CN109731137A - Preparation method of albumin coating with biological antifouling function and material with biological antifouling function - Google Patents
Preparation method of albumin coating with biological antifouling function and material with biological antifouling function Download PDFInfo
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- CN109731137A CN109731137A CN201910189300.1A CN201910189300A CN109731137A CN 109731137 A CN109731137 A CN 109731137A CN 201910189300 A CN201910189300 A CN 201910189300A CN 109731137 A CN109731137 A CN 109731137A
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- albumin
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- biological functions
- acid
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- 108010088751 Albumins Proteins 0.000 title claims abstract description 117
- 102000009027 Albumins Human genes 0.000 title claims abstract description 117
- 239000000463 material Substances 0.000 title claims abstract description 71
- 239000011248 coating agent Substances 0.000 title claims abstract description 62
- 238000000576 coating method Methods 0.000 title claims abstract description 62
- 238000002360 preparation method Methods 0.000 title claims abstract description 43
- 230000003373 anti-fouling effect Effects 0.000 title abstract 6
- 238000003618 dip coating Methods 0.000 claims abstract description 20
- 239000000758 substrate Substances 0.000 claims abstract description 19
- 239000007800 oxidant agent Substances 0.000 claims abstract description 15
- 239000003638 chemical reducing agent Substances 0.000 claims abstract description 14
- 238000006243 chemical reaction Methods 0.000 claims abstract description 13
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 13
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 13
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 12
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 238000002791 soaking Methods 0.000 claims abstract description 4
- 230000008827 biological function Effects 0.000 claims description 40
- 239000005862 Whey Substances 0.000 claims description 23
- 102000007544 Whey Proteins Human genes 0.000 claims description 23
- 108010046377 Whey Proteins Proteins 0.000 claims description 23
- 239000007864 aqueous solution Substances 0.000 claims description 14
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims description 12
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 claims description 12
- 230000001131 transforming effect Effects 0.000 claims description 9
- XTEGARKTQYYJKE-UHFFFAOYSA-M Chlorate Chemical compound [O-]Cl(=O)=O XTEGARKTQYYJKE-UHFFFAOYSA-M 0.000 claims description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 8
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 8
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethyl mercaptane Natural products CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 claims description 8
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 claims description 8
- 108010024636 Glutathione Proteins 0.000 claims description 6
- 229960003180 glutathione Drugs 0.000 claims description 6
- KMUONIBRACKNSN-UHFFFAOYSA-N potassium dichromate Chemical compound [K+].[K+].[O-][Cr](=O)(=O)O[Cr]([O-])(=O)=O KMUONIBRACKNSN-UHFFFAOYSA-N 0.000 claims description 6
- 239000012286 potassium permanganate Substances 0.000 claims description 6
- 230000002829 reductive effect Effects 0.000 claims description 6
- PNYYBUOBTVHFDN-UHFFFAOYSA-N sodium bismuthate Chemical compound [Na+].[O-][Bi](=O)=O PNYYBUOBTVHFDN-UHFFFAOYSA-N 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 6
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 6
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 claims description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 4
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 4
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- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 4
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 claims description 4
- 229940098773 bovine serum albumin Drugs 0.000 claims description 4
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- 229910017604 nitric acid Inorganic materials 0.000 claims description 4
- JRKICGRDRMAZLK-UHFFFAOYSA-L peroxydisulfate Chemical compound [O-]S(=O)(=O)OOS([O-])(=O)=O JRKICGRDRMAZLK-UHFFFAOYSA-L 0.000 claims description 4
- 210000002966 serum Anatomy 0.000 claims description 4
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 claims description 3
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 claims description 3
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- SXDBWCPKPHAZSM-UHFFFAOYSA-N bromic acid Chemical compound OBr(=O)=O SXDBWCPKPHAZSM-UHFFFAOYSA-N 0.000 claims 1
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- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 2
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Landscapes
- Materials For Medical Uses (AREA)
Abstract
本发明公开了具有生物抗污功能的白蛋白涂层的制备方法及具有生物抗污功能的材料，涉及医用材料技术领域。该具有生物抗污功能的白蛋白涂层的制备方法包括：将白蛋白、溶剂和蛋白结构转化剂混合后形成浸涂混合液；将待改性材料在浸涂混合液中浸泡；其中，蛋白结构转化剂为强氧化剂或强还原剂。具有生物抗污功能的材料包括改性基材和在改性基材上形成白蛋白涂层，白蛋白为β折叠结构。材料制备方法简便易行，且涂层与基材的结合强度高，具有长期抗凝血性能和抗菌性能，能够抑制血管细胞和组织的粘附。
The invention discloses a preparation method of an albumin coating with biological antifouling function and a material with biological antifouling function, and relates to the technical field of medical materials. The preparation method of the albumin coating with biological antifouling function includes: mixing albumin, a solvent and a protein structure conversion agent to form a dip coating mixture; soaking the material to be modified in the dip coating mixture; wherein, the protein Structural conversion agents are strong oxidizing agents or strong reducing agents. The material with biological antifouling function includes a modified substrate and an albumin coating formed on the modified substrate, and the albumin has a β-sheet structure. The material preparation method is simple and feasible, and the coating has high bonding strength with the substrate, has long-term anticoagulant properties and antibacterial properties, and can inhibit the adhesion of vascular cells and tissues.
Description
Technical field
The present invention relates to medical material tech fields, and in particular to have the system of the albumin coating of biological functions
Preparation Method and material with biological functions.
Background technique
The device that artificial heart, pacemaker, ventricular assist device etc. are contacted with blood of human body meeting in process of clinical application
With thrombus and infection problems, the increase of patient morbidity, the death rate are directly resulted in.Prevention infection and thrombus in clinic
Means mainly use heparin or antibiotic to carry out drug therapy, and such application method increases patient's bleeding, and is easy to make liver
The generation of plain induced thrombocytopenia.In addition, antibiotic can cause many serious adverse reactions after being also applied to human body.
In order to reduce the complication of clinical application, the mode of functionalizing material surface is come into being, and is to solve infection and blood
The effective means of the problems such as bolt.Polyethylene glycol (PEG) grafting/coating, polyethylene glycol (PEG) grafting/coating, at super hydrophobic surface
The surface modifying methods such as reason, surface heparinization and surface micro-nano structure processing are mostly all cumbersome, and generally requiring will be to material
It is handled to introduce functional group, to realize the fixation of functional molecular, operating process is sufficiently complex.
Albumin is primarily present in the whey of blood and mammal, if albumin is adsorbed on the surface of material in advance, just
The absorption of other albumen can be effectively prevented, and then there is anticoagulant, the function of antibacterial.But with albumin to material surface
For modified method mainly by the way of covalence graft, effect is often limited to the fixed amount of surface albumin.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation methods of albumin coating with biological functions, using one
The mode of step dip-coating more easily carries out the fixation of albumin molecule.
Another object of the present invention is to provide a kind of material with biological functions, preparation method is easy easily
Row has the anti-physical performance of biology well.
The present invention solves its technical problem and adopts the following technical solutions to realize.
The invention proposes a kind of preparation methods of albumin coating with biological functions, include the following steps:
Coating is formed on material to be modified using dip-coating mixed liquor；
Wherein, dip-coating mixed liquor is mixed by albumin solution and protein structure transforming agent, and protein structure transforming agent is
Strong oxidizer or strong reductant.
The present invention also proposes a kind of material with biological functions, including substrate modified and is formed on substrate modified
Albumin coating, albumin are β-pleated sheet structure；
Preferably, there is the material of biological functions to be prepared by above-mentioned preparation method.
The embodiment of the present invention provides a kind of beneficial effect of the preparation method of albumin coating with biological functions
It is: by mixing albumin and protein structure transforming agent, using strong oxidizer or strong reductant by the cystine linkage oxygen of albumin
Change, is changed from αhelix to β-pleated sheet structure, this give the adhesion characteristics of albumin and material, can be in almost institute
There is the material surface of shape and type to be modified, and there is good bond strength with substrate surface.
The present invention also provides a kind of materials with biological functions, including substrate modified and the shape on substrate modified
At albumin coating, albumin is β-pleated sheet structure, and such material preparation method is simple and easy to do, and the combination of coating and substrate is strong
Degree is high, has long-term anticoagulation performance and anti-microbial property, is able to suppress the adherency of vascular cell and tissue.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 be the embodiment of the present invention in stainless steel material before modified after comparison diagram；
Fig. 2 be the embodiment of the present invention in silica gel material before modified after and comparative example in modified material comparison diagram；
Fig. 3 is the fungistatic effect figure that modified material is prepared in the embodiment of the present invention and comparative example.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
The preparation method to the albumin coating with biological functions provided in an embodiment of the present invention and have below
The material of biological functions is specifically described.
The preparation method of a kind of albumin coating with biological functions provided in an embodiment of the present invention comprising such as
Lower step:
The preparation of S1, dip-coating mixed liquor
Dip-coating mixed liquor is formed after albumin, solvent and protein structure transforming agent are mixed；Wherein, protein structure transforming agent
For strong oxidizer or strong reductant.The cystine linkage in albumin is aoxidized using strong oxidizer or strong reductant, and then makes white egg
White to be changed from αhelix to β-pleated sheet structure, β-pleated sheet structure albumin has the performance of " step " self assembly.
Specifically, in dip-coating mixed liquor, the concentration of albumin is 0.002-90mg/mL, preferably 0.1-30mg/mL,
More preferably 0.5-15mg/mL.The dosage of albumin will affect the anticoagulation function of coating, while also will affect coating and material
Between bond strength.
In general, albumin include bovine serum albumin(BSA), human serum albumins, in whey albumin any one or
It is a variety of.It is suitable for being modified using strong reductant or strong oxidizer using above several albumin, utilizes the β-pleated sheet of generation
Structure assigns albumin self-assembly ability.
Specifically, strong reductant includes one of mercaptoethanol, dithiothreitol (DTT) and reductive glutathione or a variety of.
Concentration of the strong reductant in dip-coating mixed liquor is 0.002-20 mg/mL, preferably 0.1-10mg/mL.Strong reductant selection with
It is above several that the αhelix of albumin can be made to be changed into β-pleated sheet structure, and reaction rate can be realized the quick of coating fastly
Molding.The used in amounts of strong reductant will be controlled, and prevent its dosage from excessively albumin being caused to be denaturalized, and it is very few to be also prevented from its dosage
Keep albumin reaction insufficient.
Specifically, strong oxidizer include trivalent cobalt salt, chlorate, potassium permanganate, persulfate, potassium bichromate, the concentrated sulfuric acid,
Hydrochloric acid, nitric acid, hydrobromic acid, hydroiodic acid, perchloric acid, ozone, hydrogen peroxide, fluorine gas, chlorine, sodium bismuthate, periodic acid, Na2Fe04 and
Any one or more in brown lead oxide.Above several strong oxidizers can make the transformation of albumin recurring structure, and change
Efficiency is also quickly.
Further, in dip-coating mixed liquor, trivalent cobalt salt, potassium permanganate, potassium bichromate, sodium bismuthate, persulfate, height
The concentration of acid iodide, Na2Fe04 and brown lead oxide is 0.001-20 mg/mL；Preferably 0.2-10mg/mL.In dip-coating mixed liquor
In, the quality of the concentrated sulfuric acid, chlorate, hydroiodic acid, nitric acid, hydrobromic acid, perchloric acid, hydrochloric acid, chlorine, hydrogen peroxide, fluorine gas and ozone
Score is 0.02-50%；Preferably 0.1-30%.Equally, the used in amounts of strong oxidizer will carry out stringent control, white to prevent
The denaturation of albumen or albumin reaction are insufficient.
Preferably, the preparation process of dip-coating mixed liquor is to be obtained by mixing albumin aqueous solution and protein structure transforming agent,
Raw material can be made more fully to mix by the way of solution mixing, prevent local concentration excessively high.
S2, material, which impregnate, to be modified
Material to be modified is impregnated in dip-coating mixed liquor, soaking time is -8 days 2 seconds, and reaction temperature is 0-100 DEG C.It is excellent
Selection of land, soaking time 10-240min, reaction temperature are 15-45 DEG C.Material to be modified is impregnated the very short time can
Functional coating is formed on the surface of the material, and the bond strength between coating and material is high, is suitable for industrial applications.
Preferably, after having reacted material to be modified in dip-coating mixed liquor, modified sample is cleaned simultaneously
It is dry.Water-washing process is conducive to the unreacted raw material of removal adherency on the surface of the material, improves the purity of product.
In the embodiment of the present invention the advantages of preparation method: (1) it is easy to operate, without the special of complicated reaction process and valuableness
With equipment, chemical reagent etc., reactive functionality is introduced without carrying out pretreatment, process costs are low.(2) β-pleated sheet structure can make
Albumin adheres to well with material surface, suitable for the material of nearly all shape and material, is suitable for promoting and applying.(3)
Coating uniform, the thickness of albumin are easily controllable, and the bond strength between coating and material is high, can be used as long-term anticoagulation
Material, anti-biotic material use, and also have the function of inhibiting vascular cell and tissue adhension.
The embodiment of the invention also provides a kind of material with biological functions, including substrate modified and in modified base
Albumin coating is formed on material, albumin is β-pleated sheet structure；Preferably, have the material of biological functions by above-mentioned preparation
Method is prepared.The material has good anticoagulation, anti-microbial property, and the bond strength between coating and substrate is high.
Substrate modified (material to be modified) in the embodiment of the present invention can be existing any material for film forming, several
It is not limited to shape and material.Can specifically include: (1) metal material: stainless steel, titanium and its alloy, cobalt-base alloys, NiTi close
Golden, magnesium and its alloy, zinc and its alloy, iron and its alloy etc..(2) inorganic material: silica, titanium dioxide, carbon materials
(C), inorganic material such as silicon, titanium dioxide, titanium oxide, titanium nitride etc..(3) high molecular material: terylene (PET), polyvinyl alcohol
(PVALC), polyethylene (PE), polytetrafluoroethylene (PTFE) (PTFE), polyvinyl chloride (PVC), polystyrene (PS), polyurethane (PU), poly-
The high molecular materials such as propylene (PP), polycarbonate (PC), polyacrylic acid (PAA) and its derivative, polyethylene glycol and its derivative
Deng.(4) bio-medical micro-and nano-particles: zinc oxide nano-particle (quantum dot), Nano particles of silicon dioxide (quantum dot), oxidation
Titanium nanoparticle (quantum dot).(5) natural biologic material: plasticity starch-based material (PSM), sodium alginate (sodium
Alginate) collagen (collagen), fibrin (fibrous protein), Sodium Hyaluronate (sodium
Hyaluronate), gelatin (gelatin) etc..(6) artificial synthetic polypeptide class hydrogel material: poly- L- Glutamic Acid, poly- L- rely ammonia
Acid etc..The type of base material is not limited to the above material, or the mixing material of the above different materials.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment 1
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After Bovine Serum Albumin in Aqueous Solution is mixed with the concentrated sulfuric acid, water is added to be adjusted to the dense of bovine serum albumin(BSA) in mixed liquor
Degree is 0.002mg/L, and the mass fraction of the concentrated sulfuric acid is about 0.02%.
Polyvinyl chloride (PVC) extracorporal circulatory system conduit to be modified is placed in mixed liquor, is impregnated under the conditions of 0 DEG C of temperature
About 2s.Sample is cleaned, is dried.
Embodiment 2
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After human serum albumins aqueous solution is mixed with perchloric acid, water is added to be adjusted to the dense of human serum albumins in mixed liquor
Degree is 90mg/L, and the mass fraction of perchloric acid is about 50%.
Nitinol inferior vena cava filter to be modified is placed in mixed liquor, impregnates about 5 under the conditions of 100 DEG C of temperature
It.Sample is cleaned, is dried.
Embodiment 3
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with potassium permanganate, water is added to be adjusted to the concentration of whey albumin in mixed liquor
For 0.1mg/L, the concentration of potassium permanganate is about 0.001mg/L.
Silica gel central venous catheter to be modified is placed in mixed liquor, impregnates about 10min under the conditions of 15 DEG C of temperature.
Sample is cleaned, is dried.
Embodiment 4
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with potassium bichromate, water is added to be adjusted to the concentration of whey albumin in mixed liquor
For 30mg/L, the concentration of potassium bichromate is about 20mg/L.
Polysulfone hemodialyzer film to be modified is placed in mixed liquor, is impregnated about under the conditions of 15 DEG C of temperature
240min.Sample is cleaned, is dried.
Embodiment 5
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with sodium bismuthate, the concentration for adding water to be adjusted to whey albumin in mixed liquor is
0.5mg/L, the concentration of sodium bismuthate are about 0.2mg/L.
Polysulfone hemodialyzer film to be modified is placed in mixed liquor, impregnates about 10min under the conditions of 45 DEG C of temperature.
Sample is cleaned, is dried.
Embodiment 6
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with Na2Fe04, water is added to be adjusted to the concentration of whey albumin in mixed liquor
For 15mg/L, the concentration of Na2Fe04 is about 10mg/L.
Silica gel central venous catheter to be modified is placed in mixed liquor, impregnates about 15min under the conditions of 35 DEG C of temperature.
Sample is cleaned, is dried.
Embodiment 7
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with mercaptoethanol, water is added to be adjusted to the concentration of whey albumin in mixed liquor
For 0.01mg/L, the concentration of mercaptoethanol is about 0.002mg/L.
Silica gel central venous catheter to be modified is placed in mixed liquor, impregnates about 15min under the conditions of 35 DEG C of temperature.
Sample is cleaned, is dried.
Embodiment 8
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with dithiothreitol (DTT), water is added to be adjusted to the dense of whey albumin in mixed liquor
Degree is 10mg/L, and the concentration of dithiothreitol (DTT) is about 20mg/L.
Silica gel central venous catheter to be modified is placed in mixed liquor, impregnates about 15min under the conditions of 35 DEG C of temperature.
Sample is cleaned, is dried.
Embodiment 9
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with reductive glutathione, water is added to be adjusted to whey albumin in mixed liquor
Concentration be 0.1mg/L, the concentration of reductive glutathione is about 0.1 mg/L.
Silica gel central venous catheter to be modified is placed in mixed liquor, impregnates about 15min under the conditions of 35 DEG C of temperature.
Sample is cleaned, is dried.
Embodiment 10
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with reductive glutathione, water is added to be adjusted to whey albumin in mixed liquor
Concentration be 10mg/L, the concentration of reductive glutathione is about 10 mg/L.
Silica gel central venous catheter to be modified is placed in mixed liquor, impregnates about 15min under the conditions of 35 DEG C of temperature.
Sample is cleaned, is dried.
Embodiment 11
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with mercaptoethanol, water is added to be adjusted to the concentration of whey albumin in mixed liquor
For 0.1mg/L, the concentration of mercaptoethanol is about 0.05mg/L.
Stainless steel material is placed in mixed liquor, about 5 min are impregnated under the conditions of 25 DEG C of temperature.Sample is carried out clear
It washes, dry.
Embodiment 12
The present embodiment provides a kind of preparation methods of albumin coating with biological functions comprising following step
It is rapid:
After whey albumin aqueous solution is mixed with mercaptoethanol, water is added to be adjusted to the concentration of whey albumin in mixed liquor
For 0.1mg/L, the concentration of mercaptoethanol is about 0.05mg/L.
Silica gel material is placed in mixed liquor, impregnates about 5min under the conditions of 25 DEG C of temperature.Sample is cleaned, is done
It is dry.
Comparative example 1
This comparative example provides a kind of method for introducing albumin on the surface of the material, specific to walk using the method for covalence graft
It is rapid: (1) Tris of the 1.2mg distilled water of 1mL to be dissolved, obtain the buffer solution that pH is 8.5 or so, be then added 2mg's
Dopamine, it is to be dissolved sufficiently after, stainless steel sample is added, by reaction in 12 hours, is obtained on stainless steel sample surface poly- more
Bar amine coating.(2) albumin of 5mg is dissolved in Tris buffer (as above), it is to be dissolved sufficiently after, be added step 1 in obtain
The coating modified stainless steel sample of the poly-dopamine obtained carries out surface albumin covalence graft, and after grafting 12 hours, sample is successively
It is cleaned with phosphate buffer and distilled water, is dry, that is, obtaining the sample that albumin is covalently fixed on surface.
Test example 1
State before impregnating in embodiment 11 to stainless steel material and after impregnating compares, as shown in Figure 1.Explanation
Embodiment 11 successfully deposited albumin coating in stainless steel surface, and the coating is shown and the firm binding force of substrate.
Test example 2
The material being prepared in embodiment 12 and comparative example 1 is scanned Electronic Speculum characterization, as a result such as Fig. 2.
As shown in Figure 2, the embodiment of the present invention is deposited in Silica Surface albumin coating and white using conventional covalence graft
The surface of protein molecular is compared, and albumin of the present invention has the function of inhibiting platelet adhesion reaction and activation more excellently.By altogether
Valence is grafted the surface of albumin, since albumin covalence graft amount is limited to the reactive functional groups density limit of material surface introducing
System, surface has still adhered to more blood platelet, and activation still occurs in part blood platelet, some blood platelets even occur
The case where content outflow.And the coating modified sample of albumin of the invention, not only quantity is few for surface blood platelet, and all presents
For the spherical shape of tranquillization state.
Test example 3
The anti-microbial property of product is obtained in testing example 12 and comparative example 1, test result is shown in Fig. 3.
From the figure 3, it may be seen that in the albumin coating of Silica Surface deposition and using conventional covalence graft in the embodiment of the present invention
The surface of albumin molecule is compared, and the albumin coating formed in the embodiment of the present invention can more effectively inhibit the shape of bacterial plaque
At with more excellent ground antibacterial functions.
Test example 4
The anti-microbial property of material is prepared in testing example 1-10, test method: modified material is simulated in PBS
It is persistently impregnated one month in body fluid, tests modified material respectively to the bacteriostasis rate of staphylococcus aureus and Escherichia coli.
The results show that material obtained in embodiment 1-10 to the bacteriostasis rate of staphylococcus aureus successively are as follows: 91.3%,
96.2,99.6,99.8,99.4,99.8,92.2,98.2,99.3 and 99.4.Successively to the bacteriostasis rates of Escherichia coli are as follows:
89.2%, 94.3,99.4,99.7,99.6,99.4,93.7,99.1,99.5 and 99.8.
It can be seen that the modified material tool with albumin coating that the preparation method in the embodiment of the present invention is prepared
There is good bacteriostasis property, and the dosage of each component has certain influence to fungistatic effect in reaction process, using of the invention preferred
Amounts of components when fungistatic effect can further increase.
In conclusion a kind of preparation method of albumin coating with biological functions provided by the invention, passes through
Albumin and protein structure transforming agent are mixed, aoxidized the cystine linkage of albumin using strong oxidizer or strong reductant, by α spiral shell
It revolves structure to change to β-pleated sheet structure, this give the adhesion characteristics of albumin and material, have with substrate surface and combine well
Intensity.
The present invention also provides a kind of materials with biological functions, including substrate modified and the shape on substrate modified
At albumin coating, albumin is β-pleated sheet structure, and the bond strength height of such material floating coat and substrate has long-term anticoagulant
Hemorrhagic energy and anti-microbial property, are able to suppress the adherency of vascular cell and tissue.
Embodiments described above is a part of the embodiment of the present invention, instead of all the embodiments.Reality of the invention
The detailed description for applying example is not intended to limit the range of claimed invention, but is merely representative of selected implementation of the invention
Example.Based on the embodiments of the present invention, obtained by those of ordinary skill in the art without making creative efforts
Every other embodiment, shall fall within the protection scope of the present invention.
Claims (10)
1. a kind of preparation method of the albumin coating with biological functions, which comprises the steps of:
Coating is formed on material to be modified using dip-coating mixed liquor；
Wherein, the dip-coating mixed liquor is mixed by albumin solution and protein structure transforming agent, the protein structure conversion
Agent is strong oxidizer or strong reductant.
2. the preparation method of the albumin coating with biological functions according to claim 1, which is characterized in that
In the dip-coating mixed liquor, the concentration of the albumin is 0.002-90mg/mL, preferably 0.1-30mg/mL, more preferably
0.5-15mg/mL。
3. the preparation method of the albumin coating with biological functions according to claim 2, which is characterized in that institute
Stating albumin includes bovine serum albumin(BSA), human serum albumins, any one or more in whey albumin.
4. the preparation method of the albumin coating with biological functions according to claim 1, which is characterized in that institute
Stating strong reductant includes one of mercaptoethanol, dithiothreitol (DTT) and reductive glutathione or a variety of.
5. the preparation method of the albumin coating with biological functions according to claim 4, which is characterized in that institute
Stating concentration of the strong reductant in the dip-coating mixed liquor is 0.002-20mg/mL, preferably 0.1-10mg/mL.
6. the preparation method of the albumin coating with biological functions according to claim 1, which is characterized in that institute
Stating strong oxidizer includes trivalent cobalt salt, chlorate, potassium permanganate, persulfate, potassium bichromate, the concentrated sulfuric acid, hydrochloric acid, nitric acid, hydrogen
In bromic acid, hydroiodic acid, perchloric acid, ozone, hydrogen peroxide, fluorine gas, chlorine, sodium bismuthate, periodic acid, Na2Fe04 and brown lead oxide
Any one or more.
7. the preparation method of the albumin coating with biological functions according to claim 6, which is characterized in that institute
State strong oxidizer select the trivalent cobalt salt, the potassium permanganate, the potassium bichromate, the sodium bismuthate, the persulfate,
When any one or more in the periodic acid, the Na2Fe04 and the brown lead oxide, the concentration of the strong oxidizer
For 0.001-20mg/mL；Preferably 0.2-10mg/mL；
The strong oxidizer selects the concentrated sulfuric acid, chlorate, the hydroiodic acid, the nitric acid, the hydrobromic acid, described
When any one or more in perchloric acid, the hydrochloric acid, the chlorine, the hydrogen peroxide, the fluorine gas and the ozone, institute
The mass fraction for stating strong oxidizer is 0.02-50%；Preferably 0.1-30%.
8. the preparation method of the albumin coating with biological functions according to claim 1, which is characterized in that institute
The preparation process for stating dip-coating mixed liquor is to be obtained by mixing albumin aqueous solution and protein structure transforming agent.
9. the preparation method of the albumin coating with biological functions according to claim 1, which is characterized in that will
The material to be modified impregnates in the dip-coating mixed liquor to form coating；
Soaking time is -8 days 2 seconds, preferably 10-240min；
Reaction temperature is 0-100 DEG C, preferably 15-45 DEG C.
10. a kind of material with biological functions, which is characterized in that including substrate modified and the shape on the substrate modified
At albumin coating, the albumin is β-pleated sheet structure；
Preferably, the material with biological functions is prepared by preparation method of any of claims 1-9
And it obtains.
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