Nanometer enzyme bleeding stopping agent with functions of bleeding stopping and sterilization, and applications thereof
The technology of a hemostatic agent and nano-enzyme is applied in the medical application field of hemostatic antibacterial agent and nano material, which can solve the problems of long time-consuming hemostasis, adhesion, no antibacterial effect, etc., and achieves the effect of simple preparation method and broad-spectrum sterilization.
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[0039] Example 1 Polyacrylic acid (PAA), chitosan modified Fe 3 O 4 Synthesis and Characterization of Nanozyme Particles
[0040] In this example, PAA-modified Fc 3 O 4 The synthesis of the nanozyme particles was performed by a hydrothermal method. The details are as follows: 3g FeCl 3 Add 80 mL of ethylene glycol and mix well. After fully dissolving, add 6 g of anhydrous sodium acetate and 0.6 g of PAA, mix thoroughly for 30 minutes, then put the sample into the reaction kettle and react at 200°C for 14 hours. After cooling, magnetic separation, washing with ethanol, drying at 60°C, and storage in aliquots. Fe synthesized by this method 3 O 4 The surface of the nanozyme is modified with carboxyl groups.
[0041] In this example, chitosan-modified Fc 3 O 4 The nanozyme particle synthesis method is also a hydrothermal method. The details are as follows: 3gFeCl 3Add 80 mL of ethylene glycol and mix well. After fully dissolving, add 6 g of anhydrous sodium acetate and...
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[0042] Example 2Fe 3 O 4 Glucose Oxidase Modification of Nanozyme Particles
[0043] In this example, glucose oxidase was modified to Fe 3 O 4 The NHS-EDC method was used on the surface of the nanozyme. Details are as follows: 5 mg of EDC and NHS were dissolved in 1 mL of deionized water and vortexed. Then, 5 mg of the above-synthesized PAA-modified Fc was 3 O 4 Nanozyme was added to the solution and incubated at room temperature for 30 minutes. Carboxyl-activated Fe 3 O 4 Nanozyme particles were washed twice with deionized water by magnetic enrichment, and then 0.1 mg / mL of glucose oxidase was added. The reaction buffer was 50 mM sodium acetate at pH 6. After vortexing, the reaction system was incubated at 4°C overnight. Finally, glucose oxidase-modified Fe 3 O 4 Nanozyme particles were washed twice with PBS by magnetic enrichment, and finally stored in PBS buffer system.
[0044] Fe modified by glucose oxidase 3 O 4 Nanozyme particles and chitosan-modified Fe...
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[0045] Example 3 Fc modified by glucose oxidase 3 O 4 Coagulation effect of nanozyme particles and their comparison with chitosan-modified nanozymes
[0046] Chitosan has now been widely used as a hemostatic material. In this example, in order to quantitatively compare pure glucose oxidase-modified Fe 3 O 4 The coagulation ability of nanozymes, we choose in the synthesis process, namely Fe with chitosan modified 3 O 4 Nanozyme was used as a control, and in order to compare whether the modification of glucose oxidase affects Fe 3 O 4 Coagulation ability of nanozyme, select Fe without any modification 3 O 4 Nanozyme served as a positive control. In this example, the blood sample was derived from the experimental mouse C57BL / 6. The details are as follows: Freshly collected mouse blood is anticoagulated with citric acid anticoagulant. The principle of citric acid anticoagulation is mainly to chelate Ca in the blood 2+ ions, because the participation of calcium ions is ...
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