Decoupling the Amplitude and Wavelength of Anisotropic Topography and the Influence on Osteogenic Differentiation of Mesenchymal Stem Cells Using a High-Throughput Screening Approach

Abstract

High-throughput screening (HTS) methods based on anisotropically topography gradients have been broadly used to investigate the interactions between cells and biomaterials. However, few studies focus on the optimum parameters of topography for osteogenic differentiation because the structures of topography are complex with multiple combinations of parameters. In this study, we developed polydimethylsiloxane (PDMS)-based wrinkled topography gradients (amplitudes between 144 and 2854 nm and wavelengths between 0.91 and 13.62 μm) and decoupled the wavelength and amplitude via imprinting lithography and shielded plasma oxidation. The PDMS wrinkle gradient was then integrated with the bottomless 96-well plate to constitute the wrinkled HTS platform, which consists of 70 different wrinkle parameters. From the in vitro culture of bone marrow stem cells, it was observed that aligned topography has an important influence on the macroscopic cell behavior (i.e., cell area, elongation, and nucleus area). Furthermore, the optimum wrinkle parameter (wavelength: 1.91 μm; amplitude: 360 nm) for osteogenic differentiation of stem cells was determined via this screening plate approach. This screening platform is not only beneficial for a better understanding of the interactions between topography and biomaterials but also advances the development of bone tissue engineering developments.

Synergistic Effect of Cell-Derived Extracellular Matrices and Topography on Osteogenesis of Mesenchymal Stem Cells

“…That is, the difference for W0.5 is less efficient than that for W3, which might be due to the addition of the CDM, thereby masking to some degree the topographical stimulation of the cell, which is still clearly visible for the W3 + CDM (Figure ). We recently showed that topographies, both the amplitude and wavelength, play an important role but overcrowding the topography too much will certainly make the stimulus less pronounced.…”

“…That is, the difference for W0.5 is less efficient than that for W3, which might be due to the addition of the CDM, thereby masking to some degree the topographical stimulation of the cell, which is still clearly visible for the W3 + CDM (Figure 2). We recently showed that topographies, both the amplitude and wavelength, play an important role 63 but overcrowding the topography too much will certainly make the stimulus less pronounced. Until now, although some researchers have elucidated that for proliferation, migration, and differentiation on topography 64 and stiffness 65 of the substrate, YAP-dependent mechanotransduction is required, few studies investigated the mechanotransduction of cells cultured on CDMs.…”

Dual anisotropicity comprising 3D printed structures and magnetic nanoparticle assemblies: towards the promotion of mesenchymal stem cell osteogenic differentiation

“…However, using other physical factors, e.g., anisotropicity, which is also universally present in living tissue and organs, to induce directional MCS osteogenic differentiation, has been relatively less extensively explored and utilized. There are multiple reasons for this fact, and one might be that the specificity of anisotropicity in the induction of MCS osteogenesis is not sufficiently high, although various studies have reported that anisotropicity of the substrate supports MCS osteogenic differentiation 15,22,23 .…”

High-Throughput Methods in the Discovery and Study of Biomaterials and Materiobiology

“…Microfluidics-based HTS platforms may result in more breakthrough discoveries in both fundamental research and clinical application. In addition, array/grad-wellplates (array or gradient surfaces integrated into commercial well plate technology) were developed, which consists of a standard well plate with bottoms containing arrays or gradients. − The array/grad-wellplate offers several key merits compared to the prior technologies, including preventing cross-communication of cells and cross-contamination of soluble factors, incorporate high throughput assays such as ELISA, as well as allowing for robotic liquid handling and implementation of multiwell plate-based instrumentation. Importantly, the array/grad-wellplate enables straightforward studying of synergetic effects of drugs/biomolecular (gene, protein, enzyme) and material properties, to identify the optimal conditions.…”