Steering the osteoclast through the demineralization-collagenolysis balance.

There is a lot of interest for how and how much osteoclasts resorb bone. However, little is known about the mechanism which controls the orientation and the duration of a resorptive event, thereby determining the specific geometry of a cavitation. Here we show that the relative rate of collagenolysis vs. demineralization plays a critical role in this process. First we observed that when culturing osteoclasts on bone slices, excavations appeared either as round pits containing demineralized collagen, or as elongated trenches without demineralized collagen. This suggests that round pits are generated when collagen degradation is slower than demineralization, and trenches when collagen degradation is as fast as demineralization. Next we treated the osteoclasts with a low dose of a carbonic anhydrase inhibitor to slightly decrease the rate of demineralization, thereby allowing collagen degradation to proceed as fast as demineralization. This resulted in about a two-fold increase of the proportion of trenches, thus supporting our hypothesis. The same result was obtained if facilitating collagen degradation by pre-treating the bone slices with NaOCl. In contrast, when decreasing the rate of collagenolysis vs. demineralization by the addition of a cathepsin K specific inhibitor, the proportion of trenches fell close to 0%, and furthermore the round pits became almost half as deep. These observations lead to a model where the osteoclast resorption route starts perpendicularly to the bone surface, forming a pit, and continues parallel to the bone surface, forming a trench. Importantly, we show that the progress of the osteoclast along this route depends on the balance between the rate of collagenolysis and demineralization. We propose that the osteocytes and bone lining cells surrounding the osteoclast may act on this balance to steer the osteoclast resorptive activity in order to give the excavations a specific shape.