Abstract
The dynamic response of cells when subjected to mechanical impact has become increasingly relevant for accurate assessment of potential blunt injuries and elucidating underlying injury mechanisms. When exposed to mechanical impact, a biological system such as the human skin, brain, or liver is rapidly accelerated, which could result in blunt injuries. For this reason, an acceleration of greater than > 150 g is the most commonly used criteria for head injury. To understand the main mechanism(s) of blunt injury under such extreme dynamic threats, we have developed an innovative experimental method that applies a well-characterized and -controlled mechanical impact to live cells cultured in a custom-built in vitro setup compatible with live cell microscopy. Our studies using fibroblast cells as a model indicate that input acceleration ({a}_{in}) alone, even when it is much greater than the typical injury criteria, e.g., {a}_{in}>1{,}000 g, does not result in cell damage. On the contrary, we have observed a material-dependent critical pressure value above which a sudden decrease in cell population and cell membrane damage have been observed. We have unambiguously shown that (1) this critical pressure is associated with the onset of cavitation bubbles in a cell culture chamber and (2) the dynamics of cavitation bubbles in the chamber induces localized compressive/tensile pressure cycles, with an amplitude that is considerably greater than the acceleration-induced pressure, to cells. More importantly, the rate of pressure change with time for cavitation-induced pressure is significantly faster (more than ten times) than acceleration-induced pressure. Our in vitro study on the dynamic response of biological systems due to mechanical impact is a crucial step towards understanding potential mechanism(s) of blunt injury and implementing novel therapeutic strategies post-trauma.
Highlights
The dynamic response of cells when subjected to mechanical impact has become increasingly relevant for accurate assessment of potential blunt injuries and elucidating underlying injury mechanisms
We investigate damage mechanisms of live cells to address our key question of “what are the key injury mechanisms, e.g., acceleration or pressure, and injury criteria associated with mechanical impact?” Towards this fundamental question, we have developed and utilized a new experimental approach for the application of well-controlled impact to live cell populations in vitro
One possible implication of these results is that the effect of shock waves on mild blunt injury is dampened because the human body is protected by soft skin, which would behave as the thin foam layer
Summary
We present a new technique for studying cell injury mechanisms by applying biologically relevant mechanical impact to in vitro cell culture This new approach is for maintaining consistent in vitro conditions during experiments, accommodating multiple cell populations, and monitoring each population in real-time while the impact-induced accelerations which mimic blunt injury are quantified with regards to amplitude and time scale. These multiplexed, environmental control capabilities are critical for studying the relationships between mechanical impact and cell injury due to the complex nature of interpreting input–output relationships in multivariate biological systems.
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