in expression of Cox2. Again small elevations in Cox2 were observed in all groups after 1 hour which fell after 3 hours with the greatest variation seen in magnetic field alone and magnetic field plus Fz-MNP groups. No significant difference was observed between experimental groups at one hour after treatment. Discussion Wnt signalling is a crucial pathway controlling stem cell Relebactam biological activity behaviour and in recent years has become an attractive target for modulation with potential applications in stem cell therapies. This study has demonstrated the feasibility of using anti-body functionalised magnetic nanoparticles to specifically target and stimulate Frizzled 2 receptors expressed by hMSC for the activation of Wnt/-catenin signalling pathways. The efficiency of MNP coating with antibodies was first assessed by examining changes in MNP characteristics. The antibody coating process resulted in an increase in particle size relative to uncoated particles; this could be attributed to the added protein layer around the coated MNP. Furthermore, the relative increase in MNP surface charge could also be attributed to the presence of protein on the particle surface. Previous work has also shown that particle coating with antibodies alters particle surface charge in a similar manner. The binding specificity of Fz-MNP to Frizzled receptors was PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19763871 also assessed using a blocking study. Fz-MNP binding to cells was shown to be reduced when cells were pre-incubated with 11 / 18 Remote Activation of Wnt Signalling Fig 6. Anti-Frizzled MNP alter expression of Mechanosensitive genes. Gene expression analysis of mechano responsive genes in response to anti-frizzled particles or control particles and magnetic field stimulation was evaluated using quantitative real-time PCR after 1h and 3h treatment. NF-B gene expression was increased by Trek-MNP after 1h. Magnetic field treatment alone also increased NF-B expression after 3h. Treatment with Fz-MNP or Wnt 3A both showed similar but minor shifts in NF-B gene expression. c-Myc expression was increased after 1h treatment with magnetic field treatment alone with an added increase when used in conjunction with Trek-MNP. Treatment PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19761601 with Fz-MNP or Wnt 3A again both showed similar but negligible shifts in c-Myc expression. COX 2 
gene expression was shown to be significantly increased after 1h by magnetic field treatment alone but a decrease in expression was observed after 3h. Treatment with Fz-MNP or Wnt 3A both caused significant decreases in COX 2 expression after 3h treatment. Treatment with Fz-MNP was shown to significantly increase COX 2 expression when compared to Cells + magnet control after 3h. another Frizzled antibody. This infers that the Frizzled antibody successfully blocked the FzMNP binding sites on Frizzled, therefore reducing the available binding sites for Fz-MNP and consequently restricting particle binding to Frizzled receptors. This is an indicator that FzMNP are specific for Frizzled 2 receptors. Our results show that treatment of hMSC with Fz-MNP caused no noticeable changes in LRP5/6 phosphorylation, indicating that Fz-MNP are not activating and signalling through LRP co-receptors. The canonical Wnt signalling co-receptor Low density lipoprotein receptor-related protein 5/6 forms an active signalling complex with Frizzled and a 12 / 18 Remote Activation of Wnt Signalling Wnt ligand. Upon receptor activation, LRP is successively phosphorylated at multiple sites by kinases such as GSK, a process which is med