CN107137785A - One species specificity promotees the anticoagulation surface construction method of endothelial cell growth - Google Patents
One species specificity promotees the anticoagulation surface construction method of endothelial cell growth Download PDFInfo
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
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Abstract
The invention discloses the anticoagulation surface construction method that a species specificity promotees endothelial cell growth, belong to biomaterial surface technical field of modification, it comprises the following steps：1) biomaterial surface prepares poly-dopamine layer；2) biomaterial surface micro-contact printing fixes double amino-polyethyleneglycols；3) biomaterial surface fixes chitosan and heparin；4) biomaterial surface fixes REDV；The present invention is conducive to controlling the adhesion growth behavior of endothelial cell, realizes the complete healing of endothelial tissue；When being modified for blood vessel inner implantation material or equipment surfaces, the excellent rush endothelial cell growth performance of material and anticoagulation function can be assigned simultaneously；The coating that auto polymerization forms strong bonded can be carried out in all solids material surface because dopamine polymerize, and the coating formed has the stronger ability with the substance reaction containing amino, the surface that this method almost can be used for all biomaterials is modified, and promotees the anticoagulation surface of endothelial cell growth for building specificity.
Description
Technical field
The invention belongs to biomaterial surface technical field of modification, and in particular to a species specificity promotees endothelial cell growth
Anticoagulation surface construction method.
Background technology
Blood vessel inner implantation material or apparatus have highly important key effect in terms of the treatment of angiocardiopathy, so
And, after material or apparatus implantation, due to surface thrombosis and endothelium work(caused by blood constituent and inner skin cell function change
Energy disorder frequently results in graft failure.
Based on the understanding of interface biological respinse between material and blood of human body and endothelial cell, deposited in material surface inorganic
Coating, prepare macromolecule layer or immobilizing biologically active molecule and can improve blood compatibility and the imparting of material to a certain extent
The certain endothelium Forming ability of material, but these surface modifying methods can not completely inhibit thrombosis or promotion endothelium is cured completely
Close.
Research shows that complete natural endothelial layer is regulation blood coagulation-anti-freezing balance, inner membrance reparation-hyperplasia balance
Key.Thus material or implantation instrument surface rapid Cover endothelial layer will significantly reduce thrombus generation and smooth muscle is excessive
The degree of propagation, so significantly improve material or apparatus be implanted to power.
In order to form natural endothelial layer in material surface, method main at present has：In material surface plantation
Chrotoplast, material surface grafting extracellular matrix protein or endothelial growth factor promote cell adherence and growth, surface solid
Surely the bioactive molecule of the induction differentiation of endothelial progenitor cells, surface can be promoted to fix specificity and promote endothelial cell growth differentiation
The method such as antibody.Although these methods can remarkably promote the adhesion and growth of endothelial cell, its anticoagulation function has
Limit, and cell is in material surface random growth, therefore the endothelial layer obtained can't fully achieve normal endothelium
Function.
The content of the invention
Goal of the invention：It is an object of the invention to provide the anticoagulation surface construction that a species specificity promotees endothelial cell growth
Method, can be obviously improved the anticoagulation function of material, and can specifically promote endothelial cell ordering growth, so as to build special
The rush endothelial cell growth surface of property, is applied in terms of cardiovascular inner implantation material or apparatus.
Technical scheme：To achieve the above object, the present invention is adopted the following technical scheme that：
One species specificity promotees the anticoagulation surface construction method of endothelial cell growth, comprises the following steps：
1) biomaterial surface prepares poly-dopamine layer
Biomaterial is immersed into dopamine solution, abundant auto polymerization, is dried after cleaning, in triplicate, obtained poly-
The biomaterial surface of DOPA amine layer；
2) biomaterial surface micro-contact printing fixes double amino-polyethyleneglycols
Double amino-polyethyleneglycols solution are added dropwise on ribbon PDMS seals surface, ribbon PDMS seals surface is fully inhaled
After attached pair of amino-polyethyleneglycols, N2Dry, then by PDMS seals and step 1) biomaterial surface that has poly-dopamine layer fills
Tap is touched, and is dried after cleaning, obtains the biomaterial surface of amino-polyethyleneglycols modification；
3) biomaterial surface fixes chitosan and heparin
By step 2) obtained biomaterial is immersed into chitosan and heparin solution after fully reaction successively, and cleaning is dry
It is dry, obtain to surface and secure the biomaterial surface of chitosan and heparin；
4) biomaterial surface fixes REDV
By step 3) in biomaterial be immersed into end group be N- succinimides REDV solution in fully react, clean
After dry, obtain the biomaterial surface that REDV is contained on surface.
Step 1) in, the concentration of dopamine solution is 0.1~1mg/ml.
Step 1) in, dopamine solution pH is 8.0-8.5.
Step 1) in, the abundant auto polymerization time is 12-24 hours.
Step 2) in, the width of band and at intervals of 5~50 μm, the concentration of polyglycol solution is 0.1~1mg/ml.
Step 3) in, by step 2) to be immersed into chitosan solution fully reaction 4~12 first small for obtained biomaterial
When, after cleaning-drying, then fully absorption 4~8 hours is immersed into heparin solution.
Step 3) in, described chitosan and the concentration of heparin solution are 1~10mg/ml.
Step 4) in, the time fully reacted is 4~8 hours, and REDV concentration is 0.1~1mg/ml.
Beneficial effect：Compared with prior art, a species specificity of the invention promotees the anticoagulation surface of endothelial cell growth
Construction method, carries out surface modification to angiocarpy implantation material by this method, possesses following advantage：
1) heparin and REDV can be fixed on material surface specific region by Microcontact printing, so that in material surface shape
Into the bioactivity topological structure of specific pattern, be conducive to controlling the adhesion growth behavior of endothelial cell, realize endothelial tissue
Healing completely；
2) REDV peptide molecules specific can promote the adhesion and growth of endothelial cell, while can also suppress smooth muscle
The growth of cell；Heparin molecule has excellent anticoagulation function, therefore, is modified for blood vessel inner implantation material or equipment surfaces
When, the excellent rush endothelial cell growth performance of material and anticoagulation function can be assigned simultaneously；
3) coating that auto polymerization forms strong bonded can be carried out in all solids material surface because dopamine polymerize, and
And the coating formed has the stronger ability with the substance reaction containing amino, therefore, the method applied in the present invention almost can be with
Surface for all biomaterials is modified, and promotees the anticoagulation surface of endothelial cell growth for building specificity.
Brief description of the drawings
Fig. 1 is to build the schematic diagram that specificity promotees the anticoagulation surface of endothelial cell growth in biomaterial surface；
Fig. 2 grows nonparasitically upon another plant for the typical inner skin cell viscosity of biomaterial surface and grows figure；
Fig. 3 is typical platelet adhesion reaction figure after biological material surface modifying.
Embodiment
In order to further illustrate the present invention, endothelial cell is promoted to the species specificity that the present invention is provided with reference to embodiments
The anticoagulation surface construction method of growth.It should be understood that these embodiments are merely to illustrate the present invention rather than the limitation present invention
Scope.
Furthermore, it is to be understood that after the content of the invention lectured has been read, those skilled in the art can be to present invention work
Various changes or modification, these equivalent form of values equally fall within the application appended claims limited range.
As shown in figure 1, a species specificity promotees the anticoagulation surface construction method of endothelial cell growth, first using dopamine
Then the method for autohemagglutination fixes amino-polyethyleneglycols using Microcontact printing, enters one in material surface formation poly-dopamine layer
Step fixes chitosan and heparin in material surface, will finally promote arginine-glutamic acid-day of endothelial cell growth with specificity
Winter propylhomoserin-valine (REDV) is polypeptide grafted in PEG regions, obtains the anticoagulation surface that specificity promotees endothelial cell growth；Tool
Body comprises the following steps：
1) biomaterial surface prepares poly-dopamine layer
Biomaterial is immersed into the dopamine solution that pH is 8.0-8.5, abundant auto polymerization 12-24 hours, after cleaning
Dry, in triplicate, obtain the biomaterial surface of poly-dopamine layer；
2) biomaterial surface micro-contact printing fixes double amino-polyethyleneglycols
The polyglycol solution that double (3- aminopropyls) is blocked is added dropwise in banded micrographics dimethyl silicone polymer
(PDMS) seal surface, banded micrographics dimethyl silicone polymer seal surface is fully adsorbed after double amino-polyethyleneglycols,
N2Dry, then by PDMS seals and step 1) biomaterial surface that has poly-dopamine layer fully contacts 2 minutes, dry after cleaning
It is dry, obtain the biomaterial surface of amino-polyethyleneglycols modification；
3) biomaterial surface fixes chitosan and heparin
By step 2) obtained biomaterial is immersed into chitosan and heparin solution after fully reaction successively, and cleaning is dry
It is dry, obtain to surface and secure the biomaterial surface of chitosan and heparin；
4) biomaterial surface fixes REDV
By step 3) in biomaterial be immersed into end group be N- succinimides REDV solution in fully react, clean
After dry, obtain the biomaterial surface that REDV is contained on surface.
Step 1) in, the concentration of dopamine solution is 0.1~1mg/ml.
Step 2) in, the width of band and at intervals of 5~50 μm, the concentration of polyglycol solution is 0.1~1mg/ml.
Step 3) in, by step 2) to be immersed into chitosan solution fully reaction 4~12 first small for obtained biomaterial
When, after cleaning-drying, then fully absorption 4~8 hours is immersed into heparin solution, is dried after cleaning, wherein, chitosan and heparin
The concentration of solution is 1~10mg/ml.
Step 4) in, the time fully reacted is 4~8 hours, and REDV concentration is 0.1~1mg/ml.
The biomaterial that the present invention is designed includes the biomaterial of various solid kinds, as long as dopamine can be allowed to adhere to,
For example, titanium alloy material, polyurethane material, titanium deoxid film.
The biomaterial used in embodiment 1-3 is titanium alloy material.
Embodiment 1 prepares the width of band and at intervals of 5 μm of biomaterial
One species specificity promotees the anticoagulation surface construction method of endothelial cell growth, specifically includes following steps：
1) biomaterial surface prepares poly-dopamine layer
Biomaterial is immersed into the dopamine solution that pH is 8.0, abundant auto polymerization 12 hours is dried after cleaning, weight
Multiple three times, the concentration of dopamine solution is 0.1mg/ml, obtains the biomaterial surface of poly-dopamine layer；
2) biomaterial surface micro-contact printing fixes double amino-polyethyleneglycols
The polyglycol solution that double (3- aminopropyls) is blocked is added dropwise in banded micrographics dimethyl silicone polymer
(PDMS) seal surface, banded micrographics dimethyl silicone polymer seal surface is fully adsorbed after double amino-polyethyleneglycols,
N2Dry, then by PDMS seals and step 1) biomaterial surface that has poly-dopamine layer fully contacts 2 minutes, dry after cleaning
It is dry, the biomaterial surface of amino-polyethyleneglycols modification is obtained, wherein, the width of band and at intervals of 5 μm, polyglycol solution
Concentration be 0.1mg/ml；
3) biomaterial surface fixes chitosan and heparin
By step 2) obtained biomaterial is immersed into chitosan solution fully reaction 4 hours first, after cleaning-drying,
Fully absorption 4 hours is immersed into heparin solution again, obtains to surface and secures the biomaterial surface of chitosan and heparin, and shell gathers
The concentration of sugar and heparin solution is 1mg/ml；
4) biomaterial surface fixes REDV
By step 3) in biomaterial to be immersed into fully reaction in the REDV solution that end group is N- succinimides 4 small
When, dried after cleaning, obtain the biomaterial surface that REDV is contained on surface, wherein, REDV concentration is 0.1mg/ml.
Embodiment 2 prepares the width of band and at intervals of 25 μm of biomaterial
One species specificity promotees the anticoagulation surface construction method of endothelial cell growth, specifically includes following steps：
1) biomaterial surface prepares poly-dopamine layer
Biomaterial is immersed into the dopamine solution that pH is 8.5, abundant auto polymerization 24 hours is dried after cleaning, weight
Multiple three times, the concentration of dopamine solution is 1mg/ml, obtains the biomaterial surface of poly-dopamine layer；
2) biomaterial surface micro-contact printing fixes double amino-polyethyleneglycols
The polyglycol solution that double (3- aminopropyls) is blocked is added dropwise in banded micrographics dimethyl silicone polymer
(PDMS) seal surface, banded micrographics dimethyl silicone polymer seal surface is fully adsorbed after double amino-polyethyleneglycols,
N2Dry, then by PDMS seals and step 1) biomaterial surface that has poly-dopamine layer fully contacts 2 minutes, dry after cleaning
It is dry, the biomaterial surface of amino-polyethyleneglycols modification is obtained, wherein, the width of band and at intervals of 25 μm, polyethylene glycol is molten
The concentration of liquid is 1mg/ml；
3) biomaterial surface fixes chitosan and heparin
By step 2) obtained biomaterial is immersed into chitosan solution fully reaction 12 hours first, it is cleaned and dried
Afterwards, then be immersed into heparin solution fully absorption 8 hours, obtain to surface and secure the biomaterial surface of chitosan and heparin,
The concentration of chitosan and heparin solution is 10mg/ml；
4) biomaterial surface fixes REDV
By step 3) in biomaterial to be immersed into fully reaction in the REDV solution that end group is N- succinimides 8 small
When, dried after cleaning, obtain the biomaterial surface that REDV is contained on surface, wherein, REDV concentration is 1mg/ml.
Embodiment 3 prepares the width of band and at intervals of 50 μm of biomaterial
One species specificity promotees the anticoagulation surface construction method of endothelial cell growth, specifically includes following steps：
1) biomaterial surface prepares poly-dopamine layer
Biomaterial is immersed into the dopamine solution that pH is 8.5, abundant auto polymerization 20 hours is dried after cleaning, weight
Multiple three times, the concentration of dopamine solution is 0.5mg/ml, obtains the biomaterial surface of poly-dopamine layer；
2) biomaterial surface micro-contact printing fixes double amino-polyethyleneglycols
The polyglycol solution that double (3- aminopropyls) is blocked is added dropwise in banded micrographics dimethyl silicone polymer
(PDMS) seal surface, banded micrographics dimethyl silicone polymer seal surface is fully adsorbed after double amino-polyethyleneglycols,
N2Dry, then by PDMS seals and step 1) biomaterial surface that has poly-dopamine layer fully contacts 2 minutes, dry after cleaning
It is dry, the biomaterial surface of amino-polyethyleneglycols modification is obtained, wherein, the width of band and at intervals of 50 μm, polyethylene glycol is molten
The concentration of liquid is 0.5mg/ml；
3) biomaterial surface fixes chitosan and heparin
By step 2) obtained biomaterial is immersed into chitosan solution fully reaction 8 hours first, after cleaning-drying,
Fully absorption 6 hours is immersed into heparin solution again, obtains to surface and secures the biomaterial surface of chitosan and heparin, and shell gathers
The concentration of sugar and heparin solution is 5mg/ml；
4) biomaterial surface fixes REDV
By step 3) in biomaterial to be immersed into fully reaction in the REDV solution that end group is N- succinimides 6 small
When, dried after cleaning, obtain the biomaterial surface that REDV is contained on surface, wherein, REDV concentration is 0.5mg/ml.
The biomaterial that embodiment 1-3 is obtained, under equivalent assay conditions, obtains biomaterial table as shown in Figure 2
The typical inner skin cell viscosity in face is grown nonparasitically upon another plant long figure, the width of band and at intervals of 5 μm in Fig. 2 (a), in Fig. 2 (b) width of band and
At intervals of 25 μm, the width of band and at intervals of 50 μm in Fig. 2 (c).Cell random random growth on 5 μm of figure, works as bar
The width of band and interval increase, cell show regular arrangement, show oriented growth behavior.
The biomaterial that embodiment 2-3 is obtained, under equivalent assay conditions, obtains biomaterial table as shown in Figure 3
The modified typical platelet adhesion reaction figure in face, Fig. 3 (a) is control sample, Fig. 3 (b) and Fig. 3 (c) be respectively band width and
At intervals of 25 μm and 50 μm of sample, it can be seen that there is substantial amounts of platelet adhesion reaction on control sample surface, and is modified by surface
Afterwards, platelet adhesion reaction quantity greatly reduces, and shows that blood compatibility is significantly improved.
Claims (8)
1. a species specificity promotees the anticoagulation surface construction method of endothelial cell growth, it is characterised in that comprise the following steps：
1) biomaterial surface prepares poly-dopamine layer
Biomaterial is immersed into dopamine solution, abundant auto polymerization, is dried after cleaning, in triplicate, obtained poly- DOPA
The biomaterial surface of amine layer；
2) biomaterial surface micro-contact printing fixes double amino-polyethyleneglycols
Double amino-polyethyleneglycols solution are added dropwise on ribbon PDMS seals surface, ribbon PDMS seals surface is fully adsorbed double
After amino-polyethyleneglycols, N2Dry, then by PDMS seals and step 1) biomaterial surface that has poly-dopamine layer fully connects
Touch, dried after cleaning, obtain the biomaterial surface of amino-polyethyleneglycols modification；
3) biomaterial surface fixes chitosan and heparin
By step 2) obtained biomaterial is immersed into chitosan and heparin solution after fully reaction, is cleaned and dried, obtains successively
The biomaterial surface of chitosan and heparin is secured to surface；
4) biomaterial surface fixes REDV
By step 3) in biomaterial be immersed into end group be N- succinimides REDV solution in fully react, after cleaning do
It is dry, obtain the biomaterial surface that REDV is contained on surface.
2. a species specificity according to claim 1 promotees the anticoagulation surface construction method of endothelial cell growth, its feature
It is：Step 1) in, the concentration of dopamine solution is 0.1~1mg/ml.
3. a species specificity according to claim 1 promotees the anticoagulation surface construction method of endothelial cell growth, its feature
It is：Step 1) in, dopamine solution pH is 8.0-8.5.
4. a species specificity according to claim 1 promotees the anticoagulation surface construction method of endothelial cell growth, its feature
It is：Step 1) in, the abundant auto polymerization time is 12-24 hours.
5. a species specificity according to claim 1 promotees the anticoagulation surface construction method of endothelial cell growth, its feature
It is：Step 2) in, the width of band and at intervals of 5~50 μm, the concentration of polyglycol solution is 0.1~1mg/ml.
6. a species specificity according to claim 1 promotees the anticoagulation surface construction method of endothelial cell growth, its feature
It is：Step 3) in, by step 2) obtained biomaterial is immersed into chitosan solution fully reaction 4~12 hours first,
After cleaning-drying, then fully absorption 4~8 hours is immersed into heparin solution.
7. a species specificity according to claim 1 promotees the anticoagulation surface construction method of endothelial cell growth, its feature
It is：Step 3) in, described chitosan and the concentration of heparin solution are 1~10mg/ml.
8. a species specificity according to claim 1 promotees the anticoagulation surface construction method of endothelial cell growth, its feature
It is：Step 4) in, the time fully reacted is 4~8 hours, and REDV concentration is 0.1~1mg/ml.
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| CN107823725B (en) * | 2017-10-11 | 2020-10-09 | 上海君联医疗设备有限公司 | Thrombus filter capable of being recycled in delayed manner and manufacturing method thereof |
| CN109745580A (en) * | 2019-02-28 | 2019-05-14 | 天津大学 | Anticoagulant peptides and the co-modified small-caliber artificial blood vessel and preparation method of cell adhesion polypeptide |
| CN109966565A (en) * | 2019-04-23 | 2019-07-05 | 西南交通大学 | A kind of coating, material, the preparation method of material and medical supplies promoting endangium reparation |
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