{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T06:05:03Z","timestamp":1790229903232,"version":"4.1.0"},"reference-count":50,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell Dev. Biol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Porous titanium alloy scaffolds offer multiple advantages in treating large-area bone defects; however, their poor bioactivity limits their application. Low-intensity pulsed ultrasound (LIPUS) serves as a non-invasive method to enhance bone regeneration in porous titanium-6aluminium-4vanadium (Ti6Al4V) alloy scaffolds. This study aimed to examine the biological behavior and metabolism of murine osteoblastic MC3T3-E1 cells in porous Ti6Al4V (pTi64) scaffolds under LIPUS.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      MC3T3-E1 cells were incubated in pTi64 scaffolds and stimulated with LIPUS. Cell morphology, viability, and osteogenic-related gene expression were evaluated\n                      <jats:italic>in vitro<\/jats:italic>\n                      . Intracellular metabolites of MC3T3-E1 cells incubated in pTi64 scaffolds and stimulated with LIPUS for 7 days were analyzed using an ultraperformance liquid chromatography coupled with a triple quadrupole-linear ion trap mass spectrometer (UPLC-QTRAP-MS)-based cellular metabolomics approach.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      <jats:italic>In vitro<\/jats:italic>\n                      , MC3T3-E1 cells in the pTi64+LIPUS group exhibited enhanced morphology, viability, and osteogenic-related gene expression compared with the pTi64 group, suggesting that LIPUS enhances cell morphology, reduces apoptosis, and promotes osteogenic differentiation in pTi64 scaffolds. LIPUS treatment altered the cellular metabolic network; 86 differential metabolites were identified. Alterations in metabolic pathways were associated with the one-carbon pool by folate, pantothenate and coenzyme A biosynthesis, as well as the metabolism of amino acids, glycerolipids, and arachidonic acid.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>These findings offer novel insights into the osteogenic mechanisms of MC3T3-E1 cells in pTi64 scaffolds under LIPUS.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcell.2026.1945138","type":"journal-article","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T05:44:46Z","timestamp":1790228686000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Cellular metabolomics reveals the osteogenic mechanism of MC3T3-E1 cells in porous Ti6Al4V scaffolds under low-intensity pulsed ultrasound"],"prefix":"10.3389","volume":"14","author":[{"given":"Xinyun","family":"Li","sequence":"first","affiliation":[{"name":"Department of Stomatology, Affiliated Hospital of Guangdong Medical University","place":["Zhanjiang, Guangdong, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Stomatology, Shengjing Hospital of China Medical University","place":["Shenyang, Liaoning, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,24]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1361","DOI":"10.1016\/j.joca.2019.05.010","article-title":"GC-MS method for metabolic profiling of mouse femoral head articular cartilage reveals distinct effects of tissue culture and development","volume":"27","author":"Batushansky","year":"2019","journal-title":"Osteoarthr. Cartil."},{"key":"B2","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1016\/j.msec.2017.05.078","article-title":"Effect of low-intensity pulsed ultrasound on the biological behavior of osteoblasts on porous titanium alloy scaffolds: an in vitro and in vivo study","volume":"80","author":"Cao","year":"2017","journal-title":"Mat. Sci. Eng. C Mat. Biol. Appl."},{"key":"B3","doi-asserted-by":"publisher","first-page":"117647","DOI":"10.1016\/j.bone.2025.117647","article-title":"LIPUS promotes osteogenesis via the mcirc_9393-miR-217\/326-SIRT1 axis in porous Ti6Al4V scaffolds","volume":"201","author":"Cao","year":"2025","journal-title":"Bone"},{"key":"B4","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1016\/s0003-9993(03)00293-4","article-title":"Coupling agents in therapeutic ultrasound: acoustic and thermal behavior","volume":"85","author":"Casarotto","year":"2004","journal-title":"Arch. Phys. Med. Rehabil."},{"key":"B5","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1016\/j.plefa.2006.10.001","article-title":"Effects of arachidonic acid, docosahexaenoic acid and prostaglandin E(2) on cell proliferation and morphology of MG-63 and MC3T3-E1 osteoblast-like cells","volume":"76","author":"Coetzee","year":"2007","journal-title":"Prostagl. Leukot. Essent. Fat. Acids"},{"key":"B6","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1002\/cbf.1526","article-title":"Effects of arachidonic acid and docosahexaenoic acid on differentiation and mineralization of MC3T3-E1 osteoblast-like cells","volume":"27","author":"Coetzee","year":"2009","journal-title":"Cell biochem. Funct."},{"key":"B7","doi-asserted-by":"publisher","first-page":"1558","DOI":"10.1002\/jcp.26058","article-title":"Osteogenic commitment and differentiation of human mesenchymal stem cells by low-intensity pulsed ultrasound stimulation","volume":"233","author":"Costa","year":"2018","journal-title":"J. Cell. Physiol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"e3926","DOI":"10.1002\/cbf.3926","article-title":"One carbon metabolism and its implication in health and immune functions","volume":"42","author":"Dang","year":"2024","journal-title":"Cell biochem. Funct."},{"key":"B9","doi-asserted-by":"publisher","first-page":"1673","DOI":"10.1093\/humrep\/deae127","article-title":"Evaluating the safety of high-intensity focused ultrasound treatment for rectal endometriosis: results from a French prospective multicentre study including 60 patients","volume":"39","author":"Dubernard","year":"2024","journal-title":"Hum. Reprod."},{"key":"B10","doi-asserted-by":"publisher","first-page":"434","DOI":"10.3390\/metabo11070434","article-title":"Metabolomics in bone research","volume":"11","author":"Fan","year":"2021","journal-title":"Metabolites"},{"key":"B11","doi-asserted-by":"publisher","first-page":"71","DOI":"10.22074\/cellj.2015.513","article-title":"Different methylation patterns of RUNX2, OSX, DLX5 and BSP in osteoblastic differentiation of mesenchymal stem cells","volume":"17","author":"Farshdousti Hagh","year":"2015","journal-title":"Cell J."},{"key":"B12","first-page":"1070","article-title":"Three-dimensional-printed titanium alloy porous scaffold combined with trans-cinnamaldehyde for repairing osteonecrosis of the femoral head in a dog model","volume":"12","author":"Gao","year":"2020","journal-title":"Am. J. Transl. Res."},{"key":"B13","doi-asserted-by":"publisher","first-page":"68","DOI":"10.1186\/s13018-022-02960-6","article-title":"3D-printed porous Ti6Al4V scaffolds for long bone repair in animal models: a systematic review","volume":"17","author":"Gu","year":"2022","journal-title":"J. Orthop. Surg."},{"key":"B14","doi-asserted-by":"publisher","first-page":"8583","DOI":"10.18632\/aging.103168","article-title":"Gut microbiota and metabolite alterations associated with reduced bone mineral density or bone metabolic indexes in postmenopausal osteoporosis","volume":"12","author":"He","year":"2020","journal-title":"Aging"},{"key":"B15","doi-asserted-by":"publisher","first-page":"145407","DOI":"10.1016\/j.gene.2021.145407","article-title":"Electron transfer flavoprotein and its role in mitochondrial energy metabolism in health and disease","volume":"776","author":"Henriques","year":"2021","journal-title":"Gene"},{"key":"B16","doi-asserted-by":"publisher","first-page":"1117188","DOI":"10.3389\/fneur.2023.1117188","article-title":"Transcranial low-intensity ultrasound stimulation for treating central nervous system disorders: a promising therapeutic application","volume":"14","author":"Hu","year":"2023","journal-title":"Front. Neurol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1038\/nrm.2016.25","article-title":"Metabolomics: beyond biomarkers and towards mechanisms","volume":"17","author":"Johnson","year":"2016","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"e1008942","DOI":"10.1371\/journal.pcbi.1008942","article-title":"The landscape of metabolic pathway dependencies in cancer cell lines","volume":"17","author":"Joly","year":"2021","journal-title":"PLoS Comput. Biol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"22151","DOI":"10.1021\/acsomega.1c02786","article-title":"Data-independent-acquisition-based proteomic approach towards understanding the acclimation strategy of oleaginous microalga microchloropsis gaditana CCMP526 in hypersaline conditions","volume":"6","author":"Karthikaichamy","year":"2021","journal-title":"ACS Omega"},{"key":"B20","doi-asserted-by":"publisher","first-page":"9591","DOI":"10.26355\/eurrev_202009_23047","article-title":"Metabolomics and its application in the mechanism analysis on diabetic bone metabolic abnormality","volume":"24","author":"Liang","year":"2020","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"B21","doi-asserted-by":"publisher","first-page":"2198","DOI":"10.3390\/molecules29102198","article-title":"Metabolomics for clinical biomarker discovery and therapeutic target identification","volume":"29","author":"Lin","year":"2024","journal-title":"Molecules"},{"key":"B22","doi-asserted-by":"publisher","first-page":"45855","DOI":"10.31083\/FBL45855","article-title":"Odaph promotes osteoblast proliferation and differentiation by targeting the AMPK\/mTOR signaling axis to activate autophagy","volume":"31","author":"Lin","year":"2026","journal-title":"Front. Biosci."},{"key":"B23","doi-asserted-by":"publisher","first-page":"1922","DOI":"10.1016\/j.sjbs.2019.07.007","article-title":"Establishment of axon regeneration regulatory network and the role of low intensity pulsed ultrasound in the network","volume":"26","author":"Liu","year":"2019","journal-title":"Saudi J. Biol. Sci."},{"key":"B24","doi-asserted-by":"publisher","first-page":"113992","DOI":"10.1016\/j.ab.2020.113992","article-title":"1H-NMR-based urinary metabolomic analysis for the preventive effects of gushudan on glucocorticoid-induced osteoporosis rats","volume":"610","author":"Liu","year":"2020","journal-title":"Anal. Biochem."},{"key":"B25","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1007\/s11306-020-01745-4","article-title":"Comprehensive metabolomic profiling of osteosarcoma based on UHPLC-HRMS","volume":"16","author":"Lv","year":"2020","journal-title":"Metabolomics Off. J. Metabolomic Soc."},{"key":"B26","first-page":"2901","article-title":"Low-intensity pulsed ultrasound inhibits RANKL-Induced osteoclast formation via modulating ERK-c-Fos-NFATc1 signaling cascades","volume":"10","author":"Meng","year":"2018","journal-title":"Am. J. Transl. Res."},{"key":"B27","doi-asserted-by":"publisher","first-page":"345","DOI":"10.1038\/s41467-024-55036-w","article-title":"Pantothenate kinase 4 controls skeletal muscle substrate metabolism","volume":"16","author":"Miranda-Cervantes","year":"2025","journal-title":"Nat. Commun."},{"key":"B28","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1016\/j.ultrasmedbio.2006.07.026","article-title":"Relative transmissivity of ultrasound coupling agents commonly used by therapists in the UK","volume":"33","author":"Poltawski","year":"2007","journal-title":"Ultrasound Med. Biol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"e70583","DOI":"10.1096\/fj.202403395RR","article-title":"Low-intensity pulsed ultrasound promotes osteogenesis in porous titanium alloys through miR-1187\/BMP4 pathway","volume":"39","author":"Qin","year":"2025","journal-title":"FASEB J. Off. Publ. Fed. Am. Soc. Exp. Biol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"1335","DOI":"10.1016\/j.cmet.2024.04.010","article-title":"Dietary intake and glutamine-serine metabolism control pathologic vascular stiffness","volume":"36","author":"Rachedi","year":"2024","journal-title":"Cell Metab."},{"key":"B31","doi-asserted-by":"publisher","first-page":"893159","DOI":"10.3389\/fnagi.2022.893159","article-title":"Coenzyme A-Dependent tricarboxylic acid cycle enzymes are decreased in alzheimer\u2019s disease consistent with cerebral pantothenate deficiency","volume":"14","author":"Sang","year":"2022","journal-title":"Front. Aging Neurosci."},{"key":"B32","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1016\/j.ijpharm.2016.11.035","article-title":"Development of hydroxyapatite nanoparticles loaded with folic acid to induce osteoblastic differentiation","volume":"516","author":"Santos","year":"2017","journal-title":"Int. J. Pharm."},{"key":"B33","doi-asserted-by":"publisher","first-page":"108","DOI":"10.1016\/j.ultrasmedbio.2019.08.025","article-title":"Therapeutic effects of low-intensity pulsed ultrasound on osteoporosis in ovariectomized rats: intensity-dependent study","volume":"46","author":"Sun","year":"2020","journal-title":"Ultrasound Med. Biol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1186\/s40364-023-00487-4","article-title":"Targeting serine-glycine-one-carbon metabolism as a vulnerability in cancers","volume":"11","author":"Sun","year":"2023","journal-title":"Biomark. Res."},{"key":"B35","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1146\/annurev.nutr.012809.104810","article-title":"Compartmentalization of mammalian folate-mediated one-carbon metabolism","volume":"30","author":"Tibbetts","year":"2010","journal-title":"Annu. Rev. Nutr."},{"key":"B36","doi-asserted-by":"publisher","first-page":"e73914","DOI":"10.1371\/journal.pone.0073914","article-title":"Enhancement of osteogenic differentiation and proliferation in human mesenchymal stem cells by a modified low intensity ultrasound stimulation under simulated microgravity","volume":"8","author":"Uddin","year":"2013","journal-title":"PloS One"},{"key":"B37","doi-asserted-by":"publisher","first-page":"804","DOI":"10.1016\/j.ultrasmedbio.2012.11.016","article-title":"Reversal of the detrimental effects of simulated microgravity on human osteoblasts by modified low intensity pulsed ultrasound","volume":"39","author":"Uddin","year":"2013","journal-title":"Ultrasound Med. Biol."},{"key":"B38","doi-asserted-by":"publisher","first-page":"1203","DOI":"10.1039\/d2tb02129c","article-title":"A pH-neutral bioactive glass coated 3D-printed porous Ti6Al4V scaffold with enhanced osseointegration","volume":"11","author":"Wang","year":"2023","journal-title":"J. Mat. Chem. B"},{"key":"B39","doi-asserted-by":"publisher","first-page":"BSR20171072","DOI":"10.1042\/BSR20171072","article-title":"YAP\/TAZ-mediated activation of serine metabolism and methylation regulation is critical for LKB1-deficient breast cancer progression","volume":"37","author":"Wu","year":"2017","journal-title":"Biosci. Rep."},{"key":"B40","doi-asserted-by":"publisher","first-page":"1733","DOI":"10.1002\/jum.14525","article-title":"Low-intensity pulsed ultrasound accelerates traumatic vertebral fracture healing by coupling proliferation of type H microvessels","volume":"37","author":"Wu","year":"2018","journal-title":"J. Ultrasound Med. Off. J. Am. Inst. Ultrasound Med."},{"key":"B41","doi-asserted-by":"publisher","first-page":"267","DOI":"10.4252\/wjsc.v16.i3.267","article-title":"Low-intensity pulsed ultrasound reduces alveolar bone resorption during orthodontic treatment via lamin A\/C-Yes-associated protein axis in stem cells","volume":"16","author":"Wu","year":"2024","journal-title":"World J. Stem Cells"},{"key":"B42","doi-asserted-by":"publisher","first-page":"268","DOI":"10.1210\/en.2015-1690","article-title":"IGF-I and IGFBP-2 stimulate AMPK activation and autophagy, which are required for osteoblast differentiation","volume":"157","author":"Xi","year":"2016","journal-title":"Endocrinology"},{"key":"B43","doi-asserted-by":"publisher","first-page":"8060","DOI":"10.1038\/s41598-017-08868-0","article-title":"PHF20 positively regulates osteoblast differentiation via increasing the expression and activation of Runx2 with enrichment of H3K4me3","volume":"7","author":"Yang","year":"2017","journal-title":"Sci. Rep."},{"key":"B44","doi-asserted-by":"publisher","first-page":"817","DOI":"10.1016\/j.drudis.2020.12.006","article-title":"Folate-mediated one-carbon metabolism: a targeting strategy in cancer therapy","volume":"26","author":"Yang","year":"2021","journal-title":"Drug Discov. Today"},{"key":"B45","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1186\/s13018-019-1489-y","article-title":"Mechanical behavior of a titanium alloy scaffold mimicking trabecular structure","volume":"15","author":"Zhang","year":"2020","journal-title":"J. Orthop. Surg."},{"key":"B46","doi-asserted-by":"publisher","first-page":"3427430","DOI":"10.1155\/2020\/3427430","article-title":"Metabolomic analysis reveals that the mechanism of astaxanthin improves the osteogenic differentiation potential in bone marrow mesenchymal stem cells","volume":"2020","author":"Zhao","year":"2020","journal-title":"Oxid. Med. Cell. Longev."},{"key":"B47","doi-asserted-by":"publisher","first-page":"3007","DOI":"10.3390\/molecules26103007","article-title":"Bone grafts and substitutes in dentistry: a review of current trends and developments","volume":"26","author":"Zhao","year":"2021","journal-title":"Molecules"},{"key":"B48","doi-asserted-by":"publisher","first-page":"1192720","DOI":"10.3389\/fbioe.2023.1192720","article-title":"Low-intensity pulsed ultrasound regulates osteoblast-osteoclast crosstalk via EphrinB2\/EphB4 signaling for orthodontic alveolar bone remodeling","volume":"11","author":"Zhou","year":"2023","journal-title":"Front. Bioeng. Biotechnol."},{"key":"B49","doi-asserted-by":"publisher","first-page":"1296881","DOI":"10.3389\/fbioe.2023.1296881","article-title":"Recent advances of responsive scaffolds in bone tissue engineering","volume":"11","author":"Zhu","year":"2023","journal-title":"Front. Bioeng. Biotechnol."},{"key":"B50","doi-asserted-by":"publisher","first-page":"1351","DOI":"10.3390\/biom15101351","article-title":"The effects and mechanisms of low-intensity pulsed ultrasound on bone remodeling: from laboratory to clinic","volume":"15","author":"Zong","year":"2025","journal-title":"Biomolecules"}],"container-title":["Frontiers in Cell and Developmental Biology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2026.1945138\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T05:44:48Z","timestamp":1790228688000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2026.1945138\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,24]]},"references-count":50,"alternative-id":["10.3389\/fcell.2026.1945138"],"URL":"https:\/\/doi.org\/10.3389\/fcell.2026.1945138","relation":{},"ISSN":["2296-634X"],"issn-type":[{"value":"2296-634X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,24]]},"article-number":"1945138"}}