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  • Nanoscale Topography and Poroelastic Properties of Model Tissue Breast Gland Basement Membranes.

Nanoscale Topography and Poroelastic Properties of Model Tissue Breast Gland Basement Membranes.

Biophysical journal (2018-10-17)
Gloria Fabris, Alessandro Lucantonio, Nico Hampe, Erik Noetzel, Bernd Hoffmann, Antonio DeSimone, Rudolf Merkel
ABSTRACT

Basement membranes (BMs) are thin layers of condensed extracellular matrix proteins serving as permeability filters, cellular anchoring sites, and barriers against cancer cell invasion. It is believed that their biomechanical properties play a crucial role in determining cellular behavior and response, especially in mechanically active tissues like breast glands. Despite this, so far, relatively little attention has been dedicated to their analysis because of the difficulty of isolating and handling such thin layers of material. Here, we isolated BMs derived from MCF10A spheroids-three-dimensional breast gland model systems mimicking in vitro the most relevant phenotypic characteristics of human breast lobules-and characterized them by atomic force microscopy, enhanced resolution confocal microscopy, and scanning electron microscopy. By performing atomic force microscopy height-clamp experiments, we obtained force-relaxation curves that offered the first biomechanical data on isolated breast gland BMs to our knowledge. Based on enhanced resolution confocal microscopy and scanning electron microscopy imaging data, we modeled the system as a polymer network immersed in liquid and described it as a poroelastic material. Finite-element simulations matching the experimental force-relaxation curves allowed for the first quantification, to our knowledge, of the bulk and shear moduli of the membrane as well as its water permeability. These results represent a first step toward a deeper understanding of the mechanism of tensional homeostasis regulating mammary gland activity as well as its disruption during processes of membrane breaching and metastatic invasion.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Collagen Types I, II & III Antibody, clone MMCHABC, clone MMCHABC, Chemicon®, from mouse
Sigma-Aldrich
Anti-Heparan Sulfate Proteoglycan (Perlecan) Antibody, clone A7L6, clone A7L6, from rat