Single-Cell Fluidic Force Spectroscopy Reveals Dynamic Mechanical Fingerprints of Malignancy in Breast Cancer.

The interplay between cancer cell physical characteristics and metastatic potential highlights the significance of cancer cell mechanobiology. Using fluidic-based single-cell force spectroscopy (SCFS), quartz crystal microbalance with dissipation (QCM-D), and a model of cells with a spectrum of metastatic potential, we track the progression of biomechanics across the metastatic states by measuring cell-substrate and cell-to-cell adhesion forces, cell spring constant, cell height, and cell viscoelasticity. Compared to highly metastatic cells, cells in the lower spectrum of metastatic ability are found to be systematically stiffer, less viscoelastic, and larger. These mechanical transformations in cells within a cluster correlate with cells' metastatic potential but are significantly absent in single cells. Additionally, the response to chemotherapy is found to be highly dependent on cell viscoelastic properties in terms of both response time and magnitude. Shifts in cell softness and elasticity might serve as mechanoadaptive mechanisms during cancer cell metastasis, contributing to our understanding of metastasis and the effectiveness of potential therapeutic interventions.

Single-cell fluid-based force spectroscopy reveals near lipid size nano-topography effects on neural cell adhesion.

Nano-roughness has shown great potential in enhancing high-fidelity electrogenic cell interfaces, owing to its characteristic topography comparable to proteins and lipids, which influences a wide range of cellular mechanical responses. Gaining a comprehensive understanding of how cells respond to nano-roughness at the single-cell level is not only imperative for implanted devices but also essential for tissue regeneration and interaction with complex biomaterial surfaces. In this study, we quantify cell adhesion and biomechanics of single cells to nano-roughened surfaces by measuring neural cell adhesion and biomechanics via fluidic-based single-cell force spectroscopy (SCFS). For this, we introduce nanoscale topographical features on polyimide (PI) surfaces achieving roughness up to 25 nm without chemical modifications. Initial adhesion experiments show cell-specific response to nano-roughness for neuroblastoma cells (SH-SY5Y) compared to human astrocytes (NHA) around 15 and 20 nm surface roughness. In addition, our SCFS measurements revealed a remarkable 2.5-fold increase in adhesion forces (150-164 nN) for SH-SY5Y cells cultured on roughened PI (rPI) surfaces compared to smooth surfaces (60-107 nN). Our data also shows that cells can distinguish changes in nano-roughness as small 2 nm (close to the diameter of a single lipid) and show roughness dependence adhesion while favoring 15 nm. Notably, this enhanced adhesion is accompanied by increased cell elongation upon cell detachment without any significant differences in cell area spreading. The study provides valuable insights into the interplay between nano-topography and cellular responses and offers practical implications for designing biomaterial surfaces with enhanced cellular interactions.

Investigating malignancy-dependent mechanical properties of breast cancer cells.

Cancer invasiveness significantly impacts cellular mechanical properties which regulate cell motility and, subsequently, cell metastatic potential. Understanding the adhesion forces and stiffness/rigidity of cancer cells can provide better insights into their mechanical adaptability related to their degree of invasiveness. Here, we used single-cell force spectroscopy in conjunction with quartz crystal microbalance-with dissipation measurements to compare the mechanical properties of mammary epithelial cancer cells with different metastatic potentials, namely MCF-7 (non-invasive) and MDA-MB-231 (aggressive and highly invasive). Our results showed that MCF-7 exhibits larger adhesion forces, stronger intercellular forces, and a considerably stiff/rigid phenotype, contrary to MDA-MB-231. The biomechanical properties obtained are associated with the malignant potential of these cells such that the forces of adhesion and viscoelasticity are inversely proportional to cell invasiveness. This study integrates a new quantitative tool with real-time measurements to provide better insights into the mechanics of cancer cells across metastatic stages.