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Updated: Jul 12, 2025
Establishing 3-Dimensional Spheroids from Patient-Derived Tumor Samples and Evaluating their Sensitivity to Drugs
Published on: December 16, 2022
Biological analyses of the effects of TiO2 and PEG-b-PLA nanoparticles on three-dimensional spheroid-based tumor
K Koňáriková1, G M Girašková, I Žitňanová
1Institute of Medical Chemistry, Biochemistry and Clinical Biochemistry, Faculty of Medicine, Comenius University, Bratislava, Slovak Republic. katarina.konarikova@fmed.uniba.sk.
Abstract:
The aim of our study was to monitor the antiproliferative/ cytotoxic and genotoxic effects of both, poly(ethylene glycol)-block-poly(lactic acid) (PEG-b-PLA) and titanium dioxide (TiO2) nanoparticles on the tumor (HT-29, MCF-7, U118MG) and healthy (HEK-293T) cell lines during 2D cultivation and during cultivation in the spheroid form (3D cultivation). Cells or spheroids were cultivated with nanoparticles (0.01, 0.1, 1, 10, 50, and 100 ?g/ml) for 72 hours. The cytotoxic effect was determined by the MTT test and the genotoxic effect by the comet assay. We found that 2D cultivation of tumor cell lines with PEG-b-PLA and TiO2 nanoparticles had an anti-proliferative effect on human colon cancer cell line HT-29, human breast cancer cell line MCF-7, human glioma cell line U-118MG during 72h cultivation, but not on control/healthy HEK-293T cells. At the concentrations used, the tested nanoparticles caused no cytotoxic effect on tumor cell lines. Nanoparticles PEG-b-PLA induced significant damage to DNA in HT-29 and MCF-7 cells, while TiO2 nanoparticles in MCF-7 and U-118MG cells. Only PEG-b-PLA nanoparticles caused cytotoxic (IC50 = 7 mikrog/ml) and genotoxic effects on the healthy cell line HEK-293T after 72h cultivation. The cells which were cultivated in spheroid forms were more sensitive to both types of nanoparticles. After 72h cultivation, we observed the cytotoxic effect on both, the tumor and healthy cell lines.
Insights
Poly(ethylene glycol)-block-poly(lactic acid) (PEG-b-PLA) and titanium dioxide (TiO2) nanoparticles showed anti-proliferative effects on tumor cells in 2D cultures. 3D spheroid cultures were more sensitive to nanoparticle cytotoxicity.
Area of Science:
- Biomaterials Science
- Nanotechnology
- Cancer Research
Background:
- Nanoparticles are increasingly used in biomedical applications, necessitating thorough safety evaluations.
- Understanding nanoparticle interactions with different cell types and culture conditions is crucial for therapeutic development.
Purpose of the Study:
- To investigate the antiproliferative, cytotoxic, and genotoxic effects of PEG-b-PLA and TiO2 nanoparticles.
- To compare these effects in 2D cell cultures versus 3D spheroid models.
- To assess nanoparticle toxicity on both tumor and healthy cell lines.
Main Methods:
- Utilized MTT assay for cytotoxicity and comet assay for genotoxicity.
- Tested poly(ethylene glycol)-block-poly(lactic acid) (PEG-b-PLA) and titanium dioxide (TiO2) nanoparticles.
- Cultivated human tumor cell lines (HT-29, MCF-7, U118MG) and a healthy cell line (HEK-293T) in 2D and 3D spheroid formats.
Main Results:
- PEG-b-PLA and TiO2 nanoparticles exhibited anti-proliferative effects on tumor cells in 2D cultures but not on healthy cells.
- PEG-b-PLA induced DNA damage in HT-29 and MCF-7 cells; TiO2 induced damage in MCF-7 and U118MG cells.
- PEG-b-PLA showed cytotoxic and genotoxic effects on healthy HEK-293T cells at higher concentrations.
- 3D spheroid cultures demonstrated increased sensitivity to nanoparticle-induced cytotoxicity for both tumor and healthy cells.
Conclusions:
- Nanoparticle effects vary significantly between 2D and 3D culture models.
- PEG-b-PLA and TiO2 nanoparticles possess distinct genotoxic profiles.
- The heightened sensitivity of 3D spheroids suggests potential challenges for nanoparticle-based therapies in vivo.
