Fibronectin-Based Nanomechanical Biosensors to Map 3D Strains in Live Cells and Tissues

Shiwarski, Daniel J.; Tashman, Joshua W.; Tsamis, Alkiviadis; Bliley, Jacqueline M.; Blundon, Malachi A.; Aranda‐Michel, Edgar; Jallerat, Quentin; Szymanski, John M.; McCartney, Brooke M.; Feinberg, Adam W.

doi:10.1101/2020.02.11.943696

Cited by 4 publications

(13 citation statements)

References 61 publications

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“…Fabrication of the nanomechanical biosensors. A step-by-step protocol describing the fabrication process can be found at Protocol Exchange 65 . The NMBS were fabricated using an adaptation the surface-initiated assembly technique (SIA) 31,66 .…”

Section: Methodsmentioning

confidence: 99%

“…3D strain quantification and imaris strain mapping. For the image segmentation and strain quantification of the NMBS in 3D we developed an Imaris (Bitplane) and MATLAB (Mathworks) image analysis pipeline 73 . 3D image stacks obtained from confocal fluorescence microscopy were imported into Imaris for rendering and visualization.…”

Section: Methodsmentioning

confidence: 99%

“…We developed a MATLAB-based Imaris Xtension Biomechanical Analysis package to calculate 3D strain from the fluorescence NMBS images. To calculate strain from the change in length between paired NMBS nodes, coordinates for each spot in X, Y, Z were exported and read into MATLAB (XTSpotsToFilamentsWithStrain.m) 73 . An image of the NMBS in its reference state was imported and maximally intensity projected into a 2D reference image for spot overlay.…”

Section: Methodsmentioning

confidence: 99%

“…We next created an import Xtension in Imaris (XTFilamentLengthStrain.m) 73 to import each segment strain value as a filament object in Imaris that can be color coded with a strain statistic. Filaments were created for all node pairs and tracked for each time point.…”

Section: Methodsmentioning

confidence: 99%

“…To determine regional dilation and contraction within the NMBS over time, a binary mask was created from the imported Imaris filaments object for each time point and used to generate a Cell object within the Imaris area. A custom Imaris Xtension was then created to calculate regional dilation and contraction from the change area defined by a Cell within the NMBS grid region (XTCellAreaDilation.m) 73 . Changes in dilation were visualized as a color-coded dilation statistic normalized around 0, with positive dilation values corresponding to expansion and negative dilation corresponding to contraction.…”

Section: Methodsmentioning

confidence: 99%

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Fibronectin-based nanomechanical biosensors to map 3D surface strains in live cells and tissue

et al. 2020

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Mechanical forces are integral to cellular migration, differentiation and tissue morphogenesis; however, it has proved challenging to directly measure strain at high spatial resolution with minimal perturbation in living sytems. Here, we fabricate, calibrate, and test a fibronectin (FN)-based nanomechanical biosensor (NMBS) that can be applied to the surface of cells and tissues to measure the magnitude, direction, and strain dynamics from subcellular to tissue length-scales. The NMBS is a fluorescently-labeled, ultra-thin FN lattice-mesh with spatial resolution tailored by adjusting the width and spacing of the lattice from 2–100 µm. Time-lapse 3D confocal imaging of the NMBS demonstrates 2D and 3D surface strain tracking during mechanical deformation of known materials and is validated with finite element modeling. Analysis of the NMBS applied to single cells, cell monolayers, and Drosophila ovarioles highlights the NMBS’s ability to dynamically track microscopic tensile and compressive strains across diverse biological systems where forces guide structure and function.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Fibronectin-based nanomechanical biosensors to map 3D surface strains in live cells and tissue

et al. 2020

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Novel Temperature‐Sensitive Hydrogel Promotes Wound Healing Through YAP and MEK‐Mediated Mechanosensitivity

Huang

Jiang

et al. 2022

Adv Healthcare Materials

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Wound healing is a significant problem in clinical management. Various functional dressings are studied to promote wound healing through biochemical factors. They are generally expensive, complex to fabricate, and may adversely affect the wound. Mechanical forces are the critical regulators of tissue repair. Although contraction is shown to promote wound healing, the underlying mechanisms are not fully understood. In this study, a novel adhesive temperature‐sensitive mechanically active hydrogel with a simple and inexpensive preparation process is developed. The dressing is able to adhere to the wound surface and actively contract the wound in response to body temperature. This mechanical contraction enhances the proliferative activity of basal cells, reduces the inflammatory response of the wound, and promotes wound healing. Furthermore, RNA‐seq clarifies how the gene regulatory network is regulated by contraction. Finally, using pharmacological inhibitors, YAP and MEK are identified as the key signaling molecules for contraction‐mediated tissue healing in vivo.

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Microstructure-based engineering of soft biological materials

Tsamis

2022

Adv.Tech&Mat.

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Aortic disease (AoD) is a leading cause of mortality in developed countries. Two of the most common forms of AoD are aneurysm (widening) and dissection (tear in inner wall). Aneurysm and dissection often associate with bicuspid aortic valve (BAV) instead of the normal tricuspid aortic valve, and BAV aneurysms of ascending thoracic aorta have the tendency to bulge asymmetrically towards the greater curvature of aorta. Multiphoton microscopy can help us image collagen and elastin fibres, which are considered as main load-bearing constituents of the aortic wall, in order to investigate potential role of fibre microstructure in ascending thoracic aortic aneurysm or dissection. Regional differences in fibre microstructure may be driven by distinct mechanisms of vascular remodelling, and, combined with mechanical tests, could improve our understanding of the biomechanical mechanisms of aortic aneurysm and dissection potential. Should we wish to investigate the effect of microstructure in soft tissue formation and organ development, we would have to consider a rapidly growing process. In that process, the cells are the main load-bearing components, which cooperate to produce tissue-level forces that shape tissue formation. Our understanding of this phenomenon, called mechanotransduction, has advanced significantly over the past years, to the point where it is now clear that nearly every biological process is modulated by how these forces are decoded intracellularly. It is therefore important to create our own fluorescently-labeled matrix that could integrate into the tissue and enable tracking of these forces in-vivo. A new 3D optical nanomechanical biosensor (NMBS) based on fluorescent fibronectin fibres was developed based on integrated photolithography and micro-contact printing technology. NMBS was successfully validated under uniaxial tensile test of biologically relevant materials for microscopic vs. macroscopic mechanical strains. In the future, biomimetic 3D scaffolds could be fabricated by assembly of 2D fibre constructs based on the NMBS technology, in order to analyse the effect of selected set of load-bearing microstructural components on both mechanical and functional response of soft biological materials.

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Fibronectin-Based Nanomechanical Biosensors to Map 3D Strains in Live Cells and Tissues

Cited by 4 publications

References 61 publications

Fibronectin-based nanomechanical biosensors to map 3D surface strains in live cells and tissue

Fibronectin-based nanomechanical biosensors to map 3D surface strains in live cells and tissue

Novel Temperature‐Sensitive Hydrogel Promotes Wound Healing Through YAP and MEK‐Mediated Mechanosensitivity

Microstructure-based engineering of soft biological materials

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