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Cellulose-based scaffolds for fluorescence lifetime imaging-assisted tissue engineering.

Neil O'Donnell1, Irina A Okkelman1, Peter Timashev2

  • 1School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland.

Acta Biomaterialia
|September 28, 2018
PubMed
Summary

Researchers developed novel biosensor scaffolds using cellulose-binding domains (CBD) for precise pH and Ca2+ measurements in 3D cultures. This innovation aids in creating engineered tissues with controlled metabolic characteristics for cancer and stem cell research.

Keywords:
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Area of Science:

  • Biomaterials Science
  • Cell Biology
  • Microscopy Techniques

Background:

  • Quantitative pH and metabolite gradient measurement in organoids and multicellular aggregates is challenging.
  • Existing methods lack the precision needed for scaffold-based 3D cultures.
  • Developing advanced biosensing tools is crucial for understanding tissue development and disease.

Purpose of the Study:

  • To design and validate novel biosensor scaffolds for measuring pH and Ca2+ gradients in 3D cell cultures.
  • To utilize cellulose-binding domains (CBD) for efficient scaffold labeling.
  • To enable multiparametric imaging in engineered tissues using fluorescence lifetime imaging (FLIM).

Main Methods:

  • Fusion of CBD with pH-sensitive enhanced cyan fluorescent protein (CBD-ECFP).
  • Labeling of various cellulose-based scaffolds (nanofibrillar, bacterial, decellularized plant materials).
  • Utilizing FLIM for quantitative pH measurements and phosphorescent probes for oxygenation monitoring in 3D cell cultures (HCT116 cancer cells, mouse intestinal organoids).

Main Results:

  • CBD-ECFP efficiently labeled cellulose scaffolds, maintaining pH sensitivity for FLIM analysis.
  • Biosensor scaffolds detected extracellular acidification in cancer cell cultures (0.2-0.4 pH units).
  • Demonstrated multiparametric imaging combining pH and oxygenation in intestinal organoids.

Conclusions:

  • The developed CBD-based biosensor scaffolds enable precise extracellular pH monitoring in 3D engineered tissues.
  • This strategy supports the design of multiparametric FLIM assays for live cancer and stem cell-derived tissues.
  • The research facilitates controlled biofabrication of 3D tissue models with defined metabolic profiles.