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However, increasing evidence suggests that chondrocytes exhibit metabolically active behaviors, particularly under aging and pathological conditions. Here, we developed and applied an in vivo deuterium oxide (D\n                      <jats:sub>2<\/jats:sub>\n                      O)\u2013based stable isotope labeling approach to directly quantify de novo fatty\u2010acid synthesis in cartilage. Using gas chromatography\u2013mass spectrometry (GC\u2013MS), we tracked deuterium incorporation into palmitate in serum and articular cartilage from wild\u2010type (WT) and cartilage\u2010specific acetyl\u2010CoA carboxylase 1 (ACC1) knockout mice. Progressive incorporation of deuterium into palmitate confirmed active fatty\u2010acid synthesis in cartilage, with labeling kinetics distinct from those observed in serum, consistent with slower lipid turnover in the matrix\u2010dense cartilage environment. Although fractional new synthesis of palmitate was comparable between genotypes during the labeling window, ACC1\u2010deficient cartilage exhibited reduced palmitate abundance and decreased higher\u2010order isotopomer enrichment at early labeling time points, indicating attenuated lipogenic flux. These findings provide direct in vivo evidence that chondrocytes possess intrinsic lipogenic capacity and that ACC1 contributes to the maintenance of the cartilage lipid pool. Together, this study establishes D\n                      <jats:sub>2<\/jats:sub>\n                      O\u2010based isotope tracing as a robust and quantitative framework for assessing cartilage lipid metabolism in vivo and reveals de novo lipogenesis as an active metabolic process in cartilage.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Clinical Significance<\/jats:title>\n                    <jats:p>This approach offers a valuable platform for investigating lipid dysregulation in osteoarthritis and other joint diseases.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/jor.70290","type":"journal-article","created":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T06:42:23Z","timestamp":1791182543000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Stable Isotope Labeling to Track De Novo Fatty Acid Synthesis in Mouse Cartilage\u2010Enriched Joint Tissue In Vivo: A Practical Workflow"],"prefix":"10.1002","volume":"44","author":[{"given":"Shadi","family":"Moradi","sequence":"first","affiliation":[{"name":"Department of Biomedical Sciences, Heritage College of Osteopathic Medicine (HCOM) Ohio University Athens Ohio USA"},{"name":"Ohio Musculoskeletal and Neurological Institute (OMNI), Heritage College of Osteopathic Medicine (HCOM) Ohio University Athens Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6425-9570","authenticated-orcid":false,"given":"Anupama","family":"Binoy","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, Heritage College of Osteopathic Medicine (HCOM) Ohio University Athens Ohio USA"},{"name":"Ohio Musculoskeletal and Neurological Institute (OMNI), Heritage College of Osteopathic Medicine (HCOM) Ohio University Athens Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4973-1504","authenticated-orcid":false,"given":"Shouan","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, Heritage College of Osteopathic Medicine (HCOM) Ohio University Athens Ohio USA"},{"name":"Ohio Musculoskeletal and Neurological Institute (OMNI), Heritage College of Osteopathic Medicine (HCOM) Ohio University Athens Ohio USA"},{"name":"Institute of Molecular Medicine and Aging (IMMA), Heritage College of Osteopathic Medicine (HCOM) Ohio University Athens Ohio USA"},{"name":"Diabetes Institute (DI), Heritage College of Osteopathic Medicine (HCOM) Ohio University Athens Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,4]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.joca.2018.04.018"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/oby.2009.176"},{"key":"e_1_2_10_4_1","first-page":"1133","article-title":"Relation of Obesity and of Vocational and Avocational Risk Factors to Osteoarthritis","volume":"32","author":"Felson D. 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