Alendronate reduces fractures in the zebrafish OI model. (A) Outline of experimental regime for cyclical treatment of frf with 50 µg/ml alendronate and subsequent imaging of fins for spontaneous fracture occurrence. (B-D) Brightfield images of tail fins of frf fish at 11 weeks post fertilisation (wpf), either untreated and directly imaged (B), cyclically immersed in 50 µg/ml alendronate twice a week (3 h each day) for 3 weeks (D) or sham treated, where fish were immersed in normal tank water to recapitulate handling of the OI fish (C). (E) Number of spontaneous fractures observed was counted at both 11 wpf and 13 wpf (n=7 per point; Mann–Whitney test). (G) Outline of experimental regime for cyclical treatment of frf with 50 µg/ml alendronate, fracturing and subsequent imaging of fins following alternate staining with calcein (green) or Alizarin Red (red) to visualise callus growth. (H,I) Fluorescent images of frf fractures either treated with 50 µg/ml alendronate (I) or untreated (H) and imaged for calcein or Alizarin Red at the indicated time points. (J) The relative bone callus width was compared, but no difference in callus growth was noted upon treatment with alendronate (n=8 per point). *P<0.05; ANOVA with Tukey post test. Scale bars: in D, 2 mm for B-D; in I, 100 µm for H,I.
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