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The Intersecting Physical Mechanisms That Regulate Cell Viability in 3D Synthetic Hydrogels.

Nathan R Richbourg1,2,3,4, Akaansha Rampal1, Adrian Lorenzana1

  • 1The University of Massachusetts at Amherst, Amherst, Massachusetts, USA.

Advanced Materials (Deerfield Beach, Fla.)
|March 5, 2026
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Summary

Synthetic hydrogels significantly reduce cell viability due to restricted protein transport, unlike biopolymer networks. This highlights biomaterial-engineered protein permeability as crucial for 3D cell culture success.

Keywords:
confinementextracellular matrixhydrogel designpoly(ethylene glycol)transport

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Area of Science:

  • Biomaterials Science
  • Cell Biology
  • Tissue Engineering

Background:

  • Hydrogels are widely used in 3D cell culture, but their varying protein transport properties can impact cell viability.
  • Synthetic polymer networks often exhibit lower protein permeability than biopolymer networks, potentially affecting encapsulated cells.

Purpose of the Study:

  • To investigate whether reduced protein transport in synthetic hydrogels contributes to decreased cell viability.
  • To compare cell viability in poly(ethylene glycol) vinyl sulfone (PEG-VS) hydrogels versus Matrigel.
  • To quantify the effects of modulus, transport, and confinement on encapsulated cells within different hydrogel environments.

Main Methods:

  • Utilized 2D and 3D hydrogel cultures using PEG-VS and Matrigel.
  • Employed transwell experiments to decouple serum restriction from cell-gel adhesion.
  • Quantified cell viability in response to varying hydrogel properties and serum availability.

Main Results:

  • Significantly reduced cell viability was observed in PEG-VS hydrogels compared to Matrigel.
  • Serum restriction in transwell experiments mimicked the reduced cell viability seen in 3D cultures.
  • Hydrogel-restricted protein transport was identified as a key factor influencing cell viability.

Conclusions:

  • Biomaterial-designed protein transport is a critical factor for synthetic 3D cell culture systems.
  • Reduced protein and nutrient diffusion in synthetic hydrogels negatively impacts cell viability.
  • This study provides a framework for understanding microenvironmental influences on cell viability in engineered tissues.