Next Generation Cell Culture Tools Featuring Micro‐ and Nanotopographies for Biological Screening

Abstract

Cells can perceive complex mechanical cues across both the micro‐ and nanoscale which can influence their development. While causative effects between surface topography and cellular function can be demonstrated, the variability in materials used in this screening process makes it difficult to discern whether the observed phenotypic changes are indeed a result of topographical cues alone or the inherent difference in material properties. A novel approach to directly imprint micro‐ and nanoscaled topographical features into the base of conventional cell cultureware is thus developed, facilitating its compatibility with standard biological techniques and methods of analysis. The utility of this technology is demonstrated by performing high‐throughput screening across five distinct cell types to interrogate the effects of 12 surface topographies, exemplifying unique cell‐specific responses to both behavior and cell morphological characteristics. The ability of this technology to underpin new insights into how surface topographies can regulate key image descriptors to drive cell fate determination is further demonstrated. These findings will inform the future development of advanced micro‐ and nanostructured cell culture substrates that can regulate cell behavior and fate determination across the life sciences, including fundamental cell biology, drug screening, and cell therapy.

The Bumpy Road to Stem Cell Therapies: Rational Design of Surface Topographies to Dictate Stem Cell Mechanotransduction and Fate

“…This technique has been applied to pattern tissue culture plasticware, with well plates embossed with pillars, grooves, and other complex topographies of various size and spacing. 68 The MSCs, HUVECs, C2C12s, human keratinocytes (HaCaT), and L929 cells cultured demonstrated unique responses to the substrates presented in a size-dependent manner. Large-area patterning has been attained with 200 and 25 nm pillars 70 produced on 10 m long films as a means of producing a potential screening application.…”

“…Currently, photolithographically patterned microarrays and coverslips are commercially available from companies such as Cytoo, Ibidi, or Alveole, due to the demand of specific topographies for in vitro cell studies. Additionally, the application of thermal nanoimprint lithography 68 to pattern surfaces of culture-well plates could be readily scaled to provide future patterning of larger culture-ware. Current technologies to add texture to biomedical devices modify these large surface areas via nonlithographic roughening and texturing techniques mostly due to their maskless and fast production characteristics.…”

“…322,323 One strategy to combat this involves increasing throughput to enable screening of larger numbers and types of topographic patterns. This approach is relatively well-advanced, with systems such as the Topochip, 324 pore size gradients, 323 or patterning technology for standard multiwell plates 68 providing opportunities to simultaneously test hundreds of features for their effects. Future knowledge will be aided by rational experimental design, to efficiently determine how different parameters in a topography (heights, shapes, sizes, gaps, orientation, order, etc.)…”

Extracellular Vesicle-Based Coatings Enhance Bioactivity of Titanium Implants—SurfEV

“…To improve osteointegration and the longevity of orthopaedic implants, their surfaces can be modified and functionalised [3]. Currently, there are two major strategies for modifying implant surfaces with the goal of enhancing osteointegration: (a) micro-and nanostructuring of the surface to create 'cell instructive' topographies [4,5] and (b) functionalization of surfaces with bioactive molecules that enable cell adhesion and differentiation [4][5][6].…”

Role of actin cytoskeleton in cargo delivery mediated by vertically aligned silicon nanotubes

“…Rational design, engineering, and fabrication of NNs' physical geometry/architecture-either via colloidal selfassembly [52][53][54][55][56] or nanofabrication routes [42,57,58]can offer close spatial control over optimal, localized, interfacial interactions for improving the nanoinjection efficacy into target cells. Enhanced control of nanoinjection is typically achieved by engineering the physical geometry of NN arrays-their tunable topological configuration (porous, solid, hollow) and their shape, density, height, and diameter [43,45].…”