape at；Experimental group periarterial and in
Centre region has newborn collagen to be formed.
Immunohistochemistry results show: at 28 days, new vessels are only in transplanted abdominal surrounding wetting, artery in control group
Center is without blood vessel；New vessels are uniformly distributed in each layer of artery, transplanted abdominal periphery and central area number of blood vessel point in experimental group
It Wei not 48.3 ± 8.2/high power field of view, 39.3 ± 11.9/high power field of view.
The action and effect of embodiment
Related micropore takes off cell porcine aorta matrix according to the present invention, because having microcellular structure, microcellular structure
The shape of micropore be round or oval, micro-pore diameter is not of uniform size, and maximum gauge is up to 70 μm, such microcellular structure energy
Enough promote detergent intrusion organization internal to remove cell, and then the use concentration of detergent can be reduced, is effectively retained pig actively
The three-dimensional structure and biomechanical property of arteries and veins tissue, and significantly reduce the residual toxicity of detergent.In addition, such micropore knot
Structure can promote fibroblast and vascular endothelial cell infiltrative growth, to promote blood vessel and collagen newborn, so that micropore
Gap takes off cell porcine aorta matrix has good remoldability in host.Therefore, micropore takes off cell porcine aorta matrix
It is highly suitable for the transplanting of the clinical tissues engineering such as blood vessel, oesophagus.
In addition, due to this preparation method be using porcine aorta as research object, this preparation method it is low in cost,
The valuable medical resource for saving several hundred million members is engineering blood vessel, the clinical application of oesophagus and the optimization of acellular matrix
Research and development provide experimental basis and pointer.
In addition, in the above-described embodiments, because preparation micropore takes off the low concentration decontamination that cell porcine aorta matrix uses
Agent buffer is low concentration sodium lauryl sulfate buffer, includes Tris buffer, lauryl sodium sulfate in the buffer
And NaTDC, so, the cell and nuclear material of porcine aorta tissue can be effectively removed, reduce internal inflammatory reaction
And calcium deposition.
Above embodiment is preferred case of the invention, the protection scope being not intended to limit the invention.
Claims (7)
1. a kind of micropore takes off cell porcine aorta matrix, it is characterised in that:
The micropore, which takes off cell porcine aorta matrix, has microcellular structure,
The shape of the micropore of the microcellular structure is round or oval,
Wherein, the micropore, which takes off cell porcine aorta matrix, is gone by the way that porcine aorta to be placed in low concentration under vacuum conditions
It is successively molten in low concentration of detergent buffer, aseptic deionized water and phosphate again after progress thawing in dirty agent buffer
It carries out obtaining after concussion cleaning in liquid.
2. micropore according to claim 1 takes off cell porcine aorta matrix, it is characterised in that:
Wherein, the micropore size is different, and the diameter maximum of the micropore is up to 70 μm.
3. micropore according to claim 1 takes off cell porcine aorta matrix, it is characterised in that:
Wherein, the low concentration of detergent buffer includes Tris buffer, lauryl sodium sulfate and NaTDC,
The concentration range of the Tris buffer is 8~12mmol/L, and pH value is 7.4~7.8,
The lauryl sodium sulfate is 0.1%~0.5%W/V in the volume fraction range of the Tris buffer,
The NaTDC is 0~0.8%W/V in the volume fraction range of the Tris buffer.
4. micropore according to claim 1 takes off cell porcine aorta matrix, it is characterised in that:
Wherein, the porcine aorta is placed under vacuum conditions in low concentration of detergent buffer and carries out 2~10 freeze thawing, often
The cooling time of secondary freeze thawing is 2~6h.
5. micropore according to claim 1 takes off cell porcine aorta matrix, it is characterised in that:
Wherein, the porcine aortic valve after freeze thawing carries out concussion processing needs 30 in low concentration of detergent buffer
28~96h of concussion processing in~50 DEG C of shaking tables.
6. micropore according to claim 1 takes off cell porcine aorta matrix, it is characterised in that:
Wherein, 12~48h of cleaning is shaken to porcine aorta aseptic deionized water.
7. micropore according to claim 1 takes off cell porcine aorta matrix, it is characterised in that:
Wherein, 48~96h of cleaning is shaken to porcine aorta phosphate solution.
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Cited By (5)
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| CN110464877A (en) * | 2019-06-25 | 2019-11-19 | 滨州医学院 | A kind of preparation method and its effect evaluation method of acellular nerve allografts |
| CN111434358A (en) * | 2019-12-30 | 2020-07-21 | 广东泓志生物科技有限公司 | Preparation method of collagen scaffold and collagen scaffold |
| CN111905148A (en) * | 2020-07-22 | 2020-11-10 | 扬州大学 | Method for rapidly preparing rabbit acellular tracheal matrix without immunogenicity under vacuum assistance |
| CN113425905A (en) * | 2020-03-23 | 2021-09-24 | 成都中科奥格生物科技有限公司 | Blood vessel material and preparation method and application thereof |
| CN113425907A (en) * | 2020-03-23 | 2021-09-24 | 四川大学 | Pericardium material and preparation method and application thereof |
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| CN110464877A (en) * | 2019-06-25 | 2019-11-19 | 滨州医学院 | A kind of preparation method and its effect evaluation method of acellular nerve allografts |
| CN111434358A (en) * | 2019-12-30 | 2020-07-21 | 广东泓志生物科技有限公司 | Preparation method of collagen scaffold and collagen scaffold |
| CN111434358B (en) * | 2019-12-30 | 2021-09-21 | 广东泓志生物科技有限公司 | Preparation method of collagen scaffold and collagen scaffold |
| CN113425905A (en) * | 2020-03-23 | 2021-09-24 | 成都中科奥格生物科技有限公司 | Blood vessel material and preparation method and application thereof |
| CN113425907A (en) * | 2020-03-23 | 2021-09-24 | 四川大学 | Pericardium material and preparation method and application thereof |
| CN111905148A (en) * | 2020-07-22 | 2020-11-10 | 扬州大学 | Method for rapidly preparing rabbit acellular tracheal matrix without immunogenicity under vacuum assistance |
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