No abstract

The extracellular Ca2+-sensing receptor (CaR), a G protein-coupled receptor responsible for maintenance of calcium homeostasis, is implicated in regulation of skeletal metabolism. To discern the role of the osteoblast CaR in regulation of bone development and remodeling, we generated mice in which the CaR is excised in a broad population of osteoblasts expressing the 3.6 kb a1(I) collagen promoter. Conditional knockouts had abnormal skeletal histology at birth and developed progressively reduced mineralization secondary to retarded osteoblast differentiation, evident by significantly reduced numbers of osteoblasts and decreased expression of collagen I, osteocalcin and sclerostin mRNAs. Elevated expression of ankylosis protein, ectonucleotide pyrophosphatase/phosphodiesterase 1, and osteopontin mRNAs in the conditional knockout indicate altered regulation of genes important in mineralization. Knockout of the osteoblast CaR also resulted in increased expression of the receptor activator of nuclear factor kappa B ligand (RANK-L), the major stimulator of osteoclast differentiation and function, consistent with elevated osteoclast numbers in vivo. Osteoblasts from the conditional knockouts exhibited delayed differentiation, reduced mineralizing capacity, altered expression of regulators of mineralization and increased ability to promote osteoclastogenesis in co-culture experiments. We conclude that CaR signaling in a broad population of osteoblasts is essential for bone development and remodeling and plays an important role in the regulation of differentiation and expression of regulators of bone resorption and mineralization.

We determined whether quantifying neuroblastoma-associated mRNAs (NB-mRNAs) in bone marrow and blood improves assessment of disease and prediction of disease progression in patients with relapsed/refractory neuroblastoma. mRNA for CHGA, DCX, DDC, PHOX2B, and TH was quantified in bone marrow and blood from 101 patients concurrently with clinical disease evaluations. Correlation between NB-mRNA (delta cycle threshold, Δ, for the geometric mean of genes from the TaqMan Low Density Array NB5 assay) and morphologically defined tumor cell percentage in bone marrow, I-meta-iodobenzylguanidine (MIBG) Curie score, and CT/MRI-defined tumor longest diameter was determined. Time-dependent covariate Cox regression was used to analyze the relationship between Δ and progression-free survival (PFS). NB-mRNA was detectable in 83% of bone marrow (185/223) and 63% (89/142) of blood specimens, and their Δ values were correlated (Spearman = 0.67, < 0.0001), although bone marrow was 7.9 ± 0.5 stronger than blood When bone marrow morphology, MIBG, or CT/MRI were positive, NB-mRNA was detected in 99% (99/100), 88% (100/113), and 81% (82/101) of bone marrow samples. When all three were negative, NB-mRNA was detected in 55% (11/20) of bone marrow samples. Bone marrow NB-mRNA correlated with bone marrow morphology or MIBG positivity ( < 0.0001 and = 0.007). Bone marrow and blood Δ values correlated with PFS ( < 0.001; = 0.001) even when bone marrow was morphologically negative ( = 0.001; = 0.014). Multivariate analysis showed that bone marrow and blood Δ values were associated with PFS independently of clinical disease and gene status ( < 0.001; = 0.055). This five-gene NB5 assay for NB-mRNA improves definition of disease status and correlates independently with PFS in relapsed/refractory neuroblastoma. .

Altered cartilage loading is believed to be associated with osteoarthritis development. However, there are limited data regarding the influence of normal gait, an essential daily loading activity, on cartilage strains. In this study, 8 healthy subjects with no history of knee surgery or injury underwent magnetic resonance imaging of a single knee prior to and following a 20-minute walking activity at approximately 1.1 m/s. Bone and cartilage surfaces were segmented from these images and compiled into 3-dimensional models of the tibia, femur, and associated cartilage. Thickness changes were measured across a grid of evenly spaced points spanning the models of the articular surfaces. Average compartmental strains and local strains were then calculated. Overall compartmental strains after the walking activity were found to be significantly different from zero in all four tibiofemoral compartments, with tibial cartilage strain being significantly larger than femoral cartilage strain. These results provide baseline data regarding the normal tibiofemoral cartilage strain response to gait. Additionally, the technique employed in this study has potential to be used as a “stress test” to understand how factors including age, weight, and injury influence tibiofemoral cartilage strain response, essential information in the development of potential treatment strategies for the prevention of osteoarthritis.