Antibacterial bilirubin composite adsorption material for blood perfusion and preparation method of antibacterial bilirubin composite adsorption material

Abstract

The invention provides an antibacterial bilirubin composite adsorption material for blood perfusion and a preparation method thereof, and belongs to the technical field of adsorption materials, the antibacterial bilirubin composite adsorption material comprises crosslinked polystyrene-divinylbenzene and silane functionalized nano titanium dioxide, the crosslinked polystyrene-divinylbenzene is used as a carrier, and the silane functionalized nano titanium dioxide is used as an adsorbent. The silane functionalized nano titanium dioxide is used as a functional monomer, a pore-foaming agent is added, and the adsorption material PSVT is prepared through a free radical suspension polymerization method. Vinyltriethoxysilane functionalized nano titanium dioxide particles are adopted as a functional monomer to be copolymerized with crosslinked polystyrene-divinylbenzene, so that the problem of leakage of nano particles is successfully solved, the adsorption capacity of bilirubin is enhanced, and the bilirubin-containing antibacterial material has excellent antibacterial performance.

Description

Technical Field

[0001] The invention belongs to the technical field of adsorption materials, and in particular relates to an antibacterial bilirubin composite adsorption material for blood perfusion and a preparation method thereof. Background Art

[0002] Liver failure is a clinical syndrome that seriously threatens human health. When the synthesis, conversion and detoxification functions of the liver are impaired, resulting in a large amount of toxins accumulating in the body and failing to metabolize normally, liver failure will occur. Bilirubin is a hemoglobin metabolite released by damaged red blood cells and is an important indicator of liver function. Under normal circumstances, bilirubin combines with albumin to form an insoluble macromolecular complex with a diameter of about 7nm (indirect bilirubin, IBIL), which is transported to the liver through the blood circulation. Under the action of glucuronidase, bilirubin combines with glucuronic acid to form direct bilirubin (DBIL), which is excreted through the kidneys with urine. The normal level of total bilirubin (TBIL) in the blood is 1 to 10g / L. However, after liver function is damaged, bilirubin generally cannot be metabolized, resulting in a large amount of bilirubin accumulation. The imbalance of the internal environment further damages liver cells, forming a vicious cycle of hyperbilirubinemia and liver failure. Excessive bilirubin can also induce hepatic encephalopathy, causing irreversible damage to the brain, and even death in severe cases. Therefore, clearing excess bilirubin from the patient's body is of great significance for treating liver failure and saving the patient's life.

[0003] Hemoperfusion has been proven to be an important and effective strategy to remove excess bilirubin from the body and prolong patient survival. The technical focus of hemoperfusion involves the development of adsorbents.

[0004] At present, the adsorption materials for bilirubin are mainly ion exchange resins and non-ionic adsorption resins, such as BL-300 produced by Kuraray Company of Japan, BR-350 produced by Asahi Chemical of Japan, BS-300 produced by Jianfan of my country, etc., which are all anion exchange resins. These products must be used by means of plasma perfusion when applied, and there is a risk of causing electrolyte disturbance. Examples of non-ionic resins used to remove bilirubin include polymer cross-linked resins (Chinese patent number: CN104174385B), which cannot meet the application requirements for the adsorption capacity of bilirubin.

[0005] At the same time, bacterial infection during the perfusion process is also one of the reasons for poor treatment results. In severe cases, it can lead to systemic inflammatory syndrome and cause significant damage to the patient's liver. However, the development of existing adsorbent materials often ignores the antibacterial ability of adsorbent materials.

[0006] It can be seen that the development of an adsorption material with excellent bilirubin adsorption capacity, high blood compatibility and good antibacterial properties is of great significance for the treatment of patients with liver failure. Summary of the invention

[0007] The problem to be solved by the present invention is to provide an antibacterial bilirubin composite adsorption material for blood perfusion and a preparation method thereof.

[0008] In order to solve the above technical problems, the technical solution adopted by the present invention is: an antibacterial bilirubin composite adsorption material for blood perfusion, including cross-linked polystyrene-divinylbenzene and silane-functionalized nano-titanium dioxide, using the cross-linked polystyrene-divinylbenzene as a carrier, the silane-functionalized nano-titanium dioxide as a functional monomer, adding a porogen, and preparing the adsorption material-polystyrene / titanium dioxide (PSVT) by free radical suspension polymerization.

[0009] Furthermore, the porogen is a mixture of toluene and liquid wax.

[0010] Furthermore, the cross-linked polystyrene-divinylbenzene has a particle size of 0.3 to 1 mm and a specific surface area of ​​500 to 1000 m 2 / g.

[0011] Furthermore, the particle size of the silane functionalized nano titanium dioxide is 10 to 100 nm, and the added amount thereof is 0.5 to 10% of the mass of the cross-linked polystyrene-divinylbenzene.

[0012] Furthermore, the added amount of the porogen is 0.5 to 2 times the mass of the cross-linked polystyrene-divinylbenzene.

[0013] The present invention also provides a method for preparing an antibacterial bilirubin composite adsorption material for blood perfusion, comprising the following steps:

[0014] S1. Preparation of silane functionalized nano-titanium dioxide:

[0015] S11. Ultrasonic dispersion of 1-2 g of titanium dioxide in 200-500 ml of ethanol at room temperature;

[0016] S12, adding 2-10 ml of vinyl triethoxysilane and 5-20 ml of ammonia water, reacting at room temperature for 5-10 hours, and obtaining silane functionalized titanium dioxide after centrifugation and washing, thereby realizing silane functionalization of nano-titanium dioxide.

[0017] S2. Preparation of cross-linked polystyrene-divinylbenzene: cross-linked polystyrene-divinylbenzene is prepared by mixing styrene:divinylbenzene in a mass ratio of 0 to 0.5:1.

[0018] S3. Preparation of adsorption material:

[0019] S31, dissolving polyvinyl alcohol in water at room temperature to prepare a polyvinyl alcohol aqueous solution with a content of 1 to 3%;

[0020] S32, adding 0.5 to 10% of the mass of cross-linked polystyrene-divinylbenzene silane-functionalized nano-titanium dioxide and 0.5 to 2 times the mass of cross-linked polystyrene-divinylbenzene porogen, and mixing to obtain an oil phase copolymer;

[0021] S33. Mix a polyvinyl alcohol aqueous solution and an oil phase copolymer to form a mixed monomer, wherein the volume ratio of the polyvinyl alcohol aqueous solution to the oil phase copolymer is 3-4:1; use benzoyl peroxide as an initiator, and the amount used is 1-3% of the total mass of the mixed monomer; subject to suspension polymerization, slowly heat up to 70-85°C at a rate of 4-10°C / 10min, react for 2-4h, then slowly heat up to 90°C at a rate of 4-10°C / 10min, react for 3-5h, then slowly heat up to above 95°C at a rate of 4-10°C / 10min, react for 3-5h, and then filter, wash, and extract the pore-forming agent to obtain an adsorbent material - polystyrene / titanium dioxide (PSVT).

[0022] Furthermore, in step S32, 1 to 2% of the mass of cross-linked polystyrene-divinylbenzene is added with silane-functionalized nano-titanium dioxide.

[0023] Due to the adoption of the above technical solution, the present invention has the following beneficial effects:

[0024] The present invention adopts cross-linked polystyrene-divinylbenzene as a carrier, which has the advantages of good blood compatibility, simple preparation and the like compared with other adsorption materials.

[0025] The present invention adopts vinyltriethoxysilane functionalized nano titanium dioxide particles as functional monomers to copolymerize with cross-linked polystyrene-divinylbenzene, successfully overcomes the leakage problem of nanoparticles, enhances the adsorption capacity of bilirubin, and enables it to have excellent antibacterial properties.

[0026] It can be seen that the present invention is an adsorption material with rich mesopore distribution, excellent bilirubin adsorption capacity, high blood compatibility and good antibacterial properties, which can effectively remove bilirubin while avoiding bacterial infection. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The present invention will be described in detail below with reference to the accompanying drawings and in combination with examples, and the advantages and implementation methods of the present invention will become more obvious. The contents shown in the accompanying drawings are only used to explain the present invention and do not constitute any limitation to the present invention in any sense. In the accompanying drawings:

[0028] Figure 1 This is a bar graph showing the simulated serum static adsorption of bilirubin by PSVT and PS of the present invention.

[0029] Figure 2This is a graph showing the antibacterial counts of the PSVT of the present invention against Escherichia coli and Staphylococcus aureus. DETAILED DESCRIPTION

[0030] The present invention discloses an antibacterial bilirubin composite adsorption material for blood perfusion, comprising cross-linked polystyrene-divinylbenzene and silane functionalized nano-titanium dioxide, cross-linked polystyrene-divinylbenzene as a carrier, silane functionalized nano-titanium dioxide as a functional monomer, and a mixture of toluene and liquid wax as a porogen, and the adsorption material PSVT is prepared by free radical suspension polymerization. PSVT is a composite mesoporous resin microsphere, which can achieve the purpose of efficiently removing bilirubin in the patient's body and resisting bacterial infection.

[0031] Among them, the particle size of cross-linked polystyrene-divinylbenzene is 0.3-1 mm, and the specific surface area is 500-1000 m 2 / g.

[0032] The particle size of the silane functionalized nano titanium dioxide is 10-100 nm, and the added amount thereof is 0.5-10% of the cross-linked polystyrene-divinylbenzene.

[0033] The added amount of the toluene and liquid wax mixture is 0.5 to 2 times of the cross-linked polystyrene-divinylbenzene.

[0034] The present invention also provides a method for preparing an antibacterial bilirubin composite adsorption material for blood perfusion.

[0035] The preparation of PSVT1 includes the following steps:

[0036] S1. Preparation of silane functionalized nano-titanium dioxide:

[0037] S11. Ultrasonic dispersion of 1 g titanium dioxide in 250 ml ethanol at room temperature;

[0038] S12, adding 5 ml of vinyl triethoxysilane and 5 ml of ammonia water, reacting at room temperature for 8 hours, and obtaining silane functionalized titanium dioxide after centrifugation and washing, thereby realizing silane functionalization of nano-titanium dioxide.

[0039] S2. Preparation of cross-linked polystyrene-divinylbenzene: cross-linked polystyrene-divinylbenzene is prepared by mixing styrene and divinylbenzene in a mass ratio of 0.2:1.

[0040] S3. Preparation of adsorption material:

[0041] S31, dissolving polyvinyl alcohol in water at room temperature to prepare a 1% polyvinyl alcohol aqueous solution;

[0042] S32, adding 0.5% of the mass of cross-linked polystyrene-divinylbenzene silane-functionalized nano-titanium dioxide and 1.5 times the mass of the cross-linked polystyrene-divinylbenzene porogen (the ratio of toluene to liquid wax in the toluene-liquid wax mixture is 1:1), and mixing to obtain an oil phase copolymer (wherein the silane-functionalized nano-titanium dioxide is chemically grafted with the polystyrene-divinylbenzene through free radical copolymerization);

[0043] S33. Mix the polyvinyl alcohol aqueous solution and the oil phase copolymer to form a mixed monomer, wherein the volume ratio of the polyvinyl alcohol aqueous solution to the oil phase copolymer is 4:1; use benzoyl peroxide as an initiator, and the amount used is 1% of the total mass of the mixed monomer; perform suspension polymerization, slowly heat up to 80°C at a rate of 5°C / 10min, react for 3h, then slowly heat up to 90°C at a rate of 5°C / 10min, react for 4h, then slowly heat up to above 95°C at a rate of 5°C / 10min, react for 4h, and then filter, wash, and extract the porogen to obtain the adsorbent material PSVT.

[0044] The preparation of PSVT2 includes the following steps:

[0045] S1. Preparation of silane functionalized nano-titanium dioxide:

[0046] S11. Ultrasonic dispersion of 1 g titanium dioxide in 250 ml ethanol at room temperature;

[0047] S12, adding 5 ml of vinyl triethoxysilane and 5 ml of ammonia water, reacting at room temperature for 8 hours, and obtaining silane functionalized titanium dioxide after centrifugation and washing, thereby realizing silane functionalization of nano-titanium dioxide.

[0048] S2. Preparation of cross-linked polystyrene-divinylbenzene: cross-linked polystyrene-divinylbenzene is prepared by mixing styrene and divinylbenzene in a mass ratio of 0.2:1.

[0049] S3. Preparation of adsorption material:

[0050] S31, dissolving polyvinyl alcohol in water at room temperature to prepare a 1% polyvinyl alcohol aqueous solution;

[0051] S32, adding 1% of the mass of cross-linked polystyrene-divinylbenzene silane-functionalized nano-titanium dioxide and 1.5 times the mass of the cross-linked polystyrene-divinylbenzene porogen (the ratio of toluene to liquid wax in the toluene-liquid wax mixture is 1:1), and mixing to obtain an oil phase copolymer (wherein the silane-functionalized nano-titanium dioxide is chemically grafted with the polystyrene-divinylbenzene through free radical copolymerization);

[0052] S33. Mix the polyvinyl alcohol aqueous solution and the oil phase copolymer to form a mixed monomer, wherein the volume ratio of the polyvinyl alcohol aqueous solution to the oil phase copolymer is 4:1; use benzoyl peroxide as an initiator, and the amount used is 1% of the total mass of the mixed monomer; perform suspension polymerization, slowly heat up to 80°C at a rate of 5°C / 10min, react for 3h, then slowly heat up to 90°C at a rate of 5°C / 10min, react for 4h, then slowly heat up to above 95°C at a rate of 5°C / 10min, react for 4h, and then filter, wash, and extract the porogen to obtain the adsorbent material PSVT.

[0053] The preparation of PSVT3 includes the following steps:

[0054] S1. Preparation of silane functionalized nano-titanium dioxide:

[0055] S11. Ultrasonic dispersion of 1 g titanium dioxide in 250 ml ethanol at room temperature;

[0056] S12, adding 5 ml of vinyl triethoxysilane and 5 ml of ammonia water, reacting at room temperature for 8 hours, and obtaining silane functionalized titanium dioxide after centrifugation and washing, thereby realizing silane functionalization of nano-titanium dioxide.

[0057] S2. Preparation of cross-linked polystyrene-divinylbenzene: cross-linked polystyrene-divinylbenzene is prepared by mixing styrene and divinylbenzene in a mass ratio of 0.2:1.

[0058] S3. Preparation of adsorption material:

[0059] S31, dissolving polyvinyl alcohol in water at room temperature to prepare a 1% polyvinyl alcohol aqueous solution;

[0060] S32, adding 2% of the mass of cross-linked polystyrene-divinylbenzene silane-functionalized nano-titanium dioxide and 1.5 times the mass of the cross-linked polystyrene-divinylbenzene porogen (the ratio of toluene to liquid wax in the toluene-liquid wax mixture is 1:1), and mixing to obtain an oil phase copolymer (wherein the silane-functionalized nano-titanium dioxide is chemically grafted with the polystyrene-divinylbenzene by free radical copolymerization);

[0061] S33. Mix the polyvinyl alcohol aqueous solution and the oil phase copolymer to form a mixed monomer, wherein the volume ratio of the polyvinyl alcohol aqueous solution to the oil phase copolymer is 4:1; use benzoyl peroxide as an initiator, and the amount used is 1% of the total mass of the mixed monomer; perform suspension polymerization, slowly heat up to 80°C at a rate of 5°C / 10min, react for 3h, then slowly heat up to 90°C at a rate of 5°C / 10min, react for 4h, then slowly heat up to above 95°C at a rate of 5°C / 10min, react for 4h, and then filter, wash, and extract the porogen to obtain the adsorbent material PSVT.

[0062] Example:

[0063] Simulated serum static adsorption experiment of bilirubin:

[0064] Preparation of simulated high bilirubin serum: Weigh 0.75 g of bovine serum albumin and dissolve it in 50 ml of PBS buffer to prepare a simulated serum solution. Weigh 7.5 mg of bilirubin and dissolve it in 1.5 ml of dimethyl sulfoxide and 1 ml (0.1 mol / L) sodium carbonate solution, and use the simulated serum solution to make the volume to 50 ml to obtain a simulated high bilirubin serum solution.

[0065] PSVT was used as the experimental group, and ordinary cross-linked polystyrene-divinylbenzene microspheres (PS) were used as the control group. 0.5 ml of PSVT1, PSVT2, PSVT3 and PS were taken in 25 ml conical bottles, 15 ml of high bilirubin simulated serum solution was added, and the mixture was placed in a shaker away from light and incubated at 37 °C for 2 h.

[0066] The supernatant was taken to detect the bilirubin concentration, and three sets of repeated tests were performed to calculate the adsorption rate.

[0067] like Figure 1 As shown, the results show that the adsorption rate of bilirubin by PSVT2 of the present invention in the simulated serum solution is 77.54%, while the adsorption rate of bilirubin by PS is 55.92%, indicating that the adsorption capacity of the adsorbent material PSVT of the present invention for bilirubin has been significantly improved and can meet clinical requirements.

[0068] Antibacterial performance experiment of adsorption material:

[0069] Take 0.5 ml of PSVT2 in a 25 ml conical flask, add 15 ml of bacterial solution containing Escherichia coli and Staphylococcus aureus respectively, place it in a shaker, and incubate at 37°C for 2 h.

[0070] The supernatant was taken, diluted and cultured in LB medium for 24 h, and the antibacterial rate was calculated using the colony counting method.

[0071] like Figure 2As shown in Table 1, the results show that the antibacterial rate of the adsorbent material PSVT2 of the present invention against Escherichia coli is 87.16%, and the antibacterial rate against Staphylococcus aureus is 98.77%. It can be seen that PSVT has excellent antibacterial properties.

[0072] Table 1 Antibacterial rate of adsorbent material of the present invention

[0073]

[0074] It can be seen that the bilirubin clearance ability of the PSVT of the present invention is significantly higher than that of PS, especially PSVT2, whose bilirubin adsorption rate in simulated serum is 77.54%; at the same time, the antibacterial rates of the PSVT2 of the present invention against Escherichia coli and Staphylococcus aureus can reach 87.16% and 98.77%, respectively.

[0075] The embodiments of the present invention are described in detail above, but the contents are only preferred embodiments of the present invention and cannot be considered to limit the scope of implementation of the present invention. All equivalent changes and improvements made within the scope of the present invention should still fall within the scope of the present invention.

Claims

1. An antibacterial bilirubin composite adsorption material for blood perfusion, characterized in that: The invention comprises cross-linked polystyrene-divinylbenzene and silane functionalized nano titanium dioxide. The cross-linked polystyrene-divinylbenzene is used as a carrier, the silane functionalized nano titanium dioxide is used as a functional monomer, a porogen is added, and the adsorption material PSVT is prepared by a free radical suspension polymerization method.

2. The antibacterial bilirubin composite adsorption material for blood perfusion according to claim 1, characterized in that: The porogen is a mixture of toluene and liquid wax.

3. The antibacterial bilirubin composite adsorption material for blood perfusion according to claim 1, characterized in that: The cross-linked polystyrene-divinylbenzene has a particle size of 0.3 to 1 mm and a specific surface area of ​​500 to 1000 m 2 / g.

4. The antibacterial bilirubin composite adsorption material for blood perfusion according to claim 1, characterized in that: The particle size of the silane functionalized nano titanium dioxide is 10-100 nm, and the added amount thereof is 0.5-10% of the mass of the cross-linked polystyrene-divinylbenzene.

5. The antibacterial bilirubin composite adsorption material for blood perfusion according to claim 1, characterized in that: The added amount of the porogen is 0.5 to 2 times the mass of the cross-linked polystyrene-divinylbenzene.

6. A method for preparing an antibacterial bilirubin composite adsorption material for hemoperfusion, used for preparing the antibacterial bilirubin composite adsorption material for hemoperfusion according to any one of claims 1 to 5, characterized in that: The following steps are involved: S1. Preparation of silane functionalized nano-titanium dioxide: S11. Ultrasonic dispersion of 1-2 g of titanium dioxide in 200-500 ml of ethanol at room temperature; S12, adding 2-10 ml of vinyl triethoxysilane and 5-20 ml of ammonia water, reacting at room temperature for 5-10 hours, and obtaining silane functionalized titanium dioxide after centrifugation and washing, thereby achieving silane functionalization of nano-titanium dioxide; S2. Preparation of cross-linked polystyrene-divinylbenzene: mixing styrene and divinylbenzene in a mass ratio of 0 to 0.5:1 to prepare cross-linked polystyrene-divinylbenzene; S3. Preparation of adsorption material: S31, dissolving polyvinyl alcohol in water at room temperature to prepare a polyvinyl alcohol aqueous solution with a content of 1 to 3%; S32, adding 0.5 to 10% of the mass of cross-linked polystyrene-divinylbenzene silane-functionalized nano-titanium dioxide and 0.5 to 2 times the mass of cross-linked polystyrene-divinylbenzene porogen, and mixing to obtain an oil phase copolymer; S33. Mix the polyvinyl alcohol aqueous solution and the oil phase copolymer to form a mixed monomer, wherein the volume ratio of the polyvinyl alcohol aqueous solution to the oil phase copolymer is 3-4:1; use benzoyl peroxide as an initiator, and the amount used is 1-3% of the total mass of the mixed monomer; subject to suspension polymerization, slowly heat up to 70-85°C at a rate of 4-10°C / 10min, react for 2-4h, then slowly heat up to 90°C at a rate of 4-10°C / 10min, react for 3-5h, then slowly heat up to above 95°C at a rate of 4-10°C / 10min, react for 3-5h, and then filter, wash, and extract the porogen to obtain the adsorbent material PSVT.

7. The method for preparing the antibacterial bilirubin composite adsorption material for hemoperfusion according to claim 6, characterized in that: In step S32, 1-2% of the mass of cross-linked polystyrene-divinylbenzene silane-functionalized nano-titanium dioxide is added.

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