Osteogenesis Requires FAK-dependent Collagen Synthesis by Fibroblasts and Osteoblasts

Focal adhesion kinase (FAK) is critical in adhesion-dependent signaling, but its role in osteogenesis is ill defined. We deleted in fibroblasts and osteoblasts in mice bred with those expressing Cre-recombinase driven by 3.6-kb α1(I)-collagen promoter. Compared with wild-type (WT), conditional FAK-knockout (CFKO) mice were shorter (2-fold; < 0.0001) and had crooked, shorter tails (50%; < 0.0001). Microcomputed tomography analysis showed reduced bone volume (4-fold in tails; < 0.0001; 2-fold in mandibles; < 0.0001), whereas bone surface area/bone volume increased (3-fold in tails; < 0.0001; 2.5-fold in mandibles; < 0.001). Collagen density and fiber alignment in periodontal ligament were reduced by 4-fold ( < 0.0001) and 30% ( < 0.05), respectively, in CFKO mice. In cultured CFKO osteoblasts, mineralization at d 7 and mineralizing colony-forming units at d 21 were 30% ( < 0.0001) and >3-fold less than WT, respectively. Disruptions of FAK function in osteoblasts by conditional knockout, siRNA-knockdown, or FAK inhibitor reduced mRNA and protein expression of Runx2 (>30%), Osterix (>25%), and collagen-1 (2-fold). Collagen synthesis was abrogated in WT osteoblasts with Runx2 knockdown and in -null fibroblasts transfected with an FAK kinase domain mutant or a kinase-impaired mutant (Y397F). These data indicate that FAK regulates osteogenesis through transcription factors that regulate collagen synthesis.-Rajshankar, D., Wang, Y., McCulloch, C. A. Osteogenesis requires FAK-dependent collagen synthesis by fibroblasts and osteoblasts.