BMP-14 deficiency inhibits long bone fracture healing: a biochemical, histologic, and radiographic assessment.

Objectives: Bone morphogenetic proteins (BMPs) represent a distinct subset of the transforming growth factor-beta family best known for their role in joint formation and bone growth, and several recent clinical trials have begun to look at their efficacy in the augmentation of fracture healing. The goal of this research is to examine the effect of BMP-14, also known as growth differentiation factor-5 and cartilage- derived morphogenetic protein-1 (GDF-5, CDMP-1) on fracture healing by studying the long bone repair process in mice with a deficiency in this signaling peptide.

Methods: The animal model used for these studies was the BMP-14 (-/-) brachypodism (bp) mouse. Phenotypically normal heterozygous (+/-) littermates were used as controls. Closed mid shaft femur fractures were created and stabilized with intramedullary fixation in 8-week-old female mice. Forty-eight mice per genotype group were examined. On postoperative days 4, 7, 10, 14, 21, 28, 35, and 42, the mice were killed and the femurs and repair tissue were harvested for analysis. At each time point, the fracture sites were analyzed radiographically, histologically, and biochemically. For all quantitative analyses, the data were normalized and analyzed statistically using a 2-factor ANOVA test.

Results: Biochemically, peak values of normalized proteoglycan content were approximately 3 times less in the mutant fractures early in the time course of healing compared with the controls (P < 0.05). Histologically, BMP-14-deficient fractures exhibited a delay in peak area cell density, callus organization, and bone formation compared with controls. Radiographic analysis demonstrated that the peak callus was 2 weeks delayed and approximately 2 times less in the mutants compared with controls (P < 0.05). Radiographic grading of callus also demonstrated a significant difference after day 14.

Conclusions: Based on histologic, radiographic, and biochemical analysis, BMP-14-deficient mice display a short-term delay in healing of approximately 1 to 2 weeks. The observed abnormalities seem to be the result of a delay in cellular recruitment and chondrocyte differentiation in the early stages fracture repair in the absence of BMP-14. These results support the hypothesis that BMP-14 deficiency leads to a delay in fracture healing. Further studies are warranted to more closely examine the role of BMP-14 in normal fracture healing and the mechanism by which it works.