Up to 70% of the differences in human bone mass have been attributed to genetic background. These differences are associated with alterations in the biomechanical properties, micro-architecture and remodeling of bone as well as its susceptibility to fracture and its capacity for repair. In previous work it was shown that C57Bl6 mice carrying one copy of the parathyroid hormone related protein (PTHrP+/−) gene developed osteopenia by four months of age. The current study was designed to determine if the haploinsufficient phenotype was maintained on a C3H background. PTHrP+/+ and PTHrP+/− mice on C57Bl6 and C3H backgrounds were euthanised between 6 and 18 months of age. The femurs were harvested, fixed in 4% paraformaldehyde overnight and scanned on a Skyscan 1172 equipped with a 10kV X-ray source and a 10 megapixel camera at a resolution 5μm. The amount and quality of cortical and trabecular bone was quantified from 2D images and 3D reconstructions using CTAn, CTvol and CTVox software. The undecalcified specimens were embedded at low temperature in MMA, sectioned at 5 μm and stained with Von Kossa and Toluidine Blue to distinguish mineralized from unmineralized tissue.Purpose
Method
Internal fixation of fractures in the presence of osteopenia has been associated with a failure rate as high as 25%. Enhancing bone formation and osseointegration of orthopaedic hardware is a priority when treating patients with impaired bone regenerative capacity. Fibroblast Growth Factor (FGF) 18 regulates skeletal development and could therefore have applications in implant integration. This study was designed to determine if FGF 18 promotes bone formation and osseointegration in the osteopenic FGFR3−/− mouse and to examine its effect on bone marrow derived mesenchymal stem cells (MSCs). In Vivo: Intramedullary implants were fabricated from 0.4 × 10mm nylon rods coated with 300nm of titanium by physical vapour deposition. Skeletally mature, age matched female FGFR3−/− and wild type mice received bilateral intramedullary femoral implants. Left femurs received an intramedullary injection of 0.1μg of FGF 18 (Merck Serono), and right femurs received saline only. Six weeks later, femurs were harvested, radiographed, scanned by micro CT, and processed for undecalcified for histology. In Vitro: MSCs were harvested from femurs and tibiae of skeletally mature age matched FGFR3−/− and wild type mice. Cells were cultured in Alpha Modified Eagles Medium (αMEM) to monitor proliferation or αMEM supplemented with ascorbic acid and sodium beta-glycerophosphate to monitor differentiation. Proliferation was assessed through cell counts and metabolic activity at days 3, 6 and 9. Differentiation was assessed through staining for osteoblasts and mineral deposition at days 6, 9 and 12.Purpose
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