Fig. 9 | Scientific Reports

Fig. 9

From: Study of caspase-6 activity in aggressive HCT116 cells using methotrexate-encapsulated lactoferrin-conjugated solid lipid nanoparticles via in silico and in vitro approaches

Fig. 9

(a) Flow cytometry plots showing the ROS levels in untreated control cells, MTX-treated cells, MTX-SLNs-treated cells, and MTX-Lf-SLNs-treated cells. The MTX formulations, particularly MTX-Lf-SLNs, significantly decreased overproduction of ROS while maintaining a balance between oxidative stress-induced apoptosis in cancer cells and protection against ROS-mediated damage. (b) ROS levels relative to untreated control cells were presented as a quantitative bar graph. Treatment with MTX-SLNs and MTX-Lf-SLNs significantly lowered the ROS levels compared with the control (p < 0.01, ANOVA). (c) Flow cytometry plots showing MMP changes in HCT116 cells with treatments. The control cells untreated showed that the mitochondria were fully functional (high red fluorescence), whereas the MTX-treated cells had depolarized mitochondria (increased green fluorescence). MTX-SLNs partly preserved the mitochondrial function, whereas MTX-Lf-SLNs showed the highest mitochondrial preservation. (d) Bar graph summarizing the red-to-green fluorescence ratio (R/G ratio) for MMP. MTX-Lf-SLNs-treated cells exhibited the highest mitochondrial integrity (R/G ratio: 4.4 ± 0.4), significantly higher than MTX (p < 0.001) and MTX-SLNs (p < 0.05), thus confirming superior mitochondrial protection and targeted delivery efficacy of MTX-Lf-SLNs.

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