CN102560887B - Silk fibroin nano-fiber film loaded with vitamins A and E and preparation method thereof - Google Patents
Silk fibroin nano-fiber film loaded with vitamins A and E and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a silk fibroin nano-fiber film loaded with vitamins A and E and a preparation method thereof. The nano-fiber film comprises silk fibroin nano-fiber, as well as the fat-soluble vitamin A and the water-soluble vitamin E, which are loaded in the silk fibroin nano-fiber. The preparation method comprises the following steps of: (1) heating the vitamins A and E till melting, uniformly stirring, then adding deionized water at the temperature of 80-100 DEG C, and stirring for 10-20min till transparent liquid, namely an emulsion of the vitamins A and E, is formed; (2) adding silk fibroin and the deionized water into the emulsion of the vitamins A and E till the final concentration, and stirring till the complete solution of the silk fibroin so as to get a mixed solution of the three; and (3) transferring the solution into a syringe for performing electrostatic spinning to get the silk fibroin nano-fiber film. The silk fibroin nano-fiber film disclosed by the invention has the advantages of no pollution, low cost and simple operation, and the nano-fiber disclosed by the invention can be applied to artificial skin, medical dressings, hemostatic materials, skin care products and other various biomedical fields.
Description
Technical field
The invention belongs to silk fibroin nano-fiber film and preparation field thereof, particularly silk fibroin nano-fiber film of a kind of load vitamin A and E and preparation method thereof.
Background technology
Nanofiber, owing to having that yardstick is little, specific area is large and the feature such as physical property is excellent, has broad application prospects at biomedical sector.Nano fibrous membrane has little aperture and high specific area can promote hemostasis reaction and without styptic.Nano fibrous membrane loose structure is conducive to the breathing of cell and does not cause that wound is dry, and little aperture effectively stops bacterium intrusion simultaneously, has again good ventilative oxygen permeability.Change dressings brings the probability that disturbs nerve fiber regeneration.Electrostatic spinning nano fiber can be given the better self-regeneration of Skin Cell path, and from the viewpoint of organizational project, the cell conductibility that bionical nanofibrous structures has had and the compatibility of improving blood, more easily heal and skin regeneration wound.Nanofiber or the ideal material of encapsulated drug, it not only can be encapsulated into solid drugs in fiber with particle form, liquid medicine can also be encapsulated with double-deck fiber or chain pearl fibers form, thereby effectively realize the control of medicine is discharged.Because fibroin albumen is the very abundant native protein of a kind of nature, there is the excellent performances such as good biocompatibility, biodegradability, good ventilative, poisture-penetrability, non-immunogenicity, be widely used in recent years biomedicine field.
Multivitamin is found there is substantial connection with skin health, in patent CN101736430A, has just announced the preparation method that a kind of load has ascorbic silk fibroin nano-fiber, and its fiber has skin-care effect.Vitamin A is a kind of liposoluble substance, can make epithelium organ keep eubolism.The ultraviolet ray of the sun can cause the vitamin(e) A deficiency in human body skin, and may cause early ageing and the cutaneum carcinoma of skin, if can greatly reduce the destruction of ultraviolet ray to skin with vitamin A preliminary treatment skin, therefore vitamin A has the effect reducing by light aging, vitamin A also can be treated acute promyelocytic leukemia, acne and other skin skin disorders, and the skin aging that can slow down, remove wrinkle.The stability problem of vitamin A, is not solved at present at home yet.Many producers are making containing in the aqueous solution preparation process of vitamin A, taked pH value such as regulator solution, added complexing agent and remove in metal ion, finished product the measures such as filling CO 2, stabilizing vitamin A.Vitamin E is called again anti-infertility vitamin or tocopherol, is a kind of transparent faint yellow thickness grease, water insoluble, is dissolved in organic solvent, and soda acid hydrogenation process and high temperature all can not destroy vitamin E.But it is meeting eremacausis in air, and ultraviolet irradiation can make its decomposition.It can protect other easily oxidized material it is not destroyed, so it is extremely effective oxidant, in advance preventing heavy metal, produce the hepatotoxin of free radical and can cause that the various medicines that oxidant is caused injury damage.TPGS (D-α-tocopherol polyethylene glycol succinate, TPGS, Vitamin E TPGS, Tocophersolan) be the soluble derivative of vitamin E, carboxyl and polyethylene glycol (PEG) esterification by VE-succinate (VES) form, loaded < < American Pharmacopeia > >.TPGS is produced and is gone on the market by U.S. Eastman company the earliest, now be widely used in preparation research abroad, carrier as solubilizer, sorbefacient, emulsifying agent, plasticizer and slightly water-soluble and fat-soluble medicine transmission system, as the carrier Deng， U.S. of the carrier of solid dispersions, dosing eyes, the nose intracavitary administration aqueous solution that now existing TPG(mass fraction is 20%) launch is (as Mazuri
tM), as the nutritious supplementary pharmaceutical of vitamin E, folding synthesising complex E is tired and is about 77.4IUg
-1.TPGS is highly stable, and room temperature is placed and is not hydrolyzed, and almost tasteless, can be used as people's vitamin E replenishers, and particularly those are absorbed with the people of obstacle to liposoluble vitamin E.In US523469, using elephant and African elephant as animal subject, compare with taking liposoluble vitamin E, after the TPGS solution that oral mass fraction is 20%, in body, vitamin E concentration increases soon, compared with the watermiscible vitamin E of low dosage, just can reach higher blood concentration.
Summary of the invention
Technical problem to be solved by this invention is to provide silk fibroin nano-fiber film of a kind of load vitamin A and E and preparation method thereof, a plurality of biomedicine fields such as this silk fibroin nano-fiber film using artificial skin, medical dressing, hemostatic material, skin-protection product, in the whole preparation process of the present invention, do not relate to any to environment and the disagreeableness material of human health, pollution-free, cost is low, simple to operate.
The silk fibroin nano-fiber film of a kind of load vitamin A of the present invention and E, described nano fibrous membrane comprises silk fibroin nano-fiber and be carried on fat-soluble A and the watermiscible vitamin E in silk fibroin nano-fiber, and the diameter of nano fibrous membrane is 10-800nm.
Described watermiscible vitamin E is vitamin E polyethylene glycol succinic acid ester.
Described fat-soluble A is Retinol Palmitate or vitamin(e) A acetate.
The diameter of described nano fibrous membrane is 300-600nm.
The preparation method of the silk fibroin nano-fiber film of a kind of load vitamin A of the present invention and E, comprising:
(1) fat-soluble A and watermiscible vitamin E are heated to melt, stir, subsequently in 80～100 ℃ of deionized waters that add with respect to vitamin E quality 4-9 times, stir 10-20min to transparency liquid, i.e. vitamin A and E emulsion;
(2) in said vitamin A and E emulsion, add fibroin albumen and deionized water to final concentration, be stirred to fibroin albumen and dissolve completely, obtain fibroin albumen-vitamin A-vitamin E solution; Wherein, the final concentration of vitamin E is 0.1-2wt%, and the final concentration of fibroin albumen is 20-35wt%, and the final concentration of vitamin A is 0.1-8wt%;
(3) above-mentioned fibroin albumen-vitamin A-vitamin E solution is moved in syringe and carries out electrostatic spinning, obtain silk fibroin nano-fiber film.
The preparation method of the fibroin albumen in described step (2) is: by the 0.5wt%Na that goes pupa silk cocoon at 100 ℃
2cO
3in the aqueous solution, boil 2-5 time, each 30min comes unstuck, and obtains fibroin, the CaCl that is 1:2:8 by mol ratio
2, C
2h
5oH and H
2o or dissolve 1-2h at 40-60 ℃ with the 0.8g/ml LiBr aqueous solution, with distill water dialysis 3-5 days, freeze drying and get final product.
The final concentration of the vitamin E in described step (2) is 0.5-2wt%.
The final concentration of the fibroin albumen in described step (2) is 25-30wt%.
The final concentration of the vitamin A in described step (2) is 0.5-8wt%.
The process conditions of the electrostatic spinning in described step (3) are: electrostatic pressure is 16-20 kilovolt, and accepting distance is 80-220mm, and spinning speed is 0.1-1.0 ml/hour.
The present invention uses TPGS as emulsifying agent, vitamin A load entered in nanofiber, thus protection vitamin A, the utilization rate of raising medicine, reduces dosage, reduces and prevents the injury of medicine to other positions of health, reduces the burden of metabolic system simultaneously.
beneficial effect
The present invention can make VitAVitE and fibroin albumen be dissolved in water simultaneously, in the whole preparation process of the present invention, do not relate to any to environment and the disagreeableness material of human health, pollution-free, cost is low, simple to operate, and in conjunction with utilizing the good characteristics of nanofiber, vitamin A and E and fibroin albumen, load of the present invention have the nanofiber of Solvisyn-A and E can using artificial skin, a plurality of biomedicine fields such as medical dressing, hemostatic material, skin-protection product.
Accompanying drawing explanation
Fig. 1 is the scanning electron microscope (SEM) photograph of embodiment 1 prepared micro/nano fibrous membrane material;
Fig. 2 is the scanning electron microscope (SEM) photograph of embodiment 2 prepared micro/nano fibrous membrane materials;
Fig. 3 is the scanning electron microscope (SEM) photograph of embodiment 3 prepared micro/nano fibrous membrane materials;
Fig. 4 is the fiber diameter distribution profile of the micro/nano fibrous membrane material of embodiment 1 gained;
Fig. 5 is that fibroblast is spun the comparison of vitamin A-vitamin E silk fibroin nano-fiber support proliferative activity in emulsion;
Fig. 6 is that emulsion is spun the comparison of vitamin A-vitamin E nano fibrous framework cell antioxygenic property.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.In addition should be understood that those skilled in the art can make various changes or modifications the present invention after having read the content of the present invention's instruction, these equivalent form of values fall within the application's appended claims limited range equally.
Embodiment 1
(1) by the 0.5wt%Na that goes pupa silk cocoon at 100 ℃
2cO
3in the aqueous solution, boil 2 times, each 30min comes unstuck, and will come unstuck and obtain fibroin, the CaCl that is 1:2:8 by mol ratio
2, C
2h
5oH and H
2o dissolves 1.5h at 60 ℃, and freeze drying obtains loose cellular solid filament fibroin.
(2) accurately take 0.16 gram of TPGS and 0.02 gram of Retinol Palmitate, be heated to melt, magnetic agitation is even, the deionized water of 90 ℃ that adds subsequently 1.44ml, fully stir 15min, remove thermal source, be stirred to and be transparent, micro-yellow liquid, in vitamin A-vitamin E emulsion, add 1.0g fibroin albumen and deionized water, final mass is 4.0g, after sealing, continuing to be stirred to fibroin albumen dissolves completely, after standing several minutes subsequently, spinning solution is transferred in syringe, voltage is 20kV, syringe pump fltting speed is 0.3ml/h, employing aluminium foil receives, receiving range is 18cm, , obtain the silk fibroin nano-fiber film of load vitamin A and E, average diameter is 465 ± 161nm.
(1) by the 0.5wt%Na that goes pupa silk cocoon at 100 ℃
2cO
3in the aqueous solution, boil 4 times, each 30min comes unstuck, and will come unstuck and obtain fibroin, and the 0.8g/ml LiBr aqueous solution dissolves 2h at 45 ℃, and freeze drying obtains loose cellular solid filament fibroin.
(2) accurately take 0.32 gram of TPGS and 0.08 gram of Retinol Palmitate, be heated to melt, magnetic agitation is even, the deionized water of 90 ℃ that adds subsequently 1.2ml, fully stir 15min, remove thermal source, be stirred to and be transparent, micro-yellow liquid, in vitamin A-vitamin E emulsion, add 1.0g fibroin albumen and deionized water, final mass is 4.0g, after sealing, continuing to be stirred to fibroin albumen dissolves completely, after standing several minutes subsequently, spinning solution is transferred in syringe, voltage is 18kV, syringe pump fltting speed is 0.4ml/h, employing aluminium foil receives, receiving range is 18cm, , obtain the silk fibroin nano-fiber film of load vitamin A and E, average diameter is 405 ± 110nm.
(1) by the 0.5wt%Na that goes pupa silk cocoon at 100 ℃
2cO
3in the aqueous solution, boil 5 times, each 30min comes unstuck, and will come unstuck and obtain fibroin, and the 0.8g/ml LiBr aqueous solution dissolves 2h at 45 ℃, and freeze drying obtains loose cellular solid filament fibroin.
(2) accurately take 0.016 gram of TPGS and 0.04 gram of Retinol Palmitate, be heated to melt, magnetic agitation is even, the deionized water of 85 ℃ that adds subsequently 1.2ml, fully stir 15min, remove thermal source, be stirred to and be transparent, micro-yellow liquid, in vitamin A-vitamin E emulsion, add 1.5g fibroin albumen and deionized water, final mass is 4.0g, after sealing, continuing to be stirred to fibroin albumen dissolves completely, after standing several minutes subsequently, spinning solution is transferred in syringe, voltage is 18kV, syringe pump fltting speed is 0.4ml/h, employing aluminium foil receives, receiving range is 18cm, obtain the silk fibroin nano-fiber film of load vitamin A and E.
Embodiment 4
(1) by the 0.5wt%Na that goes pupa silk cocoon at 100 ℃
2cO
3in the aqueous solution, boil 5 times, each 30min comes unstuck, and will come unstuck and obtain fibroin, and the 0.8g/ml LiBr aqueous solution dissolves 1h at 40 ℃, and freeze drying obtains loose cellular solid filament fibroin.
(2) accurately take 0.32 gram of TPGS and 0.08 gram of Retinol Palmitate, be heated to melt, magnetic agitation is even, the deionized water of 90 ℃ that adds subsequently 1.2ml, fully stir 15min, remove thermal source, be stirred to and be transparent, micro-yellow liquid, in vitamin A-vitamin E emulsion, add 0.8g fibroin albumen and deionized water, final mass is 4.0g, after sealing, continuing to be stirred to fibroin albumen dissolves completely, after standing several minutes subsequently, spinning solution is transferred in syringe, voltage is 20kV, syringe pump fltting speed is 1ml/h, employing aluminium foil receives, receiving range is 22cm, obtain the silk fibroin nano-fiber film of load vitamin A and E.
Embodiment 5
(1) by the 0.5wt%Na that goes pupa silk cocoon at 100 ℃
2cO
3in the aqueous solution, boil 5 times, each 30min comes unstuck, and will come unstuck and obtain fibroin, and the 0.8g/ml LiBr aqueous solution dissolves 2h at 45 ℃, and freeze drying obtains loose cellular solid filament fibroin.
(2) accurately take 0.32 gram of TPGS and 0.02 gram of Retinol Palmitate, be heated to melt, magnetic agitation is even, the deionized water of 90 ℃ that adds subsequently 1.38ml, fully stir 15min, remove thermal source, be stirred to and be transparent, micro-yellow liquid, in vitamin A-vitamin E emulsion, add 0.8g fibroin albumen and deionized water, final mass is 4.0g, after sealing, continuing to be stirred to fibroin albumen dissolves completely, after standing several minutes subsequently, spinning solution is transferred in syringe, voltage is 16kV, syringe pump fltting speed is 1ml/h, employing aluminium foil receives, receiving range is 0.8cm, obtain the silk fibroin nano-fiber film of load vitamin A and E.
Effect embodiment 1
Biocompatibility detects
(1) material preparation and processing
As previous embodiment 1,2 method obtain solutions also adopt embodiment 1, other spinning parameters in 2 methods, adopt the circular lid slide that diameter is 14mm to receive electrostatic spinning nano fiber, tunica fibrosa is positioned in vacuum drying chamber dry, be placed in 75%(v/v) alcohol vapour 24h, carry out post processing and the sterilization of material, for cell, cultivate.
Wherein: the pure silk fibroin spinning solution that service property (quality) concentration is 25%, voltage is 20kV, and syringe pump fltting speed is 0.3ml/h, adopts the circular lid slide that diameter is 14mm to receive electrostatic spinning nano fiber, receiving range is 18cm, as a comparison the nanofiber group in embodiment 1.Adopt the circular lid slide of 14mm, as a comparison the slide group in embodiment 2.
Control group is placed in 75%(v/v equally) alcohol vapour 24h, carry out post processing and the sterilization of material, for cell, cultivate.
(2) MTT experiment
By 10
4the fibroblast suspension inoculation of individual/mL is embodiment 1,2,3 and the comparative example 1,2 through sterilizing in 24 well culture plates, every hole inoculating cell suspension 200 μ L, every group of 3 holes.Cultivate respectively after 1d, 3d, 5d and 7d, every group of 3 holes are changed and are added not containing nutrient solution 360 μ L and MTT solution (5mg/mL) the 40 μ L of serum, and incubator is cultivated 3～4h, sucks nutrient solution in hole, every hole adds 400 μ L dimethyl sulfoxide (DMSO)s, and vibration 30min dissolves crystal.Every hole is got dmso solution liquid 100 μ L and is added in 96 hole ELISA Plates, then on ELIASA, by 492nm wavelength period, surveys OD value.Establish 3 multiple holes, and repeat to test 3 times for every group.Result is as shown in Figure 5:
MTT effect:
Accompanying drawing 5 is cultivated the propagation situation of 1,3,5,7 day for being depicted as on the nano fiber scaffold of fibroblast under different vitamin As and vitamin E concentration.Compare (comparative example 2) with slide, cell has better propagation on electrostatic spinning nano fiber support.It should be noted that embodiment 1 nano fiber scaffold is because its fibre structure is more mellow and full evenly, the propagation that is more of value to cell adheres to.And embodiment 2 nanofibers are because beading quantity is more, and fibre structure is irregular, and the activity of its cell proliferation is lower than another two groups of nano fiber scaffolds (embodiment 1 and comparative example 1).Hence one can see that, and the concentration proportioning of vitamin A and vitamin E is non-obvious, and the delicate variation of component also can cause the difference of biocompatibility effect.Cultivate after 7 days, cell has in load that on vitamin and pure silk element nano fiber scaffold, (embodiment 1,2 and comparative example 1) propagation obviously faster than slide group, there is significant difference (P<0.05), and also have significant difference (P<0.05) between each nanofiber group.Hence one can see that, and emulsion is spun vitamin fibroin albumen composite nano fiber and had good biocompatibility, and the nanofiber group in embodiment 1, and biocompatibility is better than embodiment 2.
Antioxygenic property:
Cell reaches 90%(growth area covering approximately 90% by being cultured to degrees of fusion on a laboratory routine operation inoculation material), the oxidant t-BHP(50 that adds variable concentrations, 100, 200, 400 μ M), co-incubation 24 hours, use subsequently PBS buffer solution to wash 3 times, every hole adds the DMEM culture medium 360 μ L of preheating, then add the MTT40 μ L of preheating to continue to cultivate 4 hours, sucking-off culture medium, every hole adds DMSO400 μ L, in 37 ℃ of shaking tables, dissolve 20min, form purple solution, then by liquid-transfering gun draw solution, add in 96 well culture plates, every hole 100 μ L, measure light absorption value.
Anti-oxidant result:
T-BHP can induce fibroblastic cellular damage and apoptosis, is commonly used for model of oxidative, and whether the silk fibroin nano-fiber support that the present embodiment is investigated load vitamin contributes to Skin Cell to resist the oxidative damage that t-BHP causes.
Cell is cultured to the degree that is paved with that degrees of fusion reaches 90%(cell after inoculating by the method in aforementioned MTT experiment), the oxidant t-BHP(50 that adds variable concentrations, 100, 200, 400 μ M), co-incubation 24 hours, use subsequently PBS buffer solution to wash 3 times, every hole adds the DMEM culture medium 360 μ L of preheating, then add the MTT40 μ L of preheating to continue to cultivate 4 hours, sucking-off culture medium, every hole adds DMSO400 μ L, in 37 ℃ of shaking tables, dissolve 20min, form purple solution, then by liquid-transfering gun draw solution, add in 96 well culture plates, every hole 100 μ L, measure light absorption value.Result is as shown in Figure 6:
As shown in Figure 6, the SF vigor on slide control group (comparative example 2) is along with the increase of t-BHP concentration presents remarkable decline trend.The t-BHP of low concentration (50-100 μ M) spins the not infringement of cell on vitamin A-vitamin E silk fibroin nano-fiber support to the prepared emulsion of embodiment 1, increased slightly on the contrary its activity (comparing with the pure culture base group of not adding t-BHP), when t-BHP concentration is increased to 200 μ M when above, the activity of cell still remains on 85% left and right (comparing with the pure culture base group of not adding t-BHP).And pure silk element nanofiber group (comparative example 1) is poor to the tolerance of high concentration (400 μ M) t-BHP, cytoactive is only 70% left and right, shows that its antioxidant effect spins vitamin (A-E) nanofiber group not as good as emulsion.The antioxidation of vitamin E is known dawn early, and vitamin A can react with Phenoxy radical, thereby effectively plays oxidation resistant effect.Above-mentionedly show that vitamin A-vitamin E silk fibroin nano-fiber contributes to SF to anti-oxidative damage.
Claims (9)
1. a preparation method for the silk fibroin nano-fiber film of load vitamin A and E, comprising:
(1) fat-soluble A and watermiscible vitamin E are heated to melt, stir, subsequently in 80～100 ℃ of deionized waters that add with respect to vitamin E quality 4-9 times, stir 10-20min to transparency liquid, i.e. vitamin A and E emulsion;
(2) in said vitamin A and E emulsion, add fibroin albumen and deionized water to final concentration, be stirred to fibroin albumen and dissolve completely, obtain fibroin albumen-vitamin A-vitamin E solution; Wherein, the final concentration of vitamin E is 0.1-2wt%, and the final concentration of fibroin albumen is 20-35wt%, and the final concentration of vitamin A is 0.1-8wt%;
(3) above-mentioned fibroin albumen-vitamin A-vitamin E solution is moved in syringe and carries out electrostatic spinning, obtain silk fibroin nano-fiber film.
2. the preparation method of the silk fibroin nano-fiber film of a kind of load vitamin A according to claim 1 and E, is characterized in that: the fat-soluble A in described step (1) is Retinol Palmitate or vitamin(e) A acetate; Watermiscible vitamin E is vitamin E polyethylene glycol succinic acid ester.
3. the preparation method of the silk fibroin nano-fiber film of a kind of load vitamin A according to claim 1 and E, is characterized in that: the preparation method of the fibroin albumen in described step (2) is: by the 0.5wt%Na that goes pupa silk cocoon at 100 ℃
2cO
3in the aqueous solution, boil 2-5 time, each 30min comes unstuck, and obtains fibroin, the CaCl that is 1:2:8 by mol ratio
2, C
2h
5oH and H
2o or dissolve 1-2h at 40-60 ℃ with the 0.8g/ml LiBr aqueous solution, with distill water dialysis 3-5 days, freeze drying and get final product.
4. the preparation method of the silk fibroin nano-fiber film of a kind of load vitamin A according to claim 1 and E, is characterized in that: the final concentration of the vitamin E in described step (2) is 0.5-2wt%.
5. the preparation method of the silk fibroin nano-fiber film of a kind of load vitamin A according to claim 1 and E, is characterized in that: the final concentration of the fibroin albumen in described step (2) is 25-30wt%.
6. the preparation method of the silk fibroin nano-fiber film of a kind of load vitamin A according to claim 1 and E, is characterized in that: the final concentration of the vitamin A in described step (2) is 0.5-8wt%.
7. the preparation method of the silk fibroin nano-fiber film of a kind of load vitamin A according to claim 1 and E, it is characterized in that: the process conditions of the electrostatic spinning in described step (3) are: electrostatic pressure is 16-20 kilovolt, accepting distance is 80-220mm, and spinning speed is 0.1-1.0 ml/hour.
8. the preparation method of the silk fibroin nano-fiber film of a kind of load vitamin A according to claim 1 and E, is characterized in that: the diameter of described step (3) silk fibroin nano-fiber film is 10-800nm.
9. the preparation method of the silk fibroin nano-fiber film of a kind of load vitamin A according to claim 8 and E, is characterized in that: the diameter of described silk fibroin nano-fiber film is 300-600nm.
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| CN104000739B (en) * | 2014-05-19 | 2017-03-29 | 东华大学 | A kind of preparation method of the isinglass fibroin albumen composite nano-fiber membrane of load skin-protecting factor |
| CN104060401A (en) * | 2014-07-04 | 2014-09-24 | 东华大学 | Electrostatic spinning preparing method of gelatin nanofiber membrane capable of releasing vitamin A |
| CN104846555A (en) * | 2015-05-07 | 2015-08-19 | 东华大学 | Preparation method of gelatin nanofiber membrane capable of synergistically releasing vitamin A and vitamin E |
| CN105641740A (en) * | 2016-01-05 | 2016-06-08 | 重庆惠森驰恒医疗器械有限公司 | Biological nutrition dressing for thoroughly treating skin injury and preparation method thereof |
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| CN108286126A (en) * | 2018-01-24 | 2018-07-17 | 上海理工大学 | A kind of preparation method of quick-acting haemostatic powder composite cellulosic membrane |
| CN109675102A (en) * | 2018-11-22 | 2019-04-26 | 宋建康 | Load the guide tissue regeneration film and preparation method thereof of gelatin micro-nano ball |
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| CN101275291A (en) * | 2008-04-30 | 2008-10-01 | 苏州大学 | Electrostatic spinning method for polyhydroxylated polymer |
| CN101390814A (en) * | 2008-10-29 | 2009-03-25 | 东南大学 | Beauty mask based on electro spinning nano fiber |
| CN101736430A (en) * | 2010-02-05 | 2010-06-16 | 东华大学 | Method for preparing silk fibroin nano-fibre with skin-care effect |
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