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One-Step Surface Functionalization of Hydrogel-Based, Stimulus-Responsive 3D Microstructures for Human Stem Cells.

Natalie Munding1, Christina Schlagheck2, Jochen Wittbrodt2

  • 1Physical Chemistry of Biosystems, Institute of Physical Chemistry, Heidelberg University, 69120 Heidelberg, Germany.

ACS Applied Materials & Interfaces
|June 5, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces a novel one-step hydrogel functionalization method using N-hydroxysuccinimide (NHS) monomers for improved stem cell culture. This technique enhances the uniform coating of extracellular matrix proteins on 2D and 3D scaffolds, supporting cell viability and proliferation.

Keywords:
3D cellular scaffoldhuman mesenchymal stem cellpolyacrylamidesupramolecular hydrogelsurface functionalization

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Cell Biology

Background:

  • Hydrogels are crucial for stem cell culture, providing mechanical cues and 3D environments.
  • Current methods for functionalizing hydrogels with extracellular matrix (ECM) proteins are often inefficient, especially for complex 3D structures, due to limitations with photo-cross-linkers.

Purpose of the Study:

  • To develop a novel, efficient method for functionalizing hydrogels with ECM proteins.
  • To enable uniform surface modification of both 2D and 3D hydrogel scaffolds for enhanced cell culture applications.

Main Methods:

  • Integrated N-hydroxysuccinimide (NHS) monomers into copolymer hydrogels.
  • Utilized solvent miscibility to tune hydrogel stiffness and enable one-step surface functionalization.
  • Fabricated 3D microstructures using 3D printed stamps.
  • Functionalized scaffolds with fibronectin, laminin, and gelatin.

Main Results:

  • Achieved uniform, one-step surface functionalization of hydrogels with ECM proteins, bypassing photoactivation.
  • Demonstrated high viability (>80%) and maintained proliferation capacity (~60%) of human mesenchymal stem cells (hMSCs) on 2D functionalized substrates.
  • Successfully functionalized complex 3D microstructures, accommodating hMSCs within the scaffolds.
  • Observed reversible swelling/deswelling of 3D scaffolds in the presence of hMSCs.

Conclusions:

  • The developed NHS-based method provides a versatile and efficient approach for hydrogel functionalization.
  • This technique is suitable for creating advanced 2D and 3D cell culture platforms for various cell types.
  • The method overcomes limitations of traditional photo-cross-linking for uniform ECM protein presentation.