CN108314791A - A kind of method preparing matte fibroin membrane and products thereof and application - Google Patents
A kind of method preparing matte fibroin membrane and products thereof and application Download PDFInfo
- Publication number
- CN108314791A CN108314791A CN201810122565.5A CN201810122565A CN108314791A CN 108314791 A CN108314791 A CN 108314791A CN 201810122565 A CN201810122565 A CN 201810122565A CN 108314791 A CN108314791 A CN 108314791A
- Authority
- CN
- China
- Prior art keywords
- fibroin
- matte
- fibroin membrane
- silk
- membrane
- 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.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J5/00—Manufacture of articles or shaped materials containing macromolecular substances
- C08J5/18—Manufacture of films or sheets
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2389/00—Characterised by the use of proteins; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/16—Halogen-containing compounds
- C08K2003/162—Calcium, strontium or barium halides, e.g. calcium, strontium or barium chloride
Abstract
The present invention relates to a kind of methods preparing matte fibroin membrane and products thereof and application, and the specific method is as follows：Fibroin is put into dissolving in calcium chloride ethanol water ternary solvent and obtains silk fibroin solution, then it will be incubated after silk fibroin solution dialysis, centrifugation and filtering, last drying and forming-film, the present invention prepares matte fibroin membrane using the self assembling process of fibroin albumen itself, method is simple and practicable, easy to operate, and fibroin film surface obtained forms a certain size protrusion, the adherency growth for being conducive to cell, improves performance of the fibroin membrane as biomaterial.
Description
Technical field
The invention belongs to Material Fields, are related to a kind of method preparing matte fibroin membrane, further relate to obtained by this method
Product.
Background technology
Fibroin membrane is a kind of important biomaterial.The side that regenerated silk fibroin solution passes through solution-cast or rotary coating
Method can readily prepare the fibroin membrane of various thickness.Fibroin membrane is non-toxic, has no stimulation, and has good biocompatibility,
It is a kind of widely used important biomolecule material.Medicine carrying membrane and sustained release is made after being currently used primarily in fixed and cladding active material
Medicine carrying membrane, artificial skin, Wound dressing and artificial cornea etc..
As biomaterial, the surface texture of fibroin membrane grows the adherency of cell, and the deposition mineralising of inorganic matter has weight
It influences, coarse fibroin film surface is conducive to the adherency growth and biomineralization of cell.Prepare the common side of matte fibroin membrane
Method is to use freeze-drying, and the cavity that this method can be larger in fibroin film surface and internal formation is conducive to the life of cell
Long adherency.But fibroin film dynamic performance prepared by freeze-drying is very poor, to limit its use scope.Add in fibroin membrane
Other substances are added also to can reach the purpose for preparing matte fibroin membrane.Elastin laminin is added such as in fibroin, and membrane material is also adjusted
Surface roughness, this space surface structure can play the role of regulating cell growth and differentiation.But can additive very
It is good compatible with fibroin albumen without occurring being separated, it is needed to be considered.
Silk fibroin molecular chain is long-chained protein made of being inlayed by hydrophobic section and hydrophilic section, in aqueous solution, fibroin egg
Spontaneous it can be agglomerated into aggregation in vain, be embodied in beta sheet conformation of the conformation of fibroin albumen from random coil to minimum energy
Slowly transformation, this process are referred to as self assembly (Self-Assembly) process.Have much in the self assembling process of fibroin albumen
Theoretical model, such as " nucleation-growth " kinetic mechanism of the just medium proposition of Shao, which shows fibroin albumen from random coil
Conformation transition to beta sheet has the stage there are three different rates, is referred to as nucleated (initiation), growth (growth)
With adjustment (perfection)." the micelle model " etc. that also Hyoung-Joon Jin and David L.Kaplan are proposed.At present
Research surface, some external conditions such as humiture, concentration, pH value, solvent, metal ion and shearing etc. can influence fibroin molecule
Self assembling process.But how to control external condition acquisition matte fibroin membrane and have not been reported.
Invention content
In view of this, one of the objects of the present invention is to provide a kind of methods preparing matte fibroin membrane；The mesh of the present invention
Two be to provide the matte fibroin membrane made from the method；The third object of the present invention is to provide the matte fibroin
Application of the film in improving cell adhesion as biomaterial.
In order to achieve the above objectives, the present invention provides the following technical solutions：
A method of matte fibroin membrane is prepared, is as follows：It is molten that fibroin is put into calcium chloride-alcohol-water ternary
Dissolving obtains silk fibroin solution in agent, then will be incubated after silk fibroin solution dialysis, centrifugation and filtering, then drying and forming-film, obtains
Matte fibroin membrane.
Preferably, the molar ratio of calcium chloride, ethyl alcohol and water is 1 in the ternary solvent：2：8.
Preferably, the fibroin is made by following methods：Cocoon layer is cut into small pieces, it is 1 to be put into solid-to-liquid ratio：50 quality point
The Na that number is 0.5%2CO3Degumming 2 times, each 30min is boiled in boiling in aqueous solution, is then pulled the fibrous silk after degumming out, is spent
Ionized water fully washs, and 60 DEG C of low temperature dryings obtain fibroin, can also use other methods degumming.
Preferably, the fibroin addition is more than 1 by solid-to-liquid ratio：5 are put into ternary solvent, it is furthermore preferred that solid-to-liquid ratio is
1：10.
It is described to be dissolved as dissolving 10~60min in 50~80 DEG C of magnetic force heating stirrings in the present invention；It is furthermore preferred that 75
DEG C magnetic force heating stirring dissolves 30min；The dialysis is at least 3d that dialyses in pure water, it is preferred that dialyse 3d in pure water；Institute
It is that 6000~8000r/min centrifuges 20~40min to state centrifugation；It is furthermore preferred that centrifuging 30min under the conditions of 8000r/min.
In the present invention, described be incubated is at 15~50 DEG C, to be incubated 12~48 hours under the conditions of 0~100r/min；It is preferred that
, at 37 DEG C, with 40-50r/min under the conditions of be incubated 24 hours.
In the present invention, the condition of the film forming is 25 DEG C ± 5 DEG C of film-forming temperature, humidity 65% ± 5%, is needed in film forming procedure
Film thickness is controlled, solution height is at least 2mm, preferably 2~5mm after pouring into film forming device with silk fibroin protein solution.
2, the matte fibroin membrane made from the method.
3, application of the matte fibroin membrane in improving cell adhesion as biomaterial.
The beneficial effects of the present invention are：The invention discloses a kind of methods preparing matte fibroin membrane, using calcium chloride-
Alcohol-water ternary solvent dissolves fibroin albumen, using the solvent dissolve regenerated fibroin albumen be easier to assemble agglomerates this
One feature is incubated fibroin albumen and forms a certain size aggregate, the aggregate is in silk by the self assembling process of fibroin molecule
Plain surface forms a certain size protrusion, to prepare matte fibroin membrane, since matte fibroin membrane is conducive to the adherency of cell
Growth improves performance of the fibroin membrane as biomaterial.
Description of the drawings
In order to keep the purpose of the present invention, technical solution and advantageous effect clearer, the present invention provides following attached drawing and carries out
Explanation：
Fig. 1 is that fibroin film surface is made in 1 method of embodiment.
Fig. 2 is that fibroin film surface is made in 2 method of embodiment.
Fig. 3 is that fibroin film surface is made in 3 method of embodiment.
Fig. 4 is fibroin membrane cell adherence result (A：Smooth fibroin membrane；B：Matte fibroin membrane).
Specific implementation mode
Below in conjunction with attached drawing, the preferred embodiment of the present invention is described in detail.
Embodiment 1
A method of matte fibroin membrane is prepared, is included the following steps：
(1) preparation of silk fibroin protein solution：Clean silk cocoon is taken, cocoon layer is cut into small pieces, it is 1 to be put into solid-to-liquid ratio:50
The Na that mass fraction is 0.5%2CO3Boiling is boiled to remove silk gum in aqueous solution, degumming twice, each 30min；By fiber after degumming
Shape silk is pulled out, is fully washed with deionized water, and 60 DEG C of low temperature dryings obtain fibroin, by the fibroin of acquisition according to 1:10 solid-liquid
Than being put into calcium chloride-alcohol-water ternary solvent, (calcium chloride-alcohol-water molar ratio is 1:2:8) in, 75 DEG C of magnetic force heating stirrings
30min is dissolved, silk fibroin solution is packed into bag filter after dissolving, is placed in pure water the 3d that dialyses, does not stop to change water therebetween, the silk after dialysis
Plain solution centrifugation (8000r/min, 30min) filtering is spare.
(2) silk fibroin protein solution is incubated：Silk fibroin solution is incubated 24 hours under the conditions of 37 DEG C, during which with about 40-50r/
The speed of min persistently stirs.
(3) it forms a film：Take drying and forming-film in the circular die for the silk fibroin solution 10mL to a diameter of 70mm being incubated, film forming temperature
25 DEG C ± 5 DEG C of degree, humidity 65% ± 5%.
The film surface formed using this method is shown in Fig. 1, as a result shows that there is a certain size protrusion on the surface of film, shows to obtain
Matte fibroin membrane.
Embodiment 2
A method of matte fibroin membrane is prepared, is included the following steps：
(1) prepared by silk fibroin protein solution：Clean silk cocoon is taken, cocoon layer is cut into small pieces, it is 1 to be put into solid-to-liquid ratio:50 matter
Measure the Na that score is 0.5%2CO3Boiling is boiled to remove silk gum in aqueous solution, degumming twice, each 30min；It will be fibrous after degumming
Silk is pulled out, is fully washed with deionized water, and 60 DEG C of low temperature dryings obtain fibroin；By fibroin according to 1:10 solid-to-liquid ratio is put into chlorine
Changing calcium-alcohol-water ternary solvent, (calcium chloride-alcohol-water molar ratio is 1:2:8) in, 75 DEG C of magnetic force heating stirring dissolvings
Silk fibroin solution is packed into bag filter by 30min after dissolving, is placed in pure water the 3d that dialyses, and does not stop to change water therebetween, the fibroin after dialysis is molten
It is spare that liquid centrifuges (8000r/min, 30min) filtering.
(2) silk fibroin protein solution is incubated：Silk fibroin solution is incubated 24 hours under the conditions of 45 DEG C, during which with about 40-50r/
The speed of min persistently stirs.
(3) it forms a film：Take drying and forming-film in the circular die for the silk fibroin solution 10mL to a diameter of 50mm being incubated, film forming temperature
25 DEG C ± 5 DEG C of degree, humidity 65% ± 5%.
The film surface formed using this method is shown in Fig. 2, as a result shows that there is a certain size protrusion on the surface of film, shows to obtain
Matte fibroin membrane.
Embodiment 3 (comparative examples)
A method of fibroin membrane is prepared, is included the following steps：
(1) prepared by silk fibroin protein solution：Clean silk cocoon is taken, cocoon layer is cut into small pieces, it is 1 to be put into solid-to-liquid ratio:50 matter
Measure the Na of score 0.5%2CO3Boiling is boiled to remove silk gum in aqueous solution, degumming twice, each 30min, by fibrous silk after degumming
It pulls out, is fully washed with deionized water, 60 DEG C of low temperature dryings obtain fibroin；By fibroin according to 1:10 solid-to-liquid ratio is put into 9M's
In lithium bromide, 75 DEG C of magnetic force heating stirrings dissolve 30min.Silk fibroin solution is packed into bag filter after dissolving, is placed in pure water and dialyses
3d does not stop to change water therebetween, and silk fibroin solution centrifugation (8000r/min, the 30min) filtering after dialysis is spare.
(2) silk fibroin protein solution is incubated：Silk fibroin solution is incubated 24 hours under the conditions of 37 DEG C, during which with about 40-50r/
The speed of min persistently stirs.
(3) it forms a film：Take drying and forming-film in the circular die for the silk fibroin solution 10mL to a diameter of 50mm being incubated, film forming temperature
25 DEG C ± 5 DEG C of degree, humidity 65% ± 5%.
The film surface formed using this method is shown in Fig. 3, as a result shows that the surface of film is smooth, shows that this method does not obtain matte
Fibroin membrane.
To prove the performance of matte fibroin membrane, through smooth silk made from matte fibroin membrane made from embodiment 1 and embodiment 3
Plain film carries out cell adhesion experiments, is as follows：
Fibroin membrane is cut into the square of 5mm*5mm, will be put into 24 porocyte culture plates after its ultraviolet sterilization, is added one
Quantitative cell and cell culture fluid, are put into incubator and cultivate 2-4 hours, fibroin membrane is taken out after cell is adherent, is seen after flattening
It examines.For convenience of observing and distinguishing cell, red or other fluorescent dyes can be added, cell is dyed.
The results are shown in Figure 4.The results show that the cell adhered on matte fibroin membrane is more, show that matte fibroin membrane can carry
High cell adhesion.Therefore, the performance of biomaterial can be improved when as biomaterial.
In the embodiment of the present invention, it is dissolved as dissolving 10~60min in 50~80 DEG C of magnetic force heating stirrings；It dialyses in pure water
At least 3d；6000~8000r/min centrifuges 20~40min, and goal of the invention can be achieved.15~50 DEG C are incubated in, with 0~
Identical effect can be obtained by being incubated 12~48 hours under the conditions of 100r/min.
Finally illustrate, preferred embodiment above is merely illustrative of the technical solution of the present invention and unrestricted, although logical
It crosses above preferred embodiment the present invention is described in detail, however, those skilled in the art should understand that, can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (9)
1. a kind of method preparing matte fibroin membrane, which is characterized in that be as follows：Fibroin is put into calcium chloride-ethyl alcohol-
Dissolving obtains silk fibroin solution in water ternary solvent, then will be incubated after silk fibroin solution dialysis, centrifugation and filtering, then dries
Film forming, obtains matte fibroin membrane.
2. the method for preparing matte fibroin membrane according to claim 1, it is characterised in that：Calcium chloride in the ternary solvent,
The molar ratio of ethyl alcohol and water is 1：2：8.
3. the method for preparing matte fibroin membrane according to claim 1, it is characterised in that：The fibroin is by following methods system
：Cocoon layer is cut into small pieces, it is 1 to be put into solid-to-liquid ratio：The Na that 50 mass fraction is 0.5%2CO3Degumming 2 is boiled in boiling in aqueous solution
Secondary, then each 30min pulls the fibrous silk after degumming out, fully washed with deionized water, and 60 DEG C of low temperature dryings obtain
Fibroin.
4. the method for preparing matte fibroin membrane according to claim 1, it is characterised in that：The fibroin addition presses solid-to-liquid ratio
More than 1：5 are put into ternary solvent.
5. the method for preparing matte fibroin membrane according to claim 1, it is characterised in that：It is described to be dissolved as in 50~80 DEG C of magnetic
Power heating stirring dissolves 10~60min；The dialysis is at least 3d that dialyses in pure water；The centrifugation is 6000~8000r/
Min centrifuges 20~40min.
6. the method for preparing matte fibroin membrane according to claim 1, it is characterised in that：It is described to be incubated as at 15~50 DEG C,
To be incubated 12~48 hours under the conditions of 0~100r/min.
7. the method for preparing matte fibroin membrane according to claim 7, it is characterised in that：The condition of the film forming is film forming temperature
25 DEG C ± 5 DEG C of degree, humidity 65% ± 5%.
8. the matte fibroin membrane made from any one of claim 1~7 the method.
9. application of the matte fibroin membrane described in claim 8 in improving cell adhesion as biomaterial.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810122565.5A CN108314791A (en) | 2018-02-07 | 2018-02-07 | A kind of method preparing matte fibroin membrane and products thereof and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810122565.5A CN108314791A (en) | 2018-02-07 | 2018-02-07 | A kind of method preparing matte fibroin membrane and products thereof and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108314791A true CN108314791A (en) | 2018-07-24 |
Family
ID=62902972
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810122565.5A Pending CN108314791A (en) | 2018-02-07 | 2018-02-07 | A kind of method preparing matte fibroin membrane and products thereof and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108314791A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102847198A (en) * | 2012-09-17 | 2013-01-02 | 浙江星月生物科技股份有限公司 | Silk fibroin film insoluble in water, and preparation and application of silk fibroin film |
| CN103131194A (en) * | 2011-12-01 | 2013-06-05 | 上海纳米技术及应用国家工程研究中心有限公司 | Infusible fibroin membrane with controllable crystallinity and preparing method thereof |
| US20130337227A1 (en) * | 2010-11-26 | 2013-12-19 | Tokyo Institute Of Technology | Non-fibrogenesis collagen material and a manufacturing method thereof |
| CN104667349A (en) * | 2015-02-06 | 2015-06-03 | 福州大学 | Growth factor-loading silk fibroin/collagen bracket material and preparation method thereof |
| CN106279749A (en) * | 2016-09-22 | 2017-01-04 | 大连工业大学 | A kind of alginate and the preparation method of fibroin albumen composite sponge |
-
2018
- 2018-02-07 CN CN201810122565.5A patent/CN108314791A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130337227A1 (en) * | 2010-11-26 | 2013-12-19 | Tokyo Institute Of Technology | Non-fibrogenesis collagen material and a manufacturing method thereof |
| CN103131194A (en) * | 2011-12-01 | 2013-06-05 | 上海纳米技术及应用国家工程研究中心有限公司 | Infusible fibroin membrane with controllable crystallinity and preparing method thereof |
| CN102847198A (en) * | 2012-09-17 | 2013-01-02 | 浙江星月生物科技股份有限公司 | Silk fibroin film insoluble in water, and preparation and application of silk fibroin film |
| CN104667349A (en) * | 2015-02-06 | 2015-06-03 | 福州大学 | Growth factor-loading silk fibroin/collagen bracket material and preparation method thereof |
| CN106279749A (en) * | 2016-09-22 | 2017-01-04 | 大连工业大学 | A kind of alginate and the preparation method of fibroin albumen composite sponge |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6439918B2 (en) | Method for producing three-dimensional cell structure | |
| CN106676065A (en) | Adipose tissue source exosome gel, preparation method and application | |
| CN107320780A (en) | A kind of multilayer aquagel of hollow tubular structure and preparation method and application | |
| CN103691005B (en) | A kind of micro--Na fibrous tissue engineering rack and preparation method thereof | |
| WO2018014453A1 (en) | Fermentation technique to improve production level of recombinant human collagen | |
| CN106729984A (en) | A kind of Isin glue collagen repairs sponge and preparation method thereof | |
| CN101912638A (en) | Nano silver-carrying-silicon dioxide catheter and production method thereof | |
| CN107425119B (en) | Organic biocompatible resistance-variable nerve bionic device and preparation method and application thereof | |
| CN107602884A (en) | A kind of fibroin/chitosan composite intelligent hydrogel and preparation method thereof | |
| CN103261395B (en) | Bioreactor with the lid for facilitating access for culture chamber | |
| CN106729959A (en) | A kind of fibroin fiber antiseptic dressing and preparation method thereof | |
| CN105879113A (en) | Method for preparing three-dimensional cell scaffolds on basis of silk fibroins | |
| CN104483178A (en) | Preparation method of chromosomes of adult epinephelus akaara | |
| CN102764451B (en) | Preparation method and application of porous biological cellulose gel material | |
| CN108384754A (en) | A kind of preparation method of excretion body and the excretion body prepared by this method and purposes | |
| CN106039400B (en) | Ice crystal template prepares the methods and applications of regular lamellar structure three dimensional biological bracket | |
| CN1986608A (en) | Preparing process and application of nano chitosan particle | |
| CN108314791A (en) | A kind of method preparing matte fibroin membrane and products thereof and application | |
| CN102329728B (en) | Chitosan/arginine-glycine-aspartic acid (RGD) three-dimensional porous microcarrier and preparation method and application thereof | |
| CN105734006A (en) | Preparation method of acellular sodium alginate bionic hydrogel | |
| CN111635860B (en) | System and method for stem cell culture | |
| CN103484356B (en) | Fermentation reactor and preparation method for dynamically culturing and preparing ordered bacteria cellulose | |
| CN103467778B (en) | Bacteria cellulose/Yelkin TTS matrix material and Synthesis and applications | |
| CN109295136A (en) | A method of large-scale production fetal stem cell secretin | |
| CN106620877B (en) | A kind of capillary network and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180724 |