{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T20:06:19Z","timestamp":1789589179457,"version":"build-2803163510"},"reference-count":48,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T00:00:00Z","timestamp":1789516800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T00:00:00Z","timestamp":1789516800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Lasers Surg Med"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>This exploratory study aimed to assess acute and delayed changes in relative CCK\u20108\u2010derived cellular metabolic activity across selected 1030\u2010nm ultrashort\u2010pulse laser parameters in A549 lung adenocarcinoma cells and to characterize cell\u2010death and oxidative\u2010stress responses under an empirically selected high\u2010effect condition.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      A549 cells were irradiated under different power density, repetition rate, and pulse width conditions, and CCK\u20108\u2010derived metabolic activity was assessed at 0, 9, and 24\u2009h after irradiation. Direct cell counting was additionally performed under the selected 12.5\u2009W\/cm\n                      <jats:sup>2<\/jats:sup>\n                      power\u2010density condition. Under an empirically selected high\u2010effect condition within the tested ranges, cell\u2010death phenotypes were evaluated by Annexin V\u2010FITC\/PI flow cytometry, and transcriptomic changes were explored by RNA sequencing. Temporal changes in DCF fluorescence were descriptively examined by repeated fluorescence imaging of the same individual cells at predefined time intervals. NAC pretreatment was used to examine whether reactive oxygen species (ROS) scavenging attenuated the irradiation\u2010associated DCF fluorescence response.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Higher power densities were associated with lower relative CCK\u20108\u2010derived metabolic activity, particularly at 0\u2009h, while the largest delayed reduction from 0 to 24\u2009h occurred at 12.5\u2009W\/cm\n                      <jats:sup>2<\/jats:sup>\n                      . At this power density, direct cell counting showed a progressive decrease in cell number over 24\u2009h after irradiation. Under fixed total irradiation energy, higher repetition rates and longer pulse widths were also associated with lower metabolic activity within the tested ranges. Based on relatively preserved 0\u2009h activity together with a comparatively large 0\u201324\u2009h decline, 12.5\u2009W\/cm\n                      <jats:sup>2<\/jats:sup>\n                      , 200\u2009kHz, and 1500\u2009fs were empirically selected for exploratory characterization. Under this condition, Annexin V\u2010positive and PI\u2010positive fractions increased, RNA sequencing identified irradiation\u2010associated transcriptomic changes, and single\u2010cell imaging showed increased DCF fluorescence that was attenuated by NAC pretreatment.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Within the tested ranges, 1030\u2010nm ultrashort\u2010pulse laser irradiation was associated with parameter\u2010related changes in relative CCK\u20108\u2010derived metabolic activity in A549 cells. Direct cell counting under the 12.5\u2009W\/cm\n                      <jats:sup>2<\/jats:sup>\n                      power\u2010density condition also showed a progressive reduction in cell number over 24\u2009h. An empirically selected representative high\u2010effect condition showed a comparatively large delayed reduction in metabolic activity, together with changes in Annexin V\/PI\u2010defined cell\u2010death phenotypes and irradiation\u2010associated transcriptomic responses. In addition, repeated DCFH\u2010DA imaging revealed time\u2010dependent increases in single\u2010cell DCF fluorescence, which were reduced by NAC pretreatment. These preliminary descriptive findings provide a basis for further mechanistic investigation of ultrashort\u2010pulse laser\u2013cell interactions.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/lsm.70203","type":"journal-article","created":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T19:40:56Z","timestamp":1789587656000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["1030\u2010nm Ultrashort\u2010Pulse Laser Irradiation Is Associated With a Delayed Reduction in Cellular Metabolic Activity in A549 Cells"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-8781-6534","authenticated-orcid":false,"given":"Pengcheng","family":"Zhang","sequence":"first","affiliation":[{"name":"Hunan Provincial Key Laboratory of Intelligent Laser Manufacturing Hunan University Changsha China"},{"name":"College of Mechanical and Vehicle Engineering Hunan University Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziqian","family":"Tang","sequence":"additional","affiliation":[{"name":"Clinical Medical Research Center of Major Skin Diseases and Skin Health of Hunan Province Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liwen","family":"Bai","sequence":"additional","affiliation":[{"name":"Key Laboratory of High\u2010Performance Ship Technology of Ministry of Education (Wuhan University of Technology) Wuhan Hubei China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"He","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemo\/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio\u2010Nanotechnology and Molecular Engineering of Hunan Province Hunan University Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"YuChen","family":"Wu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemo and Biosensing, College of Biology Hunan University, Key Laboratory for Bio\u2010Nanotechnology and Molecule Engineering of Hunan Province Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Zhou","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemo and Biosensing, College of Biology Hunan University, Key Laboratory for Bio\u2010Nanotechnology and Molecule Engineering of Hunan Province Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Zhou","sequence":"additional","affiliation":[{"name":"Hunan Provincial Key Laboratory of Intelligent Laser Manufacturing Hunan University Changsha China"},{"name":"College of Mechanical and Vehicle Engineering Hunan University Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Genyu","family":"Chen","sequence":"additional","affiliation":[{"name":"Hunan Provincial Key Laboratory of Intelligent Laser Manufacturing Hunan University Changsha China"},{"name":"College of Mechanical and Vehicle Engineering Hunan University Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weisheng","family":"Chen","sequence":"additional","affiliation":[{"name":"Hunan Provincial Key Laboratory of Intelligent Laser Manufacturing Hunan University Changsha China"},{"name":"College of Mechanical and Vehicle Engineering Hunan University Changsha China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,16]]},"reference":[{"issue":"8","key":"e_1_2_9_2_1","doi-asserted-by":"crossref","DOI":"10.1038\/lsa.2016.133","article-title":"Ultrafast Laser Processing of Materials: From Science to Industry","volume":"5","author":"Malinauskas M.","year":"2016","journal-title":"Light, Science & Applications"},{"issue":"4","key":"e_1_2_9_3_1","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1038\/nphoton.2008.47","article-title":"Femtosecond Laser Micromachining in Transparent Materials","volume":"2","author":"Gattass R. 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