The plasminogen/plasmin proteolytic cascade plays an important role in extracellular matrix remodeling. The presence of the two plasminogen activators (PAs), tissue‐type plasminogen activator (tPA), and urokinase‐type plasminogen activator (uPA), and their inhibitor type 1 (PAI‐1) in bone cells, suggests a role in one or more aspects of bone resorption such as osteoclast formation, mineral dissolution, and degradation of the organic matrix. These different processes were assayed in vitro using cells derived from mice with either tPA (tPA−/−), uPA (uPA−/−), PAI‐1 (PAI‐1−/−) inactivation or with a combined inactivation (tPA−/−:uPA−/−) and compared with wild‐type mice (WT). First, osteoclast formation, assessed by investigating the number and characteristics of tartrate‐resistant acid phosphatase–positive multinucleated cells formed in cocultures of primary osteoblasts and bone marrow cells treated with 1α,25‐dihydroxyvitamin D₃, was not different between the different cell types. Second, dentine resorption, an assay for osteoclast activity, was not affected by the combined deficiency of both tPA and uPA. Finally, the ability to degrade nonmineralized bone‐like matrix was however, significantly reduced in tPA−/−:uPA−/− cells compared with WT cells (28.1 ± 0.6%, n = 6 vs. 56.4 ± 3.1%, n = 6, respectively, p < 0.0001). Surprisingly, collagen proteolysis by bone cells was not dependent on the presence of plasmin as suggested by degradation assays performed on type I³H‐collagen films. Taken together, these data suggest that the plasminogen activator/plasmin system is not required for osteoclast formation, nor for the resorption of the mineral phase, but is involved in the removal of noncollagenous proteins present in the nonmineralized bone matrix.