Affiliations: [a] Department of Cell Biology and Tissue Engineering, Institute of Biotechnology, Brandenburg University of Technology Cottbus-Senftenberg, Universitaetsplatz, Senftenberg, Germany
| [b] Center for Orthopaedics and Trauma Surgery, Brandenburg Hospital, Brandenburg Medical School Theodor Fontane, Hochstrasse, Brandenburg/Havel, Germany
Corresponding authors: Anne-Helen Lutter and Ursula Anderer, Department of Cell Biology and Tissue Engineering, Institute of Biotechnology, Brandenburg University of Technology Cottbus-Senftenberg, Universitaetsplatz 1, 01968 Senftenberg, Germany. Tel.: +49 3573 85923; Fax: +49 3753 85809; E-mail: [email protected] (Anne-Helen Lutter) and [email protected] (Ursula Anderer).
Abstract: BACKGROUND:Tissue engineering has become a major field of research in biotechnology and biomedicine. As a consequence, cell-based therapeutic approaches are entering the hospitals, especially for skeletal regeneration. Traumatic injuries of cartilage are treated with autologous cell suspensions or in vitro generated cartilage tissues, but there is actually no therapy available for degenerative cartilage defects. However, Osteoarthritis (OA) is a major public health problem in the world affecting 240 million people globally. OBJECTIVE:To develop suitable in vitro tissues, the properties of chondrogenic spheroids should be optimized via co-culture with cells naturally occurring as joint neighbours. METHODS:Human chondrocytes were isolated from condyles and propagated in monolayer culture. Scaffold-free spheroids were generated and co-cultured with joint-specific partner cells (osteoblast-like osteosarcoma cells, fibroblasts). Morphology and differentiation was analyzed using histochemistry (Alcian blue, Safranin O) and immunohistochemistry for cartilage markers (collagen type II, Sox9, proteoglycan), proliferation-associated protein (Ki67) and markers of connective tissue (collagen type I and actin). RESULTS:The provision of a more natural microenvironment in vitro via co-culture of chondrocyte-based aggregates with osteoblast-like Saos-2 cells enhanced the differentiation potential of chondrogenic spheroids towards hyaline cartilage. CONCLUSIONS:The study showed the positive influence of Saos-2 cells on the differentiation potential of human chondrocytes in co-culture.
Keywords: Human chondrocytes, Saos-2, HFF-1, co-culture, spheroid, scaffold-free