CN102600508A - Porcine arterial vacuum lyophilization acellular matrix, as well as preparation method and application thereof - Google Patents
Porcine arterial vacuum lyophilization acellular matrix, as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN102600508A CN102600508A CN2012100319696A CN201210031969A CN102600508A CN 102600508 A CN102600508 A CN 102600508A CN 2012100319696 A CN2012100319696 A CN 2012100319696A CN 201210031969 A CN201210031969 A CN 201210031969A CN 102600508 A CN102600508 A CN 102600508A
- Authority
- CN
- China
- Prior art keywords
- blood vessel
- lyophilizing
- pbs solution
- drying
- porcine aorta
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Abstract
The invention provides a porcine arterial vacuum lyophilization acellular matrix, as well as a preparation method and application thereof. The preparation method of the porcine arterial vacuum freeze-drying acellular matrix comprises the following steps: performing vacuum lyophilization to the porcine arterial blood vessels to obtain the best pore by adopting an optimal pre-freezing velocity; rehydrating the matrix with low-concentration detergent and performing acellular treatment; and lyophilizing the matrix and storing at normal temperature. The matrix has the advantages that the porcine aorta is used as a studying object, and a lyophilized acellular blood vessel with proper porosity, mechanical performance and immunogenicity is produced by virtue of optimized preparation conditions; animal in-vivo experiments prove the practicality of three-dimensional blood vessel clinical vascular transplantation, and experimental basis is provided to further clinical application; the preparation method has extremely low cost, so that precious medical resources reach billions of RMB can be saved assuredly; and experimental basis and guidance are provided to clinical application of tissue engineering arterial blood vessels and optimizing research and development of acellular blood vessels.
Description
Technical field
The present invention relates to field of tissue engineering technology, specifically, is a kind of porcine aorta vacuum freeze-drying acellular matrix.
Background technology
Annual newborn 150,000 the children with congenital heart disease of China.And need a large amount of blood vessels or biological sticking patch substitute in the congenital heart disease operation.Its prospect of the only domestic medical market of preresearch estimates is about more than one hundred million units every year.And clinical artificial blood vessel commonly used and freezing preservation blood vessel of the same race do not have growth property and cause narrow calcification pogoniasis operative treatment once more easily, cause the waste of a large amount of medical resources when bringing greatest misery to the patient.And mostly artificial blood vessel commonly used or biological sticking patch be import, and price is extremely expensive.Since the eighties in 20th century, along with the development of biomedical engineering technology, various generation vascular graft be used in the cardiovascular disease surgical intervention.People are seeking the best substitute of blood vessel for a long time always, comprise artificial material, biomaterial and composite etc., but have repulsion, thrombosis, endotheli ocytosis, break, shortcoming such as calcification, its therapeutic effect still is very limited.Old friend worker still can not replace the status and the effect of people's freezing preservation blood vessel of the same race fully for blood vessel graft.
Chinese patent document CN101327338A discloses a kind of porous bladder acellular matrix and method for preparing that remains with bioactie agent; Get the bladder of pig, rabbit, Canis familiaris L. or cattle; Through Digestive system; After hypotonic buffer liquid, the high osmotic buffer that contains surfactant, the buffer that contains nuclease and deionized water etc. are handled, after freezing, lyophilizing and sterilization, obtain aseptic bladder acellular matrix again.Chinese patent document CN101474426A discloses a kind of vascular stroma that removes vascular tissue's inner cell and preparation method thereof; A kind of structural intergrity that can remove the cell component in the vascular tissue and at utmost protect acellular matrix is provided, has comprised and take off cell, remove detergent and carry out the enzyme processing.Chinese patent document CN2860403Y discloses a kind of biological type artificial blood vessel, by through the crosslinked fixing and formed base material of animal blood vessels that goes antigen to handle of fixative, and is bonded in the surface layer composition that contains the anticoagulation component on the base material inner surface.But about a kind of porcine aorta vacuum freeze-drying acellular matrix and its production and application also do not appear in the newspapers at present.
Summary of the invention
The objective of the invention is provides a kind of porcine aorta vacuum freeze-drying acellular matrix to deficiency of the prior art.
One purpose more of the present invention is that a kind of method for preparing of porcine aorta vacuum freeze-drying acellular matrix is provided.
Another purpose of the present invention is that a kind of application of porcine aorta vacuum freeze-drying acellular matrix is provided.
For realizing above-mentioned purpose; The technical scheme that the present invention takes is: a kind of porcine aorta vacuum freeze-drying acellular matrix; The method for preparing of described porcine aorta vacuum freeze-drying acellular matrix may further comprise the steps: (1) lyophilizing pretreatment: adopt pre-freeze speed 1K/min, final temperature-42.9 ℃ is carried out the lyophilizing pretreatment to the porcine aorta blood vessel; (2) take off cellization: the lyophilizing blood vessel is with 0.25% trypsin PBS solution effects, and 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature vibrates then, PBS solution vibration flushing; (3) condition that adopts step (1) is the described substrate of vacuum freeze-drying once more, and room temperature is preserved.
The method for preparing of described porcine aorta vacuum freeze-drying acellular matrix may further comprise the steps: (1) lyophilizing pretreatment: adopt pre-freeze speed 1K/min, final temperature-42.9 ℃ is carried out the lyophilizing pretreatment to the porcine aorta blood vessel; (2) take off cellization: the lyophilizing blood vessel uses the 0.25% trypsin PBS solution that contains EDTA 0.04% at 37 ℃, 5%CO
2Effect is 12 hours in the incubator, and the vibration of 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature is 4 hours then, PBS solution vibration flushing 3 times, each 30 minutes; (3) condition that adopts step (1) is the described substrate of vacuum freeze-drying once more, and room temperature is preserved.
For realizing above-mentioned second purpose; The technical scheme that the present invention takes is: a kind of method for preparing of porcine aorta vacuum freeze-drying acellular matrix; The method for preparing of described porcine aorta vacuum freeze-drying acellular matrix is following: (1) lyophilizing pretreatment: adopt pre-freeze speed 1K/min, final temperature-42.9 ℃ is carried out the lyophilizing pretreatment to the porcine aorta blood vessel; (2) take off cellization: the lyophilizing blood vessel is with 0.25% trypsin PBS solution effects, and 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature vibrates then, PBS solution vibration flushing; (3) condition that adopts step (1) is the described substrate of vacuum freeze-drying once more, and room temperature is preserved.
Described method for preparing may further comprise the steps: (1) lyophilizing pretreatment: adopt pre-freeze speed 1K/min, final temperature-42.9 ℃ is carried out the lyophilizing pretreatment to the porcine aorta blood vessel; (2) take off cellization: the lyophilizing blood vessel uses the 0.25% trypsin PBS solution that contains EDTA 0.04% at 37 ℃, 5%CO
2Effect is 12 hours in the incubator, and the vibration of 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature is 4 hours then, PBS solution vibration flushing 3 times, each 30 minutes; (3) condition that adopts step (1) is the described substrate of vacuum freeze-drying once more, and room temperature is preserved.
For realizing above-mentioned the 3rd purpose, the technical scheme that the present invention takes is: the application of described porcine aorta vacuum freeze-drying acellular matrix in the biological blood vessel of preparation.
The invention has the advantages that:
1, the present invention with porcine aorta as object of study; Through optimizing preparation condition; Make have suitable porosity, mechanical property and immunogenic lyophilizing take off the cell blood vessel; Through the feasibility of animal, for further being applied to the clinical experimental basis that provides in the clinical blood vessel transplantation of body experiment proof three-dimensional structure blood vessel;
2, method for preparing cost of the present invention is extremely cheap, can save the medical resource of the preciousness of several hundred million units undoubtedly;
3, for the clinical practice of organizational project aorta vessel and the optimization research and development of taking off the cell blood vessel experimental basis and pointer are provided.
Description of drawings
Temperature variation curve when accompanying drawing 1 is blood vessel pre-freeze.
Accompanying drawing 2 is different rate of temperature fall hematochezia pipe analysis of porosity.
Accompanying drawing 3 is blood vessel gray value analyses under the different rate of temperature fall.
Accompanying drawing 4 is blood vessel α value change curves under the different pre-freeze speed.
Accompanying drawing 5 is immunogenic variations before and after the lyophilizing of porcine aorta blood vessel.
Accompanying drawing 6 is that the young pig of lyophilizing rehydration blood vessel is in the body transplantation experiments.
Accompanying drawing 7 is that lyophilizing pretreatment raising porcine aorta blood vessel takes off cell effect.
Accompanying drawing 8 is that the pretreated cell porcine aorta blood vessel Electronic Speculum of taking off of lyophilizing detects photo.
The specific embodiment
Below in conjunction with accompanying drawing the specific embodiment provided by the invention is elaborated.
Embodiment 1
1. the experiment measuring of blood vessel pre-freeze minimum temperature
The exploration that technology is preserved in the blood vessel lyophilization is to carry out the key issue that preparation research needs to be resolved hurrily.The present invention adopts lift Control Program for Microcomputers [Digital Research] cooling instrument that blood vessel is lowered the temperature with fixed rate; Mainly be in order to investigate of the influence of different pre-freeze speed, to be intended to explore the process control parameter and the flow process that obtain a suitable blood vessel lyophilizing preservation to blood vessel lyophilizing preservation quality.
Purpose is in order to measure the minimum temperature that blood vessel can reach during pre-freeze in freeze dryer, and this temperature that measures is exactly the final temperature that when using the Control Program for Microcomputers [Digital Research] cooling instrument to give blood vessel pre-freeze, is provided with.The temperature that can be known the dry run medium vessels by experiment is minimum for-42.9 ℃, sees Fig. 1, temperature variation curve when Fig. 1 is blood vessel pre-freeze.
2. different pre-freeze speed hematochezia pipe porositys, grey scale change figure.
See Fig. 2 and Fig. 3, Fig. 2, Fig. 3 are respectively porosity, the grey scale change figure of whole freeze-drying process under the different pre-freeze speed.Pre-freeze speed is 0.5K/min, 1K/min, and the rate of change of porosity is respectively 11.9%, 16.6%, 29.7% before and after the blood vessel lyophilizing of 2K/min; The rate of change of average gray value is respectively 2.27%, 3.64%, and 6.36%.Rate of temperature fall is big more, changes obviously more.
3. different pre-freeze speed are to the vascular mechanics Effect on Performance.
The mensuration of vascular mechanics performance.Arteries uses matter structure appearance that it is carried out tension test, the test of puncturing of the square of getting width and be 7mm.The power of 500N is adopted in the load of matter structure appearance in the test, with the speed of 1mm/min sample is punctured and stretches.And the blood vessel after the vacuum lyophilization soaked in normal saline 2 hours, after fully rehydration drained, the arteries of getting the same terms carried out tension test and puncture test.The pre-freeze speed lyophilization front and back vascular mechanics changes of properties that relative analysis is different.
See Fig. 4, Fig. 4 is the α value change curve of blood vessel under the different pre-freeze speed.Time puncture stress value and circumferential tension stress value increasing degree are respectively 20.10% and 32.03% to pre-freeze speed for 1K/min, and axial tensile stress reduces the amplitude minimum, is-19.84%.Think after the analysis have best hole when pre-freeze speed is for 1K/min, the performance of porosity and mechanical property.
The result shows through the rate of temperature fall of control during lyophilization, can control the pore size and the porosity of internal blood vessel, also can keep the mechanical property preferably of blood vessel simultaneously.
4. immunogenic variation before and after the lyophilizing of porcine aorta blood vessel.
See Fig. 5, Fig. 5 is immunogenic variation before and after the lyophilizing of porcine aorta blood vessel.IIF detects MHC I, class change of Expression in lyophilizing blood vessel and the fresh blood vessel respectively.A wherein: the fresh pig aorta, before handling, lyophilizing sees substantially; B: immunofluorescence detects fresh blood vessel MHC class situation; C: immunofluorescence detects fresh blood vessel MHC class situation; D: the fresh pig aorta, after handling, lyophilizing sees substantially; E: immunofluorescence detects MHC class situation in the lyophilizing blood vessel; F: immunofluorescence detects MHC class situation in the lyophilizing blood vessel.The result shows that MHC-I after the blood vessel lyophilizing, class level obviously descend.
5. aortic graft operation and the transplanting back radiography of blood vessel intervention are regularly observed the angiogenic growth situation
Under the quiet combined anesthesia of gas, lyophilizing is taken off the blood vessel of cell processing and implant young pig descending aorta place.The 4th intercostal advances breast along the left side, tremulous pulse between ligation 1-2 root, and blocking-up descending aorta proximal part and distal end excise one section descending aorta, implant the lyophilizing radiation rehydration arteries about 2cm, sew up two anastomotic stoma continuously, aerofluxus, open aortic occlusion clamp.Hemostasis, the flushing thoracic cavity, the drum lung closes breast.The success of operation is the key of later stage research.Regularly repeatedly carry out blood vessel after the transplant operation and get involved radiography observation angiogenic growth situation.
For whether the mechanical strength of verifying the blood vessel after lyophilizing is handled can satisfy application requirements, we in the pig body, substitute the part aorta with freeze dried blood vessel transplantation, and the result finds that pig can long-term surviving, show that lyophilizing handles and vasoinert application.Can bear long-term life test though the blood vessel mechanical property after the confirmation lyophilizing is handled decreases, the equal long-term surviving of young pig is more than 7 months.Though lyophilizing processing back blood vessel immunogenicity descends to some extent but still has calcification trend, also will combine to take off the cell processing and further reduce its immunogenicity.See Fig. 6, Fig. 6 is that the young pig of lyophilizing rehydration blood vessel is in the body transplantation experiments.A wherein: cryodesiccated porcine aorta is handled through rehydration again; B: the blood vessel transplantation repair deficiency that lyophilizing is handled; C: the pig survival is good after the blood vessel transplantation after lyophilizing is handled; D: in the time of three months, the row conduit detects, and transplanting place still keeps unimpeded (the arrow indication is the grafting vessel place).
6. blood vessel takes off cellization with low concentration detergent rehydration simultaneously after the lyophilizing pretreatment
The lyophilizing blood vessel with 0.25% trypsin PBS solution (containing EDTA 0.04%) at 37 ℃, 5%CO
2Effect is 12 hours in the incubator, and the vibration of 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature is 4 hours then, PBS solution vibration flushing 3 times, each 30 minutes.
Lyophilizing is handled increases the blood vessel hole; When improving vascular permeability; Also more help taking off cell solution and enter in the tube wall performance and take off cytosis, and do not compare through the blood vessel in the frozen village, the blood vessel after frozen takes off cell efficient and improves greatly; It is very desirable to take off cell effect, even the inner cell composition of tube wall also can remove totally fully.Please with reference to accompanying drawing 7, accompanying drawing 7 is that lyophilizing pretreatment raising porcine aorta blood vessel takes off cell effect.A wherein: the porcine aorta of handling without lyophilizing takes off HE dyeing before the cell; B: HE dyeing before the porcine aorta that lyophilizing is handled takes off cell; C: the porcine aorta of handling without lyophilizing is through taking off HE dyeing behind the cell, and ductus arteriosus wall inside is still residual to have the part cell; D: the porcine aorta through lyophilizing is handled dyes through HE after taking off cell, and cell residue is not seen in ductus arteriosus wall inside; E: the porcine aorta of handling without lyophilizing is through taking off Hoechst dyeing behind the cell, and ductus arteriosus wall inside is still residual to have the part cell; F: the porcine aorta through lyophilizing is handled dyes through Hoechst after taking off cell, and cell residue is not seen in ductus arteriosus wall inside; The G:DNA detection by quantitative shows after blood vessel is handled through lyophilizing takes off cell again, and residual DNA content obviously reduces.
7. blood vessel electromicroscopic photograph after the cellization is taken off in fresh blood vessel, lyophilizing blood vessel and lyophilizing
See Fig. 8, Fig. 8 is that the pretreated cell porcine aorta blood vessel Electronic Speculum of taking off of lyophilizing detects photo.A wherein: fresh blood vessel transmission electron microscope shows that a large amount of cells exist; B: through after the lyophilizing, transmission electron microscope shows still has cell to exist, but dead; C: after 12 hours, transmission electron microscope shows the acellular existence of internal blood vessel to the lyophilizing blood vessel through 0.25% trypsin treatment; D: fresh blood vessel scanning electron microscope shows that a large amount of cells exist; E: through after the lyophilizing, scanning electron microscope shows still has cell to exist, but dead; F: the lyophilizing blood vessel through 0.25% trypsin treatment 12 as a child after, scanning electron microscope shows the acellular existence of internal blood vessel.
The Electronic Speculum testing result shows that through after the lyophilizing, transmission electron microscope shows still has cell to exist, but dead; And the lyophilizing blood vessel is through 0.25% trypsin treatment after 12 hours, and transmission electron microscope and scanning electron microscope all show the acellular existence of internal blood vessel.
The method for preparing of embodiment 2 porcine aorta vacuum freeze-drying acellular matrixes
1. lyophilizing pretreatment: adopt pre-freeze speed 1K/min, final temperature-42.9 ℃ is carried out the lyophilizing pretreatment to the porcine aorta blood vessel;
2. take off cellization: adopt low concentration detergent rehydration substrate simultaneously row take off cell and handle, the lyophilizing blood vessel with 0.25% trypsin PBS solution (containing EDTA 0.04%) at 37 ℃, 5%CO
2Effect is 12 hours in the incubator, and the vibration of 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature is 4 hours then, PBS solution vibration flushing 3 times, each 30 minutes;
3. the condition that adopts step 1 is this substrate of vacuum freeze-drying once more, and room temperature is preserved.
As object of study, making up the blood vessel clinical transplantation with solid is long-term goal and the porcine aorta blood vessel is carried out vacuum lyophilization take off cell lyophilizing and the sequential rehydration of cell suspension are handled again with porcine aorta in the present invention.Handle the porcine aorta blood vessel through the vacuum freeze-drying method for removing cells, be intended to take off the preparation of cell arteritis substrate, preserve and transplant the back repair mechanism from multiple research field discussion such as heat and mass, low-temperature biological, cytology, molecular biology.Through optimizing preparation condition; Make lyophilizing blood vessel with suitable porosity and mechanical property; Associating enzyme, chemical detergent further reduce the immunogenicity of blood vessel; The Related Mechanism and the three-dimensional operating process that makes up of cell processes taken off in the lyophilizing of research blood vessel in experiment, through the feasibility of animal in the clinical blood vessel transplantation in the future of the three-dimensional structure of body experimentation blood vessel, for further being applied to the clinical experimental basis that provides.The method cost is extremely cheap, can save the medical resource of the preciousness of several hundred million units undoubtedly.
The outer three-dimensional formulation that makes up biological blood vessel flow process of cell porcine aorta vascular stroma preparation flow and this matrix body is taken off in lyophilizing, and the optimization research and development that can be the clinical practice of organizational project aorta vessel and take off the cell blood vessel provide experimental basis and pointer.
The above only is a preferred implementation of the present invention; Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the inventive method; Can also make some improvement and replenish, these improvement and replenish and also should be regarded as protection scope of the present invention.
Claims (5)
1. porcine aorta vacuum freeze-drying acellular matrix; It is characterized in that; The method for preparing of described porcine aorta vacuum freeze-drying acellular matrix may further comprise the steps: (1) lyophilizing pretreatment: adopt pre-freeze speed 1K/min, final temperature-42.9 ℃ is carried out the lyophilizing pretreatment to the porcine aorta blood vessel; (2) take off cellization: the lyophilizing blood vessel is with 0.25% trypsin PBS solution effects, and 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature vibrates then, PBS solution vibration flushing; (3) condition that adopts step (1) is the described substrate of vacuum freeze-drying once more, and room temperature is preserved.
2. porcine aorta vacuum freeze-drying acellular matrix according to claim 1; It is characterized in that; The method for preparing of described porcine aorta vacuum freeze-drying acellular matrix may further comprise the steps: (1) lyophilizing pretreatment: adopt pre-freeze speed 1K/min, final temperature-42.9 ℃ is carried out the lyophilizing pretreatment to the porcine aorta blood vessel; (2) take off cellization: the lyophilizing blood vessel uses the 0.25% trypsin PBS solution that contains EDTA 0.04% at 37 ℃, 5%CO
2Effect is 12 hours in the incubator, and the vibration of 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature is 4 hours then, PBS solution vibration flushing 3 times, each 30 minutes; (3) condition that adopts step (1) is the described substrate of vacuum freeze-drying once more, and room temperature is preserved.
3. the method for preparing of a porcine aorta vacuum freeze-drying acellular matrix; It is characterized in that; The method for preparing of described porcine aorta vacuum freeze-drying acellular matrix is following: (1) lyophilizing pretreatment: adopt pre-freeze speed 1K/min, final temperature-42.9 ℃ is carried out the lyophilizing pretreatment to the porcine aorta blood vessel; (2) take off cellization: the lyophilizing blood vessel is with 0.25% trypsin PBS solution effects, and 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature vibrates then, PBS solution vibration flushing; (3) condition that adopts step (1) is the described substrate of vacuum freeze-drying once more, and room temperature is preserved.
4. method for preparing according to claim 3 is characterized in that, described method for preparing may further comprise the steps: (1) lyophilizing pretreatment: adopt pre-freeze speed 1K/min, final temperature-42.9 ℃ is carried out the lyophilizing pretreatment to the porcine aorta blood vessel; (2) take off cellization: the lyophilizing blood vessel uses the 0.25% trypsin PBS solution that contains EDTA 0.04% at 37 ℃, 5%CO
2Effect is 12 hours in the incubator, and the vibration of 0.5%Tritox X-100 PBS solution chamber relaxing the bowels with purgatives of warm nature is 4 hours then, PBS solution vibration flushing 3 times, each 30 minutes; (3) condition that adopts step (1) is the described substrate of vacuum freeze-drying once more, and room temperature is preserved.
5. according to claim 1 or the application of 2 arbitrary described porcine aorta vacuum freeze-drying acellular matrixes in the biological blood vessel of preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210031969.6A CN102600508B (en) | 2012-02-14 | 2012-02-14 | Porcine arterial vacuum lyophilization acellular matrix, as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210031969.6A CN102600508B (en) | 2012-02-14 | 2012-02-14 | Porcine arterial vacuum lyophilization acellular matrix, as well as preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102600508A true CN102600508A (en) | 2012-07-25 |
| CN102600508B CN102600508B (en) | 2014-03-19 |
Family
ID=46518520
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210031969.6A Expired - Fee Related CN102600508B (en) | 2012-02-14 | 2012-02-14 | Porcine arterial vacuum lyophilization acellular matrix, as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102600508B (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103416394A (en) * | 2012-11-01 | 2013-12-04 | 上海理工大学 | Method for drying and storage of blood vessels |
| CN108904884A (en) * | 2018-08-07 | 2018-11-30 | 中国人民解放军第二军医大学 | A kind of preparation method of porcine aorta acellular matrix |
| CN108949565A (en) * | 2018-09-26 | 2018-12-07 | 中国科学技术大学 | Device and method for red blood cell load freeze drying protectant |
| CN109069263A (en) * | 2016-12-16 | 2018-12-21 | 厦门大开生物科技有限公司 | Porcine cornea method for removing cells and its dry cornea application method of de- cell cornea and plate layer |
| CN109078226A (en) * | 2018-08-07 | 2018-12-25 | 中国人民解放军第二军医大学 | A kind of de- cell porcine aorta matrix in micropore |
| CN111330080A (en) * | 2020-03-31 | 2020-06-26 | 江苏白衣缘生物工程有限公司 | Biomembrane for guiding regeneration of oral cavity bone and preparation method thereof |
| CN111330079A (en) * | 2020-03-31 | 2020-06-26 | 江苏白衣缘生物工程有限公司 | Artificial dura mater composite patch |
| US11604026B2 (en) | 2019-03-14 | 2023-03-14 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11634257B2 (en) | 2017-10-09 | 2023-04-25 | Terumo Bct Biotechnologies, Llc | Lyophilization container and method of using same |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2860403Y (en) * | 2005-07-29 | 2007-01-24 | 广东冠昊生物科技有限公司 | Biotype artificial blood vessel |
| WO2008097882A1 (en) * | 2007-02-02 | 2008-08-14 | Manh-Dan Ngo | Treatment of soft tissue to increase porosity |
| CN101327338A (en) * | 2008-08-01 | 2008-12-24 | 南京大学医学院附属鼓楼医院 | Lacunaris bladder acellular matrix preserving biological activity factor and preparation |
| CN101385870A (en) * | 2008-11-03 | 2009-03-18 | 中国人民解放军第四军医大学 | Method for improving de-cellular system engineering valve/blood vessel stent |
| CN101474426A (en) * | 2009-01-14 | 2009-07-08 | 首都医科大学宣武医院 | Vascular substrate without cell in vascular tissue and preparation method thereof |
| US20110046732A1 (en) * | 2006-04-21 | 2011-02-24 | Dyke Mark E Van | Structurally modified acellular tissue engineering scaffolds and methods of production |
-
2012
- 2012-02-14 CN CN201210031969.6A patent/CN102600508B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2860403Y (en) * | 2005-07-29 | 2007-01-24 | 广东冠昊生物科技有限公司 | Biotype artificial blood vessel |
| US20110046732A1 (en) * | 2006-04-21 | 2011-02-24 | Dyke Mark E Van | Structurally modified acellular tissue engineering scaffolds and methods of production |
| WO2008097882A1 (en) * | 2007-02-02 | 2008-08-14 | Manh-Dan Ngo | Treatment of soft tissue to increase porosity |
| CN101327338A (en) * | 2008-08-01 | 2008-12-24 | 南京大学医学院附属鼓楼医院 | Lacunaris bladder acellular matrix preserving biological activity factor and preparation |
| CN101385870A (en) * | 2008-11-03 | 2009-03-18 | 中国人民解放军第四军医大学 | Method for improving de-cellular system engineering valve/blood vessel stent |
| CN101474426A (en) * | 2009-01-14 | 2009-07-08 | 首都医科大学宣武医院 | Vascular substrate without cell in vascular tissue and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 《中国组织工程研究与临床康复》 20101217 殷猛等 "真空冷冻干燥过程中猪主动脉血管的力学性能" 第9539-9544页 1-5 第14卷, 第51期 * |
| 殷猛等: ""真空冷冻干燥过程中猪主动脉血管的力学性能"", 《中国组织工程研究与临床康复》 * |
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103416394A (en) * | 2012-11-01 | 2013-12-04 | 上海理工大学 | Method for drying and storage of blood vessels |
| CN109069263A (en) * | 2016-12-16 | 2018-12-21 | 厦门大开生物科技有限公司 | Porcine cornea method for removing cells and its dry cornea application method of de- cell cornea and plate layer |
| US11634257B2 (en) | 2017-10-09 | 2023-04-25 | Terumo Bct Biotechnologies, Llc | Lyophilization container and method of using same |
| CN108904884A (en) * | 2018-08-07 | 2018-11-30 | 中国人民解放军第二军医大学 | A kind of preparation method of porcine aorta acellular matrix |
| CN109078226A (en) * | 2018-08-07 | 2018-12-25 | 中国人民解放军第二军医大学 | A kind of de- cell porcine aorta matrix in micropore |
| CN108949565A (en) * | 2018-09-26 | 2018-12-07 | 中国科学技术大学 | Device and method for red blood cell load freeze drying protectant |
| US11609042B2 (en) | 2019-03-14 | 2023-03-21 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
| US11604026B2 (en) | 2019-03-14 | 2023-03-14 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11609043B2 (en) | 2019-03-14 | 2023-03-21 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
| US11740019B2 (en) | 2019-03-14 | 2023-08-29 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11747082B2 (en) | 2019-03-14 | 2023-09-05 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
| US11815311B2 (en) | 2019-03-14 | 2023-11-14 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
| CN111330080B (en) * | 2020-03-31 | 2021-12-07 | 江苏白衣缘生物工程有限公司 | Biomembrane for guiding regeneration of oral cavity bone and preparation method thereof |
| CN111330079A (en) * | 2020-03-31 | 2020-06-26 | 江苏白衣缘生物工程有限公司 | Artificial dura mater composite patch |
| CN111330080A (en) * | 2020-03-31 | 2020-06-26 | 江苏白衣缘生物工程有限公司 | Biomembrane for guiding regeneration of oral cavity bone and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102600508B (en) | 2014-03-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102600508B (en) | Porcine arterial vacuum lyophilization acellular matrix, as well as preparation method and application thereof | |
| Sierad et al. | Design and testing of a pulsatile conditioning system for dynamic endothelialization of polyphenol-stabilized tissue engineered heart valves | |
| JP5050197B2 (en) | Production method of biological scaffold | |
| Syed et al. | Evaluation of decellularization protocols for production of tubular small intestine submucosa scaffolds for use in oesophageal tissue engineering | |
| US9642937B2 (en) | Preparation method for implantable medical biological materials of animal origin | |
| CN106190949B (en) | Dry animal-derived collagen fiber tissue material, preparation method thereof and bioprosthesis | |
| Kajbafzadeh et al. | Determining the optimal decellularization and sterilization protocol for preparing a tissue scaffold of a human-sized liver tissue | |
| Sheridan et al. | Optimum parameters for freeze-drying decellularized arterial scaffolds | |
| CN105392506B (en) | Implant and manufacture the method for implant by making tissue decellularization under negative pressure | |
| CN110665061A (en) | Acellular scaffold solution-GelMA hydrogel composite material and preparation method thereof | |
| WO2009049568A3 (en) | Anisotropic implant and its method of production | |
| JP6091414B2 (en) | Method for preparing decellularized tissue product and graft comprising decellularized tissue product | |
| CN103272274B (en) | Biological repair tablet for herniae and preparation method thereof | |
| Tuan-Mu et al. | On the decellularization of fresh or frozen human umbilical arteries: implications for small-diameter tissue engineered vascular grafts | |
| CN104640577B (en) | The hydrophilic dehydration containing phosphate groups and partially purified skeleton displacement material | |
| Butter et al. | Evolution of graft morphology and function after recellularization of decellularized rat livers | |
| CN109078226A (en) | A kind of de- cell porcine aorta matrix in micropore | |
| CN105664260A (en) | Method for preparing bone tissue engineering three-dimensional porous support based on graphene/silk fibroin | |
| CN101259292A (en) | Construction method of tissue engineering blood vessel | |
| CN202458781U (en) | Lyophilized acellular swine aorta vessel matrix | |
| CN104971386B (en) | Silk-fibroin timbering material and preparation method thereof | |
| CN106552294B (en) | Biological patch material for heart repair | |
| CN107812239A (en) | A kind of preparation method of tussah silk peptide collagen compound rest | |
| CN103272275B (en) | Biological repair tablet for endocranium and preparation method thereof | |
| CN103272276B (en) | Anal fistula suppository and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140319 Termination date: 20180214 |