Proliferating osteoblasts are necessary for maximal bone anabolic response to loading in mice

Following mechanical loading, osteoblasts may arise via activation, differentiation, or proliferation to form bone. Our objective was to ablate proliferating osteoblast lineage cells in order to investigate the importance of these cells as a source for loading-induced bone formation. We utilized 3.6Col1a1-tk mice in which replicating osteoblast lineage cells can be ablated in an inducible manner using ganciclovir (GCV). Male and female mice were aged to 5- and 12-months and subjected to 5 days of tibial compression. Experimental mice were tk-positive, treated with GCV; control mice were either tk-negative treated with GCV, or tk-positive treated with PBS. We confirmed that experimental mice had a decrease in tk-positive cells that arose from proliferation. Next, we assessed bone formation after loading to low (7N) and high (11N) forces and observed that periosteal bone formation rate in experimental mice was reduced by approximately 70% for both forces. Remarkably, woven bone formation induced by high-force loading was blocked in experimental mice. Loading-induced lamellar bone formation was diminished but not prevented in experimental mice. We conclude that osteoblast proliferation induced by mechanical loading is a critical source of bone forming osteoblasts for maximal lamellar formation and is essential for woven bone formation.