CN119733482A - Chitosan-based blood perfusion adsorbent and its preparation method and application - Google Patents
Chitosan-based blood perfusion adsorbent and its preparation method and application Download PDFInfo
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Abstract
本申请涉及材料领域，具体公开了一种基于壳聚糖的血液灌流吸附剂及其制备方法和应用。该血液灌流吸附剂以改性壳聚糖微球为载体，在表面接枝菊苣酸后，固载二氧化锰纳米酶，形成复合吸附剂。本申请提供的这种血液灌流吸附剂，由于表面固载了二氧化锰纳米酶，能够高效清除活性氧，同时能清除多种自由基；且其本身具有高吸附能力，固载到壳聚糖微球表面后，能够与壳聚糖微球协同性地有效吸附血液中的炎症因子，提高血液净化的效果，同时具有血液相容性和安全性，可用于炎症患者的血液净化治疗。The present application relates to the field of materials, and specifically discloses a chitosan-based blood perfusion adsorbent, and its preparation method and application. The blood perfusion adsorbent uses modified chitosan microspheres as carriers, and after grafting chicoric acid on the surface, it immobilizes manganese dioxide nanozymes to form a composite adsorbent. The blood perfusion adsorbent provided by the present application, due to the manganese dioxide nanozymes immobilized on the surface, can efficiently remove active oxygen and a variety of free radicals; and it itself has a high adsorption capacity. After being immobilized on the surface of chitosan microspheres, it can synergistically and effectively adsorb inflammatory factors in the blood with chitosan microspheres, improve the effect of blood purification, and at the same time has blood compatibility and safety, and can be used for blood purification treatment of inflammatory patients.
Description
Technical Field
The application relates to the field of medical instruments, in particular to a chitosan-based blood perfusion adsorbent, and a preparation method and application thereof.
Background
The blood perfusion technology is an extracorporeal blood purifying method, which is to draw the blood of the patient out of the body, pass through a perfusion device with specific adsorbent to remove the harmful substances in the blood, and then return the purified blood to the body of the patient. This technique has great potential in the treatment of a variety of diseases, particularly inflammatory diseases, such as severe hepatitis, psoriasis, sepsis, and the like.
However, existing blood perfusion adsorbents have limitations in removing inflammatory factors from the blood. The blood of patients with inflammatory diseases often contains high concentrations of inflammatory factors, such as tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), etc., which are key mediators of the inflammatory response. The existing adsorbents have limited removal efficiency of the inflammatory factors, so that the treatment effect is poor. At the same time, inflammatory patients often experience increased oxidative stress due to the inflammatory response, resulting in elevated levels of free radicals and reactive oxygen species in the blood, which cause damage to the nuclear tissue, for which existing adsorbents are often forensic.
In view of this, it is necessary to develop a blood perfusion adsorbent for inflammatory patients to improve the blood perfusion purifying effect of inflammatory patients.
Disclosure of Invention
In order to improve the clearance rate of inflammatory factors, free radicals and active oxygen in the blood perfusion process and reduce the hemolysis and the safety, the application provides a chitosan-based blood perfusion adsorbent, and a preparation method and application thereof.
The application adopts the following technical scheme:
In a first aspect, the application provides a chitosan-based blood perfusion adsorbent, which takes modified chitosan microspheres as a carrier, and after chicoric acid is grafted on the surface of the modified chitosan microspheres, manganese dioxide nano enzyme is immobilized on the modified chitosan microspheres to form a composite adsorbent for removing inflammatory factors in blood of inflammatory patients.
According to the technical scheme, chitosan microspheres are used as carriers through optimization of material selection, surface modification and modification of the adsorbent, the chitosan microspheres have good biocompatibility and mechanical strength, chicoric acid is grafted after surface modification, chicoric acid is used as a connecting arm, and manganese dioxide nano-enzyme is immobilized on the surfaces of the chicoric acid.
The manganese dioxide nano enzyme has three types of antioxidant enzyme activities, including activities such as superoxide dismutase, catalase and glutathione peroxidase, can efficiently remove active oxygen and simultaneously remove various free radicals, has high adsorption capacity, and can effectively adsorb inflammatory factors in blood in cooperation with chitosan microspheres after being immobilized on the surfaces of the chitosan microspheres, thereby improving the blood purification effect. Meanwhile, 2 adjacent carboxyl groups and 4 phenolic hydroxyl groups are contained in chicoric acid molecules, so that the chicoric acid molecules can serve as an intermediate bridge to provide stable chemical bond for connecting chitosan microsphere carriers and manganese dioxide nano enzyme ligands. And the hydrophilicity of the surface of the adsorbent is enhanced through the modification of the chicoric acid, which is helpful for improving the blood compatibility and reducing the risks of blood coagulation and platelet activation.
Further, in a preferred embodiment of the present application, the modified chitosan microsphere is obtained by subjecting monodisperse chitosan microsphere to epoxidation modification. The surface of the chitosan microsphere is subjected to epoxidation modification, so that the structural stability of the chitosan microsphere can be improved, and meanwhile, the chemical reaction activity of the surface of the microsphere can be improved, and further grafting of chicoric acid is facilitated.
Preferably, in a preferred embodiment of the present application, the chitosan microsphere has a particle size of 100-200 μm and a coefficient of variation CV <5%. The chitosan microsphere has strong monodispersity, is not easy to agglomerate in the subsequent chemical modification process, can provide more consistent adsorption surface area and void structure in the blood perfusion process, and is beneficial to improving the blood perfusion efficiency.
Further, in a preferred embodiment of the present application, the modification method of the modified chitosan microsphere includes:
(1) Dispersing chitosan microspheres in alkali liquor, and carrying out ultrasonic treatment for 20-30min to obtain chitosan microsphere dispersion;
(2) 2, 3-epoxypropyl trimethyl ammonium chloride and ferrous sulfate are added into the chitosan microsphere dispersion liquid, and the mixture is placed at the temperature of 40-60 ℃ to react for 7-9h under the condition of stirring, and then the mixture is washed.
Preferably, the ultrasonic time is 24-26min;
Preferably, the reaction temperature is from 45 to 55℃for 7.5 to 8.5 hours.
Further, in the preferred embodiment of the present application, the mass ratio of the chitosan microsphere to the 2, 3-epoxypropyl trimethyl ammonium chloride is 20-30:1, preferably 23-26:1.
In a second aspect, the present application provides a method for preparing the above-mentioned hemoperfusion adsorbent, comprising:
(1) Performing epoxidation modification on the chitosan microsphere to obtain a modified chitosan microsphere;
(2) Adding the modified chitosan microsphere into an ethanol water solution containing chicoric acid, carrying out ultrasonic treatment for 40-50min, adding stannous chloride, and reacting at 70-80 ℃ for 3-4h to obtain a chicory acidified chitosan microsphere;
Stannous chloride is used as a catalyst, which helps to accelerate the reaction rate. Preferably, the reaction temperature is 73-77 ℃ and the reaction time is 3.5-4h.
1. Dispersing the chicory acidified chitosan microspheres into a glycol solution containing manganese nitrate hexahydrate, adding alkali liquor, performing pyrolysis reaction, centrifuging and washing to obtain the hemoperfusion adsorbent.
Manganese nitrate hexahydrate (Mn (NO 3)2·6H2 O) is heated under alkaline conditions to decompose and form MnO 2, mnO 2 which is free in the solution is captured by chitosan microspheres acidified by chicory in the process of gradual formation and is immobilized on the chitosan microspheres, so that the blood perfusion adsorbent is formed.
Further, in the preferred embodiment of the present application, the mass ratio of the modified chitosan microsphere to chicoric acid is 15-20:1, preferably 16-18:1.
Further, in the preferred embodiment of the application, the mass ratio of the modified chitosan microsphere to the manganese nitrate hexahydrate is 6-8:1, preferably 6.5-7.5:1.
Further, in the preferred embodiment of the present application, the reaction temperature of the above-mentioned antipyretic reaction is 50 to 80 ℃, the reaction time is 3 to 6 hours, preferably, the reaction temperature is 60 to 70 ℃, and the reaction time is 4 to 5 hours.
In a third aspect, the present application provides an application of the above-mentioned blood perfusion adsorbent in preparing a blood perfusion apparatus.
In summary, the application has the following beneficial effects:
1. The blood perfusion adsorbent provided by the application takes the chitosan microsphere as a carrier, has good biocompatibility and mechanical strength, can increase the structural stability of the chitosan microsphere after surface modification, can improve the chemical reaction activity of the microsphere surface, and is favorable for further grafting chicoric acid.
2. The blood perfusion adsorbent provided by the application takes chicoric acid as a connecting arm. As chicoric acid contains 2 adjacent carboxyl groups and 4 phenolic hydroxyl groups in the molecule, the chicoric acid can serve as an intermediate bridge to provide stable chemical bond for connecting the chitosan microsphere carrier and the manganese dioxide nano enzyme ligand. And the hydrophilicity of the surface of the adsorbent is enhanced through the modification of the chicoric acid, which is helpful for improving the blood compatibility and reducing the risks of blood coagulation and platelet activation.
3. The application provides a blood perfusion adsorbent, which takes manganese dioxide nano-enzyme as a ligand. The manganese dioxide nano enzyme has three types of antioxidant enzyme activities, including activities such as superoxide dismutase (SOD), catalase (CAT), glutathione Peroxidase (GPX) and the like, can effectively remove active oxygen and simultaneously remove various free radicals, has high adsorption capacity, and can effectively adsorb inflammatory factors in blood in cooperation with chitosan microspheres after being immobilized on the surfaces of the chitosan microspheres, thereby improving the blood purification effect.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to the following examples, which are to be construed as merely illustrative and not limitative of the scope of the invention, but are not intended to limit the scope of the invention to the specific conditions set forth in the examples, either as conventional or manufacturer-suggested, nor are reagents or apparatus employed to identify manufacturers as conventional products available for commercial purchase.
The following describes specific embodiments of the present invention in detail. It should be understood that the detailed description and specific examples, while indicating and illustrating the invention, are not intended to limit the invention.
Examples
Example 1
The embodiment provides a chitosan-based hemoperfusion adsorbent, and the preparation method thereof comprises the following steps:
1. Dispersing 50g of chitosan microsphere with the particle size of 100-200 μm (coefficient of variation CV <5%, purchased from Shanghai Pengpan biotechnology company) in 250ml of 0.05mol/L NaOH solution, and performing ultrasonic treatment for 25min to obtain a chitosan microsphere dispersion liquid, 2, adding 2, 3-epoxypropyl trimethyl ammonium chloride (2 g) into the chitosan microsphere dispersion liquid, stirring uniformly, adding a catalytic amount of ferrous sulfate, stirring under a water bath at 50 ℃ for reaction for 8 hours, and then washing with ethanol and deionized water to obtain modified chitosan microspheres;
3. Adding 35g of modified chitosan microspheres obtained in the step (2) into chicoric acid solution, carrying out ultrasonic treatment for 45min, adding catalytic stannous chloride, placing the mixed reaction solution into a water bath at a temperature of 72 ℃ for heating reaction for 4h, filtering, and washing with ethanol and deionized water in sequence to obtain chicory acidified chitosan microspheres;
4. Dissolving 5.3g of manganese nitrate hexahydrate crystal in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution, then adding the chicory-acidified chitosan microspheres obtained in the step (3), performing ultrasonic dispersion for 30min, placing in a water bath at 60 ℃, adding 4mL of ammonia water under stirring at the rotating speed of 250rmp, continuing to react for 5h, centrifuging, washing with ethanol, washing with deionized water to be neutral, and performing freeze vacuum drying.
Example 2
The present embodiment provides a chitosan-based blood perfusion adsorbent, and the preparation method thereof is different from embodiment 1 in that step (2):
2, 3-epoxypropyl trimethyl ammonium chloride (2.5 g) is added into the chitosan microsphere dispersion liquid, after uniform stirring, a catalytic amount of ferrous sulfate is added, and the mixture is stirred and reacted for 8 hours in a water bath at 50 ℃, and then ethanol and deionized water are used for washing, so as to obtain the modified chitosan microsphere.
Example 3
The present embodiment provides a chitosan-based blood perfusion adsorbent, and the preparation method thereof is different from embodiment 1 in that step (2):
2, 3-epoxypropyl trimethyl ammonium chloride (1.6 g) is added into chitosan microsphere dispersion liquid, after uniform stirring, a catalytic amount of ferrous sulfate is added, and the mixture is stirred and reacted for 8 hours in a water bath at 50 ℃, and then ethanol and deionized water are used for washing, so as to obtain the modified chitosan microsphere.
Example 4
The present embodiment provides a chitosan-based blood perfusion adsorbent, and the preparation method thereof is different from embodiment 1 in that step (2):
2, 3-epoxypropyl trimethyl ammonium chloride (2 g) is added into chitosan microsphere dispersion liquid, after uniform stirring, a catalytic amount of ferrous sulfate is added, and the mixture is stirred and reacted for 7 hours in a water bath at 60 ℃, and then washed by ethanol and deionized water, so as to obtain the modified chitosan microsphere.
Example 5
The present embodiment provides a chitosan-based blood perfusion adsorbent, and the preparation method thereof is different from embodiment 1 in that step (2):
2, 3-epoxypropyl trimethyl ammonium chloride (2 g) is added into chitosan microsphere dispersion liquid, after uniform stirring, a catalytic amount of ferrous sulfate is added, and the mixture is stirred and reacted for 9 hours in a water bath at the temperature of 40 ℃, and then ethanol and deionized water are used for washing, so that the modified chitosan microsphere is obtained.
Example 6
The present embodiment provides a chitosan-based blood perfusion adsorbent, and the preparation method thereof is different from embodiment 1 in that step (3):
Dissolving chicoric acid (2 g) in 100mL ethanol solution with the volume fraction of 70% to obtain chicoric acid solution, adding 40g of modified chitosan microspheres obtained in the step (2) into the chicoric acid solution, carrying out ultrasonic treatment for 45min, adding catalytic amount of stannous chloride, placing the mixed reaction solution into a water bath at the temperature of 72 ℃ for heating reaction for 4h, filtering, and washing with ethanol and deionized water in sequence to obtain chicory acidified chitosan microspheres.
Example 7
The present embodiment provides a chitosan-based blood perfusion adsorbent, and the preparation method thereof is different from embodiment 1 in that step (3):
Dissolving chicoric acid (2 g) in 100mL ethanol solution with the volume fraction of 70% to obtain chicoric acid solution, adding 30g of modified chitosan microspheres obtained in the step (2) into the chicoric acid solution, carrying out ultrasonic treatment for 45min, adding catalytic amount of stannous chloride, placing the mixed reaction solution into a water bath at the temperature of 72 ℃ for heating reaction for 4h, filtering, and washing with ethanol and deionized water in sequence to obtain chicory acidified chitosan microspheres.
Example 8
The present embodiment provides a chitosan-based blood perfusion adsorbent, and the preparation method thereof is different from embodiment 1 in that step (3):
Dissolving chicoric acid (2 g) in 100mL ethanol solution with the volume fraction of 70% to obtain chicoric acid solution, adding 35g of modified chitosan microspheres obtained in the step (2) into the chicoric acid solution, carrying out ultrasonic treatment for 45min, adding catalytic amount of stannous chloride, placing the mixed reaction solution into water bath at the temperature of 80 ℃ for heating reaction for 3h, filtering, and washing with ethanol and deionized water in sequence to obtain chicory acidified chitosan microspheres.
Example 9
The present embodiment provides a chitosan-based blood perfusion adsorbent, and the preparation method thereof is different from embodiment 1 in that step (3):
Dissolving chicoric acid (2 g) in 100mL ethanol solution with the volume fraction of 70% to obtain chicoric acid solution, adding 35g of modified chitosan microspheres obtained in the step (2) into the chicoric acid solution, carrying out ultrasonic treatment for 45min, adding catalytic amount of stannous chloride, placing the mixed reaction solution into a water bath at the temperature of 70 ℃ for heating reaction for 4h, filtering, and washing with ethanol and deionized water in sequence to obtain chicory acidified chitosan microspheres.
Example 10
The present example provides a chitosan-based blood perfusion adsorbent, which is different from example 1 in the preparation method thereof in the step (4):
Dissolving 4.3g of manganese nitrate hexahydrate crystal in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution, then adding the chicory-acidified chitosan microspheres obtained in the step (3), performing ultrasonic dispersion for 30min, placing in a water bath at 60 ℃, adding 4mL of ammonia water under stirring at the rotating speed of 250rmp, continuing to react for 5h, centrifuging, washing with ethanol, washing with deionized water to be neutral, and performing freeze vacuum drying.
Example 11
The present example provides a chitosan-based blood perfusion adsorbent, which is different from example 1 in the preparation method thereof in the step (4):
Dissolving 5.8g of manganese nitrate hexahydrate crystal in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution, then adding the chicory-acidified chitosan microspheres obtained in the step (3), performing ultrasonic dispersion for 30min, placing in a water bath at 60 ℃, adding 4mL of ammonia water under stirring at the rotating speed of 250rmp, continuing to react for 5h, centrifuging, washing with ethanol, washing with deionized water to be neutral, and performing freeze vacuum drying.
Example 12
The present example provides a chitosan-based blood perfusion adsorbent, which is different from example 1 in the preparation method thereof in the step (4):
Dissolving 5.3g of manganese nitrate hexahydrate crystal in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution, then adding the chicory-acidified chitosan microspheres obtained in the step (3), performing ultrasonic dispersion for 30min, placing in a water bath at 50 ℃, adding 4mL of ammonia water under stirring at the rotating speed of 250rmp, continuing to react for 6h, centrifuging, washing with ethanol, washing with deionized water to be neutral, and performing freeze vacuum drying.
Example 13
The present example provides a chitosan-based blood perfusion adsorbent, which is different from example 1 in the preparation method thereof in the step (4):
Dissolving 5.3g of manganese nitrate hexahydrate crystal in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution, then adding the chicory-acidified chitosan microspheres obtained in the step (3), performing ultrasonic dispersion for 30min, placing in a water bath at 80 ℃, adding 4mL of ammonia water under stirring at the rotating speed of 250rmp, continuing to react for 3h, centrifuging, washing with ethanol, washing with deionized water to be neutral, and performing freeze vacuum drying.
Comparative example
Comparative example 1
The comparative example provides a chitosan-based adsorbent, the preparation method of which comprises:
(1) Dissolving chicoric acid (2 g) in 100mL ethanol solution with volume fraction of 70% to obtain chicoric acid solution, adding 35g chitosan microsphere (with variation coefficient CV <5%, purchased from Shanghai Pengpan biotechnology company) into chicoric acid solution, ultrasonically treating for 45min, adding catalytic amount of stannous chloride, heating the mixed reaction solution in water bath at 72 ℃ for reaction for 4h, filtering, and washing sequentially with ethanol and deionized water to obtain chicory acidified chitosan microsphere;
(2) Dissolving 5.3g of manganese nitrate hexahydrate crystal in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution, then adding the chicory-acidified chitosan microspheres, performing ultrasonic dispersion for 30min, adding 4mL of ammonia water under stirring, placing in a water bath at 60 ℃ for reaction for 5h, centrifuging, washing with ethanol, washing with deionized water to be neutral, and performing freeze vacuum drying.
Comparative example 2
The comparative example provides a chitosan-based adsorbent, the preparation method of which comprises:
1. Dispersing 50g of chitosan microsphere with the particle size of 100-200 μm (coefficient of variation CV <5%, purchased from Shanghai Pengpan biotechnology company) in 250ml of 0.05mol/L NaOH solution, and performing ultrasonic treatment for 25min to obtain a chitosan microsphere dispersion liquid, 2, adding 2, 3-epoxypropyl trimethyl ammonium chloride (2 g) into the chitosan microsphere dispersion liquid, stirring uniformly, adding a catalytic amount of ferrous sulfate, stirring under a water bath at 50 ℃ for reaction for 8 hours, and then washing with ethanol and deionized water to obtain modified chitosan microspheres;
3. dissolving 5.3g of manganese nitrate hexahydrate crystal in 200mL of ethylene glycol to obtain manganese nitrate hexahydrate solution, then adding the modified chitosan microsphere obtained in the step (2), performing ultrasonic dispersion for 30min, adding 4mL of ammonia water under stirring, placing in a water bath at 60 ℃ for reaction for 5h, centrifuging, washing with ethanol, washing with deionized water to be neutral, and performing freeze vacuum drying.
Comparative example 3
The comparative example provides a chitosan-based adsorbent, the preparation method of which comprises:
1. Dispersing 50g of chitosan microsphere with the particle size of 100-200 μm (coefficient of variation CV <5%, purchased from Shanghai Pengpan biotechnology company) in 250ml of 0.05mol/L NaOH solution, and performing ultrasonic treatment for 25min to obtain a chitosan microsphere dispersion liquid, 2, adding 2, 3-epoxypropyl trimethyl ammonium chloride (2 g) into the chitosan microsphere dispersion liquid, stirring uniformly, adding a catalytic amount of ferrous sulfate, stirring under a water bath at 50 ℃ for reaction for 8 hours, and then washing with ethanol and deionized water to obtain modified chitosan microspheres;
3. Dissolving chicoric acid (2 g) in 100mL ethanol solution with the volume fraction of 70% to obtain chicoric acid solution, adding 35g of modified chitosan microspheres obtained in the step (2) into the chicoric acid solution, carrying out ultrasonic treatment for 45min, adding catalytic amount of stannous chloride, placing the mixed reaction solution into a water bath at the temperature of 72 ℃ for heating reaction for 4h, filtering, and washing with ethanol and deionized water in sequence to obtain chicory acidified chitosan microspheres.
Performance test
Detection method/test method
1. Blood compatibility experiments:
A hemolysis test was performed on the adsorbents provided in examples 1 to 13 and comparative examples 1 to 3 according to GB/T16886.4-2003 medical instrument biological evaluation section 4 and blood interaction test selection.
The haemolysis rate was calculated according to the formula:
Hemolysis ratio = (a-B)/(C-B) ×100%, where a is the absorbance of the sample group, B is the absorbance of the negative control group, and C is the absorbance of the positive control group.
Experimental results show that the hemolysis rate of the adsorbents provided in the examples 1-13 and the comparative examples 1-3 is less than 3%, and the adsorbents meet the national standard (the requirement is less than 5%). Therefore, the adsorbent provided by the application has good biocompatibility.
2. Clearance experiments on inflammatory factors:
The adsorption materials provided in examples 1 and 6 and comparative examples 1 to 3 were subjected to wet column packing (φ 1X 10 cm), pre-washed with physiological saline, followed by addition of 30mL of bovine plasma to which TNF- α and IL-6 were added (wherein the concentration of TNF- α was 1ng/mL and the concentration of IL-6 was 0.5 ng/mL), dynamic adsorption was circulated for 3 hours, and the inflammatory factor TNF- α and IL-6 contents in the bovine plasma before and after adsorption were measured, and the clearance was calculated, and the results are shown in Table 1.
Table 1.
As can be seen from Table 1, the adsorbents provided in examples 1 to 13 of the present application have a clearance rate of more than 80% for both TNF-. Alpha.and IL-6. In contrast, comparative examples 1 and 2 have weaker clearance rates for TNF- α and IL-6, mainly because the chitosan microspheres are not activated in a modified manner (comparative example 1) or because the chitosan microspheres are not grafted with chicoric acid (comparative example 2), resulting in that only a small amount of manganese dioxide nanoenzyme is immobilized on the surface of the chitosan microspheres by electrostatic adsorption in the following, thereby affecting the adsorption efficiency thereof. In contrast, in comparative example 3, the adsorption capacity of the chitosan microsphere itself was only relied on because the surface of the adsorbent does not carry manganese dioxide nano-enzyme, so that the clearance of both TNF-alpha and IL-6 was weaker.
3. Free radical scavenging experiments:
The adsorbents prepared in example 1 and comparative examples 1 to 3 were used as test subjects to determine their ability to scavenge hydroxyl radicals using a hydroxyl radical assay kit (purchased from Nanjing's institute of biosciences). The absorbance of the reaction solution at 550nm was measured by an enzyme-labeled instrument, and the clear rate of each adsorbent for hydroxyl radicals was calculated, and the results are shown in table 2:
table 2.
As can be seen from Table 2, the adsorbent provided in example 1 of the present application has a very strong scavenging rate for hydroxyl radicals, since the manganese dioxide nanoenzyme is immobilized on the surface of chitosan microspheres through chicoric acid with very stable bonding. Comparative examples 1-2 showed limited scavenging of hydroxyl radicals due to the low immobilized amount of manganese dioxide nanoenzyme. In contrast, in comparative example 3, the removal of hydroxyl radicals was very weak because of the absence of immobilized manganese dioxide nanoenzyme.
The present embodiment is only for explanation of the present application and is not to be construed as limiting the present application, and modifications to the present embodiment, which may not creatively contribute to the present application as required by those skilled in the art after reading the present specification, are all protected by patent laws within the scope of claims of the present application.
Claims (10)
1. A chitosan-based blood perfusion adsorbent is characterized in that modified chitosan microspheres are used as carriers of the blood perfusion adsorbent, and manganese dioxide nanoenzyme is immobilized after chicoric acid is grafted on the surfaces of the modified chitosan microspheres to form a composite adsorbent for removing inflammatory factors in blood of inflammatory patients.
2. The chitosan-based hemoperfusion adsorbent of claim 1, wherein the modified chitosan microspheres are obtained by subjecting monodisperse chitosan microspheres to epoxidation modification.
3. The chitosan-based hemoperfusion adsorbent of claim 1, wherein the chitosan microspheres have a particle size of 100-200 μm and a coefficient of variation CV < 5%.
4. The chitosan-based hemoperfusion adsorbent of claim 1, wherein the modification method of the modified chitosan microsphere comprises:
dispersing chitosan microspheres in alkali liquor, and carrying out ultrasonic treatment for 20-30min to obtain chitosan microsphere dispersion;
2, 3-epoxypropyl trimethyl ammonium chloride and ferrous sulfate are added into the chitosan microsphere dispersion liquid, and the mixture is placed at the temperature of 40-60 ℃ to react for 7-9h under the condition of stirring, and then the mixture is washed.
5. The chitosan-based hemoperfusion adsorbent of claim 4, wherein the mass ratio of the chitosan microspheres to the 2, 3-epoxypropyltrimethylammonium chloride is 20-30:1.
6. A method of preparing a hemodynamic adsorbent as claimed in any one of claims 1 to 5, which comprises:
performing epoxidation modification on the chitosan microsphere to obtain a modified chitosan microsphere;
Adding the modified chitosan microsphere into an ethanol water solution containing chicoric acid, carrying out ultrasonic treatment for 40-50min, adding stannous chloride, and reacting at 70-80 ℃ for 3-4h to obtain a chicory acidified chitosan microsphere;
dispersing the chicory acidified chitosan microspheres into a glycol solution containing manganese nitrate hexahydrate, adding alkali liquor, performing pyrolysis reaction, centrifuging and washing to obtain the hemoperfusion adsorbent.
7. The method for preparing the hemoperfusion adsorbent of claim 6, wherein the mass ratio of the modified chitosan microsphere to chicoric acid is 15-20:1.
8. The method for preparing a hemoperfusion adsorbent as claimed in claim 6 wherein the mass ratio of modified chitosan microsphere to manganese nitrate hexahydrate is 6-8:1.
9. The method for preparing a hemoperfusion adsorbent according to claim 6, wherein the reaction temperature of the antipyretic reaction is 50-80 ℃ and the reaction time is 3-6 hours.
10. Use of a hemodynamic adsorbent as claimed in any one of claims 1-5 in the manufacture of a hemodynamic apparatus.
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