Fig. 8 | Scientific Reports

Fig. 8

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. 8

(a) AO/PI fluorescence staining assay: Typical fluorescence images of HCT116 cells exposed to control, MTX, MTX-SLNs, and MTX-Lf-SLNs. Green fluorescence indicates live cells, while orange/red fluorescence signifies apoptotic cells. Control cells primarily displayed green fluorescence, indicating a significant level of viability. MTX led to heightened red/orange fluorescence, thus indicating apoptosis. Cells treated with MTX-SLNs exhibited increased apoptosis. In the scenario of MTX-Lf-SLNs treatment, apoptosis reached its highest level; the orange/red fluorescence was prevalent. (b) The bar chart displays fluorescence intensity readings of cells at 30 and 60 min. MTX-Lf-SLNs showed a notable decrease in viable cells while exhibiting a threefold rise in apoptotic cells. A statistically significant decline was observed in the viable cell populations (p < 0.05) for MTX-Lf-SLNs, while its apoptotic population increased in comparison to both MTX and MTX-SLNs (p < 0.001). Thus, it validated the successful transport of MTX via the targeted Lf ligand. These findings highlight that MTX-Lf-SLNs were more effective at inducing apoptosis owing to enhanced uptake and receptor-mediated targeting facilitated by lactoferrin.

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