An Osteochondral Culture Model to Study Mechanisms Involved in Articular Cartilage Repair

• Published in: Tissue engineering. Part C, Methods Vol. 18; no. 1; pp. 45 - 53
• Main Authors: de Vries-van Melle, Marloes L., Mandl, Erik W., Kops, Nicole, Koevoet, Wendy J.L.M., Verhaar, Jan A.N., van Osch, Gerjo J.V.M.
• Format: Journal Article
• Language: English
• Published: United States Mary Ann Liebert, Inc 01.01.2012
• Subjects: Acid phosphatase; Alkaline phosphatase; Animals; Biopsy; Bone (subchondral); Bone and Bones - metabolism; Calcein; Cartilage; Cartilage (articular); Cartilage, Articular - pathology; Cattle; Cell Culture Techniques; Cell Survival; Cells, Cultured; Chondrocytes; Chondrocytes - cytology; Excretion; Explants; Extracellular matrix; Extracellular Matrix - metabolism; Genetics; Integration; L-Lactate dehydrogenase; L-Lactate Dehydrogenase - metabolism; Mechanical properties; Models, Biological; mRNA; Osteoblasts; Osteoblasts - cytology; Osteoclasts; Ribonucleic acid; RNA; RNA, Messenger - metabolism; Seeding; Tissue engineering; Wound Healing
• ISSN: 1937-3384; 1937-3392
• DOI: 10.1089/ten.tec.2011.0339

Although several treatments for cartilage repair have been developed and used in clinical practice the last 20 years, little is known about the mechanisms that are involved in the formation of repair tissue after these treatments. Often, these treatments result in the formation of fibrocartilaginous tissue rather than normal articular cartilage. Because the repair tissue is inferior to articular cartilage in terms of mechanical properties and zonal organization of the extracellular matrix, complaints of the patient may return. The biological and functional outcome of these treatments should thus be improved. For this purpose, an in vitro model allowing investigation of the involved repair mechanisms can be of great value. We present the development of such a model. We used bovine osteochondral biopsies and created a system in which cartilage defects of different depths can be studied. First, our biopsy model was characterized extensively: we studied the viability by means of lactate dehydrogenase (LDH) excretion over time and we investigated expression of cartilage-related genes in osteochondral biopsies and compared it with conventional cartilage-only explants. After 28 days of culture, LDH was detected at low levels and mRNA could be retrieved. The expression of cartilage-related genes decreased over time. This was more evident in cartilage-only explants, indicating that the biopsy model provided a more stable environment. We also characterized the subchondral bone: osteoclasts and osteoblasts were active after 28 days of culture, which was indicated by tartrate acid phosphatase staining and alkaline phosphatase measurements, respectively, and matrix deposition during culture was visualized using calcein labeling. Second, the applicability of the model was further studied by testing two distinct settings: (1) implantation of chondrocytes in defects of different depths; (2) two different seeding strategies of chondrocytes. Differences were observed in terms of volume and integration of newly formed tissue in both settings, suggesting that our model can be used to model distinct conditions or even to mimic clinical treatments. After extensive characterization and testing of our model, we present a representative and reproducible in vitro model that can be used to evaluate new cartilage repair treatments and study mechanisms in a controlled and standardized environment.