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Directionality was determined by the sign of the mean difference. Quantitative data for the in vivo experiments were expressed as the mean?��?SD, and statistical comparisons between femurs made using a paired Student's t-test. A probability of p?Anticancer Compound Library screening components. The response of donor MSCs isolated from FGFR3+/+ marrow to implant grade titanium was first characterized ex vivo. Figure 2 shows that MSC isolated by adherence to TCP and plated on titanium formed characteristic condensations that stained positive for ALP (A and B) and supported mineral deposition (D). The quantitative data shown in Table 1 indicate the time-dependent maturation of cultures grown on titanium was similar Alectinib in vitro to that of MSC grown on TCP. Cell differentiation, as evidenced by in situ ALP staining, increased over time while ALP enzyme activity declined at day 10 concomitant with mineral deposition. Metabolism increased until day 10 and then slowed down as the cells reached the end of their lifespan once the matrix mineralized. RT-PCR analysis of RNA harvested at the same time points showed similar expression patterns for recognized markers of osteoblast differentiation, when normalized to GAPDH expression, in MSC grown on TCP or on titanium discs (data not shown). In previous work we showed that osteopenia in young adult FGFR3?/? mice was due in large part to aberrant maturation of MSC into osteoblasts.13 In the current work we depleted endogenous MSC in FGFR3?/? mice using sub-lethal irradiation to condition the mice for transplant. Figure 3 shows the apparatus used to shield the upper body while 13.5?Gy of photon irradiation was delivered to the hind limbs (A). This dose of radiation reduced the adherent MSC population by 80% (B) compared with that from age-matched non-irradiated FGFR3?/? mice (Irradiated: 3.35?��?1.05?��?105 compared to Non-irradiated: 18.08?��?4.92?��?105). When equal numbers of irradiated Sitaxentan FGFR3?/? and control FGFR3+/+ cells were re-plated on TCP there were sixfold fewer irradiated FGFR3?/? cells compared with non-irradiated FGFR3+/+ cells at day 9 (C) of culture, indicating a severe impairment in their replication capacity. Bone growth around intra-femoral implants was quantified using micro CT 6 weeks after transplantation of 105 MSC from FGFR3+/+ donor mice into the RIGHT femoral canal of irradiated FGFR3?/? recipient mice. Figure 4 shows the location of the implants in situ (A and B) and the 2?mm?��?0.