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Archive > Volume 30 Issue 6 > 2023,30(6):464-472. DOI:10.3872/j.issn.1007-385X.2023.06.002 Prev Next
Basic Research
Preparation of liposome coated hollow copper sulfide nano-enzyme and their combined laser irradiation to against melanoma B16-F10 cells
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Abstract:
Objective: To construct a hollow copper sulfide nano-enzyme lipid composite nanocarrier CuS@LIP, and to further investigate the effect and mechanism of CuS@LIP combined laser irradiation on killing melanoma B6-F10 cells. Methods: (2, 3-dioleoyloxy-propyl) -3-trimethylammonium-propane (chloride salt) (DOTAP) cationic liposomes (LIPs) were applied to encapsulate copper sulfide (CuS) to construct cationic liposome-coated copper sulfide nanocarriers (CuS@LIP). The photothermal properties and stability of CuS and CuS@LIP at different concentrations were studied under 1 064 nm laser irradiation. The peroxidase-like activity of CuS@LIP was verified by a catalytic activity detecting system of hydrogen peroxide (H2O2) and 3, 3', 5, 5' -tetramethylbenzidine(TMB). B16-F10 cells were treated respectively with a series of mass concentration gradients of CuS or CuS@LIP with or without laser. The cell viability of B16-F10 cells was determined via CCK-8 assay. Calcein-AM/PI staining and Annexin Ⅴ-FITC/PI staining combined with flow cytometry detected the effects of 20 μg/mL CuS or CuS@LIP on the cell viability and apoptosis of B16-F10 cells under laser or non-laser irradiation, respectively. Results: The average particle size of successfully prepared CuS@LIP was (178.23±6.46) nm and the average Zeta potential was (20.47±0.93) mV. The highest temperature of 80 μg/mL CuS@LIP under laser irradiation was 65.4 ℃, which was higher than that of CuS at 63.4 ℃. The endpoint temperature of CuS@LIP was basically unchanged through the cyclic test of three laser on/off switches. In addition, CuS@LIP had the same peroxidase-like catalytic activity as CuS. CuS@LIP had no significant effect on the proliferation activity of B16-F10 cells in vitro when it was less than 20 μg/mL (P>0.05), but the survival rate was significantly reduced by 58.18% after laser irradiation ([29.76±3.60]% vs [87.95±8.18]%, P<0.000 1). The apoptosis rate was significantly increased ([19.34±4.41]% vs [13.36±0.86]% , P<0.01). Conclusion: The prepared CuS@LIP has physical and chemical properties, good photothermal properties and excellent peroxidase-like catalytic activity that meet the design requirements,and it shows a better effect of killing B16-F10 cells when combined with laser irradiation.
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