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Spheroid Assay to Measure TGF-β-induced Invasion
Published on: November 16, 2011
Transforming growth factor beta (TGF-β) isomers influence cell detachment of MG-63 bone cells
Farshid Sefat1, Seyed Ali Khaghani2, Touraj Nejatian3
1School of Engineering, Design and Technology-Medical Engineering, University of Bradford, Bradford, West Yorkshire, UK; School of Engineering, Department of Biomedical Engineering, King Faisal University, Al-Hofuf, Al-Ahsa, Saudi Arabia; Tissue Engineering Group, Department of Chemistry, Chemical Biology and Biomedical Engineering, Stevens Institute of Technology, Hoboken, NJ, USA.
Abstract:
Bone repair and wound healing are modulated by different stimuli. There is evidence that Transforming Growth Factor-beta (TGF-β) super-family of cytokines have significant effects on bone structure by regulating the replication and differentiation of chondrocytes, osteoblasts and osteoclasts. There is also significant evidence that interactions with extracellular matrix molecules influence cell behaviour. In this study cell surface attachment was examined via a trypsinization assay using various TGF-β isomers in which the time taken to trypsinize cells from the surface provided a means of assessing the strength of attachment. Three TGF-β isomers (TGF-β1, 2 and 3), four combined forms (TGF-β(1+2), TGF-β(1+3), TGF-β(2+3) and TGF-β(1+2+3)) along with four different controls (BSA, HCl, BSA/HCl and negative control) were investigated in this study. The results indicated that treatment with TGF-β1, 2, 3 and HCl decreased cell attachment, however, this effect was significantly greater in the case of TGF-β3 (p<0.001) indicating perhaps that TGF-β3 does not act alone in cell detachment, but instead functions synergistically with signalling pathways that are dependent on the availability of hydrogen ions. Widefield Surface Plasmon Resonance (WSPR) microscope was also used to investigate cell surface interactions.
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