Transforming growth factor beta (TGF-β) isomers influence cell detachment of MG-63 bone cells

• We examine the effect of TGF-β isomers on bone cell detachment.
• Treatment with TGF-β1, TGF-β2, TGF-β3 and HCl decreased cell attachment.
• TGF-β3 significantly enhances cell detachment.
• Cells treated with TGF-β(2 + 3) induced greatest detachment compared to other TGF-β combinations.
• Imaging with the WSPR system enables us to further investigate the degree of attachment by looking at the focal contact.

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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