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Updated: Aug 6, 2025
Cardiac Spheroids as in vitro Bioengineered Heart Tissues to Study Human Heart Pathophysiology
Published on: January 23, 2021
Demonstration of doxorubicin's cardiotoxicity and screening using a 3D bioprinted spheroidal droplet-based system
Raven El Khoury1,2, Salma P Ramirez1,2, Carla D Loyola1,2
1Inspired Materials & Stem-Cell Based Tissue Engineering Laboratory (IMSTEL), The University of Texas at El Paso El Paso TX 79968 USA bjoddar@utep.edu.
Abstract:
Doxorubicin (DOX) is a highly effective anthracycline chemotherapy agent effective in treating a broad range of life-threatening malignancies but it causes cardiotoxicity in many subjects. While the mechanism of its cardiotoxic effects remains elusive, DOX-related cardiotoxicity can lead to heart failure in patients. In this study, we investigated the effects of DOX-induced cardiotoxicity on human cardiomyocytes (CMs) using a three-dimensional (3D) bioprinted cardiac spheroidal droplet based-system in comparison with the traditional two-dimensional cell (2D) culture model. The effects of DOX were alleviated with the addition of N-acetylcysteine (NAC) and Tiron. Caspase-3 activity was quantified, and reactive oxygen species (ROS) production was measured using dihydroethidium (DHE) staining. Application of varying concentrations of DOX (0.4 μM-1 μM) to CMs revealed a dose-specific response, with 1 μM concentration imposing maximum cytotoxicity and 0.22 ± 0.11% of viable cells in 3D samples versus 1.02 ± 0.28% viable cells in 2D cultures, after 5 days of culture. Moreover, a flow cytometric analysis study was conducted to study CMs proliferation in the presence of DOX and antioxidants. Our data support the use of a 3D bioprinted cardiac spheroidal droplet as a robust and high-throughput screening model for drug toxicity. In the future, this 3D spheroidal droplet model can be adopted as a human-derived tissue-engineered equivalent to address challenges in other various aspects of biomedical pre-clinical research.
Insights
This study shows a 3D bioprinted cardiac model effectively screens doxorubicin cardiotoxicity, outperforming 2D cultures. Antioxidants like NAC and Tiron alleviate drug-induced damage in this advanced model.
Area of Science:
- Biomedical Engineering
- Cardiology
- Toxicology
Background:
- Doxorubicin (DOX) is a vital chemotherapy drug but causes significant cardiotoxicity, potentially leading to heart failure.
- The precise mechanisms of DOX-induced cardiotoxicity are not fully understood.
- Current cell culture models may not accurately replicate the complex cardiac environment.
Purpose of the Study:
- To investigate doxorubicin (DOX) cardiotoxicity using a novel 3D bioprinted cardiac spheroidal droplet system.
- To compare the efficacy of the 3D model against traditional 2D cell cultures in assessing DOX effects.
- To evaluate the protective potential of antioxidants, N-acetylcysteine (NAC) and Tiron, against DOX-induced damage.
Main Methods:
- Human cardiomyocytes (CMs) were cultured in 2D and a 3D bioprinted cardiac spheroidal droplet system.
- Cells were exposed to varying concentrations of DOX (0.4–1 μM).
- Caspase-3 activity, reactive oxygen species (ROS) production (via DHE staining), and cell viability were quantified. Flow cytometry assessed CM proliferation.
Main Results:
- DOX exhibited a dose-dependent cytotoxic effect, with 1 μM causing maximum toxicity.
- The 3D model demonstrated significantly lower viable cells (0.22 ± 0.11%) compared to 2D cultures (1.02 ± 0.28%) after 5 days.
- NAC and Tiron mitigated DOX-induced cardiotoxicity, reducing ROS and caspase-3 activity.
Conclusions:
- The 3D bioprinted cardiac spheroidal droplet model is a robust platform for high-throughput drug toxicity screening.
- This model offers a more physiologically relevant system for studying cardiotoxicity compared to 2D cultures.
- The findings support the potential of this 3D model as a human-derived tissue-engineered equivalent for preclinical research.
