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Therefore, we aimed to evaluate the safety of tDCS-iTBS in an animal model after brain stimulations for 1 h and 4 weeks. Thirty-one Sprague Dawley rats were divided into two groups: (1) short-term stimulation for 1 h\/session (sham, low, and high) and (2) long-term for 30 min, 3 sessions\/week for 4 weeks (sham and high). The anodal stimulation applied over the primary motor cortex ranged from 2.5 to 4.5 mA\/cm2. The brain biomarkers and scalp tissues were assessed using ELISA and histological analysis (H&amp;E staining) after stimulations. The caspase-3 activity, cortical myelin basic protein (MBP) expression, and cortical interleukin (IL-6) levels increased slightly in both groups compared to sham. The serum MBP, cortical neuron-specific enolase (NSE), and serum IL-6 slightly changed from sham after stimulations. There was no obvious edema or cell necrosis seen in cortical histology after the intervention. The short- and long-term stimulations did not induce significant adverse effects on brain and scalp tissues upon assessing biomarkers and conducting histological analysis.<\/jats:p>","DOI":"10.3390\/ijms23126850","type":"journal-article","created":{"date-parts":[[2022,6,21]],"date-time":"2022-06-21T01:43:27Z","timestamp":1655775807000},"page":"6850","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Safety of Special Waveform of Transcranial Electrical Stimulation (TES): In Vivo Assessment"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3681-6453","authenticated-orcid":false,"given":"Muhammad","family":"Adeel","sequence":"first","affiliation":[{"name":"School of Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 110, Taiwan"},{"name":"International PhD Program in Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 110, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4146-5678","authenticated-orcid":false,"given":"Chun-Ching","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Interaction Design, College of Design, National Taipei University of Technology, Taipei 106, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0498-3190","authenticated-orcid":false,"given":"Bor-Shing","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Computer Science and Information Engineering, National Taipei University, New Taipei City 237, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8510-7437","authenticated-orcid":false,"given":"Hung-Chou","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Physical Medicine and Rehabilitation, School of Medicine, College of Medicine, Taipei Medical University, Taipei 110, Taiwan"},{"name":"Department of Physical Medicine and Rehabilitation, Shuang Ho Hospital, Taipei Medical University, New Taipei City 235, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jian-Chiun","family":"Liou","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 110, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu-Ting","family":"Li","sequence":"additional","affiliation":[{"name":"Taiwan Instrument Research Institute, National Applied Research Laboratories, Hsinchu 30261, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9744-4094","authenticated-orcid":false,"given":"Chih-Wei","family":"Peng","sequence":"additional","affiliation":[{"name":"School of Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 110, Taiwan"},{"name":"International PhD Program in Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 110, Taiwan"},{"name":"School of Gerontology Health Management, College of Nursing, Taipei Medical University, Taipei 110, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2008","DOI":"10.1016\/j.clinph.2009.08.016","article-title":"Safety, ethical considerations, and application guidelines for the use of transcranial magnetic stimulation in clinical practice and research","volume":"120","author":"Rossi","year":"2009","journal-title":"Clin. 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