We present a new method for generating protein-based tissue engineering 3D scaffolds with adaptable mechanical properties. Multiphoton lithography allowed fabrication of 2/3D structures with feature sizes below 200 nm lateral and below 600 nm axial with Young’s moduli down to 40 kPa. The viability and mechanotransduction of mesenchymal stem cells within the scaffolds were shown by imaging of the cellular actin cytoskeleton and the vinculin protein density via 3D direct stochastic optical reconstruction microscopy. Surface functionalization of acrylate scaffolds was shown at the single-molecule level using Laser Assisted Protein Adsorption by Photobleaching (LAPAP). This pioneering research creates promising opportunities for customized tissue engineering applications.
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