Fig 1.
Treatment with SWT-α, SWT-β, and SWT-γ does not affect the viability of HT-29, Caco-2, HCT-116, or Vero E6 cells across 24–72 h.
HT-29, Caco-2, HCT-116 and Vero E6 were treated with SWT-α (A), SWT-β (B), and SWT-γ (C) at concentrations ranging from 25–1000 µM for 24, 48, and 72 h. Cell viability was assessed using the trypan blue exclusion assay and expressed as the percentage of viable cells relative to DMSO-treated control (Ctrl). No significant differences were observed among the tested concentrations at 24, 48 and 72 h for each cell line, compared with the DMSO control, using all SWT sources (p > 0.05). Data are presented as mean ± SD from three independent experiments, each performed in triplicate (n = 3).
Fig 2.
SWT-α, SWT-β, and SWT-γ fail to induce cytotoxicity in HT-29, Caco-2, HCT-116, and Vero E6 cells following 24, 48, and 72 hours of treatment.
HT-29, Caco-2, HCT-116 and Vero E6 were treated with SWT-α (A), SWT-β (B), and SWT-γ (C) at concentrations ranging from 25–1000 µM for 24, 48, and 72 h. Cytotoxicity was evaluated using the MTT assay, and values were expressed as % cytotoxicity relative to DMSO-treated control (Ctrl). No significant differences were observed among the tested concentrations at 24, 48 and 72 h for each cell line, compared with the DMSO control, using all SWT sources (p > 0.05). Data are presented as mean ± SD from three independent experiments, each performed in triplicate (n = 3).
Fig 3.
5-FU induce cytotoxicity in HT-29, Caco-2 and HCT-116 cells following 24, 48, and 72 hours of treatment.
HT-29 (A), Caco-2 (B) and HCT-116 (C) were treated with increasing concentrations of 5-FU (10–100 μM) for 24, 48, and 72 h. Cytotoxicity was evaluated using the MTT assay, and values were expressed as % cytotoxicity relative to untreated controls. Statistical significance was observed between treated (5-FU) and control (DMSO) cells as follows: HT-29, 24 h (p > 0.05) and 48–72 h (****p < 0.0001); Caco-2, 24 h (***p ≤ 0.001) 48-–72 h (****p < 0.0001); HCT116, 24 h (p > 0.05) and 48–72 h (****p < 0.0001). Data are presented as mean ± SD (n = 3).
Fig 4.
SWT-γ fails to potentiate the cytotoxic effect of 5-FU in HT-29 cells over 24–72 h.
HT-29 cells were treated with 5-FU, alone or with 1000 µM SWT, for 24 h (A), 48 h (B) and 72 h (C). Cytotoxicity under different treatment conditions was assessed using the MTT assay. Values are expressed as percent cytotoxicity relative to untreated controls. No statistically significant differences were observed between the combination treatment and 5-FU alone at any time point (P > 0.05). Data are presented as mean ± SD (n = 3).
Fig 5.
Transient membrane permeabilization or serum deprivation does not restore SWT-γ Cytotoxicity.
(A) HT-(A) HT-29 cells were treated with increasing concentrations of SWT-γ or DMSO for 24–72 h, with or without saponin pre-permeabilization (50 µg/mL). Cells were treated with SWT (100–1000 µM) or matched vehicle controls containing equivalent DMSO concentrations (maximum 0.4% v/v) in the presence of saponin; data were normalized to the corresponding vehicle controls. (B) HT-29, HCT-116, and Caco-2 cells were exposed to 1000 µM SWT-γ, DMSO, or serum-free media for 24 h. Cytotoxicity was assessed by MTT assay, and no significant differences were observed compared with controls (p > 0.05). Data are presented as mean ± SD (n=3).
Fig 6.
SWT-γ does not induce cytotoxicity in HepG2 cells.
HepG2 cells were treated for 72 h with various concentrations of SWT-γ (267–1068µM), 5-FU (100 µM), or DMSO (cntrl). Cytotoxicity was assessed by MTT assay and expressed as percent relative to untreated controls. SWT-γ did not induce significant cytotoxicity (P > 0.05), whereas 5-FU caused robust cytotoxicity (****P < 0.0001, student’s t test) Data are presented as mean ± SD (n = 3).
Fig 7.
Morphological assessment of CRC cells following SWT-γ treatment.
Caco-2, HT-29, and HCT-116 were either treated with DMSO (A–C) or with 1000 µM of SWT-γ (D–F). Images were captured using a 10 × objective lens and are shown with a scale bar of 100 µm.
Fig 8.
Wound closure of HT-29 cells over time following SWT-γ treatment.
(A) Representative light microscopy images of scratches in HT-29 cells at 0, 12, 24, and 48 h after being left untreated or treated with DMSO or 1000 µM SWT-γ. Images were captured using a 4x objective lens. Scale bar = 100 µm. (B) Quantification of wound closure over time, expressed as percentage closure. No statistically significant differences were observed between the untreated or treated with DMSO or 1000 µM SWT-γ at any time point (P > 0.05). Data are presented as mean ± SD (n = 3).
Fig 9.
Hydrogen peroxide (H₂O₂) scavenging activity of Ascorbic Acid and SWT-γ.
No significant difference was observed between ascorbic acid and SWT-γ (p > 0.05). Data are presented as mean ± SD (n = 3).
Table 1.
% Inhibition of E. coli and S. aureus after treatment with Amikacin at 16 h and 24h.
Table 2.
% Inhibition of E. coli and S. aureus after treatment with SWT-γ at 16 h and 24h.