It was investigated whether loading multi-wall carbon
nanotubes
(CNTs) with two natural anticancer agents: ferulic acid (FUA) and
diosgenin (DGN), may enhance the anticancer effect of these drugs.
The CNTs were functionalized with carboxylic acid (CNTCOOH) or amine
(CNTNH2), loaded with the above pro-drugs, as well as both
combined and coated with chitosan or chitosan–stearic acid.
Following physicochemical characterization, the drug-loading properties
and kinetics of the drug’s release were investigated. Their
effects on normal human skin fibroblasts and MCF-7 breast carcinoma
cells, HepG2 hepatocellular carcinoma cells, and A549 non-small-cell
lung cancer cells were evaluated in vitro. Their actions at the molecular
level were evaluated by assessing the expression of lncRNAs (HULC,
HOTAIR, CCAT-2, H19, and HOTTIP), microRNAs (mir-21, mir-92, mir-145,
and mir-181a), and proteins (TGF-β and E-cadherin)
in HepG2 cells. The release of both pro-drugs depended on the glutathione
concentration, coating, and functionalization. Release occurred in
two stages: a no-burst/zero-order release followed by a sustained
release best fitted to Korsmeyer–Peppas kinetics. The combined
nanoformulation cancer inhibition effect on HepG2 cancer cells was
more pronounced than for A549 and MCF7 cells. The combined nanoformulations
had an additive impact followed by a synergistic effect, with antagonism
demonstrated at high concentrations. The nanoformulation coated with
chitosan and stearic acid was particularly successful in targeting
HepG2 cells and inducing apoptosis. The CNT functionalized with carboxylic
acid (CNTCOOH), loaded with both FUA and DGN, and coated with chitosan–stearic
acid inhibited the expression of lncRNAs and modulated both microRNAs
and proteins. Thus, nanoformulations composed of functionalized CNTs
dual-loaded with FUA and DGN and coated with chitosan–stearic
acid are a promising drug delivery system that enhances the activity
of natural pro-drugs.