Thrombospondin-2 deficiency in growing mice alters bone collagen ultrastructure and leads to a brittle bone phenotype.
Thrombospondin-2 (TSP2) is a matricellular protein component of the bone extracellular matrix. Long bones of adult TSP2-deficient mice have increased endosteal bone thickness due to expansion of the osteoblast progenitor cell pool, and these cells display deficits in osteoblastic potential. Here, we investigated the effects of TSP2 deficiency on whole bone geometric and mechanical properties in growing 6-wk-old male and female wild-type and TSP2-knockout (KO) mice. Microcomputed tomography and mechanical testing were conducted on femora and L2 vertebrae to assess morphology and whole bone mechanical properties. In a second series of experiments, femoral diaphyses were harvested from wild-type and TSP2-KO mice. Detergent-soluble type I collagen content was determined by Western blot of right femora. Total collagen content was determined by hydroxyproline analysis of left femora. In a third series of experiments, cortical bone was dissected from the anterior and posterior aspects of the femoral middiaphysis and imaged by transmission electron microscopy to visualize collagen fibrils. Microcomputed tomography revealed minimal structural effects of TSP2 deficiency. TSP2 deficiency imparted a brittle phenotype on cortical bone. Femoral tissue mineral density was not affected by TSP2 deficiency. Instead, transmission electron microscopy revealed less intensely stained collagen fibrils with altered morphology in the extracellular matrix assembled by osteoblasts on the anterior surface of TSP2-KO femora. Femoral diaphyseal bone displayed comparable amounts of total collagen, but the TSP2-KO bones had higher levels of detergent-extractable type I collagen. Together, our data suggest that TSP2 is required for optimal collagen fibrillogenesis in bone and thereby contributes to normal skeletal tissue quality.
Department of Orthopaedic Surgery, University of Michigan Medical School, Ann Arbor, Michigan.
Eugene
E
Manley
Joseph E
JE
Perosky
Basma M
BM
Khoury
Anita B
AB
Reddy
Kenneth M
KM
Kozloff
Andrea I
AI
Alford
D000818:Animals; D003094:Collagen / Q000378:metabolism*; D005260:Female; D005269:Femur / Q000378:metabolism* / Q000648:ultrastructure; D008159:Lumbar Vertebrae / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D010641:Phenotype; D019699:Thrombospondins / Q000235:genetics*; D055114:X-Ray Microtomography; D000099262:Thrombospondin 2
D003094:Collagen; D019699:Thrombospondins; D000099262:Thrombospondin 2
4-point bending; bone μCT; collagen; thrombospondin-2