CN104988561A - Preparation method of bio-coating based on hyaluronic acid complex micelle - Google Patents

Preparation method of bio-coating based on hyaluronic acid complex micelle Download PDF

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Publication number
CN104988561A
CN104988561A CN201510427308.9A CN201510427308A CN104988561A CN 104988561 A CN104988561 A CN 104988561A CN 201510427308 A CN201510427308 A CN 201510427308A CN 104988561 A CN104988561 A CN 104988561A
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China
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hyaluronic acid
composite micelle
preparation
biological coating
albumen
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CN201510427308.9A
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潘凯
刘晓亚
孙家娣
朱叶
顾瑶
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Jiangnan University
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Jiangnan University
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Abstract

The invention provides a preparation method of bio-coating based on hyaluronic acid complex micelle. The preparation method comprises the steps that firstly, hyaluronic acid and protein are adopted as elements, and complex micelle particles are prepared in an aqueous phase in a self-assembled manner; then the complex micelle particle solution is adopted as an electrolyte, the micelle is electrically inducted to achieve film formation on the surface of medical metal based on the constant-potential electro-deposition technology, and the bio-coating based on the hyaluronic acid complex micelle is obtained. The bio-coating has the good biocompatibility, biodegradability and cytocompatibility. The method is easy and convenient to operate, mild in condition and wide in application range.

Description

A kind of biological coating preparation method based on hyaluronic acid composite micelle
Technical field
The present invention relates to a kind of biological coating preparation method based on hyaluronic acid composite micelle, belong to macromolecular material and biological coating and to combine field.
Technical background
Hyaluronic acid is a kind of straight chain line style natural macromolecular; alternately linked by N-acetyl-D-glucose amine and D-Glucose aldehydic acid; be present in the middle of the skin of people, joint and cornea etc. in a large number; there is excellent biodegradability, cell compatibility, moisture retention and film forming properties, be widely used in the fields such as organizational project, makeup, wound anti and medicament slow release.Containing a large amount of carboxylic groups and oh group on hyaluronic main chain, can be used as chemically modified site, obtain the derivatives of hyaluronic acids with functional performance, be beneficial to and prepare hyaluronic acid-based multifunctional material, expand hyaluronic Application Areas.
Compared to hyaluronan molecule chain, its self-assembled micelle particle has larger specific surface area, have excellent biocompatibility, biodegradability, moisture retention and film forming properties concurrently, be easy to practical function, in biological coating field, application prospect is extensive simultaneously.But in previous work, LBL self-assembly, dip-coating and spraying method prepare the method generally adopted based on micella particle coating material at present, as Park etc. by drug taxol load in the middle of hyaluronic acid-g-Poly(D,L-lactide-co-glycolide (HA-g-PLGA) self-assembled micelle, utilize the LBL self-assembly between polylysine molecule chain and micella particle, prepare drug-carried coat (J.Mater.Chem.2009,19 (24), 4102-4107.).These method stepss are loaded down with trivial details, controllability is not strong, narrow application range, and the intermolecular forces of coating and base material is more weak.
Electro-deposition techniques is easy and simple to handle, required equipment less investment, production cost are low, high to raw-material utilization ratio, technique simple, easy handling, mild condition, is a kind of more convenient and economic method.Control the forming process of coating by regulation and control voltage, depositing time and concentration of electrolyte, and galvanic deposit can significantly improve the bonding strength between base material and coating, is subject to the favor of numerous investigator in the preparation of biological coating material.
Thus, a kind of biological coating preparation method based on hyaluronic acid composite micelle has been invented herein.The advantage of numerator self-assembly technique and electro-deposition techniques combines by this preparation method, simple operating steps, and practicality and suitability by force, belong to originality invention.
Summary of the invention
The object of this invention is to provide a kind of biological coating preparation method based on hyaluronic acid composite micelle.The method is easy and simple to handle, mild condition, applied widely, namely electro-deposition techniques is passed through, hyaluronic acid and albumen is utilized to be primitive, in aqueous phase, composite micelle particle is prepared in self-assembly, with composite micelle particle solution for electrolytic solution, by constant potential electro-deposition techniques in medical metal surface electrical induction micella film forming, obtain the biological coating based on hyaluronic acid composite micelle.
Another object of the present invention is also by controlling final concentration and the Electrodeposition Conditions of composite micelle particle solution, can to regulate and control the character of micella film.
Technical scheme of the present invention is as follows:
Based on a biological coating preparation method for hyaluronic acid composite micelle, its preparation process is as follows:
(1) preparation of composite micelle: first configure certain density hyaluronic acid aqueous solution, add appropriate albumen, after treating that albumen dissolves completely, regulate pH=3 ~ 6 of above-mentioned solution, induction compound system generation self-assembly forms composite micelle, continue stirring 10 ~ 12 hours, fixing composite micelle pattern;
Described hyaluronic acid relative molecular weight is 1 × 10 3da ~ 1 × 10 6da, albumen used is selected from: the one in bovine hemoglobin, bovine serum albumin, sphaeroprotein and egg albumen;
Prepared hyaluronic acid aqueous solution concentration is 1 ~ 5mgmL -1, the add-on of albumen is 1 ~ 3mgmL -1, the interaction between hyaluronic acid and albumen is electrostatic interaction, hydrogen bond action;
Described composite micelle has pH responsive responsiveness, ion-sensitive responsiveness, and micella size of particles is between 100 ~ 1000nm.
(2) preparation of biological coating: first with 1200 object abrasive paper for metallograph polishing medical metals, then with dehydrated alcohol and acetone, ultrasonic cleaning is carried out to base material successively, above-mentioned pretreated medical metal is immersed in the composite micelle solution prepared by step (1), electro-deposition techniques is adopted to apply constant potential voltage, make charged micella particle to medical metal surface swimming and be deposited as micella film, obtaining the biological coating based on hyaluronic acid composite micelle.
Described medical metal is selected from: titanium or titanium alloy, cobalt and cobalt base alloy, 316L stainless steel; The Electrodeposition Conditions adopted is: constant potential voltage range is 1 ~ 100V, and electrodeposition time is 1 ~ 60min, and prepared coating has excellent biocompatibility and cell compatibility.
Beneficial effect of the present invention: the present invention innovatively with hyaluronic acid and albumen for primitive, in aqueous phase, composite micelle particle is prepared in self-assembly, by constant potential electro-deposition techniques in medical metal surface electrical induction micella film forming, prepare biocompatibility and biodegradability medical bio coated material all preferably, simple operating steps, practicality and suitability by force, belong to originality invention.
Accompanying drawing explanation
Fig. 1 is the digital photograph of compound self-assembled micelle solution in embodiment 1;
Fig. 2 is the projection electron microscope picture of composite micelle in embodiment 2;
Fig. 3 is the SEM shape appearance figure of biological coating in embodiment 3.
Embodiment
Below in conjunction with embodiment, the invention will be further described, but the present invention is not limited thereto.
Embodiment 1
Based on a biological coating preparation method for hyaluronic acid composite micelle, concrete preparation process is as follows:
(1) preparation of composite micelle: be 1 × 10 by molecular weight 4the water-soluble solution of hyaluronic acid of Da, form the hyaluronic acid solution of 1mg/mL, adding bovine hemoglobin makes albumen ultimate density in solution be 1mg/mL, after treating that bovine hemoglobin is dissolved completely, regulate the pH=4.5 of above-mentioned solution, impel system generation compound to assemble and form compound self-assembled micelle, continue stirring 11 hours, fixing composite micelle pattern, the digital photograph of sample as shown in Figure 1.
(2) preparation of biological coating: first with 1200 object abrasive paper for metallograph polishing medical titanium alloys, then with dehydrated alcohol and acetone, ultrasonic cleaning is carried out to it successively, above-mentioned pretreated medical titanium alloy is immersed in the composite micelle solution prepared by step (1) as anode, 20min is deposited under 20V constant potential voltage, make the micella particle of bear electricity to medical titanium alloy surface swimming and be deposited as micella film, obtaining the biological coating based on hyaluronic acid composite micelle.
Embodiment 2
Based on a biological coating preparation method for hyaluronic acid composite micelle, concrete preparation process is as follows:
(1) preparation of composite micelle: be 1 × 10 by molecular weight 6the water-soluble solution of hyaluronic acid of Da, form the hyaluronic acid solution of 3mg/mL, adding bovine serum albumin makes albumen ultimate density in solution be 2mg/mL, after treating that bovine serum albumin dissolves completely, regulate the pH=3 of above-mentioned solution, impel system generation compound to assemble and form compound self-assembled micelle, continue stirring 10 hours, fixing composite micelle pattern, the transmission electron micrograph of composite micelle as shown in Figure 2.
(2) preparation of biological coating: first with 1200 object abrasive paper for metallograph polishing medical magnesium alloys, then with dehydrated alcohol and acetone, ultrasonic cleaning is carried out to it successively, above-mentioned pretreated medical magnesium alloy is immersed in the composite micelle solution prepared by step (1) as anode, 5min is deposited under 90V constant potential voltage, make the micella particle of bear electricity to medical magnesium alloy surface swimming and be deposited as micella film, obtaining the biological coating based on hyaluronic acid composite micelle.
Embodiment 3
Based on a biological coating preparation method for hyaluronic acid composite micelle, concrete preparation process is as follows:
(1) preparation of composite micelle: be 1 × 10 by molecular weight 5the water-soluble solution of hyaluronic acid of Da, form the hyaluronic acid solution of 5mg/mL, adding egg albumen makes albumen ultimate density in solution be 3mg/mL, after treating that egg albumen dissolves completely, regulate the pH=5.8 of above-mentioned solution, impel system generation compound to assemble and form compound self-assembled micelle, continue stirring 12 hours, fixing composite micelle pattern.
(2) preparation of biological coating: first with 1200 object abrasive paper for metallograph polishing Medical Cobalt-Based Alloys, then with dehydrated alcohol and acetone, ultrasonic cleaning is carried out to it successively, above-mentioned pretreated Medical Cobalt-Based Alloys is immersed in the composite micelle solution prepared by step (1) as anode, 40min is deposited under 50V constant potential voltage, make the micella particle of bear electricity to Medical Cobalt-Based Alloys surface swimming and be deposited as micella film, obtain the biological coating based on hyaluronic acid composite micelle, the SEM shape appearance figure of coating as shown in Figure 3.
Above-described embodiment is used for explaining and the present invention is described, instead of limits the invention, and in the protection domain of spirit of the present invention and claim, any amendment make the present invention and change, all fall into protection scope of the present invention.

Claims (6)

1., based on a biological coating preparation method for hyaluronic acid composite micelle, its key step is:
(1) preparation of composite micelle: first configure certain density hyaluronic acid aqueous solution, add appropriate albumen, after treating that albumen dissolves completely, regulate pH=3 ~ 6 of above-mentioned solution, induction compound system generation self-assembly forms composite micelle, continue stirring 10 ~ 12 hours, fixing composite micelle pattern;
(2) preparation of biological coating: first with 1200 object abrasive paper for metallograph polishing medical metals, then with dehydrated alcohol and acetone, ultrasonic cleaning is carried out to base material successively, above-mentioned pretreated medical metal is immersed in the composite micelle solution prepared by step (1), electro-deposition techniques is adopted to apply constant potential voltage, make charged micella particle to medical metal surface swimming and be deposited as micella film, obtaining the biological coating based on hyaluronic acid composite micelle.
2. a kind of biological coating preparation method based on hyaluronic acid composite micelle according to claim 1, it is characterized in that, in step (1), hyaluronic acid relative molecular weight used is 1 × 10 3da ~ 1 × 10 6da, albumen used is selected from: the one in bovine hemoglobin, bovine serum albumin, sphaeroprotein and egg albumen.
3. a kind of biological coating preparation method based on hyaluronic acid composite micelle according to claim 1, is characterized in that, hyaluronic acid aqueous solution concentration prepared in step (1) is 1 ~ 5mgmL -1, the add-on of albumen is 1 ~ 3mgmL -1, the interaction between hyaluronic acid and albumen is electrostatic interaction, hydrogen bond action.
4. a kind of biological coating preparation method based on hyaluronic acid composite micelle according to claim 1, it is characterized in that, composite micelle prepared in step (1) has pH responsive responsiveness, ion-sensitive responsiveness, and micella size of particles is between 100 ~ 1000nm.
5. a kind of biological coating preparation method based on hyaluronic acid composite micelle according to claim 1, it is characterized in that, the medical metal described in step (2) is selected from: titanium or titanium alloy, cobalt and cobalt base alloy, 316L stainless steel.
6. a kind of biological coating preparation method based on hyaluronic acid composite micelle according to claim 1, it is characterized in that, the Electrodeposition Conditions adopted in step (2) is: constant potential voltage range is 1 ~ 100V, electrodeposition time is 1 ~ 60min, and prepared coating has excellent biocompatibility and cell compatibility.
CN201510427308.9A 2015-07-20 2015-07-20 Preparation method of bio-coating based on hyaluronic acid complex micelle Pending CN104988561A (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999045942A1 (en) * 1998-03-13 1999-09-16 Entremed, Inc. Metastatin and hyaluronate binding proteins and methods of use
CN101773689A (en) * 2010-03-29 2010-07-14 苑国忠 Surgical repairing patch
CN103357066A (en) * 2013-06-28 2013-10-23 陕西巨子生物技术有限公司 Hydrogel with bioremediation activity and outstanding degradation performance and preparation method

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999045942A1 (en) * 1998-03-13 1999-09-16 Entremed, Inc. Metastatin and hyaluronate binding proteins and methods of use
CN101773689A (en) * 2010-03-29 2010-07-14 苑国忠 Surgical repairing patch
CN103357066A (en) * 2013-06-28 2013-10-23 陕西巨子生物技术有限公司 Hydrogel with bioremediation activity and outstanding degradation performance and preparation method

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
R.MA等: ""Electrodeposition of hyaluronic acid and hyaluronic acid-bovine serum albumin films from aqueous solutions"", 《COLLOIDS AND SURFACE B :BIOINTERFACES》 *

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Application publication date: 20151021