{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T07:03:07Z","timestamp":1782975787643,"version":"3.54.5"},"reference-count":29,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T00:00:00Z","timestamp":1782950400000},"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. Bioeng. Biotechnol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      In cartilage-related studies, the existing\n                      <jats:italic>in vitro<\/jats:italic>\n                      cartilage models often fail to simulate the complex pathophysiology involving a hypoxic microenvironment, abnormal intercellular mechanical stress (microstress), and pathological crosstalk between chondrocytes and vascular endothelial cells. he aim of this study was to develop and validate a novel, simplified multi-cell-component cartilage organoid model capable of mimicking these three key OA-related features\n                      <jats:italic>in vitro<\/jats:italic>\n                      , using readily available cell lines.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Organoids were constructed using SW1353 chondrocytes and HMEC-1 endothelial cell lines within low-adhesion grid microchambers. A monosodium iodoacetate (MIA)-induced OA model was developed. Histological and immunofluorescence analyses of the organoids were conducted to examine markers of matrix metabolism, cytoskeletal and intercellular microstress, hypoxic microenvironment, and chondrocyte -endothelial crosstalk. The organoid features were compared with those of standard 2D cell cultures and\n                      <jats:italic>in vivo<\/jats:italic>\n                      cartilages.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The multi-cell-component organoids successfully recapitulated critical\n                      <jats:italic>in vivo<\/jats:italic>\n                      features. They exhibited MIA-induced F-actin reorganization and PIEZO1 upregulation, developed a hypoxic core with elevated hypoxia-inducible factor 1-alpha (HIF-1\u03b1) levels, and formed an endothelial shell that invasively disrupted the chondrocyte core upon MIA treatment. This disruption was accompanied by the activation of vascular endothelial growth factor -NOTCH receptor 1 -delta-like ligand 4 (VEGF -NOTCH1 -DLL4) signaling, mirroring the pathological chondrocyte -endothelial crosstalk. Single-cell-component organoids simulated microstress-related changes but not hypoxia or chondrocyte -endothelial crosstalk.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      The multi-cell-component cartilage organoids constructed from tumor-derived cell lines using low-adhesion grid microchambers partially simulated intercellular microstress, hypoxic microenvironment, and chondrocyte -endothelial crosstalk, which was similar to that observed in cartilages in vivo. These organoids provide a new strategy for conducting cartilage-related studies\n                      <jats:italic>in vitro<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fbioe.2026.1852010","type":"journal-article","created":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T05:35:07Z","timestamp":1782970507000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Multi-cell-component cartilage organoids simulate intercellular microstress, hypoxic microenvironment, and chondrocyte\u2013endothelial crosstalk in vitro"],"prefix":"10.3389","volume":"14","author":[{"given":"Zhengchao","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Sports Medicine, Wuhan Fourth Hospital","place":["Wuhan, China"]},{"name":"Hubei Provincial Sports Medicine Center","place":["Wuhan, China"]},{"name":"Hubei Key Laboratory of Sports Injury and Precision Therapy, Wuhan Fourth Hospital","place":["Wuhan, China"]},{"name":"Hubei Provincial Clinical Research Center for Orthopaedics","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengfei","family":"Zhu","sequence":"additional","affiliation":[{"name":"Hubei Provincial Sports Medicine Center","place":["Wuhan, China"]},{"name":"Hubei Key Laboratory of Sports Injury and Precision Therapy, Wuhan Fourth Hospital","place":["Wuhan, China"]},{"name":"Hubei Provincial Clinical Research Center for Orthopaedics","place":["Wuhan, China"]},{"name":"Department of Cardiovascular, Wuhan Fourth Hospital","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongmei","family":"Li","sequence":"additional","affiliation":[{"name":"Zibo First Hospital, Zibo Prevention and Treatment Hospital for Occupation Diseases","place":["Zibo, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingsong","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Sports Medicine, Wuhan Fourth Hospital","place":["Wuhan, China"]},{"name":"Hubei Provincial Sports Medicine Center","place":["Wuhan, China"]},{"name":"Hubei Key Laboratory of Sports Injury and Precision Therapy, Wuhan Fourth Hospital","place":["Wuhan, China"]},{"name":"Hubei Provincial Clinical Research Center for Orthopaedics","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiangxia","family":"Cheng","sequence":"additional","affiliation":[{"name":"Hubei Provincial Sports Medicine Center","place":["Wuhan, China"]},{"name":"Hubei Key Laboratory of Sports Injury and Precision Therapy, Wuhan Fourth Hospital","place":["Wuhan, China"]},{"name":"Hubei Provincial Clinical Research Center for Orthopaedics","place":["Wuhan, China"]},{"name":"Department of Anesthesiology, Wuhan Fourth Hospital","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Cai","sequence":"additional","affiliation":[{"name":"Hubei Provincial Sports Medicine Center","place":["Wuhan, China"]},{"name":"Hubei Key Laboratory of Sports Injury and Precision Therapy, Wuhan Fourth Hospital","place":["Wuhan, China"]},{"name":"Hubei Provincial Clinical Research Center for Orthopaedics","place":["Wuhan, China"]},{"name":"Department of Rehabilitation, Wuhan Fourth Hospital","place":["Wuhan, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,7,2]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"804","DOI":"10.1038\/s41467-023-36408-0","article-title":"Engraftment of allogeneic iPS cell-derived cartilage organoid in a primate model of articular cartilage defect","volume":"14","author":"Abe","year":"2023","journal-title":"Nat. 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