Patterning on Topography for Generation of Cell Culture Substrates with Independent Nanoscale Control of Chemical and Topographical Extracellular Matrix Cues

Abstract

The cell microenvironment plays an important role in many biological processes, including development and disease progression. Key to this is the extracellular matrix (ECM), a complex biopolymer network serving as the primary insoluble signaling network for physical, chemical, and mechanical cues. In vitro, the ability to engineer the ECM at the micro- and nanoscales is a critical tool to systematically interrogate the influence of ECM properties on cellular responses. Specifically, both topographical and chemical surface patterning has been shown to direct cell alignment and tissue architecture on biomaterial surfaces, however, it has proven challenging to independently control these surface properties. This protocol describes a method termed Patterning on Topography (PoT) to engineer 2D nanopatterns of ECM proteins onto topographically complex substrates, which enables independent control of physical and chemical surface properties. Applications include interrogation of fundamental cell-surface interactions and engineering interfaces that can direct cell and/or tissue function.

Protein Micropatterning in 2.5D: An Approach to Investigate Cellular Responses in Multi-Cue Environments

“…Waterkotte et al introduced an approach involving maskless lithography, chemical vapor deposition, and thermoforming to combine surface topographies with protein patterns . This method, however, is limited to materials that are suitable for thermoforming and has a relatively low pattern resolution (7.5 μm), while contact-guidance cues as small as 0.1 μm 2 have been shown to affect cell response. , Another approach was developed by Sevcik et al , who made use of microcontact printing on sacrificial poly­( N -isopropylacrylamide) (PIPAAm) layers that were subsequently used for transfer of the ink from the thermally transformed PIPAAm onto a PDMS substrate containing topographies. Unfortunately, this method is also limited in terms of pattern flexibility and resolution of the final pattern due to its need for photolithographic methods and microcontact printing.…”

Engineered substrates with imprinted cell-like topographies induce direct differentiation of adipose-derived mesenchymal stem cells into Schwann cells

“…These substrata can induce changes in cell spreading, nuclear shape, nuclear orientation, cytoskeletal architecture, and make alterations in transcript levels and protein expression. While differentiation may cause changes in cell shape, changes in cell shape could interestingly alter the differentiation of stem cells [34]. In 2004, McBeath et al demonstrated that cell shape can regulate the commitment of human mesenchymal stem cells (hMSCs) to adipocyte or osteoblast fate [35].…”

Subfascial Breast Augmentation: A Systematic Review and Meta-Analysis of Capsular Contracture

“…After collecting 22 studies with reported capsular contracture rates, the studies were subjected to a meta-analysis to determine the overall rates of capsular contracture and the weighted effects of each study. Several methods were utilized toward this end 21-33 and Microsoft Excel was used for calculation of the effects, statistical analysis of the studies as well as the visual display of the data in the form of forest plots with the reported outcomes. The overall heterogeneity was reported and a Q-test was applied as well as a calculation of I 2 which is an indicator of heterogeneity previously reported in the literature.…”