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Consistent with previous reports, we found that the larval head structures appeared largely cartilaginous (Figure 1) while in the post-metamorphosis frog, ossified bone makes up the vast majority of the head (Figure 3).[21] Recent advances in micro-computed tomography (μCT) (reviewed in Ritman 2004)[22] have allowed fine resolution three-dimensional imaging of ossification in small animals.
In addition, unlike adult long bone fragments or embryonic calvarial explants often used in other models [ 33- 35], the intact femur retains the three-dimensional, multi-cell architecture including ossified bone and hematopoietic cells, stroma and matrix tissue.
Whole skeletal stains for ossified bone and cartilage were used to characterize the entire skeleton of AtrxCol2Cre mice.
Assessing the ossified bone volume of several different bones in the skeleton, we found that the double mutants exhibited a decrease in bone volume (Tables 3 and 4) and length (data not shown) of the humerus and femur.
First, there does appear to be a deficit in bone growth in the double mutants, in that bones located in both the axial and appendicular skeleton, which form by the distinct processes of endochondral and intramembranous ossification, showed a decrease in ossified bone volume (Tables 3 and 4).
This is illustrated by the lack of ossified bone matrix in haematoxylin and eosin (H&E) stained coronal sections of mutant compared with wild-type embryos (Fig. 1C).
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Live Calcein staining of 6-month-old fish marks newly ossified bones in green.
The beads' effects on overall plate development were analyzed in live adult fish using 0.2% Calcein in aquarium water to mark newly ossified bones as previously described (Kimmel et al., 2003; Wada et al., 2010).
Thus, altered muscular forces, arising due to the deficiency in phosphate levels in the serum, may be associated with nanostructural abnormalities in intramembranously ossifying bone.
On the other hand, the bone-seeking radiotracer [99mTc]MDP may offer superior detection of lytic and ossifying bone metastases often associated with OS.
In this study we have demonstrated that degree of orientation, predominant orientation and mean mineral content increases with age in intramembranously ossifying bone.
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