The combination of titanium dioxide nanoparticles with multiwall carbon nanotubes enables the generation of a hybrid nanomaterial with improved photocatalytic activity, called TiO2-MWCNT. There is great scientific and technological interest in photocatalytic materials, which present potential for the development of new antimicrobial agents, nanocomposites, catalysts and environmental remediation. Although many papers emphasize the benefits of using nanohybrids, there are still a small number of studies about its environmental impacts and toxicity. The objective of this work was to synthesize and characterize a TiO2-MWCNT nanohybrid. As well as to evaluate its aquatic ecotoxicity through in vivo (Zebrafish embryos-Danio rerio) and in vitro (RTG-2 cell line-Oncorhynchus mykiss fibroblasts) assays. TiO2-MWCNT was synthesized by the mechanochemical method (solid state milling) and characterized using the following techniques: scanning and transmission electron microscopy, Raman spectroscopy, thermogravimetric analysis, dynamic scattering light and electrophoretic scattering light. The synthesized nanohybrid showed enhanced photocatalytic activity, confirmed by photocatalysis studies. In the in vivo assays, TiO2-MWCNT did not show acute ecotoxicity up to 100 μg mL-1 (in the presence or absence of UV radiation). However, the ingestion of the nanohybrid by the embryos (oral route) was observed through μ-probe X-ray fluorescence spectroscopy (μ-XRF). Also, no toxicity was observed in the in vitro assays up to 100 μg mL-1 ; however, the cellular internalization capacity of this material was verified. In general, colloidal aspects (aggregation / agglomeration) and the formation of protein corona presented important implications for the precise determination of ecotoxicity. The results obtained in this work support future applications of the nanohybrid TiO2-MWCNT in the area of photocatalysis and contributes proactively to its ecotoxicological evaluation.