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MXenes are novel 2-D materials which have been extensively investigated for use in advanced biocomposites, water purification, biosensors, bioimaging and antibacterial systems. However, the knowledge associated with the mechanism of cytotoxic response is still limited to lack of verification against 3D spheroid-type cultures. Herein, we present a report on the in vitro cytotoxicity of Ti₂CT_x MXene against cervical cancer cell lines (HeLa) in a form of 3D spheroid with comparison to 2D cell culture system. Ti₂CT_x MXenes can be characterized by their multi-layered structure that is produced by the efficient elimination of Al and appearance of the surface functional groups. The biological results with 2D and 3D HeLa indicated that the Ti₂CT_x MXene was moderately cytotoxic to cells and the cytotoxicity was dose dependent. Our results showed that the toxicity of MXenes is potentially due to direct contact of the Ti₂CT_x …