CN106806949A - It is a kind of to prepare the method that multilayer carries heparin redox graphene coating - Google Patents
It is a kind of to prepare the method that multilayer carries heparin redox graphene coating Download PDFInfo
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- CN106806949A CN106806949A CN201611197811.0A CN201611197811A CN106806949A CN 106806949 A CN106806949 A CN 106806949A CN 201611197811 A CN201611197811 A CN 201611197811A CN 106806949 A CN106806949 A CN 106806949A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/08—Materials for coatings
- A61L31/082—Inorganic materials
- A61L31/084—Carbon; Graphite
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/08—Materials for coatings
- A61L31/10—Macromolecular materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
- A61L33/0005—Use of materials characterised by their function or physical properties
- A61L33/0011—Anticoagulant, e.g. heparin, platelet aggregation inhibitor, fibrinolytic agent, other than enzymes, attached to the substrate
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2420/00—Materials or methods for coatings medical devices
- A61L2420/02—Methods for coating medical devices
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2420/00—Materials or methods for coatings medical devices
- A61L2420/06—Coatings containing a mixture of two or more compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2420/00—Materials or methods for coatings medical devices
- A61L2420/08—Coatings comprising two or more layers
Abstract
The method that multilayer carries heparin redox graphene coating is prepared the invention provides a kind of, is comprised the following steps：S1, prepare poly-D-lysine coating in biomaterial surface；S2, the redox graphene solution for preparing load heparin；S3, the redox graphene coating for preparing individual layer load heparin；S4, the biomaterial for preparing step S3 are alternately immersed into Poly-L-Lysine Solution and load heparin redox graphene solution, are adsorbed successively respectively, obtain the redox graphene coating that multilayer carries heparin.The beneficial effects of the invention are as follows：Realize being obviously improved for material surface anticoagulation function.
Description
Technical field
Method the present invention relates to prepare graphite oxide ene coatings, more particularly to one kind prepares multilayer and carries heparin reduction-oxidation
The method of graphite ene coatings.
Background technology
Biomaterial and Implantable Medical Device have important application at aspects such as raising health and prevention diseases, so
And, for contacting blood material and Ink vessel transfusing implanted medical device, it is solidifying that the interaction between material and blood triggers
Blood reaction may trigger a series of side reaction such as thrombosis and clinical complication, ultimately result in graft failure, therefore, material
The surface property of material often has conclusive effect to implantation success or failure, and it is the key for improving material blood compatibility that surface is modified
Technology.
Heparin is a kind of gucosamine with superior anti-coagulant performance, and excellent anticoagulation effect is respectively provided with vivo and in vitro
Really, have been widely used for contacting blood material surface be modified, material surface test tube of hepari be also anticoagulant biomaterial and
The research emphasis and focus being modified with contacting blood medical apparatus surface.
Existing material surface Heparinizing method mainly has covalent immobilization and ions binding method.
Covalent immobilization is in material list using various means (e.g., corona treatment, surface self-organization, ozonisation etc.)
Face introduce can reactive group, heparin is incorporated into material surface using the reaction of carboxyl or hydroxyl in group and heparin, but
The anti-freezing bioactivity of heparin is easily destroyed due to chemical reaction.
Ions binding method is also referred to as Electrostatic Absorption method, is to carry out surface treatment to material using various means to introduce positive electricity
Lotus, such that it is able to adsorb negatively charged heparin, but this method surface Heparin-binding is weaker, and heparin amount is also less, anti-freezing
Especially long-term anticoagulant effect is not good for effect.
Graphene oxide is a kind of nano material with huge specific area, is had in terms of biomaterial with medicine equipment
Highly important potential application, with preferable biocompatibility.Its huge specific surface area has as drug carrier material
The features such as drugloading rate is big, pharmaceutical activity is high, simultaneously as its special surface texture, can occur π-π with ad hoc structure molecule
Interaction, hydrophobic interaction etc., further, since graphene oxide generally carry negative electrical charge, can also with positive electricity
There is electrostatic interaction in the material or molecule of lotus.
Five-membered ring structure in the linear chain structure of heparin can occur hydrophobic interaction with graphenic surface, so that
Graphenic surface is set to carry heparin, and because heparin carries negative electrical charge, the Graphene of the load heparin of formation shows elecrtonegativity,
LBL self-assembly effect can occur with the material surface of positive charge, therefore material surface heparin carrying amount can be made significantly to increase
Plus, improve the anticoagulant effect of material or apparatus especially long-term anticoagulation effect.
The content of the invention
In order to solve the problems of the prior art, prepare multilayer the invention provides one kind and carry heparin redox graphene
The method of coating, the redox graphene coating that multilayer carries heparin is built by the method, significantly improves the anticoagulation of material
Effect, especially long-term anticoagulation effect.
The method that multilayer carries heparin redox graphene coating is prepared the invention provides a kind of, is comprised the following steps：
S1, poly-D-lysine coating is prepared in biomaterial surface, biomaterial is first immersed in dopamine hydrochloride solution
Middle reaction, repeatedly obtains poly-dopamine coating, and the biomaterial that then will be prepared is immersed into Poly-L-Lysine Solution
Middle reaction, obtains poly-D-lysine coating；
S2, the redox graphene solution for preparing load heparin, configure graphene oxide solution, add reducing agent fully to react,
Redox graphene solution is dispersed into after centrifugal drying, and reaction is sufficiently mixed with isometric heparin sodium aqua, centrifugation is dry
Again load heparin redox graphene solution is dispersed into after dry；
S3, the redox graphene coating for preparing individual layer load heparin, step is immersed into by the biomaterial that step S1 is prepared
Fully adsorbed in load heparin redox graphene solution in rapid S2, the reduction-oxidation graphite that individual layer carries heparin is obtained after drying
Ene coatings；
S4, the biomaterial for preparing step S3 are alternately immersed into Poly-L-Lysine Solution and carry heparin reduction-oxidation graphite
In alkene solution, adsorb successively respectively, obtain the redox graphene coating that multilayer carries heparin.
As a further improvement on the present invention, in step sl, biomaterial is first immersed in the DOPA of 1-5mg/ml
Reacted 6-24 hours in amide hydrochloride, poly-dopamine coating is obtained in triplicate, the biomaterial that then will be prepared
It is immersed into the Poly-L-Lysine Solution of 5-10mg/ml and reacts 4-24 hours, obtains poly-D-lysine coating.
As a further improvement on the present invention, in step s 2, the graphene oxide solution of 0.1-2mg/ml is configured, is added
Reducing agent fully reacts 2-12 hours, is dispersed into 0.1-2mg/ml redox graphene solution after centrifugal drying, and with etc. body
Long-pending concentration is sufficiently mixed reaction 2-8 hours for the heparin sodium aqua of 1-10mg/ml, is dispersed into 1-5mg/ after centrifugal drying again
The load heparin redox graphene solution of ml.
As a further improvement on the present invention, the reducing agent of the redox graphene in step S2 be sodium borohydride or
Hydrazine.
As a further improvement on the present invention, the addition of the reducing agent is the 10-50% of graphene oxide quality.
As a further improvement on the present invention, the temperature of the reduction reaction in step S2 is 60-120 DEG C.
As a further improvement on the present invention, in step s3, the biomaterial that step S1 is prepared is immersed into step
Suddenly fully adsorbed 0.5-2 hours in the load heparin redox graphene solution in S2, individual layer is obtained after drying and carries going back for heparin
Former graphite oxide ene coatings.
As a further improvement on the present invention, in step s 4, the biomaterial for step S3 being prepared alternately is submerged
To in Poly-L-Lysine Solution and load heparin redox graphene solution, adsorb 0.5-2 hours successively respectively, obtain multilayer load
The redox graphene coating of heparin.
As a further improvement on the present invention, the biomaterial is metal material, nonmetallic materials or macromolecule material
Any one in material.
As a further improvement on the present invention, the pH value of the dopamine hydrochloride solution in step S1 is 8-8.5.
The beneficial effects of the invention are as follows：By such scheme, the catechol group in dopamine molecule in step S1 can
Combined with forming coordinate bond with any materials substrate, and firm dopamine further formed in material surface by self-polymeric reaction
Coating, the coating has secondary response, Schiff base reaction can occur so as to be formed with the amino in Poly-L-Lysine Solution
Positively charged poly-D-lysine coating；Negatively charged load heparin graphene dispersion is formed by the technical method of step S2
Liquid, so as to be fixed on poly-D-lysine coating surface by Electrostatic Absorption；By the alternating sorbent of step S3, can be in biological material
Material surface layer self assembly obtains the load heparin graphite ene coatings of sandwich construction, so as to realize the aobvious of material surface anticoagulation function
Write lifting.
With reference to specific embodiment, the present invention is further described.
The method that multilayer carries heparin redox graphene coating is prepared in biomaterial surface the invention discloses a kind of,
Poly-D-lysine (PLL) coating is prepared in biomaterial surface first, then absorption carries the redox graphene (He- of heparin
RGO), and then alternating sorbent PLL and He-rGO, the load heparin redox graphene coating with sandwich construction is obtained.Using
The method of the present invention is surface-treated to biomaterial or medicine equipment, can assign material excellent anticoagulation function,
Contacting blood material or Ink vessel transfusing Implantable Medical Device aspect are applied.
Embodiment 1：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the dopamine of 1mg/ml
In HCI solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 24 hours under normal temperature,
The step for repeating three times, obtains poly-dopamine coating, and the poly-D-lysine that the material of preparation then is immersed into 5mg/ml is molten
Reacted 4 hours in liquid, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 0.1mg/ml is configured first,
Take 20ml graphene oxide solutions and add the hydrazine hydrates of 0.2mg mono-, 80 DEG C are sufficiently stirred for reaction 2 hours, and centrifugal drying is reduced
Graphene oxide, into the solution of 0.1mg/ml, it is the heparin sodium aqua of 1mg/ml to add isometric concentration to ultrasonic disperse again
It is sufficiently mixed 2 hours, is dispersed into load heparin redox graphene solution (He-rGO) after centrifugation again.
(3) will be adsorbed 30 minutes in the He-rGO solution with the material of PLL coatings submergence 1mg/ml, distilled water cleaning
After dry, obtain individual layer carry heparin redox graphene coating (PLL/He-rGO).
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 5mg/ml and is adsorbed 30 minutes,
Distilled water is cleaned, and is immersed into the He-rGO solution of 1mg/ml after drying, and distilled water is cleaned, dried, constantly after adsorbing 30 minutes
Repeat the above steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 2：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the dopamine of 1mg/ml
In HCI solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 12 hours under normal temperature,
The step for repeating three times, obtains poly-dopamine coating, and the poly-D-lysine that the material of preparation then is immersed into 8mg/ml is molten
Reacted 24 hours in liquid, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 0.1mg/ml is configured first, taken
20ml graphene oxide solutions add 0.6mg sodium borohydrides, and 120 DEG C are sufficiently stirred for reaction 8 hours, and centrifugal drying obtains oxygen reduction
Graphite alkene, into the solution of 0.5mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 5mg/ml fills to ultrasonic disperse again
Divide mixing 4 hours, be dispersed into load heparin redox graphene solution (He-rGO) after centrifugation again.
(3) will be adsorbed 30 minutes in the He-rGO solution with the material of PLL coatings submergence 2mg/ml, distilled water cleaning
After dry, obtain individual layer carry heparin redox graphene coating (PLL/He-rGO).
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 8mg/ml and is adsorbed 30 minutes,
Distilled water is cleaned, and is immersed into the He-rGO solution of 2mg/ml after drying, and distilled water is cleaned, dried, constantly after adsorbing 30 minutes
Repeat the above steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 3：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the dopamine of 1mg/ml
In HCI solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 24 hours under normal temperature,
The step for repeating three times, obtains poly-dopamine coating, and the poly-D-lysine that the material of preparation then is immersed into 10mg/ml is molten
Reacted 12 hours in liquid, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 0.1mg/ml is configured first, taken
20ml graphene oxide solutions add 1mg sodium borohydrides, and 120 DEG C are sufficiently stirred for reaction 8 hours, and centrifugal drying obtains reduction-oxidation
Graphene, into the solution of 2mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 5mg/ml is fully mixed to ultrasonic disperse again
Close 8 hours, be dispersed into load heparin redox graphene solution (He-rGO) after centrifugation again.
(3) will be adsorbed 1 hour in the He-rGO solution with the material of PLL coatings submergence 5mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 10mg/ml and is adsorbed 1 hour, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 5mg/ml after drying, and distilled water is cleaned, dried after adsorbing 1 hour, is constantly repeated
Above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 4：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 1mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 6 hours under normal temperature, repeat
The step for three times, obtain poly-dopamine coating, then the material of preparation is immersed into the Poly-L-Lysine Solution of 8mg/ml
Reaction 4 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 1mg/ml is configured first, taken
20ml graphene oxide solutions add the hydrazine hydrates of 2mg mono-, and 80 DEG C are sufficiently stirred for reaction 4 hours, and centrifugal drying obtains reduction-oxidation
Graphene, into the solution of 0.1mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 1mg/ml is abundant to ultrasonic disperse again
Mixing 6 hours, is dispersed into load heparin redox graphene solution (He-rGO) again after centrifugation.
(3) will be adsorbed 2 hours in the He-rGO solution with the material of PLL coatings submergence 5mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 8mg/ml and is adsorbed 2 hours, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 5mg/ml after drying, and distilled water is cleaned, dried after adsorbing 2 hours, is constantly repeated
Above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 5：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 2mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 6 hours under normal temperature, repeat
The step for three times, obtain poly-dopamine coating, then the material of preparation is immersed into the Poly-L-Lysine Solution of 5mg/ml
Reaction 12 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 0.1mg/ml is configured first, taken
20ml graphene oxide solutions add the hydrazine hydrates of 0.6mg mono-, and 60 DEG C are sufficiently stirred for reaction 6 hours, and centrifugal drying obtains oxygen reduction
Graphite alkene, into the solution of 0.1mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 5mg/ml fills to ultrasonic disperse again
Divide mixing 2 hours, be dispersed into load heparin redox graphene solution (He-rGO) after centrifugation again.
(3) will be adsorbed 30 minutes in the He-rGO solution with the material of PLL coatings submergence 1mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 5mg/ml and is adsorbed 30 minutes,
Distilled water is cleaned, and is immersed into the He-rGO solution of 1mg/ml after drying, and distilled water is cleaned, dried, constantly after adsorbing 30 minutes
Repeat the above steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 6：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 2mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 24 hours under normal temperature, weight
Multiple the step for three times, poly-dopamine coating is obtained, then the material of preparation is immersed into the Poly-L-Lysine Solution of 8mg/ml
Middle reaction 4 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 1mg/ml is configured first, taken
20ml graphene oxide solutions add 10mg sodium borohydrides, and 80 DEG C are sufficiently stirred for reaction 12 hours, and centrifugal drying obtains oxygen reduction
Graphite alkene, into the solution of 0.1mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 1mg/ml fills to ultrasonic disperse again
Divide mixing 8 hours, be dispersed into load heparin redox graphene solution (He-rGO) after centrifugation again.
(3) will be adsorbed 1 hour in the He-rGO solution with the material of PLL coatings submergence 2mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 8mg/ml and is adsorbed 1 hour, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 2mg/ml after drying, and distilled water is cleaned, dried after adsorbing 1 hour, is constantly repeated
Above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 7：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 2mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 12 hours under normal temperature, weight
Multiple the step for three times, poly-dopamine coating is obtained, then the material of preparation is immersed into the Poly-L-Lysine Solution of 8mg/ml
Middle reaction 12 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 1mg/ml is configured first, taken
20ml graphene oxide solutions add the hydrazine hydrates of 2mg mono-, and 60 DEG C are sufficiently stirred for reaction 12 hours, and centrifugal drying obtains reduction-oxidation
Graphene, into the solution of 1mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 5mg/ml is fully mixed to ultrasonic disperse again
Close 6 hours, be dispersed into load heparin redox graphene solution (He-rGO) after centrifugation again.
(3) will be adsorbed 2 hours in the He-rGO solution with the material of PLL coatings submergence 5mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 8mg/ml and is adsorbed 2 hours, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 5mg/ml after drying, and distilled water is cleaned, dried after adsorbing 2 hours, is constantly repeated
Above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 8：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the dopamine of 2mg/ml
In HCI solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 24 hours under normal temperature,
The step for repeating three times, obtains poly-dopamine coating, and the poly-D-lysine that the material of preparation then is immersed into 10mg/ml is molten
Reacted 4 hours in liquid, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 1mg/ml is configured first, taken
20ml graphene oxide solutions add 6mg sodium borohydrides, and 80 DEG C are sufficiently stirred for reaction 8 hours, and centrifugal drying obtains reduction-oxidation
Graphene, into the solution of 0.5mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 10mg/ml is abundant to ultrasonic disperse again
Mixing 6 hours, is dispersed into load heparin redox graphene solution (He-rGO) again after centrifugation.
(3) will be adsorbed 2 hours in the He-rGO solution with the material of PLL coatings submergence 2mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 10mg/ml and is adsorbed 2 hours, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 2mg/ml after drying, and distilled water is cleaned, dried after adsorbing 2 hours, is constantly repeated
Above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 9：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 5mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 6 hours under normal temperature, repeat
The step for three times, obtain poly-dopamine coating, then the material of preparation is immersed into the Poly-L-Lysine Solution of 5mg/ml
Reaction 4 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 2mg/ml is configured first, taken
20ml graphene oxide solutions add the hydrazine hydrates of 20mg mono-, and 120 DEG C are sufficiently stirred for reaction 4 hours, and centrifugal drying obtains oxygen reduction
Graphite alkene, into the solution of 2mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 10mg/ml is abundant to ultrasonic disperse again
Mixing 8 hours, is dispersed into load heparin redox graphene solution (He-rGO) again after centrifugation.
(3) will be adsorbed 30 minutes in the He-rGO solution with the material of PLL coatings submergence 1mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 5mg/ml and is adsorbed 30 minutes, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 1mg/ml after drying, and distilled water is cleaned, dried after adsorbing 30 minutes, constantly weight
Multiple above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 10：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 5mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 12 hours under normal temperature, weight
Multiple the step for three times, poly-dopamine coating is obtained, then the material of preparation is immersed into the Poly-L-Lysine Solution of 8mg/ml
Middle reaction 4 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 2mg/ml is configured first, taken
20ml graphene oxide solutions add 4mg sodium borohydrides, and 80 DEG C are sufficiently stirred for reaction 12 hours, and centrifugal drying obtains reduction-oxidation
Graphene, into the solution of 0.5mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 10mg/ml is abundant to ultrasonic disperse again
Mixing 4 hours, is dispersed into load heparin redox graphene solution (He-rGO) again after centrifugation.
(3) will be adsorbed 1 hour in the He-rGO solution with the material of PLL coatings submergence 2mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 8mg/ml and is adsorbed 1 hour, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 2mg/ml after drying, and distilled water is cleaned, dried after adsorbing 1 hour, is constantly repeated
Above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 11：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 5mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 24 hours under normal temperature, weight
Multiple the step for three times, poly-dopamine coating is obtained, then the material of preparation is immersed into the Poly-L-Lysine Solution of 8mg/ml
Middle reaction 4 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 2mg/ml is configured first, taken
20ml graphene oxide solutions add 12mg sodium borohydrides, and 120 DEG C are sufficiently stirred for reaction 2 hours, and centrifugal drying obtains oxygen reduction
Graphite alkene, into the solution of 0.1mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 5mg/ml fills to ultrasonic disperse again
Divide mixing 2 hours, be dispersed into load heparin redox graphene solution (He-rGO) after centrifugation again.
(3) will be adsorbed 2 hours in the He-rGO solution with the material of PLL coatings submergence 5mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 8mg/ml and is adsorbed 2 hours, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 5mg/ml after drying, and distilled water is cleaned, dried after adsorbing 2 hours, is constantly repeated
Above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 12：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the dopamine of 5mg/ml
In HCI solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 24 hours under normal temperature,
The step for repeating three times, obtains poly-dopamine coating, and the poly-D-lysine that the material of preparation then is immersed into 10mg/ml is molten
Reacted 24 hours in liquid, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 1mg/ml is configured first, taken
20ml graphene oxide solutions add the hydrazine hydrates of 10mg mono-, and 60 DEG C are sufficiently stirred for reaction 12 hours, and centrifugal drying obtains oxygen reduction
Graphite alkene, into the solution of 0.1mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 5mg/ml fills to ultrasonic disperse again
Divide mixing 2 hours, be dispersed into load heparin redox graphene solution (He-rGO) after centrifugation again.
(3) will be adsorbed 2 hours in the He-rGO solution with the material of PLL coatings submergence 5mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 10mg/ml and is adsorbed 2 hours,
Distilled water is cleaned, and is immersed into the He-rGO solution of 5mg/ml after drying, and distilled water is cleaned, dried after adsorbing 2 hours, constantly weight
Multiple above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 13：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 1mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 12 hours under normal temperature, weight
Multiple the step for three times, poly-dopamine coating is obtained, then the material of preparation is immersed into the Poly-L-Lysine Solution of 5mg/ml
Middle reaction 12 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 0.5mg/ml is configured first, taken
20ml graphene oxide solutions add the hydrazine hydrates of 1mg mono-, and 60 DEG C are sufficiently stirred for reaction 4 hours, and centrifugal drying obtains reduction-oxidation
Graphene, into the solution of 0.1mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 1mg/ml is abundant to ultrasonic disperse again
Mixing 4 hours, is dispersed into load heparin redox graphene solution (He-rGO) again after centrifugation.
(3) will be adsorbed 30 minutes in the He-rGO solution with the material of PLL coatings submergence 1mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 5mg/ml and is adsorbed 30 minutes,
Distilled water is cleaned, and is immersed into the He-rGO solution of 1mg/ml after drying, and distilled water is cleaned, dried, constantly after adsorbing 30 minutes
Repeat the above steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 14：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 1mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 12 hours under normal temperature, weight
Multiple the step for three times, poly-dopamine coating is obtained, then the material of preparation is immersed into the Poly-L-Lysine Solution of 8mg/ml
Middle reaction 12 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 0.5mg/ml is configured first, taken
20ml graphene oxide solutions add the hydrazine hydrates of 3mg mono-, and 60 DEG C are sufficiently stirred for reaction 4 hours, and centrifugal drying obtains reduction-oxidation
Graphene, into the solution of 0.1mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 1mg/ml is abundant to ultrasonic disperse again
Mixing 4 hours, is dispersed into load heparin redox graphene solution (He-rGO) again after centrifugation.
(3) will be adsorbed 1 hour in the He-rGO solution with the material of PLL coatings submergence 2mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 8mg/ml and is adsorbed 1 hour, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 2mg/ml after drying, and distilled water is cleaned, dried after adsorbing 1 hour, is constantly repeated
Above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
Embodiment 15：Material surface is processed according to following steps
(1) PLL coatings are prepared in biomaterial surface, the sample after cleaning-drying is first immersed in the DOPA amine salt of 2mg/ml
In acid salt solution, pH value is adjusted to 8-8.5 with NaOH, dried after fully being cleaned with distilled water after being reacted 12 hours under normal temperature, weight
Multiple the step for three times, poly-dopamine coating is obtained, then the material of preparation is immersed into the Poly-L-Lysine Solution of 8mg/ml
Middle reaction 12 hours, sample obtains PLL coatings after being fully cleaned and dried with distilled water after taking out.
(2) the redox graphene solution for carrying heparin is prepared, the graphene oxide solution of 0.5mg/ml is configured first,
Take 20ml graphene oxide solutions and add 5mg sodium borohydrides, 60 DEG C are sufficiently stirred for reaction 4 hours, and centrifugal drying obtains oxygen reduction
Graphite alkene, into the solution of 0.1mg/ml, the isometric concentration of addition is that the heparin sodium aqua of 1mg/ml fills to ultrasonic disperse again
Divide mixing 4 hours, be dispersed into load heparin redox graphene solution (He-rGO) after centrifugation again.
(3) will be adsorbed 2 hours in the He-rGO solution with the material of PLL coatings submergence 2mg/ml, after distilled water cleaning
Dry, obtain the redox graphene coating (PLL/He-rGO) that individual layer carries heparin.
(4) material with PLL/He-rGO coatings is immersed into again in the PLL solution of 8mg/ml and is adsorbed 2 hours, steamed
Distilled water is cleaned, and is immersed into the He-rGO solution of 2mg/ml after drying, and distilled water is cleaned, dried after adsorbing 2 hours, is constantly repeated
Above-mentioned steps, obtain the redox graphene coating ((PLL/He-rGO) n) that multilayer carries heparin.
A kind of method for preparing multilayer load heparin redox graphene coating that the present invention is provided has advantages below：
First, the dopamine that the present invention is used can form the chemical bonding of stabilization, its follow-up step in nearly all material surface
It is rapid also relatively simple, therefore, the surface that technology path of the invention is suitable for almost all biomaterials is modified.
2nd, all reaction systems of the invention can be carried out in aqueous, to various biomaterial surfaces nearly all
Without damage, therefore, using wide, realize easily.
3rd, the present invention takes full advantage of Graphene has this feature of huge specific surface area, significantly improves material list
The loading of face heparin, simultaneously as being hydrophobic interaction, does not destroy the anti-freezing bioactivity of heparin.
4th, the positive charge material that the present invention is used is poly-D-lysine, is acted on significant promoting growth of cell and anti-
Bacterium acts on, therefore, technical scheme can not only significantly improve the anticoagulation function of biomaterial, and can promote
Cell growth and the certain antibacterial ability of imparting material.
5th, the technical solution adopted by the present invention is LBL self-assembly, surface heparin amount can by Iy self-assembled layer numerical control system,
Heparin release behavior can be controlled by PLL coatings.
6th, because technical step of the invention is all to carry out in the solution, therefore, technical scheme is to complex shape
The biomaterial of shape also is adapted for, and the surface for going for the medicine equipment of the complicated shapes such as intravascular stent, medical catheter is modified.
Above content is to combine specific preferred embodiment further description made for the present invention, it is impossible to assert
Specific implementation of the invention is confined to these explanations.For general technical staff of the technical field of the invention,
On the premise of not departing from present inventive concept, some simple deduction or replace can also be made, should be all considered as belonging to of the invention
Protection domain.
Claims (10)
1. it is a kind of to prepare the method that multilayer carries heparin redox graphene coating, it is characterised in that to comprise the following steps：
S1, poly-D-lysine coating is prepared in biomaterial surface, biomaterial is first immersed in dopamine hydrochloride solution
Middle reaction, repeatedly obtains poly-dopamine coating, and the biomaterial that then will be prepared is immersed into Poly-L-Lysine Solution
Middle reaction, obtains poly-D-lysine coating；
S2, the redox graphene solution for preparing load heparin, configure graphene oxide solution, add reducing agent fully to react,
Redox graphene solution is dispersed into after centrifugal drying, and reaction is sufficiently mixed with isometric heparin sodium aqua, centrifugation is dry
Again load heparin redox graphene solution is dispersed into after dry；
S3, the redox graphene coating for preparing individual layer load heparin, step is immersed into by the biomaterial that step S1 is prepared
Fully adsorbed in load heparin redox graphene solution in rapid S2, the reduction-oxidation graphite that individual layer carries heparin is obtained after drying
Ene coatings；
S4, the biomaterial for preparing step S3 are alternately immersed into Poly-L-Lysine Solution and carry heparin reduction-oxidation graphite
In alkene solution, adsorb successively respectively, obtain the redox graphene coating that multilayer carries heparin.
It is 2. according to claim 1 to prepare the method that multilayer carries heparin redox graphene coating, it is characterised in that：
In step S1, biomaterial is first immersed in be reacted 6-24 hours in the dopamine hydrochloride solution of 1-5mg/ml, repeats three
Secondary to obtain poly-dopamine coating, the biomaterial that then will be prepared is immersed into the Poly-L-Lysine Solution of 5-10mg/ml
Reaction 4-24 hours, obtains poly-D-lysine coating.
It is 3. according to claim 1 to prepare the method that multilayer carries heparin redox graphene coating, it is characterised in that：
In step S2, the graphene oxide solution of 0.1-2mg/ml is configured, add reducing agent fully to react 2-12 hours, after centrifugal drying
0.1-2mg/ml redox graphene solution is dispersed into, and with isometric concentration for the heparin sodium aqua of 1-10mg/ml fills
Divide hybrid reaction 2-8 hours, be dispersed into the load heparin redox graphene solution of 1-5mg/ml after centrifugal drying again.
4. the method that the preparation multilayer according to claim 1 or 3 carries heparin redox graphene coating, its feature exists
In：The reducing agent of the redox graphene in step S2 is sodium borohydride or hydrazine.
It is 5. according to claim 4 to prepare the method that multilayer carries heparin redox graphene coating, it is characterised in that：Institute
The addition for stating reducing agent is the 10-50% of graphene oxide quality.
It is 6. according to claim 4 to prepare the method that multilayer carries heparin redox graphene coating, it is characterised in that：Step
The temperature of the reduction reaction in rapid S2 is 60-120 DEG C.
It is 7. according to claim 1 to prepare the method that multilayer carries heparin redox graphene coating, it is characterised in that：
In step S3, in the load heparin redox graphene solution that the biomaterial that step S1 is prepared is immersed into step S2
Fully absorption 0.5-2 hours, obtains the redox graphene coating that individual layer carries heparin after drying.
It is 8. according to claim 1 to prepare the method that multilayer carries heparin redox graphene coating, it is characterised in that：
In step S4, the biomaterial that step S3 is prepared alternately is immersed into Poly-L-Lysine Solution and carries heparin oxygen reduction fossil
In black alkene solution, adsorb 0.5-2 hours successively respectively, obtain the redox graphene coating that multilayer carries heparin.
It is 9. according to claim 1 to prepare the method that multilayer carries heparin redox graphene coating, it is characterised in that：Institute
It is any one in metal material, nonmetallic materials or macromolecular material to state biomaterial.
It is 10. according to claim 1 to prepare the method that multilayer carries heparin redox graphene coating, it is characterised in that：
The pH value of the dopamine hydrochloride solution in step S1 is 8-8.5.
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| US11129797B1 (en) | 2021-05-07 | 2021-09-28 | King Abdulaziz University | Catechin and gingerol loaded nanomedicine |
| US11219605B1 (en) | 2021-05-07 | 2022-01-11 | King Abdulaziz University | Metal nanoparticle medicine with a mixture of natural ligands |
| US11234940B1 (en) | 2021-05-07 | 2022-02-01 | King Abdulaziz University | Method for making a noble metal nanomedicine |
| US11311492B1 (en) | 2021-05-07 | 2022-04-26 | King Abdulaziz University | Carbon nanomedicine with natural ligands |
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