Preparation method with silk fibroin nano-fiber of skin-care effect
Technical field
The invention belongs to the preparation field of silk fibroin nano-fiber, particularly relate to a kind of preparation method with silk fibroin nano-fiber of skin-care effect.
Background technology
Fibroin albumen is to be come unstuck by silkworm, is a kind of natural protein, has excellent performance such as excellent biological compatibility, biodegradability, high ventilation, poisture-penetrability, non-immunogenicity
[1,2]Be widely used in the reparation and the regeneration of tissues such as skin, bone, cartilage, tendon, nerve trachea, blood vessel.Park etc.
[3]The silk fibroin nano-fiber film that is equipped with for the solvent electro-spinning with formic acid can promote mouse preosteoblast (MC3T3-E1) to adhere to, breed, break up, and can induce the generation of BGP, promotes bone tissue restoration and regeneration.
Fibroin albumen has outstanding skin-protection and health-care effect.Fibroin albumen contains 18 seed amino acids and polypeptide composition, and wherein glycine, the third atmosphere acid and 3 kinds of simple amino acid of serine occupy 85% of Tot Prot, and are very approaching with the skin and the hair of human body, thereby becomes the popular research material of artificial skin
[4]Fibroin albumen also has good moisture preserving property and heat insulating ability, can help skin to regulate moisture, and its mechanism is identical with the interior natural moisturizing factor of human body
[5]Research confirms that fibroin albumen also has the effect that solar radiation is resisted in the absorption ultraviolet ray, and the melanin that can effectively suppress in the skin generates
[6-8]Therefore, fibroin albumen is good skin care products base material.
Vitamin is the general name of the indispensable small molecule active material of human body, and it can not synthesize in human body, perhaps can not capacity synthesize, and must absorb from the external world.Vitamin is participated in biochemical reaction widely in vivo as the auxiliary element of some important enzymes, has determined some crucial metabolic process.Many vitamins come to light and skin health has substantial connection, thereby usually appear in some skin-protection products as additive
[9]Vitamin C has many maintenance functions to skin; Being described as " beauty treatment vitamin ", is remarkable anti-oxidant, can help to reduce the infringement of radical pair skin; Help to reduce wrinkle and improve skin texture, and the generation that suppresses dermal melanin, the melanic effect that desalination has generated are arranged
[9]
List of references
[1]Santin?M,Motta?A,Freddi?G,Cannas?M.In?vitro?evaluation?of?the?inflammatory?potential?ofthe?silk?fibroin.J?Biomed?Mater?Res?1999;46:382-389.
[2]Horan?RL,Antle?K,Collette?AL,Wang?YZ,Huang?J,Moreau?JE,Volloch?V,Kaplan?DL,Altman?GH.In?vitro?degradation?of?silk?fibroin.Biomaterials?2005;26:3385-3393.
[3]Kim?KH,Jeong?L,Park?WH,et?al.Biological?efficacy?of?silk?fibroin?nanofiber?membranesfor?guided?bone?regeneration,J?Biotechnol,2005,120:327-339.
[4] Duan Qiaoyan, Duan Xiang, Zhang Huanxiang. the application [J] of fibroin albumen in organizational project. Chinese Tissue Engineering Study and clinical rehabilitation, 2007,11 (26): 5199-20.
[5] your tinkling of pieces of jade, the Yi Huang duckweed, Chen Ping, Strategy Software Systems Co., Ltd (SSS) is prosperous. the fibroin protein latest Progress. chemical propellant and macromolecular material .2007,5 (4): 26-29
[6]Maurer-Stroh?S,Eisenhaber?B,Eisenhaber?F.N-terminal?N-myristoylation?of?Proteins:Prediction?of?substrate?Proteins?from?amino?acid?sequence.J?Mol?Biol,2002,317:541-557.
[7]winkler?S,Kaplan?DL.Silk?produced?by?engineered?bacteria?encyclopedia?of?materials:Science?and?technology.Elsevier,Amsterdam:2001.8615-8618.
[8]park?S?J,Lee?KY,Ha?WS,et?al.Structural?changes?and?their?effect?on?mechanical?propertiesof?silk?fibroin/chitosan?blends.JAPPl?Polym?Sci,1999,74:2571-2575.
[9] Li Qingren, Wang Yuemei, Ding Xuefei. the skin-care effect of vitamin and application. household chemicals science .2007,30 (1): 16.
Summary of the invention
Technical problem to be solved by this invention provides a kind of preparation method with silk fibroin nano-fiber of skin-care effect, and this method is simple, and cost is low, and is environment friendly and pollution-free, is suitable for suitability for industrialized production; The gained fiber is a kind of desirable new ideas skin care products, skin wound dressing and tissue engineering bracket.
A kind of preparation method with silk fibroin nano-fiber of skin-care effect of the present invention comprises:
(1) will go the pupa degumming silkworm cocoons to obtain fibroin, the use mol ratio is 1: 2: 8 CaCl
2, C
2H
5OH, H
2O dissolves 1.5h at 60 ℃, with distill water dialysis 3-5 days, and the cellular solid filament fibroin that freeze drying obtains loosening;
(2) with vitamin C phosphoric ester sodium powder (C
6H
6Na
3O
9P) and above-mentioned fibroin albumen with mass ratio 1-5: 100 are mixed in the aseptic deionized water, and stirring and evenly mixing obtains blend electrostatic spinning liquid;
(3) above-mentioned electrostatic spinning liquid is obtained the ascorbic silk fibroin nano-fiber of load through electrostatic spinning.
Degumming method in the said step (1) does, with the 0.5%Na that goes the pupa silk cocoon at 100 ℃
2CO
3Boil 2-5 time each 30min in the aqueous solution.
Vitamin C phosphoric ester sodium powder (C in the said step (2)
6H
6Na
3O
9P) mass ratio with fibroin albumen is 1: 100,3: 100 or 5: 100.
Aseptic deionized water in the said step (2) is that deionized water is handled through autoclave sterilization by the generation of Alpha-pure 15 type pure water appearance again.
Electrostatic spinning parameter in the said step (3): voltage: 12-20 kilovolt; Electric field distance: 60-200 millimeter; Spinning speed, the 0.3-1.0 milliliter/hour.
The method that the present invention adopts static to spin prepares nanofiber; Nanofiber has been owing to provide than the bigger specific area of traditional micron order material, and the The Nomenclature Composition and Structure of Complexes that the ascorbic fibroin albumen composite nano fiber of load that is equipped with through pure green electro-spinning can better anthropomorphic dummy's body cell epimatrix makes fibroin albumen with vitamin and Skin Cell more intimate contacting arranged; The bionical physiological environment of the best is provided for cell growth and regeneration; Thereby widened this two types of material application aspect skin-protection and health-care greatly, both can be used as cosmetics and skincare product, for example; Facial mask, the dressing and the man-made support that also can be used as skin wound use.
Beneficial effect
(1) preparation method of the present invention is simple, and cost is low, and is environment friendly and pollution-free, is suitable for suitability for industrialized production;
(2) the present invention is mainly from bionic angle; Utilize the preparation method of electrostatic spinning that vitamin C and good biology performance and the skin-care effect of fibroin albumen are combined; Not only bionical on the structure; And it is bionical from forming; And then reach bionical on the physiological function; For the cell growth provides best physiological environment, be a kind of desirable new ideas skin care item, skin wound dressing and tissue engineering bracket;
(3) the present invention is in whole process of preparation, do not relate to any environment and the disagreeableness material of human health to have been realized pure green electrostatic spinning, prepares the green product of pure natural;
(4) China has abundant fibroin albumen resource, and electrostatic spinning be the preparation nanofiber the most simply, method the most easily, easy realization of industrial production has stronger using value.
Description of drawings
Fig. 1 is the stereoscan photograph of the ascorbic fibroin albumen electrostatic spinning nano fiber of load;
Fig. 2 is the ascorbic fibroin albumen electrostatic spinning nano fiber of a load film photo.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction scope of the present invention.Should be understood that in addition those skilled in the art can do various changes or modification to the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Embodiment 1
(1) will go the pupa silk cocoon at 100 ℃ 0.5%Na
2CO
3Boil 2-5 time in the aqueous solution, each 30min, the use mol ratio is 1: 2: 8 CaCl
2, C
2H
5OH, H
2O dissolves 1.5h at 60 ℃, with distill water dialysis 3-5 days, and the cellular solid filament fibroin that freeze drying obtains loosening;
(2) above-mentioned fibroin albumen 3.344 grams are dissolved in are stirred to dissolving fully in 9 milliliters of deionized aqua sterilisas; 0.03344 gram vitamin C phosphoric ester sodium (mass ratio of vitamin C phosphoric ester sodium and fibroin albumen is 1: 100) is dissolved in 1 milliliter of aseptic deionized water; Be stirred to fully dissolving, above two kinds of good solution (silk fibroin protein solution and vitamin C phosphoric ester sodium solution) of dissolving are mixed and are stirred to even; Mixed solution is carried out electrostatic spinning, spinning condition: voltage, 20 kilovolts; Electric field distance, 180 millimeters; Spinning speed, obtains the composite nano-fiber membrane that average diameter is 496 ± 98 nanometers by 0.3 milliliter/hour.
Embodiment 2
(1) will go the pupa silk cocoon at 100 ℃ 0.5%Na
2CO
3Boil 2-5 time in the aqueous solution, each 30min, the use mol ratio is 1: 2: 8 CaCl
2, C
2H
5OH, H
2O dissolves 1.5h at 60 ℃, with distill water dialysis 3-5 days, and the cellular solid filament fibroin that freeze drying obtains loosening;
(2) above-mentioned fibroin albumen 3.344 grams are dissolved in are stirred to dissolving fully in 9 milliliters of deionized aqua sterilisas; 0.10032 gram vitamin C phosphoric ester sodium (mass ratio of vitamin C phosphoric ester sodium and fibroin albumen is 3: 100) is dissolved in 1 milliliter of aseptic deionized water; Be stirred to fully dissolving, above two kinds of good solution (silk fibroin protein solution and vitamin C phosphoric ester sodium solution) of dissolving are mixed and are stirred to even; Mixed solution is carried out electrostatic spinning, spinning condition: voltage, 20 kilovolts; Electric field distance, 180 millimeters; Spinning speed, obtains the composite nano-fiber membrane that average diameter is 480 ± 90 nanometers by 0.3 milliliter/hour.
Embodiment 3
(1) will go the pupa silk cocoon at 100 ℃ 0.5%Na
2CO
3Boil 2-5 time in the aqueous solution, each 30min, the use mol ratio is 1: 2: 8 CaCl
2, C
2H
5OH, H
2O dissolves 1.5h at 60 ℃, with distill water dialysis 3-5 days, and the cellular solid filament fibroin that freeze drying obtains loosening;
(2) above-mentioned fibroin albumen 3.344 grams are dissolved in are stirred to dissolving fully in 9 milliliters of deionized aqua sterilisas; 0.1672 gram vitamin C phosphoric ester sodium (mass ratio of vitamin C phosphoric ester sodium and fibroin albumen is 5: 100) is dissolved in 1 milliliter of aseptic deionized water; Be stirred to fully dissolving, above two kinds of good solution (silk fibroin protein solution and vitamin C phosphoric ester sodium solution) of dissolving are mixed and are stirred to even; Mixed solution is carried out electrostatic spinning, spinning condition: voltage, 20 kilovolts; Electric field distance, 180 millimeters; Spinning speed, obtains the composite nano-fiber membrane that average diameter is 466 ± 80 nanometers by 0.3 milliliter/hour.