Mesenchymal stem cells (MSCs) reside around blood vessels in all organs. This reservoir of progenitors can be ‘recruited’ in response to injury. The ability to manipulate stem cells therapeutically within injured tissue provides an attractive alternative to transplantation. Stem cells are regulated by neighbouring cells. We hypothesized that endothelial cells (ECs) influence MSC differentiation into bone and fat. MSCs were sorted from fat using fluorescent activated sorting. Their capacity to differentiate into bone, fat and cartilage was used to confirm MSC phenotype. MSCs and ECs were cultured in two-dimensions (standard culture dishes) and three-dimensions (vascular networks suspended in gel). Cocultures were exposed to osteogenic and adipogenic media. The role of EC-released factors on MSC differentiation was determined using a system in which cells share media but do not contact. Wnt pathway modulators were used to investigate the role of Wnt signalling. MSCs differentiated into bone, fat and cartilage. MSCs and ECs integrated in two- and three-dimensions. MSCs and ECs formed vessel-like structures in three-dimensions. When cultured with ECs, MSC differentiation to bone was accelerated while differentiation to fat was inhibited. This effect on osteogenesis was maintained when cells shared media but did not contact. Coculture with Wnt modulators confirmed that this effect is in part, mediated through Wnt signalling. Our data suggest that ECs influence MSC differentiation. Therapeutic targeting of EC-MSCs signalling may enable manipulation of MSCs in vivo avoiding the need for cell transplantation. This could enable trauma and orthopaedic patients who have healthy resident stem cells to self-repair

Improving periprosthetic bone is essential for implant fixation and reducing peri-implant fracture risk. This studied examined the individual and combined effects of iPTH and mechanical loading at the cellular, molecular, and tissue level for periprosthetic cancellous bone. Adult rabbits had a porous titanium implant inserted bilaterally on the cancellous bone beneath a mechanical loading device on the distal lateral femur. The right femur was loaded daily, the left femur received a sham loading device, and half of the rabbits received daily PTH. Periprosthetic bone was processed up to 28 days for qPCR, histology, and uCT analysis. We observed an increase in cellular and molecular markers of osteoblast activity and decrease in adipocytic markers for both treatments, with small additional effects in the combined group. Loading and iPTH led to a decrease and increase, respectively, in osteoclast number, acting through changes in RANKL/OPG expression. Changes in SOST and beta-catenin mRNA levels suggested an integral role for the Wnt pathway. We observed strong singular effects on BV/TV of both loading (1.53 fold) and iPTH (1.54 fold). Combined treatment showed a small additive effect on bone volume. In conclusion, loading and iPTH act through a pro-osteoblastic/anti-adipocytic response and through control of bone turnover via changes in the RANKL/OPG pathway. These changes led to a small additional, but not synergistic, increase in bone volume with the combined therapy

Introduction. Canonical Wnt inhibitor Sclerostin (SOST) may be a key mechanotransduction regulator. Methods. Unloading/loading 10 week old Sost−/− and WT mice. Unloading: Quads and calf muscles injected each with 0.5U botulinum toxin (BTX, Allergan) caused tibial unloading. Loading: 1200 cycles of tibial axial loading, 1200μe on mid-shaft, 4Hz, 5 days/week. Treated and control tibiae μCT scanned (Skyscan 1174) at 2 weeks. Results. Unloading the WT tibiae significantly decreased cortical bone volume (−5%) and thickness (−7%) compared to WT control (p<0.01). Larger bone volume loss (−25%) was seen in the trabecular compartment (p<0.01), along with 10% and 22% decreases in trabecular thickness and number (p<0.01). These parameters were not altered between unloaded and control Sost−/− tibiae. Tibial loading increased cortical bone volume in WT (18%) and Sost−/− (25%) mice (p<0.01). Cortical thickness was also increased in WT (19%) and Sost−/− (17%) mice (p<0.01). The trabeculae of the WT loaded tibiae showed significant thickening (15%, p<0.01) not seen in the Sost−/− tibiae. Metaphyseal cortical bone volume increased in both loaded WT (13%) and Sost−/− (31%) tibiae compared to their controls (p<0.01), suggestive of metaphyseal corticalisation. Conclusion. SOST knockout inhibited unloading-induced bone loss, but not loading-induced bone gain. SOST may have an important role in bones response to unloading, but may not be essential for the response to loading

Objectives

The biomembrane (induced membrane) formed around polymethylmethacrylate (PMMA) spacers has value in clinical applications for bone defect reconstruction. Few studies have evaluated its cellular, molecular or stem cell features. Our objective was to characterise induced membrane morphology, molecular features and osteogenic stem cell characteristics.