Cartilage turnover in embryonic chick tibial explant cultures

Department of Animal Science, Michigan State University, East Lansing 48824-1225, USA. orthm@pilot.msu.edu

Growth plate cartilage regulates the rate of growth and ultimate length of several bones in the skeleton. Chondrocytes within the growth plate proliferate, differentiate, enlarge, and die. The extracellular matrix undergoes synthesis, reorganization, and eventually degradation. The majority of research in growth plate physiology has focused on the proliferation and differentiation of chondrocytes as well as proteins they produce for the extracellular matrix. However, little is known about the transition from hypertrophic to apoptotic chondrocytes or the regulation of terminal degradation of cartilage prior to bone formation. An explant culture has been developed to study cartilage differentiation using 12-d-old embryonic chick tibiae. We have modified the explant culture and are using it to further elucidate mechanisms involved in the regulation of growth plate cartilage turnover. In our cultures, chondrocytes mature and then die, completely degrading the cartilage in approximately 16 d. The matrix undergoes a predictable pattern of degradation in which proteoglycans followed by collagen are removed. Increases in matrix metalloproteinase activity and nitric oxide production are detected in cartilage concurrently with release of proteoglycans into media. Inhibitors of nitric oxide inhibit nitric oxide production and proteoglycan degradation, suggesting that nitric oxide, at least in part, regulates growth plate cartilage turnover in the explant culture. Information gained from using this explant culture will aid in understanding the regulation of growth plate cartilage turnover in vivo and potentially help determine the cause of bone growth diseases such as tibial dyschondroplasia.