Antitumor Effects of Melittin-Functionalized Fe3O4 Magnetic Nanoparticles in Collagen-I-Based Caco-2/BJ/HUVEC Spheroids Enhanced by Magnetic Hyperthermia
- 1Department of Cell and Molecular Biology, Faculty of Medicine, “Iuliu Hațieganu” University of Medicine and Pharmacy, 6 Louis Pasteur St., 400349 Cluj-Napoca, Romania
- 2Department of Physiology, Faculty of Medicine, “Iuliu Hațieganu” University of Medicine and Pharmacy, 1–3 Clinicilor St., 400012 Cluj-Napoca, Romania
- 3Department of Pharmaceutical Physics-Biophysics, Faculty of Pharmacy, “Iuliu Hațieganu” University of Medicine and Pharmacy, 6 Louis Pasteur St., 400349 Cluj-Napoca, Romania
Abstract
This study evaluated the integrated biochemical and ultrastructural response of three-dimensional colorectal cancer spheroids to magnetic hyperthermia (MH) mediated by melittin-functionalized Fe3O4 magnetic nanoparticles (MNPs-Mel). The spheroids comprised human Caco-2 colorectal adenocarcinoma cells, dermal fibroblasts (BJ), and human umbilical vein endothelial cells (HUVECs) in a ratio of 3:1:1 in a collagen-I matrix. They were assigned to control, alternating magnetic field (AMF) control (Control-MH), free melittin (Mel), unloaded MNPs, MNPs-MH, MNPs-Mel, and MNPs-Mel-MH groups and assessed at 24 and 72 h. MTS-reducing activity (colorimetry) and ATP-associated luminescence were quantified as viability markers, while VEGF, HIF-1α, and caspase-9 concentrations measured by ELISA were used to assess angiogenesis and apoptosis induction. MNPs’ size, shape and internalization and treatment-associated ultrastructural changes were examined by transmission electron microscopy (TEM). Treatment, time, and their interaction significantly affected all biochemical endpoints. Every intervention reduced MTS absorbance relative to control at both time points. MNPs-Mel produced lower MTS values than unloaded MNPs, whereas MNPs-Mel-MH generated the lowest ATP signal at both time points and the lowest MTS signal at 72 h. The combined treatment also reduced VEGF and increased caspase-9, maximal at 72 h, while the increase in HIF-1α at 24 h was not maintained at 72 h. TEM showed intracellular MNPs-Mel and a more restricted distribution within spheroids than in the separately examined monocultures. Free Mel and unloaded MNPs produced time-dependent ultrastructural changes, while MH accentuated the changes associated with MNP exposure. MNPs-Mel-MH led to pronounced mitochondrial polymorphism, cristae loss, matrix rarefaction, autophagic-lysosomal alterations, endoplasmic reticulum expansion, reduced intercellular adherence, and extensive disruption of Caco-2 cells, particularly at 72 h. However, biochemical endpoints represent the response of the whole spheroid and do not distinguish effects on Caco-2 cells from those on BJ fibroblasts or HUVECs. The molecular and ultrastructural findings identify MNPs-Mel-MH as the treatment associated with the broadest response in this tri-cellular spheroid model, linking sustained metabolic suppression and reduced pro-angiogenic signaling with marked mitochondrial and cellular injury.
Related
- Impact of shape modification of cobalt-ferrite nanoparticles on their magnetic and magnetothermal properties for magnetic hyperthermia
- Optimization of Doxorubicin Loading into Citrus Lemon-Derived Nanoparticles and Evaluation of Their Antitumor Effects on Tumor Cell Lines
- Electrochemical Sensing of Dopamine with a Nafion-Coated Reduced Graphene Oxide/Polypyrrole-Functionalized Magnetic Nanoparticles Composite
- Oligoglycine-functionalized gold nanoparticles: physicochemical characterisation and serum protein interactions
- Thermo-photo catalytic partial oxidation of methane on hematite nanosheets functionalized with silver nanoparticles
BibTeX
@article{campian2026antitumor,
title = {{Antitumor Effects of Melittin-Functionalized Fe3O4 Magnetic Nanoparticles in Collagen-I-Based Caco-2/BJ/HUVEC Spheroids Enhanced by Magnetic Hyperthermia}},
author = {Alex Câmpian and Ioana Bâldea and Mara Muntean and Cristian Iacoviță and Nicoleta Decea and Adrian Florea},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27209302},
url = {https://www.mdpi.com/1422-0067/27/20/9302},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Antitumor Effects of Melittin-Functionalized Fe3O4 Magnetic Nanoparticles in Collagen-I-Based Caco-2/BJ/HUVEC Spheroids Enhanced by Magnetic Hyperthermia
AU - Alex Câmpian
AU - Ioana Bâldea
AU - Mara Muntean
AU - Cristian Iacoviță
AU - Nicoleta Decea
AU - Adrian Florea
PY - 2026
DA - 2026/10/20
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27209302
UR - https://www.mdpi.com/1422-0067/27/20/9302
AB - This study evaluated the integrated biochemical and ultrastructural response of three-dimensional colorectal cancer spheroids to magnetic hyperthermia (MH) mediated by melittin-functionalized Fe3O4 magnetic nanoparticles (MNPs-Mel). The spheroids comprised human Caco-2 colorectal adenocarcinoma cells, dermal fibroblasts (BJ), and human umbilical vein endothelial cells (HUVECs) in a ratio of 3:1:1 in a collagen-I matrix. They were assigned to control, alternating magnetic field (AMF) control (Control-MH), free melittin (Mel), unloaded MNPs, MNPs-MH, MNPs-Mel, and MNPs-Mel-MH groups and assessed at 24 and 72 h. MTS-reducing activity (colorimetry) and ATP-associated luminescence were quantified as viability markers, while VEGF, HIF-1α, and caspase-9 concentrations measured by ELISA were used to assess angiogenesis and apoptosis induction. MNPs’ size, shape and internalization and treatment-associated ultrastructural changes were examined by transmission electron microscopy (TEM). Treatment, time, and their interaction significantly affected all biochemical endpoints. Every intervention reduced MTS absorbance relative to control at both time points. MNPs-Mel produced lower MTS values than unloaded MNPs, whereas MNPs-Mel-MH generated the lowest ATP signal at both time points and the lowest MTS signal at 72 h. The combined treatment also reduced VEGF and increased caspase-9, maximal at 72 h, while the increase in HIF-1α at 24 h was not maintained at 72 h. TEM showed intracellular MNPs-Mel and a more restricted distribution within spheroids than in the separately examined monocultures. Free Mel and unloaded MNPs produced time-dependent ultrastructural changes, while MH accentuated the changes associated with MNP exposure. MNPs-Mel-MH led to pronounced mitochondrial polymorphism, cristae loss, matrix rarefaction, autophagic-lysosomal alterations, endoplasmic reticulum expansion, reduced intercellular adherence, and extensive disruption of Caco-2 cells, particularly at 72 h. However, biochemical endpoints represent the response of the whole spheroid and do not distinguish effects on Caco-2 cells from those on BJ fibroblasts or HUVECs. The molecular and ultrastructural findings identify MNPs-Mel-MH as the treatment associated with the broadest response in this tri-cellular spheroid model, linking sustained metabolic suppression and reduced pro-angiogenic signaling with marked mitochondrial and cellular injury.
SN - 1422-0067
ER -