Mechanical Phenotyping of MG63s Following Vibrational Stimulation.

Nanovibration, a kilohertz-frequency, nano-amplitude mechanical stimulation, has been shown to drive osteogenesis; however, the mechanisms remain unclear. Mechanotransduction has been proposed with limited cell population-level evidence. We propose the use of a high-throughput mechanical phenotyping technique, Real-Time Deformability Cytometry (RT-DC), to observe mechanical changes in an osteogenic model, MG63s. We have demonstrated that MG63 cells respond to the nanovibrational mechanical stimulation by changing their cytoskeletal morphology and showing higher expression of osteocalcin than respective controls. We have also demonstrated the first use of Real-Time Deformability Cytometry (RT-DC) to reliably phenotype whole-cell population mechanical response to this stimulus. The use of high-throughput microfluidic techniques such as RT-DC is proving invaluable to more accurately assay population morphological changes compared to other established techniques, with potential application in mechanobiology, cellular quality control, and diagnostic scenarios. With consideration for osteogenic changes, RT-DC also poses potential use in the assessment of in vitro and ex vivo bone cell samples, highlighting clinical relevance for conditions such as osteoporosis and bone fracture.