Focal adhesion dynamics in cellular function and disease.

The effects of substrate morphology by regulating pseudopods formation on cell directional alignment and migration

Based on the concept that microenvironment and physical stimuli regulate the cell behaviors like proliferation, migration, and differentiation, this study was conducted to investigate whether nanoscale spacing by stacked graphene film affect osteogenic differentiation of human adipose-derived mesenchymal stromal cells (hADSCs). The graphene films were synthesized by a chemical vapor deposition method, followed by etching and rinsing process to fabricate single or 3-, 5-, and 7-multilayers. The height and width of wrinkles of the graphene were confirmed by SEM and atomic force microscopy (AFM), ranging from 1.5 to 12.5 nm and from 30 to 100 nm, respectively. Osteogenic differentiation was significantly (p < 0.0001) promoted as the stacking layer increased. Immunofluorescent imaging and osteogenesis-related gene expression showed which increment was saturated from three layers. The calcium deposits and expression of osteogenesis-related genes (Runt-related transcription factor 2 and Osteocalcin) were highest in the three layers. In the hADSCs cultured on the three layers, the intensity of protein expression levels of filamentous actin (F-actin) was significantly increased (p = 0.0319) and focal adhesion kinase/extracellular signal-regulated kinase signal related genes were concomitantly activated. These results demonstrated that multilayer-stacked graphene creating nanoscale spaces promotes calcium deposit and cytoskeletal integrity in hADSC-related, in vitro-osteogenesis.

https://iopscience.iop.org/article/10.1088/2053-1583/abe105/pdf

Stacked Graphene with Nanoscale Wrinkles Supports Osteogenic Differentiation of Human Adipose-derived Stromal Cells

Cells sense extracellular matrix‐induced biophysical signals, which are transduced into intracellular signaling cascades, and trigger a series of cell responses, including adhesion, migration, and lineage commitment. Traditionally, in in vitro context, monofactorial approaches are employed to control cell fate, despite the fact that in vivo cells are exposed simultaneously to a diverse range of signals. Herein, an overview of key mechanotransduction pathways is first provided. Conventional single‐factor and contemporary multifactorial methodologies, based on substrate rigidity and surface topography, are then reviewed to recapitulate in vitro the in vivo niche, in an attempt to elucidate the underlying mechanisms involved in human mesenchymal stromal cell‐material interactions. 

It Takes Two to Tango: Controlling Human Mesenchymal Stromal Cell Response via Substrate Stiffness and Surface Topography

In vivo cell niches are complex architectures that provide a wide range of biochemical and mechanical stimuli to control cell behavior and fate. With the aim to provide in vitro microenvironments mimicking physiological niches, microstructured substrates have been exploited to support cell adhesion and to control cell shape as well as three dimensional morphology. At variance with previous methods, we propose a simple and rapid protein subtractive soft lithographic method to obtain microstructured polydimethylsiloxane substrates for studying stem cell adhesion and growth. The shape of adult renal stem cells and nuclei is found to depend predominantly on micropatterning of elastomeric surfaces and only weakly on the substrate mechanical properties. Differently, focal adhesions in their shape and density but not in their alignment mainly depend on the elastomer stiffness almost regardless of microscale topography. Local surface topography with concave microgeometry enhancing adhesion drives stem cells in a quasi-three dimensional configuration where stiffness might significantly steer mechanosensing as highlighted by focal adhesion properties.

Quasi-3D morphology and modulation of focal adhesions of human adult stem cells through combinatorial concave elastomeric surfaces with varied stiffness

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Quasi-3D morphology and modulation of focal adhesions of human adult stem cells through combinatorial concave elastomeric surfaces with varied stiffness