Table 6.
Examples of selected nanomedicines discussed within the manuscript, pre-clinical models used in their evaluation, and the main readouts in vitro and in vivo.
| Nanomedicine | In vitro model(s) | Most relevant parameters in vitro | In vivo model(s) | Most relevant parameters in vivo | REF |
|---|---|---|---|---|---|
| Nanomedicines in Clinics / Clinical Trials | |||||
| Doxil® | 2D: MDA-MB-231-BR (brain-seeking subclone) | Cell toxicity | Experimental metastasis by MDA-MB-231-BR intracerebrally injected in athymic nude mice. | Antitumor efficacy, biodistribution, pharmacokinetics | [149] |
| Doxil® | – | – | GEMM FVB/n strain carrying a transgene for C3(1)-T-antigen (C3-TAg) and tumors derived from BALB/c T11/TP53Null orthotopically transplanted into BALC/c mice. | Antitumor efficacy, pharmacokinetics, macrophage levels, microvessel density | [6] |
| Unloaded CPC634 (Cristal Therapeutics) | – | – | 4 T1 orthotopically injected in BALB/c | Biodistribution, tumor accumulation, stromal distribution, uptake, ex vivo imaging | [185] |
| Ferumoxytol® (Feraheme®, AMAG Pharmaceuticals Inc., Cambridge, MA, USA) | 2D: MDA-MB-468, HUVEC 2D co-culture: RAW264.7 + MMTV-PyMT-derived cells // murine bone-marrow-derived macrophages + MMTV-PyMT-derived cells |
Cell toxicity, ROS generation, apoptosis | MMTV-PyMT-derived cancer cells orthotopically injected on female FVB/N mice. Experimental metastasis with intravenously injected KPI-GFP-Luc in NOD.Cg-PrkdcscidIL2rgtm1Wjl/SzJ mice. Metastatic to lung and liver. |
Antitumor efficacy, macrophages population study, metastasis | [69] |
| Myocet® | – | – | Domestic pigs | Cardiac safety |
[262] |
| Inorganic Systems | |||||
| MnO2-Platinum-Cobalt nanozymes | 2D: 4 T1, mouse embryonic fibroblasts NIH3T3 3D: 4 T1 cells on agarose-precoated wells |
Cell toxicity (2D and 3D), uptake (2D), ROS generation (2D and 3D), apoptosis, lipid peroxidation (2D), DNA double-strand breaks (2D), lysosomal disruption (2D) | 4 T1 subcutaneously injected in BALB/c | Antitumor efficacy, biodistribution, safety, pharmacokinetics | [82] |
| Titanium dioxide, silica, and gold NPs | 2D: Human microvascular endothelial cells, Human mammary microvascular endothelial cells, MDA-MB-231, MCF7 | Cell toxicity, migration assays, adhesion, uptake | MDA-MB-231-Luc orthotopically injected in NSG. Metastasis in lungs, liver, bone, and spleen | Antitumor efficacy, biodistribution, metastasis | [167] |
| PEGylated gold NPs | – | – | 4 T1 orthotopically injected in BALB/c, U87-MG subcutaneously injected in CD1 Nude, MMTV-PyMT GEMM in FVC/N strain, and TNBC PDX orthotopically injected in NOD-SCID | Tumor vasculature analysis, EPR effect, biodistribution, | [265] |
| Gold nanorod-assisted plasmonic photothermal therapy | – | – | Pet cats and dogs with spontaneous breast cancer tumors | Antitumor efficacy | [251] |
| Targeted inorganic systems | |||||
| Trastuzumab-labeled amine-functionalized starch-coated bionized nanoferrite nanoparticles | 2D: MCF7, MCF7/Neo, MCF/HER2, HCC1954, MDA-MB-231, BT474, SKBR3, RAW264.7 | Uptake, macrophage activation | MDA-MB-231, MCF7/Neo, MCF7/HER2, BT474, or HCC1954 cells orthotopically injected in athymic nude or NSG mice. Transgenic HuHER2-FVB/N mice-derived cells orthotopically implanted in athymic nude, NSG, or FVB/N mice. |
Antitumor efficacy, tumor accumulation | [151] |
| SPIO NPs containing a pH-responsive peptide H7K(R2)2 and paclitaxel | 2D: MDA-MB-231 | Cell toxicity, uptake | MDA-MB-231 subcutaneously injected in BALB/c nude mice | Antitumor efficacy, biodistribution | [192] |
| Magnetic IO NP-dually targeted peptides to Wnt/LRP5/6 and uPAR | 2D: MDA-MB-231 | Cell uptake, cell proliferation assay, invasion assay, cell cycle analysis, Western blotting | Orthotopic human breast cancer PDX in SCID mice | Antitumor efficacy, biodistribution | [98], [203] |
| PEG-coated carbon dots conjugated to folic acid encapsulating doxorubicin (CDs-PEG-FA/Dox) | 2D: HUVECs, T47D, and BT549 Microfluidics: HUVEC monolayer on channel A, T47D, or BT549 spheroids on channel C |
Cell toxicity, uptake, | – | – | |
| Liposomes | |||||
| Liposome containing doxorubicin and coated with anti-ICAM1 and anti- EGFR antibody | 2D: MDA-MB-231, MDA-MB-436, MDA-MB-157 and MCF10A | Cell toxicity, uptake, wound healing | MDA-MB-231-Luc orthotopically injected in nude. Experimental metastasis by MDA-MB-231-Luc lateral tail vein injection in nude mice, resulting in lung metastasis. |
Antitumor efficacy, safety, biodistribution, metastasis | [146] |
| HER2 peptide-targeted liposomal doxorubicin | 2D: BT-474, SK-BR-3, and MCF7 | Cell toxicity, uptake | Breast tumors generated by MMTV-neu mice were transplanted orthotopically in NOD-SCID mice. | Antitumor efficacy, biodistribution | [221] |
| Anginex-conjugated liposomal nanoparticles (nanobins) carrying arsenic trioxide and cisplatin | 3D: 4T1mCherry in hanging drop method. 3D co-culture: 4T1mCherry + C166-GFP + MEFs in hanging drop method (TTA: tumor tissue analogs). |
Cell toxicity, uptake | 3D 4T1mCherry or TTA orthotopically injected in athymic nude mice. Metastasis in the lungs. | Antitumor efficacy, tumor accumulation, metastasis | [95] |
| Anti-MUC1/CD44 dual-aptamer-conjugated liposomes encapsulating Dox | 2D: MCF7, MCF7-CSC. 3D: MCF7 on Matrigel layer. |
Cell toxicity (2D and 3D), Uptake (2D and 3D) | Experimental metastasis by MCF7-CSC and MCF7 lateral tail vein injection in athymic nude mice. Metastasis in lung and liver. | Metastasis | [148] |
| Polymeric Nanoparticles | |||||
| FA-PEG-PLGA-paclitaxel@indocyanine Green-Perfluorohexane NPs | 2D: MDA-MB-231, HUVEC | Cell toxicity, uptake | MDA-MB-231 orthotopically injected in BALC/c nude mice | Antitumor efficacy, biodistribution, safety | [193] |
| Rod-shaped PLGA nanoparticles encapsulating docetaxel | 2D: MD-MBA-231 | Cell toxicity | Taxane-resistant GEMMs: FVN/NJ strain carrying a transgene for C3(1) SV40 T-antigen | Antitumor efficacy, biodistribution, pharmacokinetics, dosing | [218] |
| Polymeric Micelles | |||||
| Lapatinib-loaded polycarbonate-doxorubicin conjugate micelles | 2D: 4 T1 | Cell toxicity, uptake, wound healing assay, invasion assay | 4 T1 subcutaneously injected in BALB/c | Antitumor efficacy, biodistribution, safety, | [75] |
| Polymeric micelles loaded with Zileuton® | 2D: MCF7 and MDA-MB-231. 3D: MCF7 and MDA-MB-231 in low attachment plates. |
Cell toxicity, CSC-resistance assay, cell transformation assay | MCF7 and MDA-MB-231 orthotopically injected in NOD-SCID mice | Antitumor efficacy, biodistribution, safety | [169] |
| Doxorubicin-loaded mixed micelles composed of TPGS and Tetronic® T1107 | – | – | Wild-type adult zebrafish | Biodistribution, safety | [238] |
| Polymer Conjugates | |||||
| PEG‐pLA‐pTBPC-Dox | 2D: MDA‐MB‐231, MCF7, HCC70, MDA‐MB‐468, MCF‐10A. 3D: MDA‐MB‐231 spheroids hanging drop method. |
Uptake (2D and 3D) Cytotoxicity (2D and 3D) | MDA-MB-231 orthotopically injected in CD-1 Nude mice | Antitumor efficacy, biodistribution, histology | [94] |
| PGA-Dox-(Glycine-AGM) | 2D: MCF7 and 4 T1 | Cell toxicity, Uptake | 4 T1 orthotopically injected in BALB/c | Antitumor efficacy, biodistribution, pharmacokinetics, | [182] |
| PGA-(Hydrazone-Dox)-(Glycine-AGM) | 2D: 4 T1 | Cell toxicity | 4 T1 orthotopically injected in BALB/c | Antitumor efficacy, metastasis, | [184] |
| High molecular weight dextran conjugated to a fluorophore (FITC-Dextran) | – | – | MDA-MB-231 orthotopically and subcutaneously injected in NSG | Tumor volume, Tumor accumulation, Vascularization, EPR effect | [132] |
| Additional Systems | |||||
| Chitosan-histidine-arginine/ plasmid DNA | 2D co-culture: MCF7 + primary normal human dermal fibroblasts | Uptake | – | – | [67] |
| Sterically stabilized mixed phospholipid nanomicelle encapsulating paclitaxel and decorated with VIP | 2D: MCF7 | Cell toxicity | Sprague Dawley rats intravenously injected with NMU | Antitumor efficacy, biodistribution, systemic arterial blood pressure |
[228] |
Abbreviations: AGM = aminoglutethimide; DOX = doxorubicin; EPR = enhanced permeability and retention; FA = folic acid; FITC = fluorescein isothiocyanate; GEMM = genetically modified mouse model; HPMA = hydroxypropyl methacrylamide; IHC = immunohistochemistry; IO = iron oxide; MEF = mouse embryonic fibroblasts; MMTV = mouse mammary tumor virus; NOD = non-obese diabetic; NP = nanoparticle; NSG = NOD scid gamma; pDNA = plasmid DNA; PDX = patient-derived xenograft; PEG = polyethylene glycol; PGA = poly-L-glutamic acid; pLA = poly(lactide); PLGA = poly(lactide-co-glycolic acid); PyMT = polyoma middle tumor-antigen; pTBPC = poly(2‐((tert‐butoxycarbonyl)amino)‐3‐propyl carbonate; SCID = severe combined immunodeficient; SPIO = Superparamagnetic iron oxide; TPGS = D-α-tocopheryl poly-ethylene glycol 1000 succinate