In vitro cellular interaction of drug-loaded liposomes with 2D and 3D cell culture of U87-MG cell line
Tasneem Alsheleh, Manar Zraikat, Fadwa Daoud, Dana A. Alqudah, Sharif Abdelghany, Ahmed Abu Siniyeh, Walhan Alshaer
Abstract
The distinctive physiological and physical properties of 3D cultures that mimic tumor microenvironments in vivo make them more suitable for assessing the efficacy of drugs and nanoparticles compared to 2D culture models. Therefore, this study aims to examine and contrast how liposomes interact with cell cultures in both 2D and 3D models. Hanging drop technique was used to generate 3D spheroids. Cellular toxicity of Doxorubicin and Doxil®-liposomes was tested using an MTT assay. Cellular uptake of Doxil®-liposomes was investigated in 3D and 2D cell culture models using flow cytometry and confocal microscopy. Finally, migration and invasion assays were used to investigate the Doxil®-liposomes interaction with the two models 2D model and 3D model, respectively.
Introduction
Cancer is characterized by uncontrolled growth and proliferation, leading to the formation of malignant tumors that can invade adjacent tissues. Conventional chemotherapy lacks specificity, causing severe damage to both tumor and normal cells. This necessitates the development of more effective delivery systems to reduce the side effects and increase the efficiency of the antitumor drugs [1]. The use of nanotechnology in medicine, particularly for drug delivery, has rapidly expanded in recent years [2]. Nanoparticles have been used to improve the delivery of poorly water-soluble drugs by enabling faster dissolution in the bloodstream [3].
Materials and method
2.1. Cell lines and reagents
Human glioblastoma cell line (U-87MG) (ATCC HTB-14) and Human dermal fibroblast cell line (HDF) (ATCC PCS-201-012) were obtained from ATCC. Collagen type I and Doxorubicin were purchased from Sigma-Aldrich (Poole, UK). Doxil® 20mg\10ml was purchased from Johnson & Johnson (USA).
Results and discussion
3.1. Spheroids formation
Tumor spheroids were formed by hanging drop technique after three days of incubation; each drop in the petri dish contained the same uniform size of spheroids. Cells of 2D culture models lack cell-cell and cell-matrix interactions, which are presented in native tumors. In contrast, 3D culture models have an opportunity to culture cancer cells alone or with different cell types. Thus, it can closely mimic the native environment of tumors [14]. U87-MG cells were co-cultured with HDF and successfully formed a compacted spheroid structure to investigate tumor behavior and interaction in vitro.
Conclusion
Our study involved the use of 2D and 3D cell culture models to evaluate the efficiency of nanoparticle-based drug delivery systems. We developed a 3D model replicating the in vivo conditions of tumor structure and extracellular matrix to assess the delivery of liposomal nanoparticles to spheroids through a collagen matrix. This 3D model proved to be more informative than traditional 2D models as it provided a better understanding of nanoparticle interactions. We compared the interactions of liposomal-Doxil® with cellular targets in both 2D and 3D models and found that the interaction of nanoparticles with 2D cells was more straightforward than that with 3D models.
Citation: Alsheleh T, Zraikat M, Daoud F, Alqudah DA, Abdelghany S, Abu Siniyeh A, et al. (2025) In vitro cellular interaction of drug-loaded liposomes with 2D and 3D cell culture of U87-MG cell line. PLoS ONE 20(3): e0320374. https://doi.org/10.1371/journal.pone.0320374
Editor: B. H. Jaswanth Gowda, Queen’s University Belfast, UNITED KINGDOM OF GREAT BRITAIN AND NORTHERN IRELAND
Received: December 3, 2023; Accepted: February 18, 2025; Published: March 25, 2025
Copyright: © 2025 Alsheleh et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Data Availability: All relevant data are within the manuscript file.
Funding: This work was supported by Deanship of Scientific Research, The University of Jordan (grant no. 2446).
Competing interests: The authors have declared that no competing interests exist.
