Application of PEG-b-PAA-g-SNO-IR780 NPs nanoparticles in preparation of antibacterial material
A technology of 1. peg-b-paa-g-sno, antibacterial materials, applied in the application field of preparing antibacterial materials, can solve the problems of simple, efficient and controllable release of peroxynitrite, etc., and achieve good blood flow. Compatibility, low cytotoxicity, effect of enhancing bactericidal effect
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Embodiment 1
[0043] Embodiment 1: in vitro antibacterial concentration
[0044] The effective antibacterial concentration of PSNO@IR780 NPs against Staphylococcus aureus and Escherichia coli was determined by CFU counting method. The fresh cultures of Staphylococcus aureus and Escherichia coli were first washed 3 times with PBS and diluted to 10 6 and 10 8 CFU / mL. The diluted bacterial suspension (200 μL) was added to the 96-well plate. Add PSNO@IR780 NPs into corresponding wells, so that the final concentration of IR780 photosensitizer in E. coli suspension is 10 μg / mL, 5 μg / mL, 2.5 μg / mL, 1.25 μg / mL; The final concentration of photosensitizer was 10 μg / mL, 2 μg / mL, 1 μg / mL, 0.5 μg / mL. After culturing for 0.5 h, the bacterial suspension was irradiated or not irradiated with 808nm (1 W / cm2) laser for 2 min. Dilute the treated bacterial solution, and apply 100 µL of the diluted solution on the solid medium. Incubate at 37°C for 12 h, count and calculate the survival rate of bacteria. ...
Embodiment 2
[0045] Embodiment 2: in vitro antibacterial activity evaluation
[0046] The antibacterial activity of PSNO@IR780 NPs against Staphylococcus aureus and Escherichia coli was determined by CFU counting method. First culture Staphylococcus aureus and Escherichia coli overnight in LB medium at 37°C, wash 3 times with PBS, and dilute to 10 6 and 10 8 CFU / mL. The diluted bacterial suspension (200 μL) was added to the 96-well plate. PBS, PSNO NPs, PEC@IR780NPs, and PSNO@IR780 NPs were respectively added to the corresponding wells and incubated for 0.5 h, and then 808 nm (1 W / cm 2 ) laser irradiation or not irradiating the bacterial suspension for 2 min. The IR780 concentration added to the Staphylococcus aureus suspension was 2 μg / ml, and the NO concentration was 10 μM; the IR780 concentration in the Escherichia coli suspension was 5 μg / ml, and the NO concentration was 50 μM. Dilute the treated bacterial solution, and apply 100 µL of the bacterial solution on the solid medium. ...
Embodiment 3
[0048] Example 3: Bacterial life and death staining
[0049] The treated bacteria were stained with bacterial dead-and-alive staining solution, and the survival of the bacteria was photographed under a laser confocal microscope. The green color was live bacteria, and the red color was dead bacteria. Staphylococcus aureus and Escherichia coli suspension (10 7 CFU / mL, 1 mL) and different nanoparticles (5 μg / mL), after co-incubating for 0.5 h, the bacterial suspension was irradiated or not irradiated with 808 nm (1 W / cm2) laser for 2 min. Bacteria were then isolated and washed three times by repeated centrifugation with PBS. The obtained bacteria were treated with STYO 9 / PI Live / Deadbacterial alive Kits (Thermo Fisher L7012) for 15 min, and finally the bacteria were imaged with a confocal laser scanning microscope (Nikon A1). According to the corresponding fluorescent photographs (Escherichia coli Figure 4 ) and (Staphylococcus aureus Figure 5 ) It can be seen that in the ...
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