Hierarchical polymer niches for enhanced cell attachment
Abstract
Research context / Theoretical framework Controlling and understanding adhesion of cells on artificial surfaces remains as a critical topic in materials and life sciences. In this regard, combination of top-down (contact printing) and bottom-up approaches (ATRP polymerization + layer-by-layer adsorption of polyelectrolytes and proteins) appears as a promising strategy for the design and fabrication of cell-appealing interfaces. Interestingly, this methodology allows going from 2D to 3D-like hierarchical structures of hybrid content (niches) that influence a subsequent cell attachment on top, by better exposing the specific binding sites (RGD, IKVAV moieties) towards target membrane receptors (i.e. integrins, CD44). Complementary use of Atomic Force Microscopy (AFM), with a living cell as probe, together with Quartz Crystal Microbalance with Dissipation (QCM-D), will enable an early-stage analysis and quantification of these cell-substrate interactions on the nanoscale. Hypotheses/ Research questions / Objectives The main hypotheses of the project are the following: i) Combination of substrate-anchored polymer brushes and layer-by-layer deposited polyelectrolyte chains give rise to soft 3D niches for the enhanced adsorption of ECM proteins. The transformation of 2D interfaces into 3D-like architectures will, in turn, enhance cell attachment and proliferation of cells, with particular impact on both cell morphology and the number of cell-substrate connections formed; ii) The use of Contact-Printing techniques before the grafting-from of the brushes allows the fabrication of localized individual 3D attachment points. The localized presence of specific molecules will influence the cell-substrate affinity with final impact on cell morphology and the establishment of a different number of cell-surface contacts; iii) Single-Cell Probe Force Spectroscopy (SCPFS) technique is sensitive enough to identify early stage attachment events in cell-substrate contacts. The use of a living cell acting as indenting probe will determine events taking place on the nano- and microscale. Approach / Methods The following methods will be used to study substrate preparation and cell adhesive behaviour: Atomic force microscopy (AFM) in SCPFS mode, (confocal) fluorescence microscopy, quartz crystal microbalance with dissipation (QCMD), scanning electron microscopy (SEM), and cell culture protocols.
Fluorescence and atomic force microscopy Extracellular matrix Polymeric niches Cell adhesion Quartz crystal microbalance with dissipation
Publikationen
Project staff
José Luis Toca-Herrera
Univ.Prof. Dr. José Luis Toca-Herrera
jose.toca-herrera@boku.ac.at
Tel: +43 1 47654-80311
Project Leader
01.11.2022 - 31.10.2026