阅读说明：本技术 抗蛋白吸附涂层修饰基材及其制备方法 (Anti-protein adsorption coating modified substrate and preparation method thereof ) 是由 严拓 陈希明 于 2021-06-15 设计创作，主要内容包括：本发明涉及抗蛋白吸附涂层修饰基材及其制备方法。该制备方法包括：将目标基材清洗并干燥；利用化学气相沉积法,将3-氨基丙基三甲氨基硅烷沉积于目标基材表面作为基层,并获得初级修饰基材；先将寡过氧化物溶解于二恶烷中形成寡过氧化物处理液,再将第一步所得初级修饰基材放入寡过氧化物处理液中反应,并获得次级修饰基材；先将OEGMA单体溶解于水中形成OEGMA水溶液,再将次级修饰基材放入OEGMA水溶液中反应,即得抗蛋白吸附涂层修饰基材。本发明可在目标基材表面修饰抗蛋白吸附涂层,使其能够抗蛋白吸附涂层,例如在用作医疗用品时,可有效抗蛋白吸附和血小板沉积。本发明制备过程简便易行,所得产品能有效抗污染。(The invention relates to a protein adsorption resistant coating modified substrate and a preparation method thereof. The preparation method comprises the following steps: cleaning and drying the target substrate; depositing 3-aminopropyl trimethyl amino silane on the surface of a target base material as a base layer by using a chemical vapor deposition method, and obtaining a primary modified base material; dissolving the oligomeric peroxide in dioxane to form an oligomeric peroxide treatment solution, and putting the primary modified substrate obtained in the first step into the oligomeric peroxide treatment solution for reaction to obtain a secondary modified substrate; and (3) dissolving an OEGMA monomer in water to form an OEGMA aqueous solution, and then putting the secondary modified substrate into the OEGMA aqueous solution for reaction to obtain the protein adsorption resistant coating modified substrate. The invention can modify the protein adsorption resistant coating on the surface of the target substrate, so that the protein adsorption resistant coating can be used for resisting protein adsorption and platelet deposition, for example, when the protein adsorption resistant coating is used as a medical article. The preparation process is simple and easy to implement, and the obtained product can effectively resist pollution.)

1. A preparation method of a protein adsorption resistant coating modified substrate is characterized by comprising the following steps:

firstly, cleaning and drying a target base material; depositing 3-aminopropyl trimethyl amino silane on the surface of a target base material as a base layer by using a chemical vapor deposition method, and obtaining a primary modified base material;

secondly, dissolving the oligoperoxide in dioxane to form oligoperoxide treatment solution, and putting the primary modified substrate obtained in the first step into the oligoperoxide treatment solution for reaction to obtain a secondary modified substrate;

and thirdly, dissolving an OEGMA monomer in water to form an OEGMA aqueous solution, and then putting the secondary modified substrate into the OEGMA aqueous solution for reaction to obtain the protein adsorption resistant coating modified substrate.

2. The method for preparing the protein adsorption resistant coating modified substrate as claimed in claim 1, wherein the process for preparing the oligoperoxide comprises:

dissolving benzene tetrachlorate in anhydrous dichloroethane, stirring and mixing, and adding tert-butyl hydroperoxide to form a mixture; dissolving pyridine in anhydrous dichloroethane to form a first pyridine solution, and adding the first pyridine solution into the mixture for mixing; adding PEG-9 into the mixture, firstly dissolving pyridine in anhydrous dichloroethane to form a second pyridine solution, and then adding the second pyridine solution into the mixture for mixing; stirring and reacting; filtering and drying the solid to obtain the oligoperoxide.

3. The method for preparing a substrate modified with a protein adsorption-resistant coating according to claim 2, wherein when the pyromellitic chloride is dissolved in anhydrous dichloroethane, the ratio of pyromellitic chloride to anhydrous dichloroethane is 4.6 ± 0.5 g: 15 plus or minus 3 ml; the molar ratio of the tert-butyl hydroperoxide to the benzene tetrachloride is 1: 1; the ratio of pyridine to anhydrous dichloroethane in the first pyridine solution is 1.1 ± 0.3 g: 10 plus or minus 2ml, and the molar ratio of pyridine to pyromellitic chloride in the first pyridine solution is 1: 1; the molar ratio of the PEG-9 to the pyromellitic chloride is 1: 1; the ratio of pyridine to anhydrous dichloroethane in the second pyridine solution is 2.2 ± 0.4 g: 10. + -. 2 ml.

4. The method of claim 2, wherein the oligomeric peroxide is prepared by cooling the mixture to a temperature of 1 ℃ to 5 ℃, adding the first pyridine solution dropwise at that temperature, maintaining the temperature after addition, and mixing for at least 1 hour; then at this temperature, PEG-9 was added to the mixture, and the second pyridine solution was added dropwise; in the stirring reaction process, the temperature is gradually increased to the ambient temperature, and the stirring reaction time is at least 3 h; the solid matter is dried at 40 +/-3 deg.c for at least 3 hr.

5. The method for preparing a substrate modified with a protein adsorption resistant coating according to claim 1, wherein in the first step, the target substrate is placed in a water or ethanol solution and cleaned by ultrasound, and the target substrate is protected by nitrogen gas during drying; the treatment temperature of the chemical vapor deposition method is 50-70 ℃, and the treatment time is 0.5-12 h; and (3) drying the target base material at 70 +/-5 ℃ overnight after the target base material is treated by a chemical vapor deposition method, thus obtaining the primary modified base material.

6. The method of claim 1, wherein in the second step, the ratio of the oligomeric peroxide to dioxane is 1 ± 0.5% g/ml; the reaction time is at least 24 h; and taking out the primary modified substrate after the reaction is finished, and washing away unreacted oligomeric peroxide by using dioxane to obtain the secondary modified substrate.

7. The method for preparing a substrate modified with a protein adsorption-resistant coating according to claim 1, wherein in the third step, the concentration of the OEGMA monomer in the aqueous solution of OEGMA is 0.1 ± 0.05 mol/L; and under the protection of nitrogen, putting the secondary modified base material into an OEGMA aqueous solution, reacting at 90 +/-5 ℃ for 2-48h, taking out after the reaction is finished, and washing away unreacted OEGMA monomers by water to obtain the protein adsorption resistant coating modified base material.

8. The method for preparing the protein adsorption resistant coating modified substrate as claimed in claim 1, wherein the target substrate is made of a polymer material or a metal material.

9. The substrate is modified by the protein adsorption-resistant coating prepared by the preparation method of any one of claims 1 to 8.

10. The preparation method of any one of claims 1 to 8, wherein the protein adsorption resistant coating is modified on the surface of the target substrate.

Technical Field

The invention relates to an anti-protein adsorption coating modified substrate and a preparation method thereof, belonging to the technical field of anti-fouling materials.

Background

Some medical supplies are made of polymer materials or metal materials, and they are easily contaminated by protein adsorption during medical use, and thus, the analysis capability is reduced in the case of medical analysis instruments, and it is difficult to maintain a longer service life in the case of medical consumables. It is highly desirable to develop a coating capable of resisting protein adsorption on the surface of a polymer material or a metal material.

The invention patent applications with application numbers CN201410067416.5 and CN103808786A disclose a preparation method of a capillary coating for inhibiting protein adsorption, which takes polymer molecular brush pOEGMA as a functional monomer and modifies the functional monomer on the inner wall of a quartz capillary tube by a chemical bonding method. The resulting pOEGMA-coated capillary tube has excellent ability to inhibit protein adsorption. This is clearly different from the results of the present invention.

Disclosure of Invention

The main purposes of the invention are: the method for preparing the protein adsorption resistant coating modified substrate overcomes the problems in the prior art, the protein adsorption resistant coating of the substrate can effectively resist pollution, and the protein adsorption resistant coating can effectively resist protein adsorption and platelet deposition when being applied to medical supplies. Meanwhile, a corresponding protein adsorption resistant coating modified substrate and a coating thereof are also provided.

The technical scheme for solving the technical problems of the invention is as follows:

a preparation method of a protein adsorption resistant coating modified substrate is characterized by comprising the following steps:

firstly, cleaning and drying a target base material; depositing 3-aminopropyl trimethyl amino silane on the surface of a target base material as a base layer by using a chemical vapor deposition method, and obtaining a primary modified base material;

secondly, dissolving the oligoperoxide in dioxane to form oligoperoxide treatment solution, and putting the primary modified substrate obtained in the first step into the oligoperoxide treatment solution for reaction to obtain a secondary modified substrate;

and thirdly, dissolving an OEGMA monomer in water to form an OEGMA aqueous solution, and then putting the secondary modified substrate into the OEGMA aqueous solution for reaction to obtain the protein adsorption resistant coating modified substrate.

The method can be used for modifying the surface of a target substrate with a protein adsorption-resistant coating, so that the protein adsorption-resistant coating can be used for effectively resisting protein adsorption and platelet deposition when being used as medical supplies. Note: 3-aminopropyl-trimethylaminosilane (3-aminopropyl) trimethoxysilane (APTMS for short), and OEGMA is oligo (ethylene glycol) methacrylate.

The technical scheme of the invention is further perfected as follows:

preferably, the preparation process of the oligoperoxide is as follows:

dissolving benzene tetrachlorate in anhydrous dichloroethane, stirring and mixing, and adding tert-butyl hydroperoxide to form a mixture; dissolving pyridine in anhydrous dichloroethane to form a first pyridine solution, and adding the first pyridine solution into the mixture for mixing; adding PEG-9 into the mixture, firstly dissolving pyridine in anhydrous dichloroethane to form a second pyridine solution, and then adding the second pyridine solution into the mixture for mixing; stirring and reacting; filtering and drying the solid to obtain the oligoperoxide.

More preferably, when the pyromellitic chloride is dissolved in anhydrous dichloroethane, the ratio of the pyromellitic chloride to the anhydrous dichloroethane is 4.6 ± 0.5 g: 15 plus or minus 3 ml; the molar ratio of the tert-butyl hydroperoxide to the benzene tetrachloride is 1: 1; the ratio of pyridine to anhydrous dichloroethane in the first pyridine solution is 1.1 ± 0.3 g: 10 plus or minus 2ml, and the molar ratio of pyridine to pyromellitic chloride in the first pyridine solution is 1: 1; the molar ratio of the PEG-9 to the pyromellitic chloride is 1: 1; the ratio of pyridine to anhydrous dichloroethane in the second pyridine solution is 2.2 ± 0.4 g: 10. + -. 2 ml.

More preferably, in the preparation of the oligoperoxide, the mixture is first cooled to 1 ℃ to 5 ℃, and then the first pyridine solution is added dropwise at this temperature, and after addition, the temperature is maintained and mixing is carried out for at least 1 h; then at this temperature, PEG-9 was added to the mixture, and the second pyridine solution was added dropwise; in the stirring reaction process, the temperature is gradually increased to the ambient temperature, and the stirring reaction time is at least 3 h; the solid matter is dried at 40 +/-3 deg.c for at least 3 hr.

After the preferable scheme is adopted, the preparation process of the oligomeric peroxide can be further optimized, so that the finally obtained coating has a better protein adsorption resistance effect.

Preferably, in the first step, the target base material is placed in water or ethanol solution and cleaned by ultrasonic wave during cleaning, and nitrogen is adopted to protect the target base material during drying; the treatment temperature of the chemical vapor deposition method is 50-70 ℃, and the treatment time is 0.5-12 h; and (3) drying the target base material at 70 +/-5 ℃ overnight after the target base material is treated by a chemical vapor deposition method, thus obtaining the primary modified base material.

By adopting the preferred scheme, the specific technical details of the first step can be further optimized to obtain better technical effects.

Preferably, in the second step, the ratio of the oligomeric peroxide to dioxane is 1 ± 0.5% g/ml; the reaction time is at least 24 h; and taking out the primary modified substrate after the reaction is finished, and washing away unreacted oligomeric peroxide by using dioxane to obtain the secondary modified substrate.

By adopting the preferred scheme, the specific technical details of the second step can be further optimized to obtain better technical effects.

Preferably, in the third step, the concentration of the OEGMA monomer in the OEGMA aqueous solution is 0.1 +/-0.05 mol/L; and under the protection of nitrogen, putting the secondary modified base material into an OEGMA aqueous solution, reacting at 90 +/-5 ℃ for 2-48h, taking out after the reaction is finished, and washing away unreacted OEGMA monomers by water to obtain the protein adsorption resistant coating modified base material.

By adopting the preferred scheme, the specific technical details of the third step can be further optimized to obtain better technical effect.

Preferably, the target substrate is made of a polymer material or a metal material.

By adopting the preferred scheme, the material of the target base material can be further optimized to obtain better modification effect.

The present invention also provides:

the protein adsorption resistant coating prepared by the preparation method modifies the substrate.

The preparation method is characterized in that the protein adsorption resistant coating is modified on the surface of the target substrate.

The invention can modify the protein adsorption resistant coating on the surface of the target substrate, so that the protein adsorption resistant coating can be used for resisting protein adsorption and platelet deposition, for example, when the protein adsorption resistant coating is used as a medical article. The preparation process is simple and easy to implement, and the obtained product can effectively resist pollution and has good market prospect.

Drawings

FIG. 1 is a diagram showing the main reaction process of the present invention. In the figure, APTMS is 3-aminopropyl trimethyl amino silane, CVD refers to chemical vapor deposition, oligoperoxides are oligoperoxides, and OEGMA is oligomeric (ethylene glycol) methacrylate.

FIG. 2 is a bovine serum albumin standard curve according to example 2 of the present invention.

FIG. 3 is a graph showing the results of the protein adsorption test in example 2 of the present invention.

Detailed Description

In specific implementation, the preparation method of the protein adsorption resistant coating modified substrate comprises the following steps:

firstly, cleaning and drying a target base material; and depositing the 3-aminopropyl trimethyl amino silane on the surface of the target base material as a base layer by using a chemical vapor deposition method, and obtaining a primary modified base material.

The target substrate is made of a high polymer material or a metal material. During cleaning, the target base material is put into water or ethanol solution and cleaned by ultrasound, and during drying, nitrogen is adopted to protect the target base material; the processing temperature of the chemical vapor deposition method is 50-70 ℃, and the processing time is 0.5-12 h; and (3) drying the target base material at 70 +/-5 ℃ overnight after the target base material is treated by a chemical vapor deposition method, thus obtaining the primary modified base material.

And secondly, dissolving the oligomeric hyperoxide in dioxane to form an oligomeric hyperoxide treatment solution, and putting the primary modified substrate obtained in the first step into the oligomeric hyperoxide treatment solution for reaction to obtain a secondary modified substrate.

Wherein, the preparation process of (1) the oligoperoxide comprises the following steps:

dissolving benzene tetrachlorate in anhydrous dichloroethane, stirring and mixing, and adding tert-butyl hydroperoxide to form a mixture; dissolving pyridine in anhydrous dichloroethane to form a first pyridine solution, and adding the first pyridine solution into the mixture for mixing; adding PEG-9 into the mixture, firstly dissolving pyridine in anhydrous dichloroethane to form a second pyridine solution, and then adding the second pyridine solution into the mixture for mixing; stirring and reacting; filtering and drying the solid to obtain the oligoperoxide.

In the preparation process: when the benzene tetrachlorate is dissolved in the anhydrous dichloroethane, the ratio of the benzene tetrachlorate to the anhydrous dichloroethane is 4.6 +/-0.5 g: 15 plus or minus 3 ml; the molar ratio of tert-butyl hydroperoxide to pyromellitic chloride is 1: 1; the ratio of pyridine to anhydrous dichloroethane in the first pyridine solution was 1.1 ± 0.3 g: 10 plus or minus 2ml, and the molar ratio of pyridine to pyromellitic chloride in the first pyridine solution is 1: 1; the molar ratio of PEG-9 to pyromellitic chloride is 1: 1; the ratio of pyridine to anhydrous dichloroethane in the second pyridine solution was 2.2 ± 0.4 g: 10. + -. 2 ml.

In the preparation process: cooling the mixture to 1-5 ℃, then dropwise adding the first pyridine solution at the temperature, keeping the temperature after adding, and mixing for at least 1 h; then at this temperature, PEG-9 was added to the mixture, and the second pyridine solution was added dropwise; in the stirring reaction process, the temperature is gradually increased to the ambient temperature, and the stirring reaction time is at least 3 h; the solid matter is dried at 40 +/-3 deg.c for at least 3 hr.

(2) The ratio of oligoperoxide to dioxane is 1 + -0.5% g/ml; the reaction time is at least 24 h; and taking out the primary modified substrate after the reaction is finished, and washing away unreacted oligomeric peroxide by using dioxane to obtain the secondary modified substrate.

And thirdly, dissolving an OEGMA monomer in water to form an OEGMA aqueous solution, and then putting the secondary modified substrate into the OEGMA aqueous solution for reaction to obtain the protein adsorption resistant coating modified substrate.

Wherein the concentration of OEGMA monomer in the OEGMA aqueous solution is 0.1 +/-0.05 mol/L; and under the protection of nitrogen, putting the secondary modified base material into an OEGMA aqueous solution, reacting at 90 +/-5 ℃ for 2-48h, taking out after the reaction is finished, and washing away unreacted OEGMA monomers by water to obtain the protein adsorption resistant coating modified base material.

The main process of the present invention is shown in fig. 1.

The invention is described in further detail below with reference to embodiments and with reference to the drawings. The invention is not limited to the examples given.

Example 1

This example illustrates the preparation of a sample according to the preparation method embodied in the invention described above.

(1) The specific preparation process of the oligoperoxide comprises the following steps:

4.6g (0.014mol) of a pyromellitic chloride solution was added to 15ml of anhydrous dichloroethane, and the mixture was stirred and mixed in a three-necked flask, and 1.26g (0.014mol) of t-butyl hydroperoxide was added to the mixture. The mixture was cooled to 5 ℃. 1.1g (0.014mol) of pyridine was dissolved in 10ml of anhydrous dichloroethane, added dropwise to the mixture at 5 ℃ and mixed for 1 hour. 5.6g (0.014mol) of PEG-9 were then added and a pyridine solution (2.2g pyridine +10ml dry dichloroethane) was added dropwise. Stir for 3h and gradually increase the temperature to ambient (i.e. room temperature). The solution was filtered and dried under vacuum at 40 ℃ for 3h to give 8.2g of oligoperoxide.

(2) The specific preparation process of this example is as follows:

i) the target substrate is a PU (polyurethane) sheet. The PU sheet is cleaned in ethanol solution (99%) by ultrasonic for 20min and dried under the protection of nitrogen. Then, simultaneously placing the clean PU sheet and APTMS in chemical vapor deposition equipment, and treating for 0.5-12h at the treatment temperature of 50-70 ℃; and after the treatment is finished, drying the sheet at 70 ℃ overnight to obtain the primary modified substrate.

ii) dissolving the oligoperoxide in dioxane to form a 1% oligoperoxide treatment solution; and (3) putting the primary modified substrate into the oligomeric peroxide treatment solution for reaction for 24 hours, taking out the primary modified substrate, and washing away unreacted oligomeric peroxide by using dioxane to obtain the secondary modified substrate.

iii) dissolving OEGMA monomer in water to form 0.1mol/L OEGMA aqueous solution; and under the protection of nitrogen, putting the secondary modified substrate into an OEGMA aqueous solution, reacting at 90 ℃ for 2-48h, taking out, washing away unreacted OEGMA monomer by water, and obtaining the protein adsorption resistant coating modified substrate which is marked as an oligo-OEGMA-PU sheet.

Example 2

This example is a comparative experiment for protein adsorption testing.

A sample to be tested: oligo-OEGMA-PU sheet obtained in example 1, clean PU sheet, OEGMA-PU sheet.

The preparation process of the clean PU sheet comprises the following steps: and (3) ultrasonically cleaning the PU sheet in an ethanol solution (99%) for 20min, and drying under the protection of nitrogen.

The preparation process of the OEGMA-PU sheet comprises the following steps:

i) the PU sheet is cleaned in ethanol solution (99%) by ultrasonic for 20min, and dried under the protection of nitrogen for later use.

ii) OEGMA monomer (19.08ml) was dissolved in a solution of water and methanol (12ml,1:4) and stirred under nitrogen for at least 45 min.

iii) putting the PU sheet obtained in the step i) into the solution, and standing for 0.5-12h at room temperature under the protection of nitrogen. And taking out the PU sheet from the solution, washing with alcohol and deionized water respectively, and drying overnight under nitrogen to obtain the OEGMA-PU sheet.

The protein adsorption test procedure of this example is as follows:

drawing a protein standard curve

1. Different amounts of bovine serum albumin are respectively weighed and added with a proper amount of deionized water to prepare bovine serum albumin solutions with protein concentrations of 0, 0.2, 0.4, 0.6, 0.8 and 1.0 mg/ml.

2. 0.1ml of each of the protein solutions of different concentrations was pipetted into each glass tube, and 5ml of Coomassie Brilliant blue solution was added and incubated for 10 min.

3. The samples were respectively taken for colorimetry at 595nm in a spectrophotometer, and a standard curve was drawn, as shown in FIG. 2.

(II) measurement of protein adsorption amount of sample

1. Cutting each sample to be tested into 1cm²The pieces of (a) are immersed in deionized water for at least 30 min.

2. Each sample to be tested was then placed in PBS (1M) and then immersed in bovine serum albumin (0.5mg/ml) at room temperature for 24 hours before being taken out.

3. The concentration of the bovine serum albumin solution before and after the sample immersion was measured according to the method for measuring the protein standard curve.

4. The amount of protein adsorbed can be calculated according to the following formula:

the results are shown in FIG. 3. The results show that the oligo-OEGMA-PU sheet prepared in example 1 has significantly less protein adsorption amount, i.e., has excellent protein adsorption resistance, compared to the clean PU sheet and OEGMA-PU sheet.

In addition to the above embodiments, the present invention may have other embodiments. All technical solutions formed by adopting equivalent substitutions or equivalent transformations fall within the protection scope of the claims of the present invention.