Biomacromolecular and Toxicity Responses of Bacteria upon the Nano–Bio Interfacial Interactions with Ti3C2Tx Nanosheets

The biomolecular responses of bacteria to 2D nanosheets that result from nano-bio interfacial interactions remain to be thoroughly examined. Herein, Fourier transform infrared (FTIR) multivariate and 2D correlation analyses were performed to assess the composition and conformational changes in bacterial biomacromolecules (lipids, polysaccharides, and carbohydrates) upon exposure to Ti3C2Tx nanosheets. General toxicity assays, 3D excitation-emission matrix fluorescence analyses, extended Derjaguin-Landau-Verwey-Overbeek theory interaction calculations, and isothermal titration calorimetry were also performed. Our results demonstrate that Ti3C2Tx nanosheets considerably impact Gram-positive bacteria (Bacillus subtilis), causing oxidative damage and inactivation by preferentially interacting with and disrupting the cell walls. The bilayer membrane structure of Gram-negative bacteria (Escherichia coli) endows them with increased resistance to Ti3C2Tx nanosheets. The unmodified nanosheets had a higher affinity to bacterial protein components with lower toxicity due to their susceptibility to oxidation. Surface modification with KOH or hydrazine (HMH), particularly HMH, induced stronger dispersion, antioxidation, and affinity to bacterial phospholipids, which resulted in severe cell membrane lipid peroxidation and bacterial inactivation. These findings provide valuable insight into nano-bio interfacial interactions, which can facilitate the development of antimicrobial and antifouling surfaces and contribute to the evaluation of the environmental risks of nanomaterials.