title: PHOSPHO1 is essential for mechanically competent mineralization and the avoidance of spontaneous fractures.
abstract: Phosphatases are essential for the mineralization of the extracellular matrix within the skeleton. Their precise identities and functions however remain unclear. PHOSPHO1 is a phosphoethanolamine/phosphocholine phosphatase involved in the generation of inorganic phosphate for bone mineralization. It is highly expressed at sites of mineralization in bone and cartilage. The bones of Phospho1(-/-) mice are hypomineralized, bowed and present with spontaneous greenstick fractures at birth. In this study we show that PHOSPHO1 is essential for mechanically competent mineralization that is able to withstand habitual load. Long bones from Phospho1(-/-) mice did not fracture during 3-point bending but deformed plastically. With dynamic loading nanoindentation the elastic modulus and hardness of Phospho1(-/-) tibiae were significantly lower than wild-type tibia. Raman microscopy revealed significantly lower mineral:matrix ratios and lower carbonate substitutions in Phospho1(-/-) tibia. The altered dihydroxylysinonorleucine/hydroxylysinonorleucine and pyridinoline/deoxypyridinoline collagen crosslink ratios indicated possible changes in lysyl hydroxylase-1 activity and/or bone mineralization status. The bone formation and resorption markers, N-terminal propeptide and C-terminal telopeptide of Type I collagen, were both increased in Phospho1(-/-) mice and this we associated with increased bone remodeling during fracture repair or an attempt to remodel a mechanically competent bone capable of withstanding physiological load. In summary these data indicate that Phospho1(-/-) bones are hypomineralized and, consequently, are softer and more flexible. An inability to withstand physiological loading may explain the deformations noted. We hypothesize that this phenotype is due to the reduced availability of inorganic phosphate to form hydroxyapatite during mineralization, creating an undermineralized yet active bone.
authors.affiliation: The Roslin Institute and Royal School of Veterinary Studies, University of Edinburgh, Edinburgh, UK. carmen.huesa@roslin.ed.ac.uk
authors.forename: Carmen
authors.initials: C
authors.lastname: Huesa
authors.forename: Manisha C
authors.initials: MC
authors.lastname: Yadav
authors.forename: Mikko A J
authors.initials: MA
authors.lastname: Finnilä
authors.forename: Simon R
authors.initials: SR
authors.lastname: Goodyear
authors.forename: Simon P
authors.initials: SP
authors.lastname: Robins
authors.forename: K Elizabeth
authors.initials: KE
authors.lastname: Tanner
authors.forename: Richard M
authors.initials: RM
authors.lastname: Aspden
authors.forename: José Luis
authors.initials: JL
authors.lastname: Millán
authors.forename: Colin
authors.initials: C
authors.lastname: Farquharson
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001696:Biomechanical Phenomena / Q000502:physiology; D016723:Bone Remodeling / Q000502:physiology; D002113:Calcification, Physiologic / Q000502:physiology*; D003094:Collagen / Q000378:metabolism; D004548:Elasticity; D005269:Femur / Q000473:pathology / Q000503:physiopathology; D005598:Fractures, Spontaneous / Q000201:enzymology / Q000503:physiopathology* / Q000517:prevention & control*; D006244:Hardness; D051379:Mice; D010744:Phosphoric Monoester Hydrolases / Q000172:deficiency / Q000378:metabolism*; D013977:Tibia / Q000473:pathology / Q000503:physiopathology; D014783:Viscosity
chemical_list: D015415:Biomarkers; D003094:Collagen; C552022:PHOSPHO1 protein, mouse; D010744:Phosphoric Monoester Hydrolases