Fig. 12

(a) Wound Healing Assay: Typical images showing scratch repair in HCT 116 colorectal cancer cells following 24 h. The control cells exhibited a notable migration rate with 80.45% wound closure. Cells treated with MTX exhibited a moderate inhibition level of 66.35%, while MTX-SLNs considerably reduced migration to 53.44%, and the greatest inhibition was noted in MTX-Lf-SLNs at 30.23%, attributed to the targeting of lactoferrin receptors. Yellow lines denote scratch limits at 0 and 24 h. (b) Quantitative Wound Healing: The percentage of wound healing for each group per treatment assessed through histograms: MTX-Lf-SLNs have resulted in the most significant wound closure. Asterisk, p. (c) Hen’s Egg Test-Chorioallantoic Membrane (CAM) Assay: Bar graphs of sample images. The negative control used was PBS, while Avastin served as the positive control, which notably suppressed VEGF-induced angiogenesis. MTX demonstrated a moderate inhibition of vessels (~ 75% of the control), while MTX-SLNs enhanced the anti-angiogenic effect (~ 55% of the control), and MTX-Lf-SLNs exhibited the strongest inhibition (~ 25% of the control), indicating that lactoferrin plays a role in interfering with VEGF and improving nanoparticle absorption via the targeted receptor. (d) Quantitative Examination of Angiogenic Metrics: Proportional decrease in vessel length, size, and junctions compared to the control. MTX demonstrated moderate inhibition (~ 25%), while MTX-SLNs resulted in a more significant effect (~ 45%), and MTX-Lf-SLNs caused the most substantial suppression (~ 70%). Analysis with ImageJ validated these results.