CN102605467B - 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
- Publication number
- CN102605467B CN102605467B CN201210052874.2A CN201210052874A CN102605467B CN 102605467 B CN102605467 B CN 102605467B CN 201210052874 A CN201210052874 A CN 201210052874A CN 102605467 B CN102605467 B CN 102605467B
- Authority
- CN
- China
- Prior art keywords
- solution
- polyanion
- polycation
- polyelectrolyte
- composite
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
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 natural products base composite polyelectrolyte tunica fibrosa.
Background technology
Vacuum Freezing & Drying Technology, is to utilize ice crystal distillation principle, and the moisture in the material that makes to freeze in advance is directly removed taking the distillation of ice state as steam without the thawing of ice, thereby obtains the technology of dried product.Phase Equilibrium theory from chemical thermodynamics: under certain pressure and temperature, reach certain balancing each other between three kinds of forms of water, obtain accordingly the phasor (Fig. 1) of water.Three phase point has shown the pressure and temperature condition of the gas, liquid, solid three-phase coexistence of water.In figure, OA line is the line of demarcation of solid phase and gas phase, is called distillation line; OB line is the line of demarcation of liquid phase and gas phase, is called vaporization line; OC line is the line of demarcation of solid phase and liquid phase, is called thawing line; O point is three phase point.Reduce according to pressure, the basic physics principle that boiling point declines, as long as pressure P is positioned at (diagram P < 646.5Pa, 0 DEG C of T <) under three phase point, the moisture in material can directly distil as steam without liquid phase from solid phase ice.
The operating process of vacuum freeze-drying technique comprises prefreezing, lyophilization, parsing-desiccation three phases, prefreezing: the Free water in solution is solidified, give after product drying with dry before have identical form, the irreversible change such as prevent from finding time when dry bubbling, concentrated, contraction and solute move occurs, and reduces Yin Wendu declines the material solubility reduction that causes and the variation of life characteristics.Lyophilization: the product after freezing is placed in to airtight vacuum tank and heats, its ice crystal will be sublimed into steam and overflows and make product dehydrate dry, and in the time that whole ice crystals are removed, lyophilization has just completed, and now can remove moisture 90% left and right.Parsing-desiccation: sublimation stage is failed to dry combination water and evaporate material.A part in biological tissue, need to be elevated to 30～35 DEG C by temperature by resolution phase and further remove to remove by desivac in conjunction with shipwreck.
Vacuum freeze drying has following advantages:
(1) freeze drying is carried out under condition of high vacuum degree, and oxygen is few, and readily oxidizable substance is protected, and because of anoxic sterilizing or suppress the vigor of some bacterium; (2) freeze drying is under low pressure carried out, the unlikely oxidation deterioration of material being dried; (3) freeze drying is carried out at low temperatures, the unlikely sex change of volatile thermal sensitivity composition or lose vigor in material; (4) dried material is loose porous, is spongy, dissolves rapidly and completely the almost original proterties of immediate recovery after adding water.Therefore, freeze drying is at present at medical industry, food industry, and scientific research departments etc. are widely used.
Sodium alginate (Sodium Algenate, SA) be the natural polymer being present in brown algae, be the natural polyanionic polysaccharide extracting from brown alga or bacterium, formed by guluronic acid and two kinds of construction units of its stereoisomer mannuronic acid.Sodium alginate possesses good biocompatibility, without subacute/chronic toxicity or carcinogenic reaction, can be used as edible food additives, also can be used as timbering material for medical usage.Hyaluronic acid (Hyaluronic acid, HA) have another name called sodium hyaluronate, to be present in a kind of acid mucopolysaccharide in extracellular matrix in biological tissue, be construction unit by β-D-N-acetylglucosamine and β-D-Glucose sulfonic acid with β-1, a kind of chain macromolecule that 4-glycosidic linkage is linked to be is a kind of polyelectrolyte with negative electrical charge.Because its good characteristic such as biocompatibility and biodegradable is widely used in the fields such as organizational project.Shitosan (Chitosan, CS) be chitin through deacetylation product after treatment, be occurring in nature exist natural alkaline polysaccharide, it is a kind of polycation natural polymer.Shitosan not only has nontoxic, the good advantage such as biocompatibility, biodegradability, also have cancer resistance, antibiotic property, hemostatic, strengthen many excellent physiological properties such as human immunological competence, be widely used in the aspects such as organizational project, drug carrier material and wound dressing.Heparin (Heparin) is a kind of by gucosamine, and L-idose aldehyde glycosides, N-Acetyl-D-glucosamine and D-Glucose aldehydic acid is the glutinous Alginic Sodium Diester of composition alternately, has highly acid, and altitudinal belt negative electrical charge.Meanwhile, heparin, as a kind of anti-coagulants, has blood coagulation resisting function in vivo and in vitro.Be mainly used in clinically thrombotic disease, myocardial infarction, operation on vessels of heart, cardia catheterization, extracorporal circulatory system, haemodialysis etc.Along with pharmacology and clinical medical progress, the application of heparin constantly expands.Gamma-polyglutamic acid-(poly-γ-glutamic acid, γ-PGA) be the water-soluble polyamino acid that the fermentation of occurring in nature microorganism produces, be a kind of special anion nature polymer, its structure is glutamic acid units forms peptide bond high molecular polymer by alpha-amido and γ-carboxyl.γ-PGA is the Main Ingredients and Appearance of composition natto viscoloid, has the effect that promotes mineral absorption.The molecular structure that γ-PGA is special, makes it have extremely strong moisture-retaining capacity, adds γ-PGA in cosmetics or skin care products, can effectively increase the moisture-retaining capacity of skin, promotes skin health.Epsilon-polylysine (ε-PL) has high molecular characteristic, has a large amount of primary amino radicals in strand, amino protonated in solution, becomes cationic polymer with a large amount of positive charges.As a kind of natural microbial metabolism product, its cost is low, and environmental friendliness can be decomposed into lysine in vivo and be digested and assimilated completely, without any toxic and side effect.Composite polyelectrolyte (polyelectrolyte complexes (PECs)) is by two kinds of mixed with polymers with opposite charges, by the electrostatic force between positive and negative charge, a kind of composite that intermolecular self assembly forms occurs.This intermolecular self assembly has potential using value at aspects such as drug loading transport, genetic engineerings.The formation of soluble poly pentalyte must meet following condition: 1) two component polyelectrolyte of composition composite polyelectrolyte wherein a certain component must contain weak ionizing group; 2) difference of the molecular weight that between two component polyelectrolyte of composition composite polyelectrolyte, existence is larger; 3) component of long-chain is excessive; 4) there is the little molecule that can play shielding action.If one or more conditions can not meet, polyelectrolyte occurs to assemble and precipitation, thereby can not obtain uniform solution.
Composite polyelectrolyte tunica fibrosa has the electrical characteristic of pH adjustable band, under acid condition, the amino of polycation is protonated and make tunica fibrosa positively charged, under alkali condition, carbonyl ionization on polyanion, and make tunica fibrosa electronegative, thisly can the electrical characteristic of pH accommodation zone there is potential using value at aspects such as bioengineering, filtration, flocculants; Meanwhile, the composite polyelectrolyte tunica fibrosa of preparation has a good application prospect at aspects such as bio-medical materials.Up to now, relevant desivac is prepared the still rarely seen report of research report of polyelectrolyte tunica fibrosa.
Summary of the invention
One of object of the present invention provides a kind of method of preparing composite polyelectrolyte tunica fibrosa.
Two of object of the present invention provides the method for multiple composite polyelectrolyte.
The method of preparing composite polyelectrolyte tunica fibrosa provided by the present invention, comprises the following steps:
(1) preparation of polyelectrolyte solution: polyanion and polycation are dissolved in respectively in solvent separately, be mixed with respectively the solution that percentage by weight is 0.001～1wt%, then solution is fully stirred, so that dissolve completely, obtain polyanion and said polycation solution.
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), gamma-polyglutamic acid-(γ-PGA) (M
w=100 000～10 000 000g/mol); Polycation is: shitosan (CS) (M
w=3000～200 000g/mol), epsilon-polylysine (ε-PL) (M
w=5000～100 000g/mol).
(2) by polyanion and said polycation solution by volume 1～9: 9～1 ratio mix with diverse ways, obtain composite polyelectrolyte solution.
Described diverse ways is: method 1. is slowly poured polyanion or said polycation solution in polycation or polyanion solution; Method 2., by having in the container of moveable bulkhead in the middle of polyanion and said polycation solution, is taken dividing plate away, allows two kinds of solution spread 1～120min; Method 3. is transferred to polyanion or the said polycation solution of preparation in step (1) in liquid nitrogen frozen device, open refrigerating plant, make prepared polyanion or said polycation solution Quick freezing in liquid nitrogen environment, then the polyanion freezing or said polycation solution are transferred in freeze drier, be under-10～-80 DEG C, the vacuum condition that is 1～600Pa in temperature, carry out frozen dried 12～48h, obtain polyanion or polycation tunica fibrosa.The polyanion of preparation or polycation tunica fibrosa are put into polycation or polyanion solution, obtain composite polyelectrolyte solution; Method 4. is poured polycation or polyanion solution on the polyanion or polycation tunica fibrosa of preparation in method 3 into, obtains composite polyelectrolyte solution.
(3) desivac is prepared composite polyelectrolyte tunica fibrosa: the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-10～-80 DEG C, the vacuum condition that is 1～600Pa in temperature, carry out frozen dried 12～48h, obtain composite polyelectrolyte tunica fibrosa.
(4) MTT cytotoxicity test: the PBS of the composite polyelectrolyte nanofibers film obtaining in step (3) is embathed to liquid and process respectively L929 cell after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and mensuration wavelength is 490nm, and calculates its P value.
(5) cell inoculation experiments: the composite polyelectrolyte based nano-fiber film obtaining in step (3) is carried out to cell inoculation experiments, and obtained composite polyelectrolyte based nano-fiber film is assessed.
There is potential using value at aspects such as bioengineering, filtration, flocculants in composite polyelectrolyte tunica fibrosa of the present invention; Meanwhile, the composite polyelectrolyte tunica fibrosa of preparation has a good application prospect at aspects such as bio-medical materials.
Brief description of the drawings:
The phasor of Fig. 1 water
Fig. 2 is the container with moveable bulkhead providing by invention step (2).
Fig. 3 is the hyaluronic acid that obtains of the technical scheme that provides by embodiment 1 and the SEM pattern of chitosan composite fiber film
Detailed description of the invention
Embodiment 1:
(1) be that the shitosan that 5000g/mol hyaluronic acid and molecular weight are 3000g/mol is dissolved in respectively deionized water by molecular weight, be made into the solution that percentage by weight is respectively 0.001wt% and 0.002wt%, then solution is fully stirred, so that dissolve completely, obtain hyaluronic acid and chitosan solution.
(2) hyaluronic acid and the chitosan solution ratio of 1: 2 are by volume mixed, its mixed method is: hyaluronic acid solution is slowly poured in chitosan solution, obtained composite polyelectrolyte solution.
(3) the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-80 DEG C, the vacuum condition that is 1Pa in temperature, carry out frozen dried 48h, obtain composite polyelectrolyte tunica fibrosa.
(4) PBS of the hyaluronic acid obtaining in step (3) and chitosan nano fiber membrane is embathed to liquid and process respectively L929 cell after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and mensuration wavelength is 490nm, and compared with blank sample, the P value of its 24 hours, 48 hours, 72 hours is all greater than 0.05, there is no significant difference, shows that obtained nano fibrous membrane does not exist toxicity.
(5) hyaluronic acid obtaining in step (3) and chitosan nano fiber membrane are carried out to cell inoculation experiments, there is excellent cell and attach and multiplication characteristic.
Embodiment 2:
(1) be that the epsilon-polylysine that 2 000 000g/mol hyaluronic acids and molecular weight are 5000g/mol is dissolved in respectively deionized water by molecular weight, be made into the solution that percentage by weight is respectively 0.001wt% and 1wt%, then solution is fully stirred, so that dissolve completely, obtain hyaluronic acid and epsilon-polylysine solution.
(2) hyaluronic acid and the epsilon-polylysine solution ratio of 3: 1 are by volume mixed, its mixed method is: epsilon-polylysine solution is slowly poured in hyaluronic acid solution, obtained composite polyelectrolyte solution.
(3) the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-10 DEG C, the vacuum condition that is 600Pa in temperature, carry out frozen dried 48h, obtain composite polyelectrolyte tunica fibrosa.
(4) PBS of the hyaluronic acid obtaining in step (3) and epsilon-polylysine nano fibrous membrane is embathed to liquid and process respectively L929 cell after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and mensuration wavelength is 490nm, and compared with blank sample, the P value of its 24 hours, 48 hours, 72 hours is all greater than 0.05, there is no significant difference, shows that obtained nano fibrous membrane does not exist toxicity.
(5) hyaluronic acid obtaining in step (3) and epsilon-polylysine nano fibrous membrane are carried out to cell inoculation experiments, there is excellent cell and attach and multiplication characteristic.
Embodiment 3:
(1) be 2 × 10 by viscosity average molecular weigh
6sodium alginate and molecular weight be that the shitosan of 200 000g/mol is dissolved in respectively in the acetic acid solution of deionized water and 2wt%, be made into the solution that percentage by weight is respectively 0.01wt% and 0.001wt%, then solution is fully stirred, so that dissolve completely, obtain sodium alginate and chitosan solution.
(2) sodium alginate and the chitosan solution ratio of 9: 1 are by volume mixed, its mixed method is: sodium alginate soln and chitosan solution are poured into respectively in container as shown in Figure 2, take dividing plate away, allow two kinds of solution spread 120min, obtain composite polyelectrolyte solution.
(3) the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-60 DEG C, the vacuum condition that is 200Pa in temperature, carry out frozen dried 24h, obtain composite polyelectrolyte tunica fibrosa.
(4) PBS of the sodium alginate obtaining in step (3) and chitosan nano fiber membrane is embathed to liquid and process respectively L929 cell after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and mensuration wavelength is 490nm, and compared with blank sample, the P value of its 24 hours, 48 hours, 72 hours is all greater than 0.05, there is no significant difference, shows that obtained nano fibrous membrane does not exist toxicity.
(5) sodium alginate obtaining in step (3) and chitosan nano fiber membrane are carried out to cell inoculation experiments, there is excellent cell and attach and multiplication characteristic.
Embodiment 4:
(1) be 2 × 10 by viscosity average molecular weigh
6sodium alginate and molecular weight be that the epsilon-polylysine of 100 000g/mol is dissolved in respectively in deionized water, be made into the solution that percentage by weight is respectively 0.001wt% and 0.1wt%, then solution is fully stirred, so that dissolve completely, obtain sodium alginate and epsilon-polylysine solution.
(2) sodium alginate and the epsilon-polylysine solution ratio of 1: 9 are by volume mixed, its mixed method is: sodium alginate soln and epsilon-polylysine solution are poured into respectively in container as shown in Figure 2, take dividing plate away, allow two kinds of solution spread 1min, obtain composite polyelectrolyte solution.
(3) the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-40 DEG C, the vacuum condition that is 500Pa in temperature, carry out frozen dried 12h, obtain composite polyelectrolyte tunica fibrosa.
(4) PBS of the sodium alginate obtaining in step (3) and epsilon-polylysine nano fibrous membrane is embathed to liquid and process respectively L929 cell after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and mensuration wavelength is 490nm, and compared with blank sample, the P value of its 24 hours, 48 hours, 72 hours is all greater than 0.05, there is no significant difference, shows that obtained nano fibrous membrane does not exist toxicity.
(5) sodium alginate obtaining in step (3) and epsilon-polylysine nano fibrous membrane are carried out to cell inoculation experiments, there is excellent cell and attach and multiplication characteristic.
Embodiment 5:
(1) heparin that is 1200g/mol by molecular weight and molecular weight are that the shitosan of 3000g/mol is dissolved in respectively deionized water, be made into the solution that percentage by weight is respectively 1wt% and 0.05wt%, then solution is fully stirred, so that dissolve completely, obtain heparin and chitosan solution.
(2) heparin solution of preparation in step (1) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared heparin solution is freezed in liquid nitrogen environment, then the heparin solution freezing is transferred in freeze drier, be under-10 DEG C, the vacuum condition that is 1Pa in temperature, carry out frozen dried 48h, obtain heparin tunica fibrosa.The heparin tunica fibrosa of preparation is put into chitosan solution, obtain composite polyelectrolyte solution.Wherein the volume ratio of two kinds of solution is: 2: 8.
(3) the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-40 DEG C, the vacuum condition that is 360Pa in temperature, carry out frozen dried 36h, obtain composite polyelectrolyte tunica fibrosa.
(4) PBS of the heparin obtaining in step (3) and chitosan nano fiber membrane is embathed to liquid and process respectively L929 cell after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and mensuration wavelength is 490nm, and compared with blank sample, the P value of its 24 hours, 48 hours, 72 hours is all greater than 0.05, there is no significant difference, shows that obtained nano fibrous membrane does not exist toxicity.
(5) heparin obtaining in step (3) and chitosan nano fiber membrane are carried out to cell inoculation experiments, there is excellent cell and attach and multiplication characteristic.
Embodiment 6:
(1) be that the heparin of 40 000g/mol and molecular weight are that the epsilon-polylysine of 50 000g/mol is dissolved in respectively deionized water by molecular weight, be made into the solution that percentage by weight is respectively 0.05wt% and 0.7wt%, then solution is fully stirred, so that dissolve completely, obtain heparin and epsilon-polylysine solution.
(2) the epsilon-polylysine solution of preparation in step (1) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared epsilon-polylysine solution is freezed in liquid nitrogen environment, then the epsilon-polylysine solution freezing is transferred in freeze drier, be under-10 DEG C, the vacuum condition that is 1Pa in temperature, carry out frozen dried 48h, obtain epsilon-polylysine tunica fibrosa.The epsilon-polylysine tunica fibrosa of preparation is put into heparin solution, obtain composite polyelectrolyte solution.Wherein the volume ratio of two kinds of solution is: 6: 4.
(3) the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-35 DEG C, the vacuum condition that is 10Pa in temperature, carry out frozen dried 24h, obtain composite polyelectrolyte tunica fibrosa.
(4) PBS of the heparin obtaining in step (3) and epsilon-polylysine nano fibrous membrane is embathed to liquid and process respectively L929 cell after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and mensuration wavelength is 490nm, and compared with blank sample, the P value of its 24 hours, 48 hours, 72 hours is all greater than 0.05, there is no significant difference, shows that obtained nano fibrous membrane does not exist toxicity.
(5) heparin obtaining in step (3) and epsilon-polylysine nano fibrous membrane are carried out to cell inoculation experiments, there is excellent cell and attach and multiplication characteristic.
Embodiment 7:
(1) be that the gamma-polyglutamic acid-of 100 000g/mol and molecular weight are that the epsilon-polylysine of 10 000g/mol is dissolved in respectively deionized water by molecular weight, be made into the solution that percentage by weight is respectively 0.01wt% and 0.09wt%, then solution is fully stirred, so that dissolve completely, obtain gamma-polyglutamic acid-and epsilon-polylysine solution.
(2) the epsilon-polylysine solution of preparation in step (1) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared epsilon-polylysine solution is freezed in liquid nitrogen environment, then the epsilon-polylysine solution freezing is transferred in freeze drier, be under-30 DEG C, the vacuum condition that is 200Pa in temperature, carry out frozen dried 24h, obtain epsilon-polylysine tunica fibrosa.Gamma-polyglutamic acid-solution is poured on the epsilon-polylysine tunica fibrosa of preparation, obtained composite polyelectrolyte solution.Wherein the volume ratio of two kinds of solution is: 5: 8.
(3) the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-55 DEG C, the vacuum condition that is 50Pa in temperature, carry out frozen dried 12h, obtain composite polyelectrolyte tunica fibrosa.
(4) PBS of the gamma-polyglutamic acid-obtaining in step (3) and epsilon-polylysine nano fibrous membrane is embathed to liquid and process respectively L929 cell after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and mensuration wavelength is 490nm, and compared with blank sample, the P value of its 24 hours, 48 hours, 72 hours is all greater than 0.05, there is no significant difference, shows that obtained nano fibrous membrane does not exist toxicity.
(5) gamma-polyglutamic acid-obtaining in step (3) and epsilon-polylysine nano fibrous membrane are carried out to cell inoculation experiments, there is excellent cell and attach and multiplication characteristic.
Embodiment 8:
(1) be that the gamma-polyglutamic acid-of 10 000 000g/mol and molecular weight are that the shitosan of 5 000g/mol is dissolved in respectively deionized water by molecular weight, be made into the solution that percentage by weight is respectively 0.001wt% and 0.5wt%, then solution is fully stirred, so that dissolve completely, obtain gamma-polyglutamic acid-and chitosan solution.
(2) the gamma-polyglutamic acid-solution of preparation in step (1) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared gamma-polyglutamic acid-solution is freezed in liquid nitrogen environment, then the gamma-polyglutamic acid-solution freezing is transferred in freeze drier, be under-65 DEG C, the vacuum condition that is 90Pa in temperature, carry out frozen dried 24h, obtain gamma-polyglutamic acid-tunica fibrosa.Chitosan solution is poured on the gamma-polyglutamic acid-tunica fibrosa of preparation, obtained composite polyelectrolyte solution.Wherein the volume ratio of two kinds of solution is: 9: 2.
(3) the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-60 DEG C, the vacuum condition that is 400Pa in temperature, carry out frozen dried 48h, obtain composite polyelectrolyte tunica fibrosa.
(4) PBS of the gamma-polyglutamic acid-obtaining in step (3) and chitosan nano fiber membrane is embathed to liquid and process respectively L929 cell after 24 hours, 48 hours, 72 hours, abandon supernatant, with PBS washing 2 times.ELIASA is measured light absorption value, and mensuration wavelength is 490nm, and compared with blank sample, the P value of its 24 hours, 48 hours, 72 hours is all greater than 0.05, there is no significant difference, shows that obtained nano fibrous membrane does not exist toxicity.
(5) gamma-polyglutamic acid-obtaining in step (3) and chitosan nano fiber membrane are carried out to cell inoculation experiments, there is excellent cell and attach and multiplication characteristic.
Claims (1)
1. desivac is prepared the method for composite polyelectrolyte tunica fibrosa, it is characterized in that comprising the following steps:
(1) preparation of polyelectrolyte solution: polyanion and polycation are dissolved in respectively in solvent, be mixed with respectively the solution that percentage by weight is 0.001～1wt%, then solution is fully stirred, so that dissolve completely, obtain polyanion and said polycation solution;
(2) by polyanion and said polycation solution by volume 1～9: 9～1 ratio mix with diverse ways, obtain composite polyelectrolyte solution;
(3) desivac is prepared composite polyelectrolyte tunica fibrosa: the composite polyelectrolyte solution of preparation in step (2) is transferred in liquid nitrogen frozen device, open refrigerating plant, prepared composite polyelectrolyte solution is freezed in liquid nitrogen environment, then the polyelectrolyte composite solution freezing is transferred in freeze drier, be under-10～-80 DEG C, the vacuum condition that is 1～600Pa in temperature, carry out frozen dried 12～48h, obtain composite polyelectrolyte tunica fibrosa
In described step (2), mixed method is:
Method 1: polyanion or the said polycation solution of preparation in step (1) are transferred in liquid nitrogen frozen device, open refrigerating plant, make prepared polyanion or said polycation solution Quick freezing in liquid nitrogen environment, then the polyanion freezing or said polycation solution are transferred in freeze drier, be under-10～-80 DEG C, the vacuum condition that is 1～600Pa in temperature, carry out frozen dried 12～48h, obtain polyanion or polycation tunica fibrosa; The polyanion of preparation or polycation tunica fibrosa are put into polycation or polyanion solution, obtain composite polyelectrolyte solution;
Method 2: polycation or polyanion solution are poured on the polyanion or polycation tunica fibrosa of preparation in method 1, obtained composite polyelectrolyte solution;
Described polyanion is: hyaluronic acid M
w=5000～2000000g/mol, sodium alginate viscosity average molecular weigh are 2 × 10
6, heparin M
w=1200～40000g/mol or gamma-polyglutamic acid-M
w=100000～10000000g/mol; Polycation is: shitosan M
w=3000～200000g/mol or epsilon-polylysine M
w=5000～100000g/mol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210052874.2A CN102605467B (en) | 2012-03-02 | 2012-03-02 | Method for preparing composite polyelectrolyte fiber membrane by utilizing freeze-drying way |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210052874.2A CN102605467B (en) | 2012-03-02 | 2012-03-02 | Method for preparing composite polyelectrolyte fiber membrane by utilizing freeze-drying way |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102605467A CN102605467A (en) | 2012-07-25 |
| CN102605467B true CN102605467B (en) | 2014-10-29 |
Family
ID=46523215
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210052874.2A Expired - Fee Related CN102605467B (en) | 2012-03-02 | 2012-03-02 | Method for preparing composite polyelectrolyte fiber membrane by utilizing freeze-drying way |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102605467B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
| 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 |
| CN105063801A (en) * | 2015-08-27 | 2015-11-18 | 常州大学 | Method for preparing crosslinked chondroitin sulfate derivative fiber |
| CN109157982B (en) * | 2018-09-14 | 2021-07-16 | 北京工业大学 | Method for preparing polyelectrolyte composite nanofiltration membrane by using freeze drying technology |
| CN109966552B (en) * | 2019-03-22 | 2020-04-07 | 宁波市医疗中心李惠利医院 | Tissue engineering scaffold material and preparation method, application and application method thereof |
| CN110644232B (en) * | 2019-09-20 | 2021-10-12 | 南通大学 | Preparation method of natural antibacterial water-retention mask |
| CN111996616B (en) * | 2020-08-20 | 2021-09-28 | 江南大学 | Sodium alginate/polylysine self-assembled fiber with controllable size |
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 (2)
| Title |
|---|
| 壳聚糖基聚电解质复合物在生物医学领域的应用;张停等;《化学通报》;20120131;第75卷(第1期);正文第46-50页 * |
| 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》.2012,第75卷(第1期),正文第46-50页. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102605467A (en) | 2012-07-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102605467B (en) | Method for preparing composite polyelectrolyte fiber membrane by utilizing freeze-drying way | |
| Fan et al. | Preparation and characterization of oxidized konjac glucomannan/carboxymethyl chitosan/graphene oxide hydrogel | |
| Assaad et al. | Injectable thermosensitive chitosan hydrogels with controlled gelation kinetics and enhanced mechanical resistance | |
| Liu et al. | Rapid gelling, self-healing, and fluorescence-responsive chitosan hydrogels formed by dynamic covalent crosslinking | |
| Jia et al. | Self-healing hydrogels of low molecular weight poly (vinyl alcohol) assembled by host–guest recognition | |
| Hu et al. | Hydrogels prepared from pineapple peel cellulose using ionic liquid and their characterization and primary sodium salicylate release study | |
| Sun et al. | Preparation and characterization of novel curdlan/chitosan blending membranes for antibacterial applications | |
| Mishra et al. | Tamarind xyloglucan: a polysaccharide with versatile application potential | |
| Lazaridou et al. | Cryogelation of cereal β-glucans: structure and molecular size effects | |
| Kim et al. | Injectable, self-healable and adhesive hydrogels using oxidized Succinoglycan/chitosan for pH-responsive drug delivery | |
| Li et al. | Injectable polysaccharide hybrid hydrogels as scaffolds for burn wound healing | |
| Li et al. | Effect of ε-polylysine addition on κ-carrageenan gel properties: Rheology, water mobility, thermal stability and microstructure | |
| CN101602814B (en) | Carboxymethyl curdlan gum physical hydrogel and preparation method thereof | |
| EP2197417B1 (en) | Pharmaceutical composition comprising s-nitrosoglutathione and polysaccharide | |
| CN105153438A (en) | Preparation method of high-strength high-swelling nanocellulose and polyving akohol composite hydrogel | |
| Zhang et al. | Study of injectable Blueberry anthocyanins-loaded hydrogel for promoting full-thickness wound healing | |
| Luo et al. | Bioactive therapeutics-repair-enabled citrate-iron hydrogel scaffolds for efficient post-surgical skin cancer treatment | |
| Xia et al. | Preparation and characterization of a poly (ethylene glycol) grafted carboxymethyl konjac glucomannan copolymer | |
| Araújo et al. | Novel hydrogels based on yeast chitin-glucan complex: Characterization and safety assessment | |
| Kim et al. | Injectable hydrogels derived from phosphorylated alginic acid calcium complexes | |
| KR101875115B1 (en) | Manufacturing method for hydrogel foam using agarose form seaweed | |
| Li et al. | Structure and properties of chitin/alginate blend membranes from NaOH/urea aqueous solution | |
| Zhou et al. | Semi-interpenetrating polymer network hydrogels based on water-soluble N-carboxylethyl chitosan and photopolymerized poly (2-hydroxyethyl methacrylate) | |
| CN101942121B (en) | Method for improving viscosity and stability of sodium alga acid | |
| Zheng et al. | Fabrication of phenylalanine amidated pectin using ultra-low temperature enzymatic method and its hydrogel properties in drug sustained release application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141029 |