CN103965310A - Self-assembling method for preparing fibroin microspheres - Google Patents
Self-assembling method for preparing fibroin microspheres Download PDFInfo
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Abstract
The invention discloses a self-assembling method for preparing fibroin microspheres. The method comprises the following steps: shearing cocoon shells of silkworms into pieces, putting the sheared cocoon shells into a Na2CO3 water solution to be subjected to degumming treatment, and cleaning with deionized water; putting the cleaned fibroin sample into the Na2CO3 water solution to be subjected to the degumming treatment again, cleaning with deionized water, then flatly spreading the fibroin sample in a baking oven, drying to obtain silkworm fibroin fiber and the like until a milk white liquid is obtained, and then freeze-drying into powder, so that a finished product can be prepared. According to the invention, the fibroin microspheres are prepared without adding any poisonous chemical cross-linking agents, the preparation technology is simple, the reaction condition is moderate, and the fact that the structural characteristics of silk fibroin is kept is facilitated, the utilization ratio of silk fibroin is increased, the production cost is reduced, and the yield of fibroin microspheres is increased. The fibroin microspheres prepared by adopting the self-assembling method can be applied to the fields of medicine carrying micro-system, cellular uptake and the like, and a new thought and a new method are provided for the development of biomedicine.
Description
Technical field
The invention belongs to a kind of preparation method of fibroin microballoon, specifically, is a kind of self-assembling method of fibroin microballoon.
Background technology
Silk fibroin is the topmost integral part of silk, account for 2/3, formed by 18 seed amino acids such as glycine, L-Ala, Serines, it is a kind of natural protein, because of features such as its good mechanical property, biocompatibility, biological degradability, ventilative oxygen-permeable and moisture-absorbing moisture-keepings, in the fields such as biomedicine, makeup, food, obtain studying more and more widely and apply ([J] Progress in Polymer Science, 2007,32:991-1007).The fibroin microballoon of preparing taking silk fibroin as raw material, has that specific surface area is large, an advantage such as good biocompatibility, biodegradable, good mobility and diffustivity, is therefore studied widely.Traditional method of preparing fibroin microballoon mainly contains emulsion process, spray-drying process, laminar jet technology, mechanical crushing method, salting-out process etc.But adopt the organic solvent removal that traditional method (as emulsion process, spray-drying process, laminar jet method) adds in preparation process thoroughly can not cause toxicity problem; The microspherulite diameter of preparing is in addition larger, and preparation condition needs high pressure-temperature to destroy the structure of protein.In recent years, self-assembly has become a kind of novel synthetic method, its reaction conditions gentleness, and self-assembly method becomes prefered method prepared by silk fibroin microballoon.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provide a kind of easy and simple to handle, reaction conditions is gentle, do not add any chemical cross-linking agent and obtain the method having compared with the silk fibroin microballoon of small particle size.
The present invention is achieved through the following technical solutions:
A kind of self-assembling method of preparing fibroin microballoon is characterised in that, comprises the steps: that (1) shreds Silkworm Cocoon Shell, puts into the Na of mass percent as 0.3%-0.7% taking the bath raio of (1:50)-(1:00)
2cO
3in the aqueous solution, carry out after degumming process 25-35min, use washed with de-ionized water 1-3 time; Clean fibroin sample is dropped into the Na that mass percent is 0.3%-0.7%
2cO
3in the aqueous solution, carry out again after degumming process 25-35min, with after washed with de-ionized water 1-3 time, fibroin sample is shakeout and is placed in the baking oven oven dry of 40-50 DEG C and obtains bombyx mori silk fibroin fiber;
(2) the bombyx mori silk fibroin fiber in step (1) is added in the LiBr solution that concentration is 7-11mol/L and is placed in shaking table 5-8h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and is obtained compared with pure silk cellulose solution with deionized water dialysis 3-5d, after natural air drying, filtration, obtain pure silk cellulose solution;
(3) the pure silk cellulose solution in step (2) is adjusted into the solution that concentration is 3%-7%;
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5-10min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is (1:5)-(1:8), continuation magnetic stirring apparatus 10-30min;
(5) solution in step (4) is placed in to-20 DEG C～-80 DEG C refrigerator and cooled and freezes 24-48h;
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
As improving further, Na in step of the present invention (1)
2cO
3the temperature of the aqueous solution is 85-105 DEG C, and boiling water bath can improve the thoroughness of coming unstuck, and improves the purity of silk fibroin, makes the ball surface of preparation Paint Gloss.
As improving further, in step of the present invention (1), silkworm silk cocoon is shredded, be added to the Na of mass percent as 0.5% taking the bath raio of 1:100
2cO
3in the aqueous solution, carry out degumming process.
As improving further, in step of the present invention (1), in degumming process 30min after, by washed with de-ionized water 2 times, then the 30min that comes unstuck, washed with de-ionized water 3 times, then to carry out temperature be that 50 DEG C of oven dry obtain bombyx mori silk fibroin fiber.Repeatedly clean sample and can thoroughly remove the interference of sericin; Temperature is suitable, can not affect the change of silk fibroin structure.
As improving further, in step of the present invention (2), the bombyx mori silk fibroin fiber in step (1) is added in the LiBr solution that concentration is 9mol/L and is placed in shaking table 6h, thus accelerate dissolution reduce molecular weight and obtain silk fibroin solution.
As improving further, in step of the present invention (2), the concentration of LiBr solution is 8-10mol/L.
As improving further, in step of the present invention (2), the concentration of LiBr solution is 9mol/L, what make that fibroin dissolves in the short period of time is more thorough, the excessive concentration of LiBr solution can make the overlong time of dialysis, and dialysis is not thorough, if too low, the treatment time that silk fibroin solution dissolves is long, and gelatin phenomenon can occur.
As improving further, it is 3% solution that the pure silk cellulose solution in step of the present invention (3) is adjusted into concentration, can make the fibroin microballoon mean particle dia of preparation be controlled at 1um left and right, and microspherulite diameter is evenly distributed.
As improving further, in step of the present invention (4), silk fibroin solution in step (3) is placed in to beaker magnetic agitation 5min, then dropwise add aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:5, continue magnetic stirring apparatus 30min, can be evenly distributed by microspherulite diameter, microballoon favorable dispersity.
The present invention compared with prior art, have the following advantages and effect: under the condition of not adding any poisonous chemical cross-linking agent, prepared fibroin microballoon, preparation technology is simple, reaction conditions is gentle, be conducive to keep silk fibroin constitutional features, improve silk fibroin utilization ratio, reduce production costs, improve the output of fibroin microballoon.Fibroin microballoon prepared by the present invention can be used for the fields such as medicine carrying micro-system, cellular uptake, for biomedical development provides new approaches and method.
Brief description of the drawings
Fig. 1 is the Electronic Speculum figure of fibroin microballoon in the embodiment of the present invention 1;
Fig. 2 is the Electronic Speculum figure of fibroin microballoon in the embodiment of the present invention 3;
Fig. 3 is the Electronic Speculum figure of fibroin microballoon in the embodiment of the present invention 5;
Fig. 4 is the Electronic Speculum figure of fibroin microballoon in the embodiment of the present invention 6.
Embodiment
Below in conjunction with accompanying drawing and by embodiment, the present invention is described in further detail, and following examples are explanation of the invention and the present invention is not limited to following examples.
Embodiment 1
In the present embodiment, the preparation method of fibroin microballoon in turn includes the following steps.
(1) silkworm silk cocoon is shredded, take 20g and put into the Na of mass percent as 0.5% taking the bath raio of 1:00
2cO
3in the aqueous solution.In the aqueous solution, carry out degumming process, after degumming process 30min, use washed with de-ionized water 2 times; Repeat again above step, with after washed with de-ionized water 3 times, fibroin sample is shakeout and is placed in the baking oven of 50 DEG C and dries and obtain bombyx mori silk fibroin fiber; Described Na
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
(2) take 10g bombyx mori silk fibroin fiber in step (1) and be added in the LiBr solution that concentration is 9mol/L and be placed in shaking table 6h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and within 4 days, is obtained compared with pure silk cellulose solution with deionized water dialysis, after natural air drying, filtration, obtain pure silk cellulose solution.The present invention can be placed in the silk fibroin in step (1) finite concentration LiBr solution dissolution process certain hour, obtains thick liquid: LiBr strength of solution used is generally 7～11M, is preferably 8-10M, and the best is 9M; Treatment time used is 5-8h, and the best is 6h.
(3) the pure silk cellulose solution in step (2) is adjusted into the solution that concentration is 3%-7%, preferably 3% concentration.
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:5, continues magnetic stirring apparatus 30min.
(5) solution in step (4) is placed in to-20 DEG C of refrigerator and cooled and freezes 24h.
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
(7), as Fig. 1, this fibroin microsphere particle diameter is 0.694 ± 0.210 μ m, microspherulite diameter distribution uniform, favorable dispersity, microsphere surface is more smooth, and it is even that spherical this that is rule illustrates that the certain condition of control can be prepared particle diameter, the fibroin microballoon of favorable dispersity.
Embodiment 2
(1) silkworm silk cocoon is shredded, take 20g and put into the Na of mass percent as 0.5% taking the bath raio of 1:00
2cO
3in the aqueous solution.In the aqueous solution, carry out degumming process, after degumming process 30min, use washed with de-ionized water 2 times; Repeat again above step, with after washed with de-ionized water 3 times, fibroin sample is shakeout and is placed in the baking oven of 50 DEG C and dries and obtain bombyx mori silk fibroin fiber; Described Na
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
(2) take 10g bombyx mori silk fibroin fiber in step (1) and be added in the LiBr solution that concentration is 9mol/L and be placed in shaking table 5h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and within 4 days, is obtained compared with pure silk cellulose solution with deionized water dialysis, after natural air drying, filtration, obtain pure silk cellulose solution.
(3) the pure silk cellulose solution in step (2) being adjusted into concentration is 3% solution.
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:8, continues magnetic stirring apparatus 30min.
(5) solution in step (4) is placed in to-20 DEG C of refrigerator and cooled and freezes 24h.
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
(7) this fibroin microsphere particle diameter is 1.192 ± 0.137 μ m.
Embodiment 3
(1) silkworm silk cocoon is shredded, take 20g and put into the Na of mass percent as 0.5% taking the bath raio of 1:00
2cO
3in the aqueous solution.In the aqueous solution, carry out degumming process, after degumming process 30min, use washed with de-ionized water 2 times; Repeat again above step, with after washed with de-ionized water 3 times, fibroin sample is shakeout and is placed in the baking oven of 50 DEG C and dries and obtain bombyx mori silk fibroin fiber; Described Na
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
(2) take 10g bombyx mori silk fibroin fiber in step (1) and be added in the LiBr solution that concentration is 9mol/L and be placed in shaking table 5h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and within 4 days, is obtained compared with pure silk cellulose solution with deionized water dialysis, after natural air drying, filtration, obtain pure silk cellulose solution.
(3) the pure silk cellulose solution in step (2) being adjusted into concentration is 5% solution.
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:5, continues magnetic stirring apparatus 30min.
(5) solution in step (4) is placed in to-20 DEG C of refrigerator and cooled and freezes 24h.
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
(7), as Fig. 2, this fibroin microsphere particle diameter is 1.519 ± 0.134 μ m.Microspherulite diameter distribution uniform, dispersiveness is not good, and most microballoons present the spherical of rule, this explanation increases after fibroin concentration, and microspherulite diameter increases, along with solution viscosity increases, part silk fibroin does not have fully and isopropanol reaction, and presents out-of-shape, dispersed not good phenomenon.
Embodiment 4
(1) silkworm silk cocoon is shredded, take 20g and put into the Na of mass percent as 0.5% taking the bath raio of 1:00
2cO
3in the aqueous solution.In the aqueous solution, carry out degumming process, after degumming process 30min, use washed with de-ionized water 2 times; Repeat again above step, with after washed with de-ionized water 3 times, fibroin sample is shakeout and is placed in the baking oven of 50 DEG C and dries and obtain bombyx mori silk fibroin fiber; Described Na
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
(2) take 10g bombyx mori silk fibroin fiber in step (1) and be added in the LiBr solution that concentration is 9mol/L and be placed in shaking table 5h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and within 4 days, is obtained compared with pure silk cellulose solution with deionized water dialysis, after natural air drying, filtration, obtain pure silk cellulose solution.
(3) the pure silk cellulose solution in step (2) being adjusted into concentration is 5% solution.
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:8, continues magnetic stirring apparatus 30min.
(5) solution in step (4) is placed in to-20 DEG C of refrigerator and cooled and freezes 24h.
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
(7) this fibroin microsphere particle diameter is 2.356 ± 0.148 μ m.
Embodiment 5
(1) silkworm silk cocoon is shredded, take 20g and put into the Na of mass percent as 0.5% taking the bath raio of 1:00
2cO
3in the aqueous solution.In the aqueous solution, carry out degumming process, after degumming process 30min, use washed with de-ionized water 2 times; Repeat again above step, with after washed with de-ionized water 3 times, fibroin sample is shakeout and is placed in the baking oven of 50 DEG C and dries and obtain bombyx mori silk fibroin fiber; Described Na
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
(2) take 10g bombyx mori silk fibroin fiber in step (1) and be added in the LiBr solution that concentration is 9mol/L and be placed in shaking table 6h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and within 5 days, is obtained compared with pure silk cellulose solution with deionized water dialysis, after natural air drying, filtration, obtain pure silk cellulose solution.
(3) the pure silk cellulose solution in step (2) being adjusted into concentration is 7% solution.
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:5, continues magnetic stirring apparatus 30min.
(5) solution in step (4) is placed in to-20 DEG C of refrigerator and cooled and freezes 48h.
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
(7), as Fig. 3, this fibroin microsphere particle diameter is 5.103 ± 0.217 μ m.Microspherulite diameter distribution uniform, but dispersiveness is not good, has stack region, occasionally has block irregularly shapedly, and this explanation silk fibroin solution concentration is larger, and microspherulite diameter is larger, and along with the increasing of solution viscosity, its dispersed variation, the stacking phenomenon of fibroin microballoon is serious.
Embodiment 6
(1) silkworm silk cocoon is shredded, take 20g and put into the Na of mass percent as 0.5% taking the bath raio of 1:00
2cO
3in the aqueous solution.In the aqueous solution, carry out degumming process, after degumming process 30min, use washed with de-ionized water 2 times; Repeat again above step, with after washed with de-ionized water 3 times, fibroin sample is shakeout and is placed in the baking oven of 50 DEG C and dries and obtain bombyx mori silk fibroin fiber; Described Na
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
(2) take 10g bombyx mori silk fibroin fiber in step (1) and be added in the LiBr solution that concentration is 9mol/L and be placed in shaking table 6h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and within 4 days, is obtained compared with pure silk cellulose solution with deionized water dialysis, after natural air drying, filtration, obtain pure silk cellulose solution.
(3) the pure silk cellulose solution in step (2) being adjusted into concentration is 7% solution.
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:8, continues magnetic stirring apparatus 30min.
(5) solution in step (4) is placed in to-20 DEG C of refrigerator and cooled and freezes 48h.
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
(7), as Fig. 4, this fibroin microsphere particle diameter is 4.083 ± 0.182 μ m.Microspherulite diameter obviously increases, dispersiveness is not good, there are adhesion phenomenon and stacking phenomenon to occur, and have irregularly shaped, this explanation, silk fibroin solution concentration and volume increase, microspherulite diameter obviously increases, the increase of solution viscosity makes self-assembling reaction insufficient, present irregularly shaped, and with adhesion and stacking phenomenon.
Embodiment 7
(1) silkworm silk cocoon is shredded, take 20g and put into the Na of mass percent as 0.5% taking the bath raio of 1:00
2cO
3in the aqueous solution.In the aqueous solution, carry out degumming process, after degumming process 30min, use washed with de-ionized water 2 times; Repeat again above step, with after washed with de-ionized water 3 times, fibroin sample is shakeout and is placed in the baking oven of 50 DEG C and dries and obtain bombyx mori silk fibroin fiber; Described Na
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
(2) take 10g bombyx mori silk fibroin fiber in step (1) and be added in the LiBr solution that concentration is 9mol/L and be placed in shaking table 7h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and within 3 days, is obtained compared with pure silk cellulose solution with deionized water dialysis, after natural air drying, filtration, obtain pure silk cellulose solution.
(3) the pure silk cellulose solution in step (2) being adjusted into concentration is 3% solution.
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:5, continues magnetic stirring apparatus 30min.
(5) solution in step (4) is placed in to-20 DEG C of refrigerator and cooled and freezes 24h.
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
(7) this fibroin microsphere particle diameter is 0.717 ± 0.255 μ m.
Embodiment 8
(1) silkworm silk cocoon is shredded, take 20g and put into the Na of mass percent as 0.5% taking the bath raio of 1:00
2cO
3in the aqueous solution.In the aqueous solution, carry out degumming process, after degumming process 30min, use washed with de-ionized water 2 times; Repeat again above step, with after washed with de-ionized water 3 times, fibroin sample is shakeout and is placed in the baking oven of 50 DEG C and dries and obtain bombyx mori silk fibroin fiber; Described Na
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
(2) take 10g bombyx mori silk fibroin fiber in step (1) and be added in the LiBr solution that concentration is 9mol/L and be placed in shaking table 5h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and within 4 days, is obtained compared with pure silk cellulose solution with deionized water dialysis, after natural air drying, filtration, obtain pure silk cellulose solution.
(3) the pure silk cellulose solution in step (2) being adjusted into concentration is 3% solution.
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:5, continues magnetic stirring apparatus 30min.
(5) solution in step (4) is placed in to-20 DEG C of refrigerator and cooled and freezes 48h.
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
(7) this fibroin microsphere particle diameter is 0.679 ± 0.199 μ m.
Embodiment 9
(1) silkworm silk cocoon is shredded, take 20g and put into the Na of mass percent as 0.5% taking the bath raio of 1:00
2cO
3in the aqueous solution.In the aqueous solution, carry out degumming process, after degumming process 30min, use washed with de-ionized water 2 times; Repeat again above step, with after washed with de-ionized water 3 times, fibroin sample is shakeout and is placed in the baking oven of 50 DEG C and dries and obtain bombyx mori silk fibroin fiber; Described Na
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
(2) take 10g bombyx mori silk fibroin fiber in step (1) and be added in the LiBr solution that concentration is 9mol/L and be placed in shaking table 5h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and within 3 days, is obtained compared with pure silk cellulose solution with deionized water dialysis, after natural air drying, filtration, obtain pure silk cellulose solution.
(3) the pure silk cellulose solution in step (2) being adjusted into concentration is 5% solution.
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:5, continues magnetic stirring apparatus 30min.
(5) solution in step (4) is placed in to-80 DEG C of refrigerator and cooled and freezes 48h.
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
(7) this fibroin microsphere particle diameter is 1.4529 ± 0.162 μ m.
What more than enumerate is only part specific embodiment of the present invention; obviously, the invention is not restricted to above embodiment, can also have many distortion; all distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention, all should think protection scope of the present invention.
Claims (9)
1. a self-assembling method of preparing fibroin microballoon, is characterized in that, comprises the steps:
(1) Silkworm Cocoon Shell is shredded, put into the Na of mass percent as 0.3%-0.7% taking the bath raio of (1:50)-(1:00)
2cO
3in the aqueous solution, carry out after degumming process 25-35min, use washed with de-ionized water 1-3 time; Clean fibroin sample is dropped into the Na that mass percent is 0.3%-0.7%
2cO
3in the aqueous solution, carry out again after degumming process 25-35min, with after washed with de-ionized water 1-3 time, fibroin sample is shakeout and is placed in the baking oven oven dry of 40-50 DEG C and obtains bombyx mori silk fibroin fiber;
(2) the bombyx mori silk fibroin fiber in step (1) is added in the LiBr solution that concentration is 7-11mol/L and is placed in shaking table 5-8h, accelerate dissolution obtains silk fibroin solution; Silk fibroin solution is placed in dialysis membrane and is obtained compared with pure silk cellulose solution with deionized water dialysis 3-5d, after natural air drying, filtration, obtain pure silk cellulose solution;
(3) the pure silk cellulose solution in step (2) is adjusted into the solution that concentration is 3%-7%;
(4) get silk fibroin solution in step (3) and be placed in beaker magnetic agitation 5-10min, then dropwise add successively aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is (1:5)-(1:8), continuation magnetic stirring apparatus 10-30min;
(5) solution in step (4) is placed in to-20 DEG C～-80 DEG C refrigerator and cooled and freezes 24-48h;
(6), after the freezing sample in step (5) is at room temperature thawed, observe and present milky white liquid, then lyophilize powdered, thus make finished product.
2. the self-assembling method of preparing fibroin microballoon according to claim 1, is characterized in that,
Na in described step (1)
2cO
3the temperature of the aqueous solution is 85-105 DEG C.
3. the self-assembling method of preparing fibroin microballoon according to claim 1, is characterized in that,
In described step (1), silkworm silk cocoon is shredded, be added to the Na of mass percent as 0.5% taking the bath raio of 1:100
2cO
3in the aqueous solution, carry out degumming process.
4. the self-assembling method of preparing fibroin microballoon according to claim 1, is characterized in that,
In described step (1), in degumming process 30min after, by washed with de-ionized water 2 times, then the 30min that comes unstuck, washed with de-ionized water 3 times, then to carry out temperature be that 50 DEG C of oven dry obtain bombyx mori silk fibroin fiber.
5. according to the self-assembling method of preparing fibroin microballoon described in claim 1 or 2 or 3 or 4, it is characterized in that,
In described step (2), the bombyx mori silk fibroin fiber in step (1) is added in the LiBr solution that concentration is 9mol/L and is placed in shaking table 6h, thus accelerate dissolution reduce molecular weight and obtain silk fibroin solution.
6. the self-assembling method of preparing fibroin microballoon according to claim 5, is characterized in that,
In described step (2), the concentration of LiBr solution is 8-10mol/L.
7. the self-assembling method of preparing fibroin microballoon according to claim 6, is characterized in that,
In described step (2), the concentration of LiBr solution is 9mol/L.
8. the self-assembling method of preparing fibroin microballoon according to claim 1, is characterized in that,
It is 3% solution that pure silk cellulose solution in described step (3) is adjusted into concentration.
9. according to the self-assembling method of preparing fibroin microballoon described in claim 1 or 2 or 3 or 4 or 6 or 7 or 8, it is characterized in that,
In described step (4), silk fibroin solution in step (3) is placed in to beaker magnetic agitation 5min, then dropwise adds aqueous isopropanol, the volume ratio that makes Virahol and silk fibroin solution is 1:5, continues magnetic stirring apparatus 30min.
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| CN104825398A (en) * | 2015-04-24 | 2015-08-12 | 中原工学院 | Preparation method and use of surface mesoporous silk fibroin microsphere drug slow-release carrier |
| CN105832682A (en) * | 2016-05-27 | 2016-08-10 | 浙江大学 | Method for preparing honeycomb silk fibroin porous microsphere sustained drug release vector |
| CN106729982A (en) * | 2016-12-29 | 2017-05-31 | 浙江大学 | A kind of preparation method of silk fibroin nanosphere |
| CN108553689A (en) * | 2018-04-13 | 2018-09-21 | 浙江大学 | A kind of fibroin porous microsphere and preparation method thereof with nanofiber micro-structure |
| CN108578373A (en) * | 2018-05-29 | 2018-09-28 | 安徽大学 | Preparation method of rifampicin-loaded silk fibroin nano-microspheres |
| CN110327307A (en) * | 2019-06-26 | 2019-10-15 | 浙江大学 | A kind of preparation method and product of medicine carrying fibroin nano-microcapsule |
| CN113663119A (en) * | 2021-07-07 | 2021-11-19 | 核工业总医院 | Preparation method of iodized oil silk fibroin embolism microsphere with perspective developing function |
| CN113666995A (en) * | 2021-08-25 | 2021-11-19 | 浙江大学 | Method for quickly and efficiently preparing native tussah silk fibroin powder |
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| CN104825398A (en) * | 2015-04-24 | 2015-08-12 | 中原工学院 | Preparation method and use of surface mesoporous silk fibroin microsphere drug slow-release carrier |
| CN105832682A (en) * | 2016-05-27 | 2016-08-10 | 浙江大学 | Method for preparing honeycomb silk fibroin porous microsphere sustained drug release vector |
| CN105832682B (en) * | 2016-05-27 | 2018-11-13 | 浙江大学 | A kind of preparation method of cellular silk fibroin porous microsphere drug slow-released carrier |
| CN106729982A (en) * | 2016-12-29 | 2017-05-31 | 浙江大学 | A kind of preparation method of silk fibroin nanosphere |
| CN108553689A (en) * | 2018-04-13 | 2018-09-21 | 浙江大学 | A kind of fibroin porous microsphere and preparation method thereof with nanofiber micro-structure |
| CN108553689B (en) * | 2018-04-13 | 2020-03-27 | 浙江大学 | Silk fibroin porous microsphere with nanofiber microstructure and preparation method thereof |
| CN108578373A (en) * | 2018-05-29 | 2018-09-28 | 安徽大学 | Preparation method of rifampicin-loaded silk fibroin nano-microspheres |
| CN108578373B (en) * | 2018-05-29 | 2019-12-03 | 安徽大学 | Preparation method of rifampicin-loaded silk fibroin nano-microspheres |
| CN110327307A (en) * | 2019-06-26 | 2019-10-15 | 浙江大学 | A kind of preparation method and product of medicine carrying fibroin nano-microcapsule |
| CN113663119A (en) * | 2021-07-07 | 2021-11-19 | 核工业总医院 | Preparation method of iodized oil silk fibroin embolism microsphere with perspective developing function |
| CN113666995A (en) * | 2021-08-25 | 2021-11-19 | 浙江大学 | Method for quickly and efficiently preparing native tussah silk fibroin powder |
| CN113666995B (en) * | 2021-08-25 | 2024-01-23 | 浙江大学 | Method for preparing native tussah silk fibroin powder |
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