Well Plate Integrated Topography Gradient Screening Technology for Studying Cell‐Surface Topography Interactions

Abstract

New high‐throughput technologies for cell–material interaction studies provide researchers with powerful tools to speed up research in the field of biomaterial–cell interactions. However, sharing technologies is often difficult due to the necessity of specific knowledge and experiences. Engineered surfaces can elucidate effects of surface topography on cell behavior, which is of critical value for gaining control over cellular processes. Here, the translation of a gradient‐based high‐throughput cell screening approach for aligned nano/micro topographies interacting with cells is presented. An aligned topography 96‐well plate is created by upscaling of highly specific gradient technology. The resulting cell culture dishes are compatible with general laboratory and imaging equipment, and the platform allows for studying cell behavior with regard to adhesion and alignment. The challenge lies in increasing the dimensions of the previous 1 × 1 cm gradient topography substrate, to be able to cover the span of a 96‐well plate and translate it into a standardized cell‐screening tool. Adhesion experiments of human bone marrow derived mesenchymal stem cells confirm the standardization, compatibility, and usability of the technology. In the process of using multi‐system imaging and analysis, it becomes apparent that future challenges need to include universally applied data analysis approaches.

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

“…The bottomless 96-well plate embedded with the PDMS membrane was prepared as described previously . Briefly, the samples for wrinkle gradients were cut into sizes of 9.6 × 7.5 cm to cover wells A1–H1 through columns 1–10 of the plate (Greiner Bio-One no.…”

“…The bottomless 96-well plate embedded with the PDMS membrane was prepared as described previously. 40 Briefly, the samples for wrinkle gradients were cut into sizes of 9.6 × 7.5 cm to cover wells A1−H1 through columns 1−10 of the plate (Greiner Bio-One no. 655101), and column 11 was sacrificed to connect the wrinkle gradient with column 12, which acts as a flat control (1.5 × 7.5 cm) (Figure S1).…”

“…In this study, we hypothesize that both wavelength and pitch contribute to topography-mediated cell stimulation. To investigate these individual contributions of topography parameters, we prepared a HTS platform consisting of 70 different combinations of wavelengths and amplitudes in one 96-well plate, following the work of der Boon et al 40 By sequentially preparing aligned topography gradients via a silicone stretch-oxidation-release method and imprinting lithography, followed by selective amplitude reduction using plasma oxidation for different durations, the amplitude and wavelength were decoupled, and the whole substrate was integrated into standardized well-plate technology. Using the multiparameter topography well plate, the optimum wavelength/amplitude combination for osteogenesis of human bone marrow-derived MSCs (hBM-MSCs) was investigated.…”

High-Throughput Routes to Biomaterials Discovery

“…The influence of surface topology on cell behavior has been studied using a “wrinkle” screening format. Zhou et al used nanometer-sized PDMS wrinkles to optimize aspect ratio parameters for osteoblast attachment, with this technology being further adaptable to 96-well plate formats (on a micrometer scale) for easy cell culture experiments (Figure ) as well as coating wrinkles in inorganic biocompounds to investigate the influence of biomolecules on cell behavior. , …”

“…Well plate integrated topography gradient screening technology for studying cell–surface topography interactions. Adapted with permission from ref . Copyright 2019 John Wiley and Sons.…”