CN112972771A - Preparation method of bioactive surface coating based on polymerization of bis (vinylsulfonyl) methane - Google Patents

Preparation method of bioactive surface coating based on polymerization of bis (vinylsulfonyl) methane Download PDF

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CN112972771A
CN112972771A CN202110373853.XA CN202110373853A CN112972771A CN 112972771 A CN112972771 A CN 112972771A CN 202110373853 A CN202110373853 A CN 202110373853A CN 112972771 A CN112972771 A CN 112972771A
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bis
methane
surface coating
vinylsulfonyl
bioactive surface
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程昉
刘波
刘鸿鑫
张强
王华楠
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Dalian University of Technology
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Abstract

The invention discloses a preparation method of a bioactive surface coating based on polymerization of bis (vinylsulfonyl) methane. The method comprises the steps of using bis (vinylsulfonyl) methane to react and polymerize on the surface of a pretreated substrate material under a catalytic condition, and carrying out damp-heat sterilization after the polymerization reaction to realize the biological activity modification of the material. The method is based on the self-polymerization of bis (vinylsulfonyl) methane, the film forming thickness is increased along with the prolonging of the reaction time, and the controllability is strong; the surface of various substrate materials can be modified, and the application range is wide; the terminal damp-heat sterilization method is adopted for sterilization, the operation is simple, the process environment requirement is low, and the influence on the prepared coating is small; the modified surface vinylsulfonyl coating is beneficial to cell adhesion and has good biocompatibility; no water-free and oxygen-free operation is needed, the reaction condition is mild, and the reproducibility is good; is a preparation method of a bioactive surface coating with great potential.

Description

Preparation method of bioactive surface coating based on polymerization of bis (vinylsulfonyl) methane
Technical Field
The invention belongs to the field of surface modification of biological materials, and particularly relates to a novel method for preparing a bioactive surface coating of various materials, in particular to a method for preparing a surface coating based on a bis (vinyl sulfone) methane polymerization modified material.
Technical Field
The biological material is a natural or artificial or semi-artificial synthetic functional material which is directly contacted with cells, tissues and organs, acts on a biological system, can diagnose and treat diseases of a life system, replace and repair damaged parts or induce regeneration of tissues of the damaged parts. The biological material is fused with multiple subjects of materials science, biology, chemistry, medicine and the like, and simultaneously covers various engineering technologies and manufacturing technologies. Depending on the biomaterial functionality, can be divided into functions of transmitting or bearing loads, such as bones, teeth, joints; to assume blood flow functions, such as blood vessels, heart valves; acoustic, optical, electrical conduction functions such as the lens, cochlea. When the biological material comes into contact with living systems, the surface of the material first interacts with the physiological environment in which it is located. The structure and the property of the material surface interface are definite, and the material surface interface has important significance for subsequent application of the material.
The biological function surface interface construction method mainly comprises two methods of physical adsorption and chemical coupling. Physical adsorption is the immobilization of functional active compound molecules on the surface of a material by van der waals forces, electrostatic interactions, hydrophobic interactions, and the like. The physical fixing method is simple and convenient to operate, but has the defects of weak adsorption, easy separation, poor repeatability and great influence on material performance. The chemical coupling is to fix functional active compound molecules on the surface of a material by utilizing chemical bonds and construct a surface interface with biological functions by modifying the chemical structure of the surface. Compared with physical adsorption, chemical coupling has three advantages: firstly, the covalent bond has much higher physical adsorption strength than Van der Waals force, electrostatic acting force and the like, the stability of the biological function surface interface of chemical coupling is high, and the service life is obviously prolonged. Secondly, the covalent bond binding sites are fixed, so that the density of the immobilized biomolecules is convenient to control, and the immobilized biomolecules have saturation. Third, the covalent bonding direction is fixed, and the immobilized biomolecules have good orientation. Therefore, chemical coupling is the most important method for constructing the interface of biological functional surface.
However, chemical coupling also presents a number of problems during the construction process: the reaction rate is difficult to regularly control, and the reaction reproducibility is poor; the surface properties of different materials are different greatly, the activation difficulty of partial materials is high, and the chemical coupling agent is difficult to achieve the universality of the materials; due to different subsequent coupling molecular species, coupling agents with different end groups are required to be used for connection, and general reactivity is difficult to achieve. Researches find that bis (vinylsulfonyl) methane can be polymerized on the surfaces of various functional materials to generate polymer films in the presence of a catalyst, the reaction conditions are mild, and unreacted alkenyl can further react with sulfydryl, amino and hydroxyl, so that the polymer film is a very potential biomolecule coupling agent.
Disclosure of Invention
The invention aims to provide a preparation method of a bioactive surface coating based on bis (vinylsulfonyl) methane polymerization, which comprises the steps of carrying out reaction polymerization on the surface of a pretreated material by using bis (vinylsulfonyl) methane under a catalytic condition, and carrying out damp-heat sterilization after the polymerization reaction to realize bioactive modification on the material. The method comprises the following steps: dissolving bis (vinylsulfonyl) methane in an aprotic polar solvent, immersing the pretreated substrate material in a reaction solution in the presence of a catalyst, adjusting the pH value to 8-9, reacting at 25-100 ℃ for 1-48 hours, and performing moist heat sterilization to prepare the bioactive surface coating.
In the above-mentioned embodiment, preferably, the aprotic polar solvent is selected from acetonitrile, acetone, N-dimethylformamide, dimethylsulfoxide, tetrahydrofuran, dioxane, dichloromethane, and chloroform.
In the above technical solutions, the catalyst is preferably sodium ethoxide, triethylamine, hexamethylenetetramine, and N, N-dimethylaminopyridine, and more preferably, the catalyst is N, N-dimethylaminopyridine.
In the above technical solution, preferably, the molar concentration of the catalyst is 1 to 10% of the molar concentration of bis (vinylsulfonyl) methane.
In the above technical solution, the substrate material is selected from inorganic silicon, metal oxide, glass, organic glass and polymer. Preferably, the metal is selected from titanium and steel, the metal oxide is selected from alumina, and the polymer is selected from polypropylene, polystyrene, polytetrafluoroethylene and polyether ether ketone.
In the above technical solution, preferably, the material pretreatment is that after the surface of the material is uniformly polished by 600-1500# sandpaper, the material is washed by ethanol and dried, and more preferably, the sandpaper is 1200 meshes.
In the above technical scheme, the reaction temperature is preferably 25-40 ℃.
In the above-mentioned technical solutions, the reaction time is preferably 6 to 12 hours.
In the above technical solution, the wet heat sterilization condition is preferably 100-.
In addition, the invention also protects the bioactive surface coating material prepared by the preparation method of the bioactive surface coating based on the polymerization of the bis (vinylsulfonyl) methane.
Has the advantages that:
compared with the traditional method, the preparation method of the bioactive surface coating based on the polymerization of the bis (vinylsulfonyl) methane has the following advantages:
(1) the controllability is strong, the method is based on the self-polymerization of bis (vinylsulfonyl) methane, and the film-forming thickness is increased along with the prolonging of the reaction time;
(2) the broad spectrum is good, the surface of various substrate materials can be modified, the application range of the substrate is wide, and the substrate is superior to a dopamine deposition method;
(3) the process environment requirement is low, no whole-process sterile operation is needed, terminal damp-heat sterilization is adopted, the operation is simple, and the coating is not damaged.
(4) The biocompatibility is good, and the modified bioactive surface coating is beneficial to cell adhesion;
(5) the reaction condition is mild, and no water or oxygen is needed; the reaction can be carried out by adopting a conventional solution, and the operation is simple and easy to implement; the reproducibility is good.
In conclusion, the method is a novel preparation method of the bioactive surface coating with great potential.
Drawings
The invention is shown in the attached drawings in the specification, wherein:
FIG. 1: static water contact angles of the surfaces of various materials before and after modification of the bis (vinylsulfonyl) methane coating.
FIG. 2: the thickness of the bis (vinylsulfonyl) methane coating on the surface of the alumina varied with the reaction time.
FIG. 3: polyether-ether-ketone surface infrared spectrograms before and after modification of the bis (vinylsulfonyl) methane coating.
FIG. 4: titanium surface cell proliferation experiments before and after modification of the bis (vinylsulfonyl) methane coating.
Detailed Description
The invention discloses a preparation method of a surface coating based on bis (vinylsulfonyl) methane polymerization, and the method is applied to the surfaces of various materials. The materials are selected from inorganic silicon, metal oxide, glass, organic glass and polymer. The metal material is preferably titanium or steel, the metal oxide is preferably alumina, and the polymer is preferably polypropylene, polystyrene, polytetrafluoroethylene or polyether ether ketone. The materials are pretreated by uniformly polishing the surfaces of the materials by 600-1500# sandpaper, washing by ethanol and drying. The catalyst is pyridine, triethylamine, 1, 5-diazabicyclo [4.3.0] non-5-ene, hexamethylenetetramine and N-methylmorpholine.
The following specific examples are intended to further illustrate the present invention and should not be construed as limiting the invention in any way.
Example 1: preparation of multiple material bioactive surface coatings by polymerization of bis (vinylsulfonyl) methane catalyzed by triethylamine
100mmol/L of bis (vinylsulfonyl) methane (BVS, Tci) and 10mmol/L of triethylamine (TEA, Aladdin) are prepared as a reaction solution, and the pH of the reaction solution is adjusted to 8.5 by using sodium hydroxide. Respectively taking 2cm multiplied by 2cm inorganic silicon chips, steel sheets, titanium sheets, glass sheets, aluminum oxide sheets, polypropylene sheets, polystyrene sheets, polytetrafluoroethylene sheets and polyether ether ketone sheets, uniformly grinding through 800# SiC abrasive paper, washing with ethanol and drying. And immersing the obtained substrate material sample into the prepared reaction liquid, and standing for reaction at 25 ℃. After reaction for 6h and 24h, respectively, the samples were taken out, washed with a large amount of acetone, dried, and sterilized by moist heat with water vapor at 121 ℃ for 20 minutes. The static water contact angle was measured with 2. mu.L of ultrapure water, and the average value was obtained at least three times for each sample, and the results are shown in FIG. 1. After the surfaces of various materials are modified by bis (vinylsulfonyl) methane, the surface contact angles all reach about 50 degrees (the polystyrene sample is partially dissolved in acetone within 24 hours). The result shows that the surface coating preparation method based on the polymerization preparation of the bis (vinylsulfonyl) methane can be applied to various substrates and has broad spectrum.
Example 2: preparation of alumina bioactive surface coating by 1, 5-diazabicyclo [4.3.0] non-5-alkene catalytic polymerization of bis (vinylsulfonyl) methane
Preparing dichloromethane solution of 100mmol/L bis (vinylsulfonyl) methane (BVS, Tci) and 1 mmol/L1, 5-diazabicyclo [4.3.0] non-5-ene (DBN, Aladdin) as reaction liquid, and adjusting the pH of the reaction liquid to 8.5 by using sodium hydroxide. And uniformly polishing an aluminum oxide sheet of 2cm multiplied by 2cm by 1200# SiC abrasive paper, washing by ethanol and drying. Immersing the obtained aluminum oxide sheet into the prepared reaction solution, and standing for reaction at 37 ℃. The aluminum oxide flakes were taken out at a predetermined timing (every 1 hour during the reaction from the start of the reaction to the end of 48 hours), washed with a large amount of methylene chloride, dried, and subjected to moist heat sterilization with steam at 121 ℃ for 30 minutes. The change in the thickness of bis (vinylsulfonyl) methane on the alumina plate was measured with an ellipsometer. At least three measurements per sample were averaged and the results are shown in figure 2. The increase in coating thickness is divided into two distinct stages with increasing bis (vinylsulfonyl) methane treatment time: the coating thickness exceeded the ellipsometric detection limit after a slow growth over the first 2 hours and then a rapid growth up to 12 hours. The results show that the thickness of the surface coating prepared based on the polymerization of bis (vinylsulfonyl) methane increases with the reaction time, and is controllable.
Example 3: preparation of polyether-ether-ketone bioactive surface coating by polymerization of bis (vinylsulfonyl) methane under catalysis of hexamethylenetetramine
100mmol/L bis (vinylsulfonyl) methane (BVS, Tci) and 8mmol/L hexamethylenetetramine (HMTA, Beijing Yinaoka Tech Co., Ltd.) were prepared as reaction solutions, and the pH of the reaction solution was adjusted to 8.5 using sodium hydroxide. And uniformly polishing a 2cm multiplied by 2cm polyetheretherketone sheet by 1500# abrasive paper, washing with ethanol, and drying. And immersing the polyether-ether-ketone sheet into the prepared reaction liquid, and standing and reacting for 24 hours at 40 ℃. The PEEK sheet was taken out, rinsed with a large amount of tetrahydrofuran, dried, and subjected to moist heat sterilization with 121 ℃ steam for 25 minutes, which was designated as sterilized PEEK. A tetrahydrofuran solution containing 100mmol/L of BVS without catalyst was designated BVS-PEEK as a control. Characterization was performed using infrared spectroscopy: after 24 hours of reaction, the polyetheretherketone sheet was taken out and dried, and its surface infrared spectrum was measured. The IR spectra were repeated after moist heat sterilization and at least three measurements were made for each sample, the results being shown in FIG. 3. When in measurement, a clean original piece of polyether-ether-ketone is taken as a background, and after the reaction is finished, under the catalysis condition of hexamethylenetetramine, the peak 1 (-1324 cm) in the infrared spectrum-1) Peak 2 (. about.1129 cm)-1) The S ═ O asymmetric and symmetric tensile vibration peaks were assigned, and the moist heat sterilization treatment did not affect the stability of the surface coating. The results show that the material surface coating modified by the polymerization of the bis (vinylsulfonyl) methane is successfully prepared.
Example 4: preparation of titanium bioactive surface coating by N-methylmorpholine catalyzed bis (vinylsulfonyl) methane polymerization
Preparing 100mmol/L bis (vinylsulfonyl) methane(BVS, Tci) and 3mmol/L of N-methylmorpholine (Miro chemical reagent, Tianjin) in dimethyl sulfoxide were used as the reaction solution, and the pH of the reaction solution was adjusted to 8.5 with sodium hydroxide. After 2cm multiplied by 2cm titanium sheets are evenly polished by 1500# SiC abrasive paper, washed by ethanol and dried. Immersing the obtained titanium sheet into the prepared reaction solution, and standing for reaction at 30 ℃. Taking out the titanium sheet at a set time, washing with a large amount of dimethyl sulfoxide, drying, and performing wet heat sterilization for 40 minutes by using water vapor at 121 ℃. Subcultured bone marrow mesenchymal cells of SD rat (primary cells were obtained from femoral medullary cavity of 3-week-old SD rat) were collected at 1.5X 105cells/cm2The cell density of the cells is inoculated to the surface of the obtained titanium plate, a CCK-8 experiment is carried out after 0-3 days of culture, the absorbance of 450nm is detected by using an enzyme-labeling instrument, and the measurement result is shown as the attached figure 4. The blank control group was the proliferation of cells seeded on α -MEM medium (37 ℃). The experimental results of the unmodified pure titanium plate group showed that the cells hardly proliferated on the pure titanium surface, and the initial cell number was smaller than that of the blank control group. The proliferation condition of the titanium sheet cells modified by the bis (vinylsulfonyl) methane is obvious, the proliferation trend is similar to that of a blank control group, and the result shows that the bioactive surface coating prepared by polymerizing the bis (vinylsulfonyl) methane has good biocompatibility and does not influence the normal growth of the cells.
It will be apparent to those skilled in the art from this disclosure that many changes and modifications can be made, or equivalents modified, in the embodiments of the invention without departing from the scope of the invention. Therefore, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention shall still fall within the protection scope of the technical solution of the present invention, unless the contents of the technical solution of the present invention are departed.

Claims (10)

1.一种基于双(乙烯砜基)甲烷聚合的生物活性表面涂层制备方法,其特征在于,所述方法包括如下步骤:将双(乙烯砜基)甲烷溶解于非质子极性溶剂,在催化剂存在条件下将经前处理的基底材料浸没在反应液中,调节pH 8-9,25-100℃反应1-48小时,再经湿热灭菌得到生物活性表面涂层。1. A method for preparing a bioactive surface coating based on bis(vinylsulfone-based) methane polymerization, wherein the method comprises the steps of: dissolving bis(vinylsulfone-based) methane in an aprotic polar solvent, and in the In the presence of a catalyst, the pretreated base material is immersed in the reaction solution, adjusted to pH 8-9, reacted at 25-100° C. for 1-48 hours, and then subjected to moist heat sterilization to obtain a biologically active surface coating. 2.根据权利要求1所述的一种基于双(乙烯砜基)甲烷合的生物活性表面涂层制备方法,其特征在于,所述的非质子极性溶剂选自乙腈、丙酮、氯仿、二噁烷、二氯甲烷、四氢呋喃、二甲基亚砜和N,N-二甲基甲酰胺。2. a kind of bioactive surface coating preparation method based on bis(vinylsulfone) methane compound according to claim 1, is characterized in that, described aprotic polar solvent is selected from acetonitrile, acetone, chloroform, Oxane, dichloromethane, tetrahydrofuran, dimethyl sulfoxide and N,N-dimethylformamide. 3.根据权利要求1所述的一种基于双(乙烯砜基)甲烷聚合的生物活性表面涂层制备方法,其特征在于,所述的催化剂选自吡啶、三乙胺、1,5-二氮杂双环[4.3.0]壬-5-烯、六亚甲基四胺和N-甲基吗啉。3. a kind of bioactive surface coating preparation method based on bis(vinylsulfone-based) methane polymerization according to claim 1, is characterized in that, described catalyst is selected from pyridine, triethylamine, 1,5-diethylamine Azabicyclo[4.3.0]non-5-ene, hexamethylenetetramine and N-methylmorpholine. 4.根据权利要求1所述的一种基于双(乙烯砜基)甲烷聚合的生物活性表面涂层制备方法,其特征在于,所述的催化剂的用量为双(乙烯砜基)甲烷摩尔浓度的1~10%。4. a kind of bioactive surface coating preparation method based on bis (vinyl sulfone) methane polymerization according to claim 1, is characterized in that, the consumption of described catalyst is bis (vinyl sulfone) methane molar concentration 1 to 10%. 5.根据权利要求1所述的一种基于双(乙烯砜基)甲烷聚合的生物活性表面涂层制备方法,其特征在于,所述的基底材料选自无机硅、金属、金属氧化物、玻璃、有机玻璃和聚合物。5. The method for preparing a bioactive surface coating based on bis(vinylsulfone-based) methane polymerization according to claim 1, wherein the base material is selected from the group consisting of inorganic silicon, metal, metal oxide, glass , plexiglass and polymers. 6.根据权利要求5所述的一种基于双(乙烯砜基)甲烷合的生物活性表面涂层制备方法,其特征在于,所述的金属选自钛、钢,所述的金属氧化物选自氧化铝,所述的聚合物选自聚丙烯、聚苯乙烯、聚四氟乙烯、聚醚醚酮。6 . The method for preparing a bioactive surface coating based on bis(vinylsulfone) methane according to claim 5 , wherein the metal is selected from titanium and steel, and the metal oxide is selected from the group consisting of titanium and steel. 7 . From alumina, the polymer is selected from polypropylene, polystyrene, polytetrafluoroethylene, polyetheretherketone. 7.根据权利要求1所述的一种基于双(乙烯砜基)甲烷聚合的生物活性表面涂层制备方法,其特征在于,所述的材料前处理为600-1500#砂纸均匀打磨材料表面后,乙醇冲洗、干燥。7. a kind of bioactive surface coating preparation method based on bis(vinylsulfone) methane polymerization according to claim 1, is characterized in that, described material pretreatment is after 600-1500# sandpaper evenly polishes the surface of material , rinsed with ethanol and dried. 8.根据权利要求1所述的一种基于双(乙烯砜基)甲烷聚合的生物活性表面涂层制备方法,其特征在于,所述的反应温度为25-40℃;反应时间6-12小时。8 . The method for preparing a bioactive surface coating based on bis(vinylsulfone-based) methane polymerization according to claim 1 , wherein the reaction temperature is 25-40° C.; the reaction time is 6-12 hours. 9 . . 9.根据权利要求1所述的一种基于双(乙烯砜基)甲烷聚合的生物活性表面涂层制备方法,其特征在于,所述的湿热灭菌为100-130℃水蒸气处理15-60分钟。9 . The method for preparing a bioactive surface coating based on bis(vinylsulfone-based) methane polymerization according to claim 1 , wherein the moist heat sterilization is 100-130° C. steam treatment for 15-60° C. 10 . minute. 10.权利要求1-9中任意一项所述的方法制备的生物活性表面涂层。10. A bioactive surface coating prepared by the method of any one of claims 1-9.
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