Application of a Novel Widefield Surface Plasmon Resonance Microscope in Cell Imaging and Wound Closure Properties of TGF-β3,BSA/HCl and HCl in Cultured Human Bone Cell Monolayer

摘要

A newly developed Widefield Surface Plasmon Resonance (WSPR) Microscope was used to investigate the morphology of MG63 bone cells and their interfacial interactions with ECM proteins. This allowed detailed imaging of cell surface coupling at lateral resolution down to ～500 nm. In this work, bone repair was investigated and modulated by different stimulus including growth factors. TGF-β3 is a cytokine known to be associated with the scarless healing of skin and it is highly probable that it may play a role in the repair of other tissues. Thus the aim of this study was to investigate the effect of TGF-β3 on closure of a model wound in cultured monolayers of the MG63 human bone cells. This in vitro work examined and compared the wound closure properties of TGF-β3, and its dosage carriers HCl and BSA/HCl. The wound healing response was investigated in TC grade culture flasks by creating a wound (with average scratch width of 300 ± 10-30 μm SD, 1.7-5 μm SEM) on confluent monolayers of MG63 human bone cells. After wounding, cultures were then treated with 50ng/ml TGF-P3 at concentration of 4 mM HCl and 1 mg/ml BSA and distilled water. Also, the same method was applied for cell cultured monolayers with no growth factor as control and with HCl/BSA and HCl only solutions. After wounding, wound width was measured every 5 h over a 30-h period. The results showed that TGF-β3 enhanced the rate of wound repair in a monolayer of MG63 bone cells. It was found that after 20 h all the culture flasks treated with TGF-β3 (with 15.5% of wound remained open), HCl (with 16% of wound remained open) and finally BSA/HCl (with 17.7% of wound remained open) had resulted in faster wound healing compared to control (with 85% of wound remained open). These results indicated that wound closure in model MG63 wound with TGF-β3 was higher than the control.