Control of Cell Alignment and Morphology by Redesigning ECM‐Mimetic Nanotopography on Multilayer Membranes

Abstract

Inspired by native extracellular matrix (ECM) together with the multilevel architecture observed in nature, a material which topography recapitulates topographic features of the ECM and the internal architecture mimics the biological materials organization is engineered. The nanopatterned design along the XY plane is combined with a nanostructured organization along the Z axis on freestanding membranes prepared by layer-by-layer deposition of chitosan and chondroitin sulfate. Cellular behavior is monitored using two different mammalian cell lines, fibroblasts (L929) and myoblasts (C2C12), in order to perceive the response to topography. Viability, proliferation, and morphology of L929 are sensitively controlled by topography; also differentiation of C2C12 into myotubes is influenced by the presence of nanogrooves. This kind of nanopatterned structure has also been associated with strong cellular alignment. To the best of the knowledge, it is the first time that such a straightforward and inexpensive strategy is proposed to produce nanopatterned freestanding multilayer membranes. Controlling cellular alignment plays a critical role in many human tissues, such as muscles, nerves, or blood vessels, so these membranes can be potentially useful in specific tissue regeneration strategies.

Cell Behavior within Nanogrooved Sandwich Culture Systems

“…This cell behavior is also in accordance with previous studies using grooved substrates. [9,10] Regarding SW conditions, the software analysis also corroborated the findings from qualitative data of the disposition of F-actin cytoskeleton (Figure 1), i.e. cells aligned according to the groove direction of the substrate on which they were seeded.…”

“…Additionally, we have mapped cellular orientation based on the alignment of actin filaments, and not the orientation of the cell body by cytoplasmic staining, as has been described in previous works. [10,21] Therefore, our study highlights relevant findings that should be considered when designing surface engineering strategies aiming to control cell behavior.…”

“…Conversely, mesenchymal stromal cells (MSCs) seeded on substrates that restrict spreading and lead to a rounded cell morphology impede the formation of mature adhesions, consequently directing adipogenic differentiation. [2] Other types of cells are also affected by contact guidance, such as pre-osteoblasts, [9] fibroblasts, [10] and myoblasts. [10] Moreover, the use of aligned substrates with different submicron groove and ridge widths has shown promising results in directing the differentiation of MSCs towards osteogenic, [11] adipogenic, [11] myogenic, [12] and chondrogenic [13] lineages.…”

“…[2] Other types of cells are also affected by contact guidance, such as pre-osteoblasts, [9] fibroblasts, [10] and myoblasts. [10] Moreover, the use of aligned substrates with different submicron groove and ridge widths has shown promising results in directing the differentiation of MSCs towards osteogenic, [11] adipogenic, [11] myogenic, [12] and chondrogenic [13] lineages. We have recently proposed nanogrooved microdiscs, "topodiscs", as substrates for cell-mediated assembly structures, [14] which could be used in bottom-up tissue engineering strategies.…”

Bioinspired multilayer membranes as potential adhesive patches for skin wound healing

“…To better control the physicochemical cell-material interactions and improve the biocompatibility, different strategies have been suggested; for instance, one can modify the chemistry of the surface with specific motifs30, increase the hydrophilicity of the surface 31 or even create a topography suitable to generate a cell response 32. In the last decades, biomimetic materials have boosted tissue engineering and regenerative medicine field 33, 34, with different suitable properties of materials being inspired in natural systems.…”