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Fabrication of Micropatterned Hydrogels for Neural Culture Systems using Dynamic Mask Projection Photolithography

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Cell-guiding microporous hydrogels by photopolymerization-induced phase separation.

Monica Z Müller¹, Margherita Bernero¹, Chang Xie^1,2

¹Institute for Biomechanics, ETH Zurich, Zurich, Switzerland.

Nature Communications

|May 27, 2025

Summary

Researchers developed novel microporous hydrogels using photopolymerization-induced phase separation (PIPS). This technique enables controlled pore formation in 3D cell cultures, enhancing cell growth and differentiation for tissue engineering applications.

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Area of Science:

Biomaterials Science
Tissue Engineering
Polymer Chemistry

Background:

Microporous scaffolds are crucial for solute transport and cell interactions in 3D cell culture.
Existing materials lack spatiotemporally controlled pore formation capabilities in aqueous environments.
Developing instructive scaffolding materials for advanced tissue engineering remains a challenge.

Purpose of the Study:

To introduce cell-guiding microporous hydrogels fabricated via photopolymerization-induced phase separation (PIPS).
To demonstrate the ability to control pore size and formation in situ during 3D cell culture.
To evaluate the efficacy of these hydrogels in supporting cell viability, morphology, and differentiation.

Main Methods:

Formulation of PIPS resins using ionic polymers (norbornene-functionalized polyvinyl alcohol, dextran sulfate), a di-thiol linker, and a photoinitiator.
Photocrosslinking to induce phase separation and micropore formation.
Tuning pore size (2-40 μm) by controlling light intensity, polymer composition, and molecular charge.

Main Results:

Achieved in situ, light-controlled micropore formation in the presence of living cells.
Demonstrated high cell viability (>95%) and supported cell spreading and 3D morphogenesis with RGD-functionalized hydrogels.
Enhanced osteogenic differentiation, matrix mineralization, and collagen secretion in human mesenchymal stromal cells.

Conclusions:

Photopolymerization-induced phase separation (PIPS) offers a novel method for creating cell-guiding microporous hydrogels.
These PIPS hydrogels are promising instructive scaffolding materials for 3D cell culture and complex tissue engineering.
The ability to control pore formation in situ facilitates advanced applications in regenerative medicine.