Macrophages (MO) are multipotential cells capable of giving rise to osteoclasts and of resorbing bone. Since both of these processes are ultimately dependent upon the attachment of cells to a mineralized bone surface, we have examined in this study the mechanism by which such attachment is achieved. The data show that elicited rat peritoneal MO bind to bone in a temperature-dependent and -saturable manner with half-maximal attachment occurring within 10 min at 37°C and reaching a plateau by ~60 min. The kinetics of binding are essentially the same whether devitalized bone particles or viable calvaria are used as a substrate. The attachment of MO to bone is inhibited by some sugars (e.g., N-acetylgalactosamine, thiogalactoside, β-lactose), fetuin and asialofetuin, and by pretreating the bone with periodate. Binding is also significantly reduced when MO are preincubated with tunicamycin and swainsonine at nontoxic concentrations sufficient to inhibit or alter glycosylation. On the other hand, exposing the cells to neuraminidase increases the capacity of MO to bind to bone. Collectively, our observations indicate that the attachment of MO to bone is a highly regulated process and is mediated, at least in part, by saccharides located on both the cell and the bone surface.