A kind of exosome-superoxide dismutase nano preparation with anti-aging effect and preparation method thereof
A technology of dismutase nano and superoxide, applied in the field of exosome-superoxide dismutase nano preparation and its preparation, can solve the problems of limited application, instability, short half-life, etc., to improve health status and enzyme activity Low-impact, long-release effects
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Embodiment 1
[0024] First, human embryonic kidney epithelial cells (HEK293) were cultured in DMEM medium containing 10% (volume percentage) fetal bovine serum (FBS) from which exosomes were removed, and the culture conditions were 37 ° C, 5% (volume Percentage) carbon dioxide environment, after the cell confluency reaches 90%, obtain 500 mL of HEK293 cell supernatant.
[0025] The above-mentioned HEK293 cell supernatant was centrifuged at 300g for 10min to remove suspended cells, centrifuged at 2000g for 10min to remove cell debris, and centrifuged at 10000g for 30min to remove larger particulate matter; the obtained supernatant was filtered through a 0.22μm filter membrane to remove particles larger than 220nm. Then, the supernatant was concentrated through an ultrafiltration centrifuge tube with a molecular weight cut-off of 100kDa, and the concentrated supernatant from the upper layer of the ultrafiltration centrifuge tube was centrifuged at 100,000g for 1.5h in an ultracentrifuge tube t...
Embodiment 2
[0029]Dilute the empty exosome solution synthesized in Example 1 and the PBS suspensions of three exosome-superoxide dismutase nano-preparations (SOD@EXO) to 1 mg / mL with PBS buffer, and spread on Copper mesh covered with 300-mesh carbon film was air-dried at room temperature. The copper grid coated with the nano-preparation was detected by a transmission electron microscope, and the accelerating voltage was 200kV. Such as figure 1 As shown, it can be clearly observed that the exosomes loaded with SOD molecules are spherical.
Embodiment 3
[0031] Dilute the exosome-loaded solution obtained in Example 1 and the PBS suspensions of the three exosome-superoxide dismutase nano-preparations (SOD@EXO) to 1 mg / mL with PBS buffer, and then dissolve In the PBS buffer filtered through a 0.22 μm filter membrane, a 10 μg / mL solution was finally prepared, and the particle size of the samples was tested with a Zetasizer Nano ZS90 particle size potentiometer. As shown in Table 1, the particle size of exosomes before and after loading SOD was between 100nm and 200nm.
[0032] Table 1: Particle size characterization data of empty exosomes synthesized in Example 1 and three nano-preparations
[0033]
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