Cartilage-Specific Matrix and Integrin Expression in Three-Dimensional Articular Chondrocyte Cultures Overexpressing Human Interleukin-10

• Published in: Clinical medicine insights. Arthritis and musculoskeletal disorders Vol. 2008; no. 1; p. CMAMD.S560
• Main Authors: Müller, Riccarda D., John, Thilo, Kohl, Benjamin, Feldner, Anja, Zreiqat, Hala, Ertel, Wolfgang, LaFace, Drake, Hutchins, Beth, Oberholzer, Andreas, Schulze-Tanzil, Gundula
• Format: Journal Article
• Language: English
• Published: London, England SAGE Publishing 01.01.2008; SAGE Publications; Sage Publications Ltd
• Subjects: alginate; chondrocyte; 3D culture; EGFP; interleukin-10; adenovirus
• ISSN: 1179-5441; 1178-1149; 1179-5441; 1178-1149
• DOI: 10.4137/CMAMD.S560

Riccarda D. Müller1, Thilo John1, Benjamin Kohl1, Anja Feldner1, Hala Zreiqat2, Wolfgang Ertel1, Drake LaFace3, Beth Hutchins3, Andreas Oberholzer4 and Gundula Schulze-Tanzil1,5 1Charité-University of Medicine, Campus Benjamin Franklin, Berlin, Dep. of Trauma and Reconstructive Surgery, FEM, Krahmerstrasse 6-10, 12207 Berlin, Germany. 2Tissue Engineering and Biomaterials Research Unit, Biomedical Engineering, School of AMMEJ07, University of Sydney, NSW 2006, Australia. 3Canji Inc., San Diego, 3525 John Hopkins Court, Ca 92121, U.S.A. 4Department of Joint and Sport Surgery, Klinik Pyramide am See, Bellerivestr. 34, 8034 Zürich. 5Centre of Anatomy, Dep. of Cell and Neurobiology, Charité-University of Medicine, Campus Benjamin Franklin, Schumannstrasse 20/21, 10098 Berlin, Germany. Abstract Interleukin (IL)-10 overexpression inhibits joint inflammation, however the effect of high local concentrations of IL-10 on chondrocyte homeostasis remains unclear. The aim of this study was to determine the effects of IL-10 overexpression on cartilage matrix production in three-dimensional (3D) chondrocyte cultures. Human articular chondrocytes were transduced with adenoviral vectors alone (adv/empty) or by vectors either overexpressing enhanced green fluorescence protein (adv/EGFP) or human IL-10 (adv/hIL-10) before their transfer to a 3D culture system. Non-transduced chondrocytes were used as controls. The expression of IL-10 or EGFP was confirmed using ELISA or flow cytometry. Chondrocytes synthesis of collagen types II and I, aggrecan, fibronectin and β1-integrin was determined over a period of 14 days post transduction using flow cytometry or immunohistochemistry. adv/EGFP or adv/IL-10 transduced chondrocytes expressed EGFP or secreted IL-10 detectable over the 2 weeks culture period. No suppression of collagen type II, aggrecan or β1-integrin synthesis by IL-10 overexpression was found and the deposition of collagen type I and fi bronectin remained unaffected compared to the controls. IL-10 overexpression does not impair key features of chondrocytes differentiated phenotype (e.g. collagen type II and aggrecan expression) suggesting the potential use of IL-10 for gene therapeutic approaches in the joint.