Table 4.
In vitro evaluation studies of the different compound-loaded liposomes.
| Formulation/Compound (No/Name) | Assay (Cell Lines) | Main Results | Reference |
|---|---|---|---|
| F1/Cuphen | MTS assay (MNT-1 and B16F10) | IC50 = 4.4 ± 0.2 µM (MNT-1) and 5.1 ± 0.1 µM (B16F10) versus 3.1 ± 0.2 µM (MNT-1) and 3.3 ± 0.3 µM (B16F10) for free Cuphen | [97] |
| Flow cytometry (MNT-1) | Loss of cell viability = 80% | ||
| Hemolytic activity assay | Hemolysis < 4% | ||
| F2/Cuphen | MTS assay (B16F10) | IC50 = 2.6 ± 0.9 µM versus 3.4 ± 0.6 µM for free Cuphen | [101] |
| F3/AN169 | MTT assay (Mel 3.0) | IC50 = 0.8 ± 0.01 µM versus 0.75 ± 0.04 µM for free compound | [178] |
| F4/4-S-CAP | Trypan blue dye-exclusion method (B16) | Relative cell number = 46.6 ± 0.9% (400 μM) | [182] |
| F5/Bis-AO | MTT (B16F10, A549, SKOV-3 and NIH3T3) | Cell viability around 18%, 85%, 22% and 85%, respectively (10 µM) | [52] |
| Flow cytometry (B16F10) | ↑ Necrotic cells accumulation | ||
| Western Blot (B16F10) | Up-regulation of RIP1 cleaved fragments Up-regulation of caspase-8 |
||
| F6/Edelfosine | MTT assay (A375) | ↑ Tumor cells death (to 48.0 ± 4.1%) | [53] |
| F7/Celecoxib + Plumbagin | MTS assay (UACC 903 and 1205 Lu) | Cell viability ~25% | [54] |
| Western Blot (UACC 903 and 1205 Lu) | ↑ COX-2 levels ↓ Protein levels of pSTAT3 (Y705) ↓ Cyclins B1, D1 and H Induction of caspase-3/7 activity |
||
| F8/C6 | MTT assay (WM-115, SK-Mel2, WM-266.4 and A-375) | Cell survival ~40% (WM-115), ~35% (SK-Mel2), ~10% (WM-266.4) and ~55% (A-375) (10 µM) | [55] |
| Colorimetric assay (WM-115) | ↑ Activity of caspse-3 and caspase-9 | ||
| Flow cytometry and ELISA assay (WM-115) | ↑ Annexin V percentage and ssDNA ELISA OD | ||
| Western Blot (WM-115 and A-375) | ↑ Protein phosphatase activity (PP1) Inhibition of Akt-mTOR signaling |
||
| F9/ATK | Hemolytic activity assay | 0.55% hemolysis versus arachidonyl trifluoromethyl ketone dissolved in ethanol (2.9%) | [56] |
| MTS assay (UACC 903 and 1205 Lu) | IC50 = 20 μmol/L | ||
| Western Blot (UACC 903 and 1205 Lu) | ↑ Caspase-3/7 activity ↓ Levels of cyclin D1 and cPLA2 activity ↑ p21 and p27, cleaved PARP and LC3B and COX-2 protein expression ↓ Levels of pAKT and pSTAT3 |
||
| F10/3N-TPI | MTT assay (A375P and B16F10) | IC50 = 372.5 ± 42.5 µM (A375P cells) and >400 µM (B16F10 cells) versus 13.0 ± 0.5 µM (A375P cells) and 33.3 ± 1.0 (B16F10 cells) for free compound | [184] |
| Hemolytic activity assay | No hemolysis at <2 mM | ||
| Cell migration assay (B16F10) | ↓ Wound healing of about 40–50% (100, 200 and 400 µM) | ||
| Western Blot (B16F10) | Inhibition of VEGF and MMP-2 activity | ||
| F11/PHO-S | MTT assay (B16F10) | IC50 = 0.8 mM versus 4.4 mM for free form | [185] |
| Flow cytometry (B16F10) | ↑ Population of cells in the G2/M phase (20.5 ± 1.2% versus 14.4 ± 1.3% for free compound) | ||
| F12/PHO-S | Flow cytometry (B16F10) | ↑ TRAIL-DR4 receptor expression 8.4 ± 0.4% Modulation of the expression of caspases 3 (11.7 ± 0.3%) and 8 (29.8 ± 5.5%) at 2 mM ↑ Free cytochrome c in the cytoplasm (4.4 ± 0.6%) at 2 mM |
[186] |
| F13/Camptothecin | Bioluminescence assay (B16F10) | Cell viability = 18% versus 32% of non-targeted liposome and 48% of free camptothecin (50 µM) | [57] |
| F14/BP | MTT assay (B16F10, K-balb) | IC50 = 12.2 and 15.3 µg/mL, respectively | [188] |
| Flow cytometry (B16F10) | ↑ Cell cycle arrest at G0/G1 phase ↓ Protein expression of RB, p-RB, CDK4 and cyclin D1 ↑ Protein expression of P53, p-P53 and P21 |
||
| TUNEL assay (B16F10) | Chromatin condensation DNA fragmentation Apoptotic bodies |
||
| Immunocytochemistry | Activation of Fas, FasL, Cleaved-Cas-8 Activation of Bax, AIF, Cleaved-Cas-9 |
||
| Western Blot (B16F10) | Activation of caspase-3, -8 and -9 | ||
| F15/Leelamine | Hemolytic activity assay | 3.3% hemolysis versus leelamine dissolved in DMSO (15.8%) | [189] |
| MTS assay (UACC 903 and 1205 Lu) | IC50 = 2.3 µM | ||
| ELISA assay (UACC 903 and 1205 Lu) | ↓ Cellular proliferation ↑ Cellular apoptosis |
||
| Flow cytometry (UACC 903 and 1205 Lu) | ↑ Sub-G0/G1 and G0/G1 cell populations | ||
| Western Blot (UACC 903 and 1205 Lu) | ↓ Activity of PI3K/Akt, STAT3 and MAPK Inhibition of Akt phosphorylation ↓ Expression of cyclin D1 ↑ Cleaved caspase-3 and PARP protein levels Inhibition of phosphorylation of STAT3 |
||
| F16/Anthocyanin | DPPH assay | Radical-scavenging activity = 64 and 76% (20 and 50 mg/mL, respectively) | [190] |
| MTT assay (A375) | Cell viability = 80% (200 mg/mL) | ||
| Melanin content assay (A375) | ↓ Melanin production (inhibitory effect of 60% versus 30% of free anthocyanin at 50 mg/mL) | ||
| Cellular tyrosinase assay (A375) | Inhibition of tyrosinase activity (58% versus 30% of free anthocyanin at 50 mg/mL) | ||
| Western Blot (A375) | Inhibition of tyrosinase and MITF expression | ||
| F17/Juglone | MTT assay (B16F10) | IC50 = 4.1 µM versus 7.8 µM for free compound | [59] |
| F18/Lactoferrin | MTS assay (B16F10) | ↓ Cell viability 10–15% regarding free compound | [191] |
| Flow cytometry (B16F10) | ↑ Cell cycle arrest at G0-G1 phase | ||
| F19/CytD | MTT assay (B16) | Relative inhibition = 73.3 ± 8.9% (7.5 μg/mL) | [60] |
| TUNEL assay (B16) | Induction of cell apoptosis | ||
| F20/natsumikan extracts | WST-1 (B16) | ↑ Inhibitory effect comparing to free extract | [192] |
| Fluorescence microscopic assay (B16) | Induction of apoptosis |