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Core-shell structured heparinoid microsphere based on graphene oxide grafted urease, and preparation method and applications of core-shell structured heparinoid microsphere

A technology of ene-grafted urease and core-shell structure, applied in the field of heparin-like microspheres with core-shell structure and its preparation, can solve problems such as poor blood compatibility, achieve inhibition of complement activation and contact activation, low hemolysis rate, urea High removal effect

Active Publication Date: 2019-12-31
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, graphene oxide has poor blood compatibility, which will cause negative effects when it comes into direct contact with blood, and free two-dimensional nanosheets cannot be easily separated from blood.

Method used

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  • Core-shell structured heparinoid microsphere based on graphene oxide grafted urease, and preparation method and applications of core-shell structured heparinoid microsphere
  • Core-shell structured heparinoid microsphere based on graphene oxide grafted urease, and preparation method and applications of core-shell structured heparinoid microsphere

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Embodiment 1

[0035] A preferred embodiment of the present invention provides a method for preparing heparin-like microspheres with a core-shell structure based on graphene oxide grafted with urease. The specific steps are as follows:

[0036] S1. Core layer preparation: disperse graphene oxide in PBS buffered saline solution with a pH of 7.4-7.6 to obtain a solution concentration of 1mg / mL; add 0.01g NHS and 0.015g EDC and mix well, then add 0.2g urease, React at 4°C for 12 hours to obtain a urease-immobilized graphene oxide solution, which should be stored at 4°C before use;

[0037] S2. Shell layer preparation: Dissolve 8g PES in 88.8g DMAC to obtain a uniform solution, then add 0.96g AA, 0.72g AMPS, 0.04g AIBN and 0.06g MBA, react under nitrogen atmosphere at 75°C for 12h, cool to room temperature to obtain a heparan-like polymer solution;

[0038] S3. Preparation of microspheres: drop the urease-immobilized graphene oxide solution prepared in step S1 into the heparan-like polymer solu...

Embodiment 2

[0040] A preferred embodiment of the present invention provides a method for preparing heparin-like microspheres with a core-shell structure based on graphene oxide grafted with urease. The specific steps are as follows:

[0041] S1. Core layer preparation: disperse graphene oxide in PBS buffered saline solution with pH 7.4-7.6 to obtain a solution concentration of 1 mg / mL; add 0.01g NHS and 0.015g EDC and mix well, then add 0.18g urease, React at 4°C for 12 hours to obtain a urease-immobilized graphene oxide solution, which should be stored at 4°C before use;

[0042] S2. Shell layer preparation: Dissolve 8g PES in 89g DMAC to obtain a uniform solution, then add 0.97g AA, 0.73g AMPS, 0.04g AIBN and 0.06g MBA, react at 75°C for 12h under nitrogen atmosphere, and cool to room temperature , to obtain a heparan-like polymer solution;

[0043] S3. Preparation of microspheres: drop the urease-immobilized graphene oxide solution prepared in step S1 into the heparan-like polymer solut...

Embodiment 3

[0045] A preferred embodiment of the present invention provides a method for preparing heparin-like microspheres with a core-shell structure based on graphene oxide grafted with urease. The specific steps are as follows:

[0046] S1. Core layer preparation: disperse graphene oxide in PBS buffered saline solution with pH of 7.4-7.6 to obtain a solution concentration of 1 mg / mL; add 0.01g NHS and 0.017g EDC and mix well, then add 0.2g urease, React at 4°C for 12 hours to obtain a urease-immobilized graphene oxide solution, which should be stored at 4°C before use;

[0047] S2. Shell layer preparation: Dissolve 8g PES in 90g DMAC to obtain a uniform solution, then add 0.97g AA, 0.75g AMPS, 0.04g AIBN and 0.07g MBA, react at 75°C for 12h under nitrogen atmosphere, and cool to room temperature , to obtain a heparan-like polymer solution;

[0048] S3. Preparation of microspheres: drop the urease-immobilized graphene oxide solution prepared in step S1 into the heparan-like polymer s...

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Abstract

The invention discloses a core-shell structured heparinoid microsphere based on graphene oxide grafted urease, and a preparation method and applications of the core-shell structured heparinoid microsphere. According to the preparation method, a graphene oxide two-dimensional nanosheet layer is taken as a base material, immobilizing urease on the surface of the base material by carbodiimide methodso as to obtain a core solution; in-situ cross-linking polymerization is adopted to prepare a heparinoid polymer solution to obtain a shell solution; and reverse-forward phase inversion method is adopted to obtain the core-shell structured heparinoid microsphere. The core-shell structured heparinoid microsphere is capable of removing urea with high efficiency in hemoperfusion, relieving patient pain, and can also be used for dialysate regeneration in hemodialysis, and provide technical support for a wearable artificial kidney.

Description

technical field [0001] The invention belongs to the field of blood purification, and in particular relates to a heparin-like microsphere with a core-shell structure based on graphene oxide grafted with urease and a preparation method and application thereof. Background technique [0002] Chronic kidney disease (CKD) is a worldwide health problem that not only causes kidney failure but also complications such as cardiovascular disease (CVD). For CKD patients, excess uremic toxins accumulate in the body and cannot be effectively excreted. Hemoperfusion is one of the common methods to remove uremic toxins. Although activated carbon, zeolite and other adsorbents have been developed to adsorb urea, due to the high solubility and stability of urea in aqueous solution, no effective physical adsorption materials have been found to remove urea. [0003] Urease is an aminohydrolase whose active site contains two nickel(II) ions, which can catalyze the decomposition of urea into ammo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/24B01J20/28B01J20/30B01D15/08
CPCB01D15/08B01J20/24B01J20/28021
Inventor 何超张珏周密赵长生李明远罗一晏冰清郝媛媛刘尚宇
Owner SICHUAN UNIV
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