CN109498829A - Fibroin wound dressing and its preparation method and application - Google Patents
Fibroin wound dressing and its preparation method and application Download PDFInfo
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- CN109498829A CN109498829A CN201811249902.3A CN201811249902A CN109498829A CN 109498829 A CN109498829 A CN 109498829A CN 201811249902 A CN201811249902 A CN 201811249902A CN 109498829 A CN109498829 A CN 109498829A
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- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
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Abstract
The invention discloses fibroin wound dressing and its preparation method and application, the dressing is the dual network structure of wire protein fiber support network and sericin hydrogel network interpenetrating；The wire protein fiber bracket is the composition or silk gum fibrous framework of fibroin and silk gum, and the hydrogel is sericin hydrogel.Fibroin wound dressing of the present invention has porous structure, good permeability；With good mechanical strength, it is convenient for operation stitching；With high-biocompatibility；Skin wound healing can be effectively facilitated, can be used as skin histology wound repair；The sericin wound dressing has good biocompatibility and cell adhesion, can long-term sertoli cell survival, proliferation and migration；The wound dressing can be using carrying medicament and growth factor etc. as delivery vector；The dressing is also a kind of three-dimensional porous sericin biological support, can be used for the growth of extracellular matrix sertoli cell and promotes nutriment exchange, can be applied to the reparation of Various Tissues damage and the treatment of disease.
Description
Technical field
The invention belongs to biomedical material technologies, are related to a kind of new silk protein material, specially silk egg
White wound dressing and its preparation method and application.
Background technique
Silk is mainly made of two kinds of albumen, and one kind is fibroin, and another kind is the silk gum for being wrapped in fibroin albumen surface.When
Before, silkworm can be divided into two major classes according to whether produced silk-fibroin contains fibroin, and one kind is common wild type cultivated silkworm breed variety, spit
Silk-fibroin out not only contains fibroin, but also contains silk gum；Another kind of is saltant type cultivated silkworm breed variety, and the silk-fibroin that such silkworm is spat is only
Containing silk gum without fibroin.For a long time due to the deficiency studied and recognized, people focus mainly on fibroin albumen research and
Using until in recent years, it has been found that sericin not only has degradable, antibacterial, anticancer, anti-oxidant, anticoagulation, hydrophilic, guarantor
Wet, non-immunogenicity and the promotion bioactivity such as cell adherence and proliferation, and be conducive to repair rich in amino, hydroxyl, carboxyl etc.
Decorations or the active group of transformation, sericin are re-recognized and obtain people and more and more paid close attention to, and have become a kind of excellent already
Different novel tissue engineering material.
It is currently based on fibroin and Various Tissues engineering material is prepared for subsequent processing techniques people by separation, purifying
Material, such as hydrogel, sericin film, silk gum sponge-like material, silk gum nanofiber and nanoparticle etc..On the other hand, this project
Group early period is developed by the spinning habit of silkworm Biotechnology in Genetic Breeding and regulation silkworm with natural structure and active silk egg
White biological support (dressing), this is that the natural sericin of development and utilization opens new approach.Although this kind of natural silk gum egg
White to have very good characteristic, however there are still some shortcomings, such as, it is difficult to carrying medicament, as wound dressing easily with wound
Mouth adhesion.
Summary of the invention
The technical issues of solution: it for overcome the deficiencies in the prior art, obtains a kind of high concentration and is easy to the silk gum of plastic
Albumen and drug loading rate are high, be not easy when being used as dressing with wound adhesion, the present invention provides fibroin wounds to apply
Material and its preparation method and application.
Technical solution: fibroin wound dressing, the dressing are wire protein fiber support network and sericin water-setting
The dual network structure of glue network interpenetrating；The wire protein fiber bracket is the composition or silk gum of fibroin and silk gum.The water-setting
Glue is sericin hydrogel.
The preparation method of the fibroin wound dressing comprising the steps of:
(1) building man silkworms spin silk platform, selects smooth stylobate, and blotting paper or absorbent cloth are covered first on stylobate, then
Cover the nylon wire or cotton gauze of one layer of 2~30 mesh；
(2) by silkworm rearing to 5 matured silkworm phases in age, after draining completely in Silkworm, Bombyx mori, step (1) building is transferred them to
On platform, the throw-in density of every batch of silkworm is 250/square metre or less；
(3) silkworm is completed after spinning and pupating, and chooses healthy silkworm pupa for changing moth, production of hybrid seeds passage；
(4) silkworm that step (3) is reserved seed for planting is raised in the way of step (2), and repeats step (3)；
(5) repeat passage 15~30 generations after, raised in the way of step (2), to family silkworms spin silk complete, by net and lower layer
Paper or cloth separate, then separate fibroin from network, can be obtained wire protein fiber bracket；
(6) take fibroin deficient mutant silk cocoon, remove impurity and pollution silk cocoon rinse repeatedly with water again after, in room temperature baking
It is dry or dry, it crushes, water is added to be placed in 100~180 DEG C, 1~10 hour, the mass ratio of silk cocoon and water was 1/100~20/100, was obtained
Obtain silk cocoon suspension；Silk cocoon suspension is placed in 100~150 DEG C, 5~600min, removal insoluble matter is molten to get sericin
Liquid；
(7) sericin solution made from part steps (6) is taken, is hydrolyzed using trypsase, alkali protease etc.,
To obtain the sericin of different molecular weight；
(8) after the sericin solution for obtaining step (6) He Buju (7) 100/0~5/100 mixing by volume, then
The wire protein fiber bracket that step poly- (5) obtain is immersed in mixed solution, is taken out after -2 hours 1 second, is placed in 0~60 DEG C, to
After sericin solution plastic, then it is placed in 0 DEG C or less freezing, and low-temperature vacuum drying, can be obtained fibroin wound dressing.
Preferably, cultivated silkworm breed variety includes wild type and fibroin deficient mutant；Wherein, wild type cultivated silkworm breed variety be cyanines pine A,
White jade, bright moon or lotus；The kind of fibroin deficient mutant silkworm be 190Nd-s, 188Nd-s, 185Nd-s, 150Nd-s,
140Nd-s, 139Nd-s or Nd.Wire protein fiber bracket in the fibroin wound dressing includes two major classes: when use
It is sericin fibrous framework that silkworm obtains when being fibroin deficiency type domestic silkworms kind；When using is wild type silkworm, then
Fibroin fiber bracket had not only contained sericin but also had contained fibroin albumen.
Preferably, the pH value of sericin solution is 1-11 in step (8).
Application of the fibroin wound dressing described above in bio-medical material.
Application of the fibroin wound dressing described above in cell carrier.
Application of the fibroin wound dressing described above in pharmaceutical carrier.
The utility model has the advantages that (1) present invention is prepared using native fibroin fibrous framework technology combination novel hydrogels for the first time
Technological development has gone out the fibroin wound dressing with dual-network, which not only possesses the natural knot of fibroin
Structure and activity, while this programme introduces fibroin network structure, provides for wound dressing carrying medicament and growth factor
It is convenient, while more fibrous fibroin bracket, surface more smooth, it is not easy and wound adhesion.The silk that this programme obtains simultaneously
Glue protein simple production process, it is low in cost, there is replicability；(2) the dual network structure that the present invention uses passes through regulation water
The ratio of gel and fibrous framework can regulate and control degradation rate, porosity, mechanical property of dressing etc.；(3) institute through the invention
Fibroin wound dressing made from method is stated with porous structure, good permeability；With good mechanical strength, convenient for operation
Suture；With high-biocompatibility；Skin wound healing can be effectively facilitated, can be used for skin histology wound repair；(4) of the invention
The fibroin wound dressing has good biocompatibility and cell adhesion, can long-term sertoli cell survival, increase
It grows and migrates；(5) dressing is also a kind of three-dimensional porous sericin biological support, can be used for extracellular matrix sertoli cell
Growth and promotion nutriment exchange, can be applied to the reparation of Various Tissues damage and the treatment of disease.
Detailed description of the invention
Fig. 1 is fibroin wound dressing structure chart, and A is fibroin (containing fibroin and silk gum) wound dressing macrostructure
Figure, B are fibroin (containing fibroin and silk gum) wound dressing micro-structure diagram；C is fibroin (containing only silk gum) wound dressing
Macrostructure figure, D are fibroin (containing only silk gum) wound dressing micro-structure diagram；
Fig. 2 is fibroin wound dressing porosity statistical chart；
Fig. 3 is sericin wound dressing swelling kinetics figure；
Fig. 4 is Mouse Skin Fibroblasts NIH3T3 cell viability detection figure；
Fig. 5 is cell migration figure in sericin wound dressing；
Fig. 6 is sericin wound dressing drug release rate figure；
Fig. 7 is that sericin wound dressing promotes skin wound healing figure.
Specific embodiment
Following embodiment further illustrates the contents of the present invention, but should not be construed as limiting the invention.Without departing substantially from
In the case where spirit of that invention and essence, to modification made by the method for the present invention, step or condition and replaces, belong to the present invention
Range.Unless otherwise specified, the conventional means that technological means used in embodiment is well known to those skilled in the art.
Embodiment 1 prepares the wound dressing of silkworm sericin
(1) cultivated silkworm breed variety is selected
Fibroin deletion form mutation cultivated silkworm breed variety is selected (to be saved by Inst. of Silkworm, Chinese Academy of Agricultural Sciences and be commercialized outward
There is provided), sericin is contained only in the silk-fibroin that such cultivated silkworm breed variety is spat and without fibroin albumen.
(2) preparation man silkworms spin silk platform
Select 6 pieces with a thickness of 1cm, length and width be 50cm cardboard, making a surface area is 1.5m2Cube paper
Case.Carton is put on a smooth table top, covers 3 layers in its upward one side with a thickness of 0.4mm through high-temperature sterilization
Newsprint.Then one layer of cotton gauze (10 mesh) is covered on paper again, and the surrounding of cotton gauze is fixed.
(3) selection and breeding of silkworm kind
Selection bombyx mori silk fibroin deficient mutant kind (190Nd-s, 188Nd-s, 185Nd-s, 150Nd-s, 140Nd-s,
139Nd-s), under standard environment (28 DEG C of 1-2 age, 25 DEG C of 3-5 age), new fresh mulberry leaf is selected to feed, using one day three Hui Yu.To
When silkworm 5 age maturation, silkworm is placed on one and is covered in the silkworm plaque of newspaper, after waiting the excretion of silkworms excrement clean, then silkworm turned
It moves on on family's silkworms spin silk platform of step (2) production.Spinning platform is in the uniform environment of light, allows silkworm on platform
Spinning, personnel aside observe, and if discovery has silkworm to fall down on platform, i.e., the silkworm to fall down are put back to platform again.Family's curley waste
It pupates on platform after complete silk.It to the end of pupating, selects to mate after pupating more normal individuation moth and reserve seed for planting, remaining is washed in a pan
It eliminates.By such method breeding 20 more than generation, it can be obtained the fibroin deficiency type domestic silkworms new varieties for being adapted to spin on platform.
(4) sericin wound dressing is prepared
Selection is raised by the cultivated silkworm breed variety after step (3) breeding, and method for breeding is the same as described in step (3).To silkworm 5
After age is mature, silkworm is transferred in the silkworm plaque for being covered with newspaper, is placed on step after their internal excrement drain clean, then by silkworm
(2) on the spinning platform made, 100 silkworms of every square metre of placement.It after 24 hours, is finished at this point, silkworm has spun, by this batch
Silkworm is removed.Then 100 silkworms that will be spun are reapposed again, and after same 24 hours, to family, silkworms spin silk finishes, and then turns
It moves, then relays 100 silkworms that will be spun.After 24 hours, silkworm is removed.Net with sericin is gently taken
Under, then net is slowly removed, can be obtained certain thickness sericin fibrous framework.
Silkworms spin silk that platform is made of three parts for family, and lower layer's stylobate is a smooth platform.Intermediate one layer for blotting paper or
Cloth, top layer are nylon wire or cotton gauze.
(5) the sericin fibrous framework that step (4) obtains is subsequently placed in room temperature and is dried in the air with ultrapure water rinsed clean
It is dry.
(6) it takes 100 grams of fresh silk cocoons of 185Nd-s to be cleaned up with ultrapure water, is placed in room temperature and dries, be then crushed to
Mixing is sufficiently stirred hereinafter, weighing 10 grams of powder again and 100ml ultrapure water being added in 0.2mm, is finally placed in 121 DEG C, 15min.
(7) the silk gum solution centrifugation removal insoluble matter obtained step (6), obtains sericin solution.
(8) the sericin fibrous framework in step (5) is immersed in the solution in step (7), room temperature keeps 10
Minute, it then takes out.
(9) sample is placed in 4 DEG C again, 24 hours, is then put in -80 DEG C, 12 hours.
(10) sample is placed in low temperature vacuum drier to the silk gum that can be obtained interpenetrating dual network structure after low temperature drying
Albumen wound dressing.
Embodiment 2 prepares Juvenile Hormone wound dressing
(1) cultivated silkworm breed variety is selected
Wild type cultivated silkworm breed variety (saved by Inst. of Silkworm, Chinese Academy of Agricultural Sciences and commercialization provides outward) is selected, it should
Not only contain sericin in the silk-fibroin that class cultivated silkworm breed variety is spat but also contain fibroin albumen.
(2) preparation man silkworms spin silk platform
Select 6 pieces with a thickness of 1cm, length and width be 50cm cardboard, making a surface area is 1.5m2Cube paper
Case.Carton is put on a smooth table top, covers 3 layers in its upward one side with a thickness of 0.4mm through high-temperature sterilization
Newsprint.Then one layer of cotton gauze (10 mesh) is covered on paper again, and four sides of cotton gauze are tightened and fixed.
(3) selection and breeding of silkworm kind
Wild type cultivated silkworm breed variety (cyanines pine or white jade) is chosen, under standard environment (28 DEG C of 1-2 age, 25 DEG C of 3-5 age), is selected
New fresh mulberry leaf feeding, using one day three Hui Yu.When silkworm 5 age maturation, silkworm is placed on one and is covered in the silkworm plaque of newspaper, etc.
After the excretion of silkworm excrement is clean, then silkworm is transferred on family's silkworms spin silk platform of step (2) production.Allow silkworm on platform
Spinning, personnel aside observe, and if discovery has silkworm to fall down on platform, i.e., the silkworm to fall down are put back to platform again.Family's curley waste
It pupates on platform after complete silk.It to the end of pupating, selects to mate after pupating more normal individuation moth and reserve seed for planting, remaining is washed in a pan
It eliminates.By such method breeding 15 more than generation, it can be obtained the fibroin deficiency type domestic silkworms new varieties for being adapted to spin on platform.
(4) sericin wound dressing is prepared
Selection is raised by the cultivated silkworm breed variety after step (3) breeding, and method for breeding is the same as described in step (3).To silkworm 5
After age is mature, silkworm is transferred in the silkworm plaque for being covered with newspaper, is placed on step after their internal excrement drain clean, then by silkworm
(2) on the spinning platform made, 100 silkworms of every square metre of placement.It after 24 hours, is finished at this point, silkworm has spun, by this batch
Silkworm moves away.Then 100 silkworms that will be spun are reapposed again, and after same 24 hours, to family, silkworms spin silk finishes, then
Transfer, then relay 100 silkworms that will be spun.After 24 hours, silkworm is removed.Net with sericin is gently taken
Under, then net is slowly removed, then can be obtained certain thickness sericin fibrous framework.
Silkworms spin silk that platform is made of three parts for family, and lower layer's stylobate is a smooth platform.Intermediate one layer for blotting paper or
Cloth, top layer are nylon wire or cotton gauze.
(5) the fibroin fiber bracket that step (4) obtains is subsequently placed in room temperature and is dried in the air with ultrapure water rinsed clean
It is dry.
(6) it takes 100 grams of fresh silk cocoons of 185Nd-s to be cleaned up with ultrapure water, is placed in room temperature and dries, be then crushed to
Mixing is sufficiently stirred hereinafter, weighing 10 grams of powder again and 100ml ultrapure water being added in 0.2mm, is finally placed in 121 DEG C, 15min.
(7) the silk gum solution centrifugation removal insoluble matter obtained rapid (six), obtains sericin solution.
(8) the sericin fibrous framework in rapid (five) is immersed in the solution in step (7), room temperature is kept for 10 points
Clock then takes out.
(9) sample is placed in 4 DEG C again, 24 hours, is then put in -80 DEG C, 12 hours.
(10) sample is placed in low temperature vacuum drier to the silk gum that can be obtained interpenetrating dual network structure after low temperature drying
Albumen wound dressing.
The wound dressing prepared to embodiment 1 and 2 method of embodiment is tested for the property, as a result as follows:
As shown in Figure 1, wherein A is the macrograph (being prepared by 2 scheme of embodiment) of fibroin wound dressing, by commonly counting
Code camera shooting；B is the micro-structure diagram (being prepared by 2 scheme of embodiment) of fibroin wound dressing, using electron microscope
It takes pictures.From B figure as it can be seen that wound dressing is interpenetrating dual network structure, one of them is the network structure of threadiness, another is silk
Glue protein hydrogel network structure.C is the macrograph (being prepared by 1 scheme of embodiment) of sericin wound dressing, by commonly counting
Code camera shooting；D is the micro-structure diagram (being prepared by 1 scheme of embodiment) of sericin wound dressing, using electron microscope
It takes pictures.
The measurement of the porosity of silk-fibroin wound dressing is using ultrapure water as substitution solution.It is w by dry weight1Sample
Product are immersed in ultrapure water, are taken out after 1 hour, after the extra water of removal sample surfaces the weight of sample is referred to as w2, then sample
Porosity be (w2-w1)/w2× 100%.As a result as shown in Figure 2.
The measurement of sericin wound dressing swelling kinetics, is soaked in different pH value (pH for sericin wound dressing
3.0, pH 7.4, pH 11.0) PBS solution in, be placed in 37 DEG C of constant incubators, daily replace a PBS solution, pre-
If time point, take out and the water for removing excess surface be re-weighed.The expansion rate formula of sericin wound dressing are as follows: (Mw-
Md)/Md× 100%, wherein MwFor the moisture of sample, MdFor the initial dry weight of sample.As a result as shown in Figure 3.
3 silkworm sericin wound dressing of embodiment is used as cell carrier
(1) mouse detects at skin fibroblasts (NIH3T3) sericin wound dressing cell viability
(1) sericin wound dressing is cut to sheet first, is then washed 3 times with sterilizing PBS, then impregnated with 75% ethyl alcohol
It 1 hour, is finally washed 1 time with the PBS that sterilized, it is spare to be placed in room temperature.
(2) cell collected from Tissue Culture Flask is suspended, is dispelled, planted in 96 porocyte culture plates, to cell
Sericin wound dressing pretreated in step (1) is transferred in culture plate containing cell after adherent, every hole places one
Piece sericin wound dressing.The cell culture well of sericin wound dressing is not placed as control.Cell culture training used
Support base be DMEM high glucose medium, cell be placed in cell incubator (37 DEG C, CO2Concentration is 5%, humidity 100%) in culture.
As shown in figure 4, using CCK8 method detection cell after planting after 1 day and 2 days cell vigor.As the result is shown:
Mouse Skin Fibroblasts (NIH3T3) sericin wound dressing group is compared with control group, without aobvious in terms of cell viability
Write sex differernce, the results showed that, sericin wound dressing has good cell compatibility.
(2) the cell migration detection of sericin wound dressing
Cell line used is Mouse Skin Fibroblasts (NIH3T3).
(1) round sheet is cut into sericin wound dressing first, is then washed 3 times with sterilizing PBS, then soaked with 75% ethyl alcohol
Bubble 1 hour, is finally washed 1 time with the PBS that sterilized, is placed in 24 porocyte culture plates.
(2) cell collected from Tissue Culture Flask is suspended, is dispelled, plantation contains sericin wound in step (1)
In the tissue culture plate of dressing, initial repopulating cell number is counted.After cell is adherent, sericin wound dressing is shifted
Into 24 new porocyte culture plates.Each remaining cell quantity in hole in former culture plate is counted again, then learns sericin wound
Initial repopulating cell quantity in dressing.
Hereafter, every other day sericin wound dressing is transferred in new tissue culture plate once, while counts former
Remaining cell number and record is performed in corresponding hole in culture plate.It is last for statistical analysis.Cell culture culture used
Base be DMEM high glucose medium, cell be put in cell incubator (37 DEG C, CO2Concentration is 5%, humidity 100%) in culture.
As a result as shown in figure 5, the cell that migrates out from sericin wound dressing of 21 days inner cells in statistics is presented
The phenomenon that sustainable growth.Show that sericin wound dressing has good cell compatibility, is capable of depositing for long-term sertoli cell
Living, proliferation and migration.
4 silkworm sericin wound dressing of embodiment is used for medicine controlled releasing, and horseradish peroxidase (HRP) is used as mode medicine
Object
(1) in 1 sericin wound dressing preparation process of embodiment, sericin solution that step 7 is obtained it is dense
Degree is adjusted to 8% (w/v), and 0.1ml HRP (concentration 1mg/ml) ratio then is added in 1ml sericin solution and mixes HRP
It is even into sericin solution, then weigh.
(2) sericin fibrous framework is cut to the circle that diameter is 1ml, weighs, is then immersed in step (1) respectively
HRP- sericin solution in, room temperature keep 10 minutes, then take out and weigh.According to HRP- sericin solution and silk gum
The variation for (not immersing before solution and after immersing solution) weight before and after azelon calculates each sample initial load HRP's
Amount.
(3) sample is placed in 4 DEG C again, 24 hours, is then put in -80 DEG C, 12 hours.
(4) sample is placed in low temperature drying in low temperature vacuum drier.
(5) sample after freeze-drying is placed in 5ml centrifuge tube, and 2ml PBS (pH7.4) is added, be placed in 37 DEG C；
It is careful at the 1st, 2,3,4,5,6,7,8,9,11,14 day respectively to take out supernatant, and rejoin 2mL PBS (pH
7.4) using the content of HRP in enzyme-linked immunization (ELISA) measurement supernatant, added up by calculating in various time points supernatant
HRP content and drugloading rate ratio i.e. obtain drug add up release rate.
As shown in fig. 6, drug (HRP) is released to 30.13% in 24 hours, reach 56.20% or so in 3 days, 14 days
For 90.34%., the result shows that, sericin wound dressing has good controlled-release function, can be used as pharmaceutical carrier.
5 silkworm sericin wound dressing of embodiment is used for mouse skin injury repair
(1) sericin wound dressing is cut into the round sheet that diameter is 1cm first, then small with the immersion 1 of 75% ethyl alcohol
When, it is finally washed 1 time with the PBS that sterilized, it is spare that drying is placed on room temperature.
(2) mouse trauma model is prepared, the animal of selection is the C57 mouse of 8 week old.First choice will be at mouse anesthesia and depilation
Reason；Then the round skin that diameter is 1cm is cut off in back of mice, the pretreated sericin wound dressing of step (1) is pasted
It is then attached on wound face again on gauze, the mouse of Bandaid dressing processing is as control.
Experimental result as shown in fig. 7, sericin dressing group compared with control group (Bandaid group) wounds in mice healing rate more
Fastly, the Bandaid group mouse skin damage surface of a wound not yet heals completely after 14 days, and the mouse skin of sericin wound dressing group damages
Hurt the surface of a wound to heal completely.Show that sericin wound dressing can effectively facilitate skin ultrastructure, is suitable for skin histology
Injury repair.
Claims (7)
1. fibroin wound dressing, which is characterized in that the dressing is wire protein fiber support network and sericin water-setting
The dual network structure of glue network interpenetrating；The wire protein fiber bracket is the composition or silk gum fibrous framework of fibroin and silk gum,
The hydrogel is sericin hydrogel.
2. the preparation method of fibroin wound dressing described in claim 1, which is characterized in that comprise the steps of:
(1) building man silkworms spin silk platform, selects smooth stylobate, blotting paper or absorbent cloth is covered first on stylobate, then cover
The nylon wire or cotton gauze of one layer of 2~30 mesh；
(2) by silkworm rearing to 5 matured silkworm phases in age, after draining completely in Silkworm, Bombyx mori, the platform of step (1) building is transferred them to
On, the throw-in density of every batch of silkworm is 250/square metre or less；
(3) silkworm is completed after spinning and pupating, and chooses healthy silkworm pupa for changing moth, production of hybrid seeds passage；
(4) silkworm that step (3) is reserved seed for planting is raised in the way of step (2), and repeats step (3)；
(5) after repeating 15~30 generations of passage, raised in the way of step (2), to family, silkworms spin silk completes, by net and lower layer's paper or
Cloth separates, then separates fibroin from network, can be obtained wire protein fiber bracket；
(6) fibroin deficient mutant silk cocoon is taken, after cleaning up, drying is simultaneously crushed, and adds water to be placed in 100~180 DEG C, 1~10 is small
When, obtain sericin solution；
(7) sericin solution made from part steps (6) is taken, is hydrolyzed using trypsase, alkali protease etc., to obtain
Obtain the sericin of different molecular weight；
(8) after the sericin solution for obtaining step (6) He Buju (7) 100/0~5/100 mixing by volume, then will step
The wire protein fiber bracket that poly- (5) obtain is immersed in mixed solution, is taken out after -2 hours 1 second, 0~60 DEG C is placed in, to silk gum
After protein solution plastic, then it is placed in 0 DEG C or less freezing, and low-temperature vacuum drying, can be obtained fibroin wound dressing.
3. the preparation method of fibroin wound dressing according to claim 2, which is characterized in that cultivated silkworm breed variety includes open country
Raw type and fibroin deficient mutant；Wherein, wild type cultivated silkworm breed variety is cyanines pine A, white jade, bright moon or lotus；Fibroin deletion mutation
The kind of type silkworm is 190Nd-s, 188Nd-s, 185Nd-s, 150Nd-s, 140Nd-s, 139Nd-s or Nd.
4. the preparation method of fibroin wound dressing according to claim 2, which is characterized in that silk gum in step (8)
The pH value of protein solution is 1-11.
5. application of the fibroin wound dressing in bio-medical material described in claim 1.
6. application of the fibroin wound dressing in cell carrier described in claim 1.
7. application of the fibroin wound dressing in pharmaceutical carrier described in claim 1.
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| CN113402733A (en) * | 2021-05-24 | 2021-09-17 | 重庆医科大学 | Composite hydrogel and preparation method and application thereof |
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