Perspectives in Nanotoxicity Studies for Carbon‐Based Nanomaterials in 3D Platform

The conventional two-dimensional (2D) in vitro cell culture system has been a staple in biological research for decades. However, due to numerous limitations, this strategy is being replaced by 3D cell culture, representing a significant advancement in cell culture techniques. The 3D culture has garnered substantial attention across various scientific fields due to its ability to replicate in vivo-like physiological conditions. In the same spirit, mimicking in vivo scenarios is crucial for assessing the adverse effects of materials, in particularly carbon-based nanomaterials. This review highlights recent advancements in cell culture techniques and various innovative 3D cell culturing strategies, and concurrently addresses the current status and limitations of cytotoxicological studies related to carbon-based nanomaterials. The application of 3D cell cultures in toxicological studies of carbon-based nanomaterials is discussed with an emphasis on the potential of these advanced techniques. Additionally, the review delves into the remaining challenges associated with strategies for assessing the toxicity of carbon nanomaterials in 3D cell culture platforms and explores future perspectives in this evolving field.