CN104695205B - Preparation method of collagen fiber membrane and application thereof - Google Patents
Preparation method of collagen fiber membrane and application thereof Download PDFInfo
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- CN104695205B CN104695205B CN201510114478.1A CN201510114478A CN104695205B CN 104695205 B CN104695205 B CN 104695205B CN 201510114478 A CN201510114478 A CN 201510114478A CN 104695205 B CN104695205 B CN 104695205B
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Abstract
The invention provides a preparation method of a collagen fiber membrane and application thereof. The collagen fiber membrane prepared by the method is outstanding in biocompatibility, relatively high in mechanical strength, applicable to human soft tissue injury repair, and meets the suturing requirement in actual use.
Description
Technical field
The invention belongs to biomedicine field, specifically, the present invention relates to a kind of collagenous fibres membrane preparation method and its
Application.
Background technology
Collagenous fibres have excellent biological safety, stronger with cellular affinity, also have certain cell simultaneously and lure
Lead ability, cell differentiation can be guided to rise in value, be a kind of unusual tissue engineering bracket material of prospect.At present with collagen as raw material
Film product mainly have acellular matrix, collagen to be lyophilized sponge, collagen electrospinning film etc., but to have itself inevitable for they
Defect.
Acellular matrix：Mechanical strength is good, and has very rational gap structure, can provide for the growth of cell
Excellent environment.But acellular matrix is by physicochemical by animal skin, pericardium, peritonaeum, little goldbeater's skin etc. for raw material
Method removes cell component and obtains, and this process, not by above-mentioned tissue dispersion, is difficult to would be possible to lead to host that immunity occurs
The material of reaction removes clean.
Collagen is lyophilized sponge：It is lyophilized and obtained by the collagen solution extracting from animal tissue.It has excellent biofacies
Capacitive, be conducive to the gap structure that cell grows into, but its intensity be relatively low, limit he as tooth film, hernia paster, brain can be sutured
The use of film etc..
Collagen electrospinning film：Obtained by electrostatic spinning by collagen solution.It has preferable biocompatibility, preferable power
Learn performance and certain gap structure, but collagen electrospinning can use the organic solvents such as hexafluoroisopropanol, its toxicity is larger, be difficult to remove
Only.In addition document report electrospinning is had can to destroy the triple helix structure of collagen.
Therefore, those skilled in the art are devoted to overcoming the existing product strength prepared for raw material with collagen low, can not
Suture is it is impossible to the defect of load.
Content of the invention
It is an object of the invention to provide a kind of collagen fiber membrane preparation method and applications.
A kind of a first aspect of the present invention, there is provided method preparing collagen fiber membrane, methods described includes step：
(1) collagenous fibres are dissolved in acid medium, obtain collagenous fibril solution；
(2) described collagenous fibril solution is poured in container, be placed under volatile base environment stifling, described collagen is obtained
Tunica fibrosa；
Wherein, the bottom of described container is filter structure；Described filter screen is the filter screen of 50-500 mesh, and described filter screen allows institute
State the water in sample solution and flow out described container, and the collagen fiber matrix that described collagenous fibril is formed is retained in described filter screen
On.
In another preference, in described step (1), in described collagenous fibril solution, the concentration of collagenous fibril is
1g/L-200g/L is it is therefore preferable to 10g/L-100g/L, more preferably 15g/L-50g/L.
In another preference, the pH of described acid medium is 0-6；Preferably pH is 0.5-5.5；More preferably pH is
1.0-5.0；Optimally pH is 2.0-4.0.
In another preference, described acid medium is aqueous acid.
In another preference, described acid medium is selected from：Aqueous hydrochloric acid solution, aqueous sulfuric acid, aqueous solution of nitric acid, acetic acid
The aqueous solution or a combination thereof.
In another preference, described volatile base is selected from the group：Ammoniacal liquor.
In another preference, described volatile base is ammoniacal liquor；Preferably, described ammonia concn be 0.1% (w/w)-
5% (w/w)；It is highly preferred that described ammonia concn is 0.5% (w/w) -2% (w/w).
In another preference, the fumigation time in described step (2) is 8h-48h；Preferably, described fumigation time is
12h-36h；It is highly preferred that described fumigation time is 18h-30h..
In another preference, in described step (1), also include dispersion steps, be homogenized using refiner so that glue
Fibrillation is substantially soluble in acid medium, forms collagenous fibril solution；Preferably, rotating speed is 100r/min-5000r/min,
It is preferably 500r/min-3000r/min.
In another preference, described Homogenization time is 1min-30min, preferably 5min-20min.
In another preference, in described step (1), also include being centrifuged de-bubble step, using centrifuge to described collagen
Fibrillation solution carries out centrifugation and removes bubble removing；Preferably, centrifugal rotational speed is 1000r/min-10000r/min, more preferably
2000r/min-8000r/min.
In another preference, described centrifugation time is 1min-30min, more preferably 5min-20min.
In another preference, the bottom of described container is filter structure；Preferably, described filter screen is the filter of 50-500 mesh
Net；More preferably described filter screen is the filter screen of 100-400 mesh；Most preferably, described filter screen is the filter screen of 150-250 mesh.
In another preference, in described step (2), described collagenous fibril solution is coated on described filter screen, so
After be placed under volatile base environment stifling.
In another preference, in described step (2), the coating thickness of described collagenous fibril solution is 2-20mm；Excellent
Selection of land coating thickness is 4-15mm；More preferably coating thickness is 5-10mm.
In another preference, in described step (2), after the completion of fumigating, described collagen fiber membrane is taken out drying.
In another preference, after described step (2), also include step：
(3) described collagen fiber membrane is placed in swelling in phosphate buffer, freeze-drying after pure water cleaning.
In another preference, methods described also includes step, and described collagen fiber membrane is carried out heat cross-linking.Described heat is handed over
The method of connection is the conventional method in this area.
In another preference, methods described also includes step, by described collagen fiber membrane sterilization treatment.
In another preference, described collagenous fibres are to be obtained with the method that acidleach carries.
A kind of a second aspect of the present invention, there is provided collagen fiber membrane, described collagen fiber membrane passes through described in claim 1
Method be prepared.
In another preference, described collagen fiber membrane tensile strength is between 5MPa-7MPa.
In another preference, the thickness of described collagen fiber membrane is 0.5mm-1.5mm.
A kind of a third aspect of the present invention, there is provided device preparing collagen fiber membrane, described device includes：Shuttle
With volatile base container, described shuttle is placed in the top of described volatile base container；
Wherein, described shuttle is arranged to be used for holding collagenous fibril sample solution, the bottom of described shuttle
Portion is filter structure.
In another preference, described filter screen is selected from：Nylon leaching net, stainless steel filtering net.
In another preference, described filter screen is the filter screen of 50-500 mesh；More preferably described filter screen is 100-400 purpose
Filter screen；Most preferably, described filter screen is the filter screen of 150-250 mesh.
In another preference, the side wall of described shuttle is made up of stainless steel material.
In another preference, described volatile base container is arranged for holding volatile base solution.
In another preference, described volatile base is ammoniacal liquor.
In another preference, described device also includes housing, and described shuttle and described volatile base container are located at
Described enclosure interior.
In another preference, described housing includes main body and lid, when described cover cap closes on the body, institute
State main body and described lid constitutes an airtight space, described shuttle and described volatile base container are located at described confined air
Interior.
It should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the present invention and having in below (eg embodiment)
Can be combined with each other between each technical characteristic of body description, thus constituting new or preferred technical scheme.As space is limited, exist
This no longer tires out one by one states.
Brief description
Fig. 1 shows collagen fiber membrane biological safety (immunogenicity) testing result.
Fig. 2 is the device result schematic diagram preparing collagen fiber membrane of better embodiment of the present invention.
Fig. 3 A, Fig. 3 B respectively illustrate the collagen fiber membrane photo of preparation in embodiment 1.
Specific embodiment
The present inventor, by extensively in-depth study, obtains a kind of preparation method of collagen fiber membrane, first by collagen
Fibrinolysis, in acidic aqueous solution, then adopt volatile base to fumigate, induction collagenous fibril forms longer collagenous fibres,
And be uniformly distributed, air-dry and obtain hard collagem membrane, be lyophilized after collagem membrane is swelling in aqueous systems and can obtain collagen fiber membrane
Finished product.If needed can will be swelling after collagen gel first do crosslinking Treatment and then be lyophilized again.The glue prepared in this way
Former film saves the activity of collagen to greatest extent, has preferable mechanical property and pore structure simultaneously, and non-collagen becomes
Divide and easily remove totally, biological safety is excellent.On this basis, complete the present invention.
Compared with traditional collagenous fibres membrane preparation method, method proposed by the invention can retain collagenous fibres length,
The mechanical strength of reserved materials and biocompatibility most possibly.Traditional collagenous fibres film build method, will pass through homogenizer powder
Broken collagenous fiber bundle, obtains collagenous fibres suspension, then does later stage process again.So longer collagenous fibres are interrupted,
Destroy the mechanical strength after its shaping, and strong mechanism has the risk of collagenous degeneration.
The method of the present invention prepares collagen solution (collagenous fibril solution, sample solution) by the way of acid treatment, so
Afterwards collagenous fibril solution is issued and is conigenous assembling in the inducing action of other external conditions such as alkaline steam, form long fibre.
Fiber solution is air-dried, then swelling, optionally crosslinked, finally sample is lyophilized packaging sterilizing or is directly packaged in containing guarantor
In the packaging material of liquid storage and sterilize.
Can be very easy to according to the method prepare the collagen fiber membrane with preferable intensity, its thickness, porosity
All can regulate and control as desired.The product finally giving can be used for dural patch, tooth film, periosteum, nerve trachea, hernia paster etc.
Product.
The preparation method of the collagen fiber membrane of the present invention, belongs to biomaterial for medical purpose field, can solve the problem that current collagen is fine
The low problem of dimension film-strength, expands the range of application of collagen fiber membrane.Using the collagenous fibres prepared by the method in the present invention
Film can be widely used in the repairing of various soft tissues in vivo, such as dural patch, hernia paster, tooth film, skin reparative membrane, nerve
Sticking patch, tendon repairing etc..
It is preferably carried out in mode one, method of operating of the present invention includes step：
1. collagenous fibres are dissolved in acid (pH0-4.8) aqueous solution, obtain homogeneous collagenous fibril solution sample
Product.
2. sample is poured in stainless steel frame, stainless steel frame bottom surface is closed with 200 mesh nylon leaching nets.
3. sample is placed in closed container, closed container bottom fills 1% ammoniacal liquor, takes when sample pH value is to 6.0 ± 0.5
Go out (about placing 20h), air-dry, obtain collagen fiber membrane.
4. will be swelling in 0.01M, the phosphate buffer of pH=7.2 for the collagen fiber membrane after air-drying, treat that thickness reaches
1mm about after, take out, cleaned up with pure water, be lyophilized, heat cross-linking can be carried out if needed.
5. pack after sample cutting, sterilize.
Present invention also offers a kind of device (as shown in Figure 2) preparing collagen fiber membrane, described device includes：Sample holds
Device 1 and volatile base container 2, described shuttle 1 is placed in the top of described volatile base container 2；
Wherein, described shuttle 1 is arranged to be used for holding collagenous fibril solution 11, the bottom of described shuttle
For filter structure；Described volatile base container 2 is arranged for holding volatile base solution 21.
In present embodiment, described shuttle 1 can hang on inside described volatile base container 2, and in setting one
Individual lid 3, when lid 3 covers on described volatile base container 2, forms an airtight space, it is to avoid outside volatile base
Let out.
In one preferably embodiment, described device also includes housing, and described shuttle and described volatile base hold
Device is located at described enclosure interior；Preferably, described housing includes main body and lid, when described cover cap closes on the body
When, described main body and described lid constitute an airtight space, and described shuttle and described volatile base container are located at described
In confined space.
In other embodiments, a lid can be set above described shuttle, when described lid covers
When above described shuttle, form an airtight space, it is to avoid volatile base leaks.
Main advantages of the present invention are：
(1) method of the present invention can retain collagenous fibres length, most possibly the mechanical strength of reserved materials and life
Thing compatibility；
(2) adopt the collagen fiber membrane of method of the present invention preparation excellent compared with collagen film dynamic performance prepared by additive method
Different, it is fully available for internal soft tissue repairing operation；
(3) collagenous fibril, in collagenous fibril extraction process, is dispersed in acid solution by the way of homogenate by the present invention
In, stifling collagenous fibril later can be self-assembly of thicker collagenous fiber bundle, collagenous fiber bundle is no longer done even
Slurry is processed, and traditional method working process follow-up for convenience, homogenized to be carried out to collagenous fiber bundle before the forming.
The method of the present invention obtains collagenous fibres by the way of induction self assembly in situ, and the collagenous fibres obtaining are evenly distributed, and keep away
Exempt from collagenous fibres and be homogenized in forming process to lead to longer collagenous fibres to be interrupted, the mechanics remaining after its shaping is strong
Degree, and avoid strong mechanism and have the risk of collagenous degeneration.
With reference to specific embodiment, the old present invention in detail further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than restriction the scope of the present invention.Unless otherwise indicated, otherwise percentage and number are calculated by weight.In following examples
Experiment material used and reagent all can obtain from commercially available channel if no special instructions.Experiment material used in the embodiment of the present invention
Material all can obtain from commercially available channel if no special instructions.
The preparation of embodiment 1 collagen fiber membrane
Take 10g collagenous fibres sample (its preparation method may be referred to United States Patent (USP) 5997895), swelling in 500ml, 0.1M
Acetum (pH2.5) in, then with refiner by collagenous fibres thoroughly dispersion so that collagenous fibres to be completely dissolved in acetic acid molten
In liquid, (this process is collagenous fibril extraction process, by collagenous fibril by the way of homogenate to obtain collagenous fibril solution
It is dispersed in acid solution, stifling collagenous fibril later can be self-assembly of thicker collagenous fiber bundle, to collagenous fibres
Shu Buzai does homogenized, and traditional method will carry out homogenized before the forming to collagenous fiber bundle, after being so just easy to
The working process in face), obtain collagenous fibril sample solution after centrifugation de-bubble.This sample solution is coated in 200 purposes not
On rust steel filter screen, THICKNESS CONTROL is in 5mm-7mm.
Filter screen is put in the sealing container of the ammoniacal liquor filling 1% and is fumigated, take out sample after 24 hours, and air-dry.
After the PBS of 0.01M is swelling, fully clean gained collagen fiber membrane, freeze-drying, then cutting sample, bag with pure water
Irradiation after dress.
The present embodiment, under conditions of ammoniacal liquor is fumigated, induction collagenous fibril is assembled into thicker collagenous fibres, assembles
The collagenous fibres arriving are thicker, and electron microscopic observation has light and shade striped, have good mechanical property, are comparatively close to internal collagenous fibres
The state of normal presence.
The biological safety (cytotoxicity) of embodiment 2 collagen fiber membrane
1st, leaching liquor makes
According to the regulation of GB/T 16886, the collagenous fibres membrane material of this test is because thickness is in below 5mm, leaching liquor
Volume is calculated according to material surface area, 6cm2/ ml is extracted.Each class sample is required to the extraction of 24h, 48h and 72h
Liquid.Extraction temperature is 37 DEG C.The super-clean bench all in cell room for the operation is carried out.Leaching liquor is to be used to cultivate containing of L929 cell
1640 culture mediums of 10%FBS.
2nd, cell culture
Propose the last week recovery and cultivated L929 cell (purchased from Shanghai Bai Li bio tech ltd), spread after digestion into 96
In orifice plate, every hole 2 × 105Individual cell.Next day suctions out the culture medium in orifice plate, adds leaching liquor, does 6 multiple holes.Control group, uses
Phenol containing 6.4%-culture medium solution cultured cells.
3rd, data monitoring
Culture 48h detection, 72h detection and corresponding 2 control group plates are detected using cck-8 method.Detection mode is
450nm absorbance is measured on ELIASA.
4th, data analysis
The computational methods of cell proliferation rate are that (experimental group absorbance-blank group absorbance)/(ordinary culture medium culture group is inhaled
Luminosity-blank group absorbance) * 100%, the rate of increase is more than or equal to 80% explanation no cytotoxicity.
Final data is calculated as follows：
Result：Positive controls cell growth rate using phenol is respectively 4.6% and 4.3%.
The testing result obtaining all shows that cytoactive is more than 80% it was demonstrated that gained leaching liquor no cytotoxicity, thus saying
Bright detected collagen fiber membrane no soluble cell toxicant residual.And partial data is much larger than 100%, hint may
There is facilitation to cell metabolism.
Embodiment 3, collagen fiber membrane biological safety (immunogenicity)
1st, plating cells：Spread in 6 × 4 square formations in 96 orifice plates and (grind science and technology purchased from upper sea valley limited into RAW264.7 cell
Company), every hole 100 μ l cell suspension, 2 × 104Individual cell, is placed in overnight incubation in 37 DEG C of cell culture incubators.
2nd, immersion liquid makes：Collagenous fibres membrane sample to be measured is taken to insert making leaching liquor in centrifuge tube.Sample weight 1.1g, plus
Enter 11ml extraction stoste.The extraction stoste that this test uses is 1640 culture mediums.
3rd, sample-adding stimulates：Leaching liquor or comparison liquid difference that 4 row are separately added into：Fiber membrane sample leaching liquor, blank are right
According to (1640 culture medium), negative control (BSA, 40ug/ul) and positive control (ConA, 40ug/ul and 100ug/ml).The same day inhales
Net cell culture fluid, clean after cell three times with DPBS, adds respective sample in arranging 8 respectively, each sample 6 hole, every hole 100
μl.It is further cultured for 4h, is then washed with DPBS 3 times, add 0.1% neutral red solution 100ul, continue culture 30min.During suction is abandoned
Property red solution, wash 3 times, add lysate (glacial acetic acid and absolute ethyl alcohol=1:1) 200ul, stands overnight.
4th, absorbance detection：After 12 hours, cell cracks completely, detects 540nm absorbance on ELIASA.
5th, research process and result：
Testing result as shown in figure 1, data display collagenous fibres membrane sample leaching liquor to the excitant of macrophage all very
Low, close to 1640 blank culture mediums, also lower than negative control BSA solution, illustrate that four kinds of detected samples all no may be used
The immunogenicity of stimulating expression of macrophage.
Embodiment 4 collagen fiber membrane intensity detection
With reference to the process of embodiment 1, according to the technical parameter of table 1 design, prepare collagem membrane, and according to national legislation
Require to measure its mechanical property (concrete assay method is as described in Example 5), its result is as shown in the table：
Table 1
Through repeatedly verifying, in other embodiments, the acid medium dissolving collagenous fibres for 2.0-4.0 using pH,
In collagenous fibril sample solution, the concentration of collagen is 10g/L-100g/L, using the filter screen of 100-400 mesh, in concentration is
Fumigated under the aqueous ammonia conditions of 0.5% (w/w) -2% (w/w), fumigation time is 12h-36h, all can prepare performance relatively
Good, meet the collagen fiber membrane of application claims.
Embodiment 5 collagenous fibres film-strength reappearance detects
Randomly select collagen fiber membrane (by the method preparation of embodiment 1) and collagen sponge (using enzyme process from ox heel string
Middle extraction collagen, is then prepared into collagen sponge, preparation method bibliography in the way of lyophilized：Extracting collagen of bovine tendon with enzyme
Research, Ye Yichun etc.,《Chinese leather》The 7th phase of volume 34) each 3, sample is cut into 10mm width and the strip of 80mm length,
It is placed on 23 DEG C of temperature, until sample is stable in the environment of humidity 50%.
Set the nominal clamp distance of Tensile Tester as 50mm ± 1mm, clamp style in clamper center.Pre-
Power is set as 2N.
Machine, with the constant elongation speed tensile sample of 10mm/min until fracture, record the strong of every piece of sample
Power-extension curve, calculates the ultimate strength (the corresponding power of curve peak) of sample.
Calculate the tensile strength of product according to below equation：
Tensile strength=ultimate strength ÷ (batten width × batten is thick)
Test result is as follows：
Above-mentioned testing result shows, its tensile strength of collagen fiber membrane prepared according to the methods of the invention is collagen sponge
Nearly 6 times, illustrate that high tensile strength is had by collagen fiber membrane prepared by the method for the present invention.
Comparative example 1
Prepare collagen fiber membrane according to the method in embodiment 1, difference is, sample solution is coated in 200 purposes not
After on rust steel filter screen, fumigate 30min using ammoniacal liquor, other conditions are essentially identical, 5 group collagem membranes are obtained.According to side in embodiment 4
Method detects gained collagen film strength, and after testing, the mean intensity of gained collagem membrane is less than 2.5Mpa.
Comparative example 2
Prepare collagen fiber membrane according to the method in embodiment 1, difference is, sample solution is coated in 200 purposes not
On rust steel filter screen, reuse ammoniacal liquor and fumigate 30min, then standing and drying 20h, other conditions are essentially identical, 5 group collagens are obtained
Film.According to method detection gained collagen film strength in embodiment 5, after testing, the mean intensity of gained collagem membrane is less than 2Mpa.
Comparative example 3
Prepare collagen fiber membrane according to the method in embodiment 1, difference is, sample solution is coated in 200 purposes not
On rust steel filter screen, standing and drying 20h, then reuse ammoniacal liquor and fumigate 30min, other conditions are essentially identical, 5 group collagens are obtained
Film.According to method detection gained collagen film strength in embodiment 5, after testing, the mean intensity of gained collagem membrane is less than
1.5Mpa.
Comparative example 4
Prepare collagen fiber membrane according to the method in embodiment 1, difference is, sample solution is coated in there is no hole
On flat board, carry out the preparation of collagen fiber membrane, other conditions are essentially identical, 5 group collagen fiber membranes are obtained.According in embodiment 5
Method detects gained collagenous fibres film strength, and after testing, the mean intensity of gained collagen fiber membrane is less than 3Mpa；Repeat to test
Find that its multiple sample fails portion's fracture in the sample, show that its product uniformity is poor, do not meet national legislation and require, simultaneously
Observe by the naked eye, compared with the method providing with the present invention, its product thickness heterogeneity obtaining, color evenness is also poor.
The all documents referring in the present invention are all incorporated as reference in this application, independent just as each document
It is incorporated as with reference to like that.In addition, it is to be understood that after the above-mentioned instruction content having read the present invention, those skilled in the art can
To make various changes or modifications to the present invention, these equivalent form of values equally fall within the model that the application appended claims are limited
Enclose.
Claims (22)
1. a kind of method preparing collagen fiber membrane is it is characterised in that methods described includes step：
(1) collagenous fibres are dissolved in acid medium, obtain collagenous fibril solution；
(2) described collagenous fibril solution is poured in container, be placed on and fumigate 8h-48h under volatile base environment, be obtained described
Collagen fiber membrane；
Wherein, the bottom of described container is filter structure；Described filter screen is the filter screen of 50-500 mesh, and described filter screen allows described sample
Water in product solution flows out described container, and the collagen fiber matrix that described collagenous fibril is formed be retained in described filter screen it
On.
2. the method for claim 1 is it is characterised in that in described step (1), collagen in described collagenous fibril solution
The concentration of fibrillation is 10g/L-100g/L.
3. the method for claim 1 is it is characterised in that in described step (1), collagen in described collagenous fibril solution
The concentration of fibrillation is 15g/L-50g/L.
4. the method for claim 1 is it is characterised in that the pH of described acid medium is 0-6.
5. the method for claim 1 is it is characterised in that the pH of described acid medium is 0.5-5.5.
6. the method for claim 1 is it is characterised in that the pH of described acid medium is 1.0-5.0.
7. the method for claim 1 is it is characterised in that the pH of described acid medium is 2.0-4.0.
8. the method for claim 1 is it is characterised in that described volatile base is ammoniacal liquor.
9. method as claimed in claim 8 is it is characterised in that described ammonia concn is 0.1% (w/w) -5% (w/w).
10. method as claimed in claim 8 is it is characterised in that described ammonia concn is 0.5% (w/w) -2% (w/w).
11. the method for claim 1 are it is characterised in that the fumigation time in described step (2) is 12h-36h.
12. the method for claim 1 are it is characterised in that the fumigation time in described step (2) is 18h-30h.
13. the method for claim 1 are it is characterised in that described filter screen is the filter screen of 100-400 mesh.
14. the method for claim 1 are it is characterised in that described filter screen is the filter screen of 150-250 mesh.
15. the method for claim 1 are it is characterised in that in described step (2), described collagenous fibril solution is applied
Overlay on described filter screen, be then placed under volatile base environment stifling.
A kind of 16. collagen fiber membranes are it is characterised in that described collagen fiber membrane is by the method described in any of the above-described claim
It is prepared.
17. collagen fiber membranes as claimed in claim 16 are it is characterised in that described collagen fiber membrane tensile strength is in 5MPa-
Between 7MPa.
18. collagen fiber membranes as claimed in claim 16 are it is characterised in that the thickness of described collagen fiber membrane is 0.5mm-
1.5mm.
A kind of 19. devices preparing collagen fiber membrane are it is characterised in that described device includes：Shuttle and volatile base hold
Device, described shuttle is placed in the top of described volatile base container；
Wherein, described shuttle is arranged to be used for holding collagenous fibril sample solution, and the bottom of described shuttle is
Filter structure；
Described device also includes housing, and described shuttle and described volatile base container are located at described enclosure interior；
Described housing includes main body and lid, when described cover cap closes on the body, described main body and described lid structure
An airtight space, described shuttle and described volatile base container is become to be located in described confined space；
Described filter screen is the filter screen of 50-500 mesh.
20. devices preparing collagen fiber membrane as claimed in claim 19 are it is characterised in that described filter screen is 100-400 mesh
Filter screen.
21. devices preparing collagen fiber membrane as claimed in claim 19 are it is characterised in that described filter screen is 150-250 mesh
Filter screen.
22. devices preparing collagen fiber membrane as claimed in claim 19 are it is characterised in that described volatile base container is set
Put for holding volatile base solution, and, described volatile base is ammoniacal liquor.
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| ES2842501T5 (en) | 2015-09-21 | 2023-04-13 | Modern Meadow Inc | Fiber Reinforced Fabric Composite Materials |
| US11542374B2 (en) | 2016-02-15 | 2023-01-03 | Modern Meadow, Inc. | Composite biofabricated material |
| CN106880872B (en) * | 2016-12-23 | 2019-10-29 | 北京大清生物技术股份有限公司 | Natural extracellular matrix biomembrane and the preparation method and application thereof |
| AU2018253595A1 (en) | 2017-11-13 | 2019-05-30 | Modern Meadow, Inc. | Biofabricated leather articles having zonal properties |
| CN113286864A (en) | 2019-01-17 | 2021-08-20 | 现代牧场股份有限公司 | Layered collagen material and preparation method thereof |
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| CN101791438B (en) * | 2010-03-16 | 2013-02-27 | 浙江大学 | Method for preparing bioactive poly(lactic-co-glycolic acid)/collagen/hydroxyapatite composite fiber membrane for bone repair |
| CN204569795U (en) * | 2015-03-16 | 2015-08-19 | 苏州景卓生物技术有限公司 | A kind of device preparing collagen fiber membrane |
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