Nano-enzyme diagnosis and treatment agent for acute kidney injury as well as preparation method and application of nano-enzyme diagnosis and treatment agent
A technology of acute kidney injury and nano-enzyme, applied in the field of biomedical materials, can solve the problems of large toxic and side effects, low utilization rate of small molecule drugs, limited curative effect, etc., and achieve good therapeutic effect, excellent biocompatibility and biosafety Effect
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Embodiment 1
[0053] Embodiment 1: synthetic platinum nanozyme diagnosis and treatment agent
[0054] Synthesis of platinum nanozyme diagnostic agent: such as figure 1 As indicated, 1 g of chloroplatinic acid hexahydrate was added to a 90 mmol / L solution of polyvinylpyrrolidone in ethylene glycol, then stirred and heated to 120°C. NaOH solution (1 mol / L, 500 μL) was added dropwise to initiate platinum nucleation and reduction reaction, and stirring was continued for 30 minutes. After cooling to room temperature, it was centrifuged by ultrafiltration and washed several times with deionized water. The black product was dissolved in water after lyophilization and quantified by ICP-MS.
[0055] figure 1 is a roadmap for the synthesis of nanozyme diagnostic agents, where H 2 PtCl 6 Represents chloroplatinic acid, PVP represents polyvinylpyrrolidone, and NaOH represents sodium hydroxide. The surface ligands in the nanozyme therapeutic agent can well stabilize platinum nanoparticles.
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Embodiment 2
[0057] Example 2: The ability of platinum nanozyme diagnostic agent to scavenge various reactive oxygen species / active nitrogen, and the effect of platinum nanozyme diagnostic agent to scavenge hydroxyl free radicals
[0058] The efficiency of scavenging hydroxyl radicals of different concentrations of platinum nanozyme diagnostic agents (0-50 μg / mL) was determined by the hydroxyl radical antioxidant capacity (HORAC) kit (Cell Biolabs, Inc., USA). Testing was performed according to the protocol provided by the manufacturer.
[0059] Such as Figure 4 As shown, the platinum nanozyme diagnostic agent can effectively scavenge hydroxyl radicals, and has a concentration-dependent characteristic.
[0060] The superoxide anion scavenging efficiency of different concentrations of platinum nanozyme diagnostic agent (0-20 μg / mL) was determined by SOD detection kit (Sigma-Aldrich, USA). Testing was performed according to the protocol provided by the manufacturer.
[0061] Such as Fi...
Embodiment 3
[0068] Example 3: Cytotoxicity of Platinum Nanozyme Therapeutic Agent and Protection of Kidney Cells by Scavenging Various Reactive Oxygen / Reactive Nitrogen The standard MTT method was used to evaluate the effect of platinum nanozyme diagnostic agent on the survival rate of 293T renal embryonic cells.
[0069] 293T cells at 1×10 per well 4 Density seeded into 96-well plate, and placed in 37 degrees, 5% CO 2 Cultivate under the condition for 12h. Next, suck out the old culture medium in the 96-well plate, and add culture medium solutions containing different concentrations of platinum nanozyme therapeutic agent respectively. After continuing to cultivate for 20 or 44 hours, suck out the old medium in the 96-well plate, add 100 μL of MTT medium solution (0.8 mg / mL) to each well, and continue to cultivate for 4 hours. Aspirate the residual medium in the 96-well plate, Add 150 μ L of DMSO solution to each well, after shaking gently, detect the OD value of each well (detection wa...
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