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Photothermal modulation is a remote and non\u2010genetic technique to control neural activity with high spatiotemporal resolution. The local heat release by photothermally active nanomaterial will change the membrane properties of the interfaced neurons during light illumination. Recently, it is demonstrated that the two\u2010dimensional Ti<jats:sub>3<\/jats:sub>C<jats:sub>2<\/jats:sub>T<jats:sub><jats:italic>x<\/jats:italic><\/jats:sub> MXene is an outstanding candidate to photothermally excite neurons with low incident energy. However, the safety of using Ti<jats:sub>3<\/jats:sub>C<jats:sub>2<\/jats:sub>T<jats:sub><jats:italic>x<\/jats:italic><\/jats:sub> for neural modulation is unknown. Here, the biosafety of Ti<jats:sub>3<\/jats:sub>C<jats:sub>2<\/jats:sub>T<jats:sub><jats:italic>x<\/jats:italic><\/jats:sub>\u2010based photothermal modulation is thoroughly investigated, including assessments of plasma membrane integrity, mitochondrial stress, and oxidative stress. It is demonstrated that culturing neurons on 25\u00a0\u00b5g\u00a0cm<jats:sup>\u22122<\/jats:sup> Ti<jats:sub>3<\/jats:sub>C<jats:sub>2<\/jats:sub>T<jats:sub><jats:italic>x<\/jats:italic><\/jats:sub> films and illuminating them with laser pulses (635\u00a0nm) with different incident energies (2\u201310\u00a0\u00b5J per pulse) and different pulse frequencies (1 pulse, 1\u00a0Hz, and 10\u00a0Hz) neither damage the cell membrane, induce cellular stress, nor generate oxidative stress. The threshold energy to cause damage (i.e., 14\u00a0\u00b5J per pulse) exceeded the incident energy for neural excitation (&lt;10\u00a0\u00b5J per pulse). This multi\u2010assay safety evaluation provides crucial insights for guiding the establishment of light conditions and protocols in the clinical translation of photothermal modulation.<\/jats:p>","DOI":"10.1002\/adhm.202302330","type":"journal-article","created":{"date-parts":[[2023,9,26]],"date-time":"2023-09-26T09:41:13Z","timestamp":1695721273000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Photothermal Excitation of Neurons Using MXene: Cellular Stress and Phototoxicity Evaluation"],"prefix":"10.1002","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9294-3705","authenticated-orcid":false,"given":"Yingqiao","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Materials Science and Engineering Carnegie Mellon University  Pittsburgh PA 15213 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5486-9997","authenticated-orcid":false,"given":"Jane E.","family":"Hartung","sequence":"additional","affiliation":[{"name":"Department of Neurobiology University of Pittsburgh  Pittsburgh PA 15260 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5390-6311","authenticated-orcid":false,"given":"Adam","family":"Goad","sequence":"additional","affiliation":[{"name":"A.J. Drexel Nanomaterials Institute and Department of Materials Science and Engineering Drexel University  Philadelphia PA 19104 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-6939-5552","authenticated-orcid":false,"given":"Mat\u00edas A.","family":"Preisegger","sequence":"additional","affiliation":[{"name":"Department of Neurobiology University of Pittsburgh  Pittsburgh PA 15260 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-3486-9010","authenticated-orcid":false,"given":"Benjamin","family":"Chacon","sequence":"additional","affiliation":[{"name":"A.J. 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