CN102605467A - Method for preparing composite polyelectrolyte fiber membrane by utilizing freeze-drying way - Google Patents
Method for preparing composite polyelectrolyte fiber membrane by utilizing freeze-drying way Download PDFInfo
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- CN102605467A CN102605467A CN2012100528742A CN201210052874A CN102605467A CN 102605467 A CN102605467 A CN 102605467A CN 2012100528742 A CN2012100528742 A CN 2012100528742A CN 201210052874 A CN201210052874 A CN 201210052874A CN 102605467 A CN102605467 A CN 102605467A
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Abstract
The invention relates to a method for preparing a composite polyelectrolyte fiber membrane by utilizing a freeze-drying way. The method comprises the steps of: dissolving polyanions and polycations respectively in a solvent to prepare a solution in percentage by weight of 0.001-1 wt%; then fully stirring the solution; mixing the solution containing the polyanions and the polycations at a volume ratio of 1-9: 9-1 by using different methods to obtain a composite polyelectrolyte solution; and finally transferring the composite polyelectrolyte solution into a liquid nitrogen refrigeration device to enable the solution to be frozen in the liquid nitrogen environment and freeze-drying the frozen solution at a temperature of -10 to -80 DEG C and at a vacuum degree of 1-600 Pa for 12-48 h so as to obtain the composite polyelectrolyte fiber membrane. The nanometer fiber prepared by the method has the characteristics of adjustable pH value and electriferous property; that is to say, hyaluronic acid is ionized to carry with negative charges under an alkali condition, and chitosan is protonized to carry with positive charges under an acid condition. Under such a pH condition, transformation with electric charges has higher application value in the field of bioengineering and biomedical materials.
Description
Technical field
The invention belongs to the preparation field of boiomacromolecule composite nano-fiber material, relate to the preparation method of the compound polyelectrolyte tunica fibrosa of natural products base.
Background technology
Vacuum Freezing & Drying Technology is to utilize ice crystal distillation principle, and making the moisture in the material that freezes in advance is that steam is removed with the distillation of ice attitude directly without the thawing of icing, thereby obtains the technology of dried product.Can know by the Phase Equilibrium theory in the chemical thermodynamics: under certain pressure and temperature, reach certain balancing each other between three kinds of forms of water, obtain the phasor (Fig. 1) of water in view of the above.Three phase point has shown the pressure and temperature condition of the gas, liquid, solid three-phase coexistence of water.The OA line is the line of demarcation of solid phase and gas phase among the figure, is called the distillation line; The OB line is the line of demarcation of liquid phase and gas phase, is called the vaporization line; The OC line is the line of demarcation of solid phase and liquid phase, is called the thawing line; The O point is a three phase point.Reduce according to pressure, the basic physics principle that boiling point descends, as long as pressure P is positioned under the three phase point (diagram P＜646.5Pa, T＜0 ℃), the moisture in the material can from solid phase ice without liquid phase directly distillation be steam.
The operating process of vacuum freeze-drying technique comprises prefreezing, lyophilization, parsing-desiccation three phases; Prefreezing: the Free water in the solution is solidified; Give behind the product drying with before the drying identical form is arranged; Irreversible changes such as bubbling when preventing to find time drying, concentrate, contraction and solute move takes place, and reduces Yin Wendu and descends that the material solubility that causes reduces and the variation of life characteristics.Lyophilization: the product after will freezing places the enclosed vacuum vessel in heating, and its ice crystal will be sublimed into the steam effusion and product is dehydrated, and when whole ice crystals were removed, lyophilization had just been accomplished, and can remove moisture about 90% this moment.Parsing-desiccation: sublimation stage is failed dry bound water evaporate material.A part of bound water in the biological tissue is difficult to remove through desivac, need temperature be elevated to 30～35 ℃ through resolution phase and further remove.
Vacuum freeze drying has advantage:
(1) freeze drying is carried out under condition of high vacuum degree, and oxygen is few, and readily oxidizable substance is protected, and sterilizes or suppress the vigor of some bacterium because of anoxic; (2) freeze drying is under low pressure carried out, the unlikely oxidation deterioration of the material that is dried; (3) freeze drying is carried out at low temperatures, the unlikely sex change of volatile thermal sensitivity composition or lose vigor in the material; (4) dried matter is loose porous, is spongy, adds that dissolving almost recovers original proterties rapidly and fully immediately behind the water.Therefore, freeze drying is at present at medical industry, food industry, and scientific research departments etc. are widely used.
(Sodium Algenate is the natural polymer that is present in the brown algae SA) to sodium alginate, is the natural polyanionic polysaccharide that from brown alga or bacterium, extracts, and is made up of guluronic acid and two kinds of construction units of its stereoisomer mannuronic acid.Sodium alginate possesses excellent biological compatibility, no subacute/reaction of chronic toxicity or carcinogenic, can be used as edible food additives, also can be used as timbering material and be used for medical usage.Hyaluronic acid (Hyaluronic acid; HA) have another name called sodium hyaluronate; Be to be present in a kind of acid mucopolysaccharide in the extracellular matrix in the biological tissue; By β-D-N-acetylglucosamine and β-D-glucose sulfonic acid be construction unit with β-1, a kind of chain macromolecule that the 4-glycosidic linkage is linked to be is a kind of polyelectrolyte that has negative electrical charge.Owing to characteristics such as its excellent biological compatibility and biodegradable are widely used in fields such as organizational project.(Chitosan is the product of chitin after deacetylation is handled CS) to shitosan, is the natural alkaline polysaccharide that occurring in nature exists, and it is a kind of polycation natural polymer.Advantages such as that shitosan not only has is nontoxic, excellent biological compatibility, biodegradability; Also have many excellent physiological properties such as cancer resistance, antibiotic property, hemostatic, enhances human body immunocompetence, be widely used in aspects such as organizational project, drug carrier material and wound dressing.Heparin (Heparin) is a kind of by gucosamine, and the glutinous polysaccharide sulfuric acid ester that L-idose aldehyde glycosides, N-acetylglucosamine and D-glucuronic acid are alternately formed has highly acid, and the altitudinal belt negative electrical charge.Simultaneously, heparin all has blood coagulation resisting function in vivo and in vitro as a kind of anti-coagulants.Be mainly used in thrombotic disease, myocardial infarction, operation on vessels of heart, cardia catheterization, extracorporal circulatory system, haemodialysis etc. clinically.Along with pharmacology and clinical medical progress, the application of heparin constantly enlarges.Gamma-polyglutamic acid-(poly-γ-glutamic acid; γ-PGA) is the water-soluble polyamino acid that the occurring in nature microbial fermentation produces; Be a kind of special anion nature polymer, its structure is the high molecular polymer that glutamic acid units is passed through alpha-amido and γ-carboxyl formation peptide bond.γ-PGA is a Main Ingredients and Appearance of forming natto viscoloid, has the effect that promotes that mineral matter absorbs.γ-PGA specific molecule structure makes it have extremely strong moisture-retaining capacity, adds γ-PGA in cosmetics or skin care products, can increase the moisture-retaining capacity of skin effectively, promotes skin health.(there are a large amount of primary amino radicals in ε-PL) have high molecular characteristic to epsilon-polylysine in the strand, amino protonated in solution, having a large amount of positive charges becomes cationic polymer.As a kind of natural microbial metabolism product, its cost is low, and environmental friendliness can be decomposed into lysine in vivo and absorbed by catapepsis, has no toxic and side effect.Compound polyelectrolyte (polyelectrolyte complexes (PECs)) is the mixed with polymers that has opposite charges with two kinds, through the electrostatic force between positive and negative charge a kind of composite that intermolecular self assembly forms takes place.This intermolecular self assembly has potential using value at aspects such as drug loading transportation, genetic engineerings.The formation of soluble poly pentalyte must be satisfied following condition: the wherein a certain component of two component polyelectrolyte of 1) forming compound polyelectrolyte must contain weak ionizing group; The difference of the molecular weight that 2) existence is bigger between the two component polyelectrolyte of the compound polyelectrolyte of composition; 3) component of long-chain is excessive; 4) there is the micromolecule that can play shielding action.If one or more conditions can not satisfy, polyelectrolyte then takes place to assemble and deposition, thereby can not obtain uniform solution.
Compound polyelectrolyte tunica fibrosa has the electrical characteristic of pH adjustable band; Polycation amino protonated under acid condition and make the tunica fibrosa positively charged; Under alkali condition; Carbonyl ionization on the polyanion, and make tunica fibrosa electronegative, but there is potential using value in the characteristic of this pH regulator charging property at aspects such as bioengineering, filtration, flocculants; Simultaneously, the compound polyelectrolyte tunica fibrosa of preparation has a good application prospect at aspects such as bio-medical materials.Up to now, relevant desivac prepares the still rarely seen report of research report of polyelectrolyte tunica fibrosa.
Summary of the invention
One of the object of the invention provides a kind of method for preparing compound polyelectrolyte tunica fibrosa.
Two of the object of the invention provides the method for multiple compound polyelectrolyte.
The method of the compound polyelectrolyte tunica fibrosa of preparation provided by the present invention may further comprise the steps:
(1) preparation of polyelectrolyte solution: polyanion and polycation are dissolved in respectively in the solvent separately; Be mixed with the solution that percentage by weight is 0.001～1wt% respectively; Then solution is fully stirred, so that dissolving fully, polyanion and said polycation solution promptly obtained.
Described polyanion is: hyaluronic acid (HA) (M
W=5000～2 000 000g/mol) (viscosity average molecular weigh is 2 * 10 to sodium alginate (SA)
6), heparin (Heparin) (M
W=1200～40 000g/mol), the gamma-polyglutamic acid-((M of γ-PGA)
W=100 000～10 000 000g/mol); Polycation is: shitosan (CS) (M
W=3000～200 000g/mol), the epsilon-polylysine ((M of ε-PL)
W=5000～100 000g/mol).
(2) with polyanion and said polycation solution by volume 1～9: 9～1 ratio mix with diverse ways, obtain compound polyelectrolyte solution.
Described diverse ways is: method 1. is slowly poured polyanion or said polycation solution in polycation or the polyanion solution; Method 2. has in the container of moveable bulkhead in the middle of polyanion and the said polycation solution, takes dividing plate away, lets two kinds of solution diffusion 1～120min; Method 3. is transferred to the polyanion or the said polycation solution of preparation in the step (1) in the liquid nitrogen frozen device; Open refrigerating plant; Polyanion or the said polycation solution prepared are freezed in liquid nitrogen environment rapidly, and polyanion that will freeze then or said polycation solution are transferred in the freeze drier, are that-10～-80 ℃, vacuum are under the condition of 1～600Pa in temperature; Carry out freeze-drying and handle 12～48h, obtain polyanion or polycation tunica fibrosa.The polyanion or the polycation tunica fibrosa of preparation are put into polycation or polyanion solution, obtain compound polyelectrolyte solution; Method 4. is poured polycation or polyanion solution on the polyanion or polycation tunica fibrosa of preparation in the method 3 into, obtains compound polyelectrolyte solution.
(3) desivac prepares compound polyelectrolyte tunica fibrosa: the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is that-10～-80 ℃, vacuum are under the condition of 1～600Pa in temperature; Carry out freeze-drying and handle 12～48h, obtain compound polyelectrolyte tunica fibrosa.
(4) MTT cytotoxicity test: the PBS of the compound polyelectrolyte nano fibrous membrane that obtains in the step (3) is embathed liquid handle the L929 cell respectively after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and the mensuration wavelength is 490nm, and calculates its P value.
(5) cell inoculation experiment: the compound polyelectrolyte based nano-fiber film that obtains in the step (3) is carried out the cell inoculation experiment, and resulting compound polyelectrolyte based nano-fiber film is assessed.
The present invention is compound, and there is potential using value in the polyelectrolyte tunica fibrosa at aspects such as bioengineering, filtration, flocculants; Simultaneously, the compound polyelectrolyte tunica fibrosa of preparation has a good application prospect at aspects such as bio-medical materials.
Description of drawings:
The phasor of Fig. 1 water
Fig. 2 is the container that has moveable bulkhead that is provided by in the invention step (2).
Fig. 3 is the hyaluronic acid that obtains by the technical scheme that embodiment 1 is provided and the SEM pattern of chitosan composite fiber film
The specific embodiment
Embodiment 1:
(1) be that 5000g/mol hyaluronic acid and molecular weight are that the shitosan of 3000g/mol is dissolved in deionized water respectively with molecular weight; Be made into percentage by weight and be respectively the solution of 0.001wt% and 0.002wt%; Then solution is fully stirred; So that dissolving promptly obtains hyaluronic acid and chitosan solution fully.
(2) with hyaluronic acid and 1: 2 by volume mixed of chitosan solution, its mixed method is: hyaluronic acid solution is slowly poured in the chitosan solution, obtained compound polyelectrolyte solution.
(3) the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is under the condition of 1Pa in temperature for-80 ℃, vacuum; Carry out freeze-drying and handle 48h, obtain compound polyelectrolyte tunica fibrosa.
(4) PBS of hyaluronic acid that obtains in the step (3) and chitosan nano fiber membrane is embathed liquid and handle the L929 cell respectively after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and the mensuration wavelength is 490nm, compares with blank, and its 24 hours, 48 hours, 72 hours P value does not have significant difference all greater than 0.05, shows that there is not toxicity in resulting nano fibrous membrane.
(5) hyaluronic acid that obtains in the step (3) and chitosan nano fiber membrane are carried out the cell inoculation experiment, have excellent cell and attach and multiplication characteristic.
Embodiment 2:
(1) be that 2 000 000g/mol hyaluronic acids and molecular weight are that the epsilon-polylysine of 5000g/mol is dissolved in deionized water respectively with molecular weight; Be made into percentage by weight and be respectively the solution of 0.001wt% and 1wt%; Then solution is fully stirred; So that dissolving promptly obtains hyaluronic acid and epsilon-polylysine solution fully.
(2) with hyaluronic acid and 3: 1 by volume mixed of epsilon-polylysine solution, its mixed method is: epsilon-polylysine solution is slowly poured in the hyaluronic acid solution, obtained compound polyelectrolyte solution.
(3) the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is under the condition of 600Pa in temperature for-10 ℃, vacuum; Carry out freeze-drying and handle 48h, obtain compound polyelectrolyte tunica fibrosa.
(4) PBS of hyaluronic acid that obtains in the step (3) and epsilon-polylysine nano fibrous membrane is embathed liquid and handle the L929 cell respectively after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and the mensuration wavelength is 490nm, compares with blank, and its 24 hours, 48 hours, 72 hours P value does not have significant difference all greater than 0.05, shows that there is not toxicity in resulting nano fibrous membrane.
(5) hyaluronic acid that obtains in the step (3) and epsilon-polylysine nano fibrous membrane are carried out the cell inoculation experiment, have excellent cell and attach and multiplication characteristic.
Embodiment 3:
(1) be 2 * 10 with viscosity average molecular weigh
6Sodium alginate and the molecular weight shitosan that is 200 000g/mol be dissolved in respectively in the acetic acid solution of deionized water and 2wt%; Be made into percentage by weight and be respectively the solution of 0.01wt% and 0.001wt%; Then solution is fully stirred; So that dissolving promptly obtains sodium alginate and chitosan solution fully.
(2) with sodium alginate and 9: 1 by volume mixed of chitosan solution; Its mixed method is: sodium alginate soln and chitosan solution are poured into respectively in the container as shown in Figure 2; Take dividing plate away, let two kinds of solution diffusion 120min, obtain compound polyelectrolyte solution.
(3) the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is under the condition of 200Pa in temperature for-60 ℃, vacuum; Carry out freeze-drying and handle 24h, obtain compound polyelectrolyte tunica fibrosa.
(4) PBS of sodium alginate that obtains in the step (3) and chitosan nano fiber membrane is embathed liquid and handle the L929 cell respectively after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and the mensuration wavelength is 490nm, compares with blank, and its 24 hours, 48 hours, 72 hours P value does not have significant difference all greater than 0.05, shows that there is not toxicity in resulting nano fibrous membrane.
(5) sodium alginate that obtains in the step (3) and chitosan nano fiber membrane are carried out the cell inoculation experiment, have excellent cell and attach and multiplication characteristic.
Embodiment 4:
(1) be 2 * 10 with viscosity average molecular weigh
6Sodium alginate and the molecular weight epsilon-polylysine that is 100 000g/mol be dissolved in the deionized water respectively; Be made into percentage by weight and be respectively the solution of 0.001wt% and 0.1wt%; Then solution is fully stirred, so that dissolving fully, sodium alginate and epsilon-polylysine solution promptly obtained.
(2) with sodium alginate and 1: 9 by volume mixed of epsilon-polylysine solution; Its mixed method is: sodium alginate soln and epsilon-polylysine solution are poured into respectively in the container as shown in Figure 2; Take dividing plate away, let two kinds of solution diffusion 1min, obtain compound polyelectrolyte solution.
(3) the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is under the condition of 500Pa in temperature for-40 ℃, vacuum; Carry out freeze-drying and handle 12h, obtain compound polyelectrolyte tunica fibrosa.
(4) PBS of sodium alginate that obtains in the step (3) and epsilon-polylysine nano fibrous membrane is embathed liquid and handle the L929 cell respectively after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and the mensuration wavelength is 490nm, compares with blank, and its 24 hours, 48 hours, 72 hours P value does not have significant difference all greater than 0.05, shows that there is not toxicity in resulting nano fibrous membrane.
(5) sodium alginate that obtains in the step (3) and epsilon-polylysine nano fibrous membrane are carried out the cell inoculation experiment, have excellent cell and attach and multiplication characteristic.
Embodiment 5:
(1) be that the heparin of 1200g/mol and shitosan that molecular weight is 3000g/mol are dissolved in deionized water respectively with molecular weight; Be made into percentage by weight and be respectively the solution of 1wt% and 0.05wt%; Then solution is fully stirred, so that dissolving fully, heparin and chitosan solution promptly obtained.
(2) heparin solution of preparation in the step (1) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The heparin solution of being prepared is freezed in liquid nitrogen environment, and the heparin solution that will freeze is then transferred in the freeze drier, is under the condition of 1Pa in temperature for-10 ℃, vacuum; Carry out freeze-drying and handle 48h, obtain the heparin tunica fibrosa.The heparin tunica fibrosa of preparation is put into chitosan solution, obtain compound polyelectrolyte solution.Wherein the volume ratio of two kinds of solution is: 2: 8.
(3) the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is under the condition of 360Pa in temperature for-40 ℃, vacuum; Carry out freeze-drying and handle 36h, obtain compound polyelectrolyte tunica fibrosa.
(4) PBS of heparin that obtains in the step (3) and chitosan nano fiber membrane is embathed liquid and handle the L929 cell respectively after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and the mensuration wavelength is 490nm, compares with blank, and its 24 hours, 48 hours, 72 hours P value does not have significant difference all greater than 0.05, shows that there is not toxicity in resulting nano fibrous membrane.
(5) heparin that obtains in the step (3) and chitosan nano fiber membrane are carried out the cell inoculation experiment, have excellent cell and attach and multiplication characteristic.
Embodiment 6:
(1) be that the heparin of 40 000g/mol and epsilon-polylysine that molecular weight is 50 000g/mol are dissolved in deionized water respectively with molecular weight; Be made into percentage by weight and be respectively the solution of 0.05wt% and 0.7wt%; Then solution is fully stirred; So that dissolving promptly obtains heparin and epsilon-polylysine solution fully.
(2) the epsilon-polylysine solution of preparation in the step (1) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The epsilon-polylysine solution of being prepared is freezed in liquid nitrogen environment, and the epsilon-polylysine solution that will freeze is then transferred in the freeze drier, is under the condition of 1Pa in temperature for-10 ℃, vacuum; Carry out freeze-drying and handle 48h, obtain the epsilon-polylysine tunica fibrosa.The epsilon-polylysine tunica fibrosa of preparation is put into heparin solution, obtain compound polyelectrolyte solution.Wherein the volume ratio of two kinds of solution is: 6: 4.
(3) the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is under the condition of 10Pa in temperature for-35 ℃, vacuum; Carry out freeze-drying and handle 24h, obtain compound polyelectrolyte tunica fibrosa.
(4) PBS of heparin that obtains in the step (3) and epsilon-polylysine nano fibrous membrane is embathed liquid and handle the L929 cell respectively after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and the mensuration wavelength is 490nm, compares with blank, and its 24 hours, 48 hours, 72 hours P value does not have significant difference all greater than 0.05, shows that there is not toxicity in resulting nano fibrous membrane.
(5) heparin that obtains in the step (3) and epsilon-polylysine nano fibrous membrane are carried out the cell inoculation experiment, have excellent cell and attach and multiplication characteristic.
Embodiment 7:
(1) be that the gamma-polyglutamic acid-of 100 000g/mol and epsilon-polylysine that molecular weight is 10 000g/mol are dissolved in deionized water respectively with molecular weight; Be made into percentage by weight and be respectively the solution of 0.01wt% and 0.09wt%; Then solution is fully stirred; So that dissolving promptly obtains gamma-polyglutamic acid-and epsilon-polylysine solution fully.
(2) the epsilon-polylysine solution of preparation in the step (1) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The epsilon-polylysine solution of being prepared is freezed in liquid nitrogen environment, and the epsilon-polylysine solution that will freeze is then transferred in the freeze drier, is under the condition of 200Pa in temperature for-30 ℃, vacuum; Carry out freeze-drying and handle 24h, obtain the epsilon-polylysine tunica fibrosa.Gamma-polyglutamic acid-solution is poured on the epsilon-polylysine tunica fibrosa of preparation, obtained compound polyelectrolyte solution.Wherein the volume ratio of two kinds of solution is: 5: 8.
(3) the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is under the condition of 50Pa in temperature for-55 ℃, vacuum; Carry out freeze-drying and handle 12h, obtain compound polyelectrolyte tunica fibrosa.
(4) PBS of gamma-polyglutamic acid-that obtains in the step (3) and epsilon-polylysine nano fibrous membrane is embathed liquid and handle the L929 cell respectively after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and the mensuration wavelength is 490nm, compares with blank, and its 24 hours, 48 hours, 72 hours P value does not have significant difference all greater than 0.05, shows that there is not toxicity in resulting nano fibrous membrane.
(5) gamma-polyglutamic acid-that obtains in the step (3) and epsilon-polylysine nano fibrous membrane are carried out the cell inoculation experiment, have excellent cell and attach and multiplication characteristic.
Embodiment 8:
(1) be that the gamma-polyglutamic acid-of 10 000 000g/mol and shitosan that molecular weight is 5 000g/mol are dissolved in deionized water respectively with molecular weight; Be made into percentage by weight and be respectively the solution of 0.001wt% and 0.5wt%; Then solution is fully stirred; So that dissolving promptly obtains gamma-polyglutamic acid-and chitosan solution fully.
(2) the gamma-polyglutamic acid-solution of preparation in the step (1) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The gamma-polyglutamic acid-solution of being prepared is freezed in liquid nitrogen environment, and the gamma-polyglutamic acid-solution that will freeze is then transferred in the freeze drier, is under the condition of 90Pa in temperature for-65 ℃, vacuum; Carry out freeze-drying and handle 24h, obtain the gamma-polyglutamic acid-tunica fibrosa.Chitosan solution is poured on the gamma-polyglutamic acid-tunica fibrosa of preparation, obtained compound polyelectrolyte solution.Wherein the volume ratio of two kinds of solution is: 9: 2.
(3) the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is under the condition of 400Pa in temperature for-60 ℃, vacuum; Carry out freeze-drying and handle 48h, obtain compound polyelectrolyte tunica fibrosa.
(4) PBS of gamma-polyglutamic acid-that obtains in the step (3) and chitosan nano fiber membrane is embathed liquid and handle the L929 cell respectively after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and the mensuration wavelength is 490nm, compares with blank, and its 24 hours, 48 hours, 72 hours P value does not have significant difference all greater than 0.05, shows that there is not toxicity in resulting nano fibrous membrane.
(5) gamma-polyglutamic acid-that obtains in the step (3) and chitosan nano fiber membrane are carried out the cell inoculation experiment, have excellent cell and attach and multiplication characteristic.
Claims (3)
1. desivac prepares the method for compound polyelectrolyte tunica fibrosa, it is characterized in that may further comprise the steps:
(1) preparation of polyelectrolyte solution: be dissolved in polyanion and polycation in the solvent respectively; Be mixed with the solution that percentage by weight is 0.001～1wt% respectively; Then solution is fully stirred, so that dissolving fully, polyanion and said polycation solution promptly obtained;
(2) with polyanion and said polycation solution by volume 1～9: 9～1 ratio mix with diverse ways, obtain compound polyelectrolyte solution;
(3) desivac prepares compound polyelectrolyte tunica fibrosa: the compound polyelectrolyte solution of preparation in the step (2) is transferred in the liquid nitrogen frozen device; Open refrigerating plant; The compound polyelectrolyte solution of being prepared is freezed in liquid nitrogen environment, and the polyelectrolyte composite solution that will freeze is then transferred in the freeze drier, is that-10～-80 ℃, vacuum are under the condition of 1～600Pa in temperature; Carry out freeze-drying and handle 12～48h, obtain compound polyelectrolyte tunica fibrosa.
2. method according to claim 1 is characterized in that, polyanion is in the step (1): hyaluronic acid M
W=5000～2 000 000g/mol, sodium alginate viscosity average molecular weigh are 2 * 10
6, heparin M
W=1200～40 000g/mol or gamma-polyglutamic acid-M
W=100 000～10 000 000g/mol; Polycation is: shitosan M
W=3000～200 000g/mol or epsilon-polylysine M
W=5000～100 000g/mol.
3. preparation method according to claim 1 is characterized in that, mixed method is in the step (2): method 1. is slowly poured polyanion or said polycation solution in polycation or the polyanion solution; Method 2. has in the container of moveable bulkhead in the middle of polyanion and the said polycation solution, takes dividing plate away, lets two kinds of solution diffusion 1～120min; Method 3. is transferred to the polyanion or the said polycation solution of preparation in the step (1) in the liquid nitrogen frozen device; Open refrigerating plant; Polyanion or the said polycation solution prepared are freezed in liquid nitrogen environment rapidly, and polyanion that will freeze then or said polycation solution are transferred in the freeze drier, are that-10～-80 ℃, vacuum are under the condition of 1～600Pa in temperature; Carry out freeze-drying and handle 12～48h, obtain polyanion or polycation tunica fibrosa; The polyanion or the polycation tunica fibrosa of preparation are put into polycation or polyanion solution, obtain compound polyelectrolyte solution; Method 4. is poured polycation or polyanion solution on the polyanion or polycation tunica fibrosa of preparation in the method 3 into, obtains compound polyelectrolyte solution.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104083798A (en) * | 2014-07-03 | 2014-10-08 | 东南大学 | Antibacterial polyelectrolyte composite nanofiber membrane and preparation method thereof |
| CN105063801A (en) * | 2015-08-27 | 2015-11-18 | 常州大学 | Method for preparing crosslinked chondroitin sulfate derivative fiber |
| CN105088413A (en) * | 2015-08-27 | 2015-11-25 | 常州大学 | Preparation method for chondroitin sulfate/chitosan derivative polyelectrolyte complex bi-crosslinked fibers |
| CN105113053A (en) * | 2015-08-27 | 2015-12-02 | 常州大学 | Preparation method of hyaluronic acid derivative /chitosan derivative compound polyelectrolyte bi-crosslinking fiber |
| CN105694120A (en) * | 2014-11-28 | 2016-06-22 | 中国海洋大学 | Preparation method of chitosan oligosaccharide-hyaluronic acid composite nanometer particle |
| CN109157982A (en) * | 2018-09-14 | 2019-01-08 | 北京工业大学 | A method of polyelectrolyte composite nanometer filtering film is prepared using Freeze Drying Technique |
| CN109966552A (en) * | 2019-03-22 | 2019-07-05 | 宁波市医疗中心李惠利医院 | Tissue engineering bracket material and preparation method thereof, application, application method |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1539514A (en) * | 2003-11-03 | 2004-10-27 | 刘永庆 | Method for preparing multifunctional biological repair material |
| CN1844215A (en) * | 2006-04-28 | 2006-10-11 | 江南大学 | Process for preparing hyaluronic acid-chitosan crosslinked biocompatible materials |
| CN101215388A (en) * | 2008-01-15 | 2008-07-09 | 东华大学 | Method for preparing bacteria cellulose composite membrane |
| CN101280467A (en) * | 2008-05-20 | 2008-10-08 | 暨南大学 | Preparation and application of chitosan-based nano-fiber |
| CN101824160A (en) * | 2010-03-30 | 2010-09-08 | 东华大学 | Preparation method of chitosan/polyvinyl alcohol/polylactic acid blended porous membrane |
-
2012
- 2012-03-02 CN CN201210052874.2A patent/CN102605467B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1539514A (en) * | 2003-11-03 | 2004-10-27 | 刘永庆 | Method for preparing multifunctional biological repair material |
| CN1844215A (en) * | 2006-04-28 | 2006-10-11 | 江南大学 | Process for preparing hyaluronic acid-chitosan crosslinked biocompatible materials |
| CN101215388A (en) * | 2008-01-15 | 2008-07-09 | 东华大学 | Method for preparing bacteria cellulose composite membrane |
| CN101280467A (en) * | 2008-05-20 | 2008-10-08 | 暨南大学 | Preparation and application of chitosan-based nano-fiber |
| CN101824160A (en) * | 2010-03-30 | 2010-09-08 | 东华大学 | Preparation method of chitosan/polyvinyl alcohol/polylactic acid blended porous membrane |
Non-Patent Citations (1)
| Title |
|---|
| 张停等: "壳聚糖基聚电解质复合物在生物医学领域的应用", 《化学通报》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104083798A (en) * | 2014-07-03 | 2014-10-08 | 东南大学 | Antibacterial polyelectrolyte composite nanofiber membrane and preparation method thereof |
| CN104083798B (en) * | 2014-07-03 | 2016-09-07 | 东南大学 | A kind of antibacterial polyelectrolyte composite nano-fiber membrane and preparation method thereof |
| CN105694120A (en) * | 2014-11-28 | 2016-06-22 | 中国海洋大学 | Preparation method of chitosan oligosaccharide-hyaluronic acid composite nanometer particle |
| CN105694120B (en) * | 2014-11-28 | 2019-07-05 | 中国海洋大学 | A kind of preparation method of chitosan oligosaccharide hyaluronic acid composite nano particle |
| CN105063801A (en) * | 2015-08-27 | 2015-11-18 | 常州大学 | Method for preparing crosslinked chondroitin sulfate derivative fiber |
| CN105088413A (en) * | 2015-08-27 | 2015-11-25 | 常州大学 | Preparation method for chondroitin sulfate/chitosan derivative polyelectrolyte complex bi-crosslinked fibers |
| CN105113053A (en) * | 2015-08-27 | 2015-12-02 | 常州大学 | Preparation method of hyaluronic acid derivative /chitosan derivative compound polyelectrolyte bi-crosslinking fiber |
| CN109157982A (en) * | 2018-09-14 | 2019-01-08 | 北京工业大学 | A method of polyelectrolyte composite nanometer filtering film is prepared using Freeze Drying Technique |
| CN109966552A (en) * | 2019-03-22 | 2019-07-05 | 宁波市医疗中心李惠利医院 | Tissue engineering bracket material and preparation method thereof, application, application method |
| CN110644232A (en) * | 2019-09-20 | 2020-01-03 | 南通大学 | Preparation method of natural antibacterial water-retention mask |
| CN111996616A (en) * | 2020-08-20 | 2020-11-27 | 江南大学 | Sodium alginate/polylysine self-assembled fiber with controllable size |
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