{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:35:40Z","timestamp":1760240140658,"version":"build-2065373602"},"reference-count":40,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2019,3,27]],"date-time":"2019-03-27T00:00:00Z","timestamp":1553644800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Gesellschaft f\u00fcr Arthroskopie und Gelenkchirurgie (AGA)","award":["26\/2007"],"award-info":[{"award-number":["26\/2007"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Cartilage defects represent an increasing pathology among active individuals that affects the ability to contribute to sports and daily life. Cell therapy, such as autologous chondrocyte implantation (ACI), is a widespread option to treat larger cartilage defects still lacking standardization of in vitro cell culture parameters. We hypothesize that mRNA expression of cytokines and proteases before and after ACI is influenced by in vitro parameters: cell-passage, cell-density and membrane-holding time. Knee joint articular chondrocytes, harvested from rabbits (n = 60), were cultured\/processed under varying conditions: after three different cell-passages (P1, P3, and P5), cells were seeded on 3D collagen matrices (approximately 25 mm3) at three different densities (2 \u00d7 105\/matrix, 1 \u00d7 106\/matrix, and 3 \u00d7 106\/matrix) combined with two different membrane-holding times (5 h and two weeks) prior autologous transplantation. Those combinations resulted in 18 different in vivo experimental groups. Two defects\/knee\/animal were created in the trochlear groove (defect dimension: \u2205 4 mm \u00d7 2 mm). Four identical cell-seeded matrices (CSM) were assembled and grouped in two pairs: One pair giving pre-operative in vitro data (CSM-i), the other pair was implanted in vivo and harvested 12 weeks post-implantation (CSM-e). CSMs were analyzed for TNF-\u03b1, IL-1\u03b2, MMP-1, and MMP-3 via qPCR. CSM-i showed higher expression of IL-1\u03b2, MMP-1, and MMP-3 compared to CSM-e. TNF-\u03b1 expression was higher in CSM-e. Linearity between CSM-i and CSM-e values was found, except for TNF-\u03b1. IL-1\u03b2 expression was higher in CSM-i at higher passage and longer membrane-holding time. IL-1\u03b2 expression decreased with prolonged membrane-holding time in CSM-e. For TNF-\u03b1, the reverse was true. Lower cell-passages and lower membrane-holding time resulted in stronger TNF-\u03b1 expression. Prolonged membrane-holding time resulted in increased MMP levels among CSM-i and CSM-e. Cellular density was of no significant effect. We demonstrated cytokine and MMP expression levels to be directly influenced by in vitro culture settings in ACI. Linearity of expression-patterns between CSM-i and CSM-e may predict ACI regeneration outcome in vivo. Cytokine\/protease interaction within the regenerate tissue could be guided via adjusting in vitro culture parameters, of which membrane-holding time resulted the most relevant one.<\/jats:p>","DOI":"10.3390\/ijms20071545","type":"journal-article","created":{"date-parts":[[2019,3,29]],"date-time":"2019-03-29T03:38:52Z","timestamp":1553830732000},"page":"1545","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Chondrocyte Culture Parameters for Matrix-Assisted Autologous Chondrocyte Implantation Affect Catabolism and Inflammation in a Rabbit Model"],"prefix":"10.3390","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0690-2892","authenticated-orcid":false,"given":"Martin","family":"Sauerschnig","sequence":"first","affiliation":[{"name":"Department of Orthopedic Sports Medicine, Klinikum rechts der Isar, Technical University of Munich, Munich 81675, Germany"},{"name":"Trauma Hospital Graz, Unfallkrankenhaus der Allgemeinen Unfallversicherungsanstalt (AUVA), Teaching Hospital Medical University Graz, Graz 8010, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Markus T.","family":"Berninger","sequence":"additional","affiliation":[{"name":"Department of Orthopedic Sports Medicine, Klinikum rechts der Isar, Technical University of Munich, Munich 81675, Germany"},{"name":"Department of Trauma Surgery, Trauma Center (BGU) Murnau, Murnau 82418, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Theresa","family":"Kaltenhauser","sequence":"additional","affiliation":[{"name":"Department of Orthopedic Sports Medicine, Klinikum rechts der Isar, Technical University of Munich, Munich 81675, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Plecko","sequence":"additional","affiliation":[{"name":"Trauma Hospital Graz, Unfallkrankenhaus der Allgemeinen Unfallversicherungsanstalt (AUVA), Teaching Hospital Medical University Graz, Graz 8010, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gabriele","family":"Wexel","sequence":"additional","affiliation":[{"name":"Department of Orthopedic Sports Medicine, Klinikum rechts der Isar, Technical University of Munich, Munich 81675, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3003-3378","authenticated-orcid":false,"given":"Martin","family":"Sch\u00f6nfelder","sequence":"additional","affiliation":[{"name":"Exercise Biology, Technical University of Munich, 80809 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valerie","family":"Wienerroither","sequence":"additional","affiliation":[{"name":"Trauma Hospital Graz, Unfallkrankenhaus der Allgemeinen Unfallversicherungsanstalt (AUVA), Teaching Hospital Medical University Graz, Graz 8010, Austria"},{"name":"Department of General Surgery, Medical University of Graz, Graz 8036, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andreas B.","family":"Imhoff","sequence":"additional","affiliation":[{"name":"Department of Orthopedic Sports Medicine, Klinikum rechts der Isar, Technical University of Munich, Munich 81675, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Philip B.","family":"Sch\u00f6ttle","sequence":"additional","affiliation":[{"name":"Department of Orthopedic Sports Medicine, Klinikum rechts der Isar, Technical University of Munich, Munich 81675, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0484-4847","authenticated-orcid":false,"given":"Elizabeth Rosado","family":"Rosado Balmayor","sequence":"additional","affiliation":[{"name":"Experimental Trauma Surgery, Klinikum rechts der Isar, Technical University of Munich, Munich 81675, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gian M.","family":"Salzmann","sequence":"additional","affiliation":[{"name":"Department of Orthopedic Sports Medicine, Klinikum rechts der Isar, Technical University of Munich, Munich 81675, Germany"},{"name":"Gelenkzentrum Rhein-Main, Wiesbaden 65183, Germany"},{"name":"Musculoskeletal Centre, Schulthess Klinik Zurich, Zurich 8008, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,3,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1056\/NEJM199410063311401","article-title":"Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation","volume":"331","author":"Brittberg","year":"1994","journal-title":"N. 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