Adhesion of osteoclasts and monocytes to developing bone

Osteoclast resorption of bone matrix during bone development is preceded by cell attachment to bone. An in vitro assay is described that measures adhesion of isolated osteoclasts and their presumed progenitor cell, the monocyte, to embryonic chick bone. Osteoclasts were isolated from day 19 chick tibia and enriched. Circulating chick hatchling monocytes were purified by Ficoll-hypaque sedimentation and attachment to plastic. Isolated cells were labeled with (32P)-KH2PO4 to quantitate cell attachment. In one series of experiments, labeled cells were inoculated into vessels containing day 19 tibias with either endosteal or periosteal bone surface exposed. Labeled cells were also inoculated into chambers containing day 6 (cartilage cores), day 12 (bone and cartilage cores), or day 19 embryonic tibias (bone). Cultures of stage 24 chick limb mesenchyme and embryonic chick skin fibroblasts served as controls. Results demonstrate that twice as many osteoclasts and monocytes adhere to bone as compared to fibroblasts and stage 24 limb mesenchymal cells. Furthermore, there does not appear to be selective adhesion to the endosteal as compared to periosteal bone surfaces. When the data is calculated on the basis of cell attachment per area of each substrate, the day 12 cores had slightly higher number of osteoclasts and monocytes attached compared to day 19 bones; day 6 cartilage cores bound few cells. These observations suggest that osteoclasts and monocytes have a high affinity to bone which seems to be influenced by the developmental age and composition of the substrate.