3D inverted opal hydrogel scaffolds with oxygen sensing capability

The measurement of local oxygen level in 3D cell culture is desired but remains as a challenge problem. We developed a 3D cell scaffold with luminescence-based oxygen sensing capability that opens the possibility of 3D mapping of oxygen level during cell growth. Hydrogel inverted opal scaffold was prepared by photo-polymerization of poly(2-hydroxyethyl methacrylate (pHEMA) and poly(methacryloyloxy)ethyl-trimethylammonium chloride (pMEATAC) monomer using close-packed bead assembly as template. Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium chloride (Ru(dpp)(3)), was coated on the pHEMA-pMEATAC 3D scaffolds by layer-by-layer (LBL) assembly. pHEMA-pMEATAC copolymer was coated on top of the Ru(dpp)(3) layer as a protection layer. The fluorescence emission of Ru(dpp)(3) can be dynamically quenched by oxygen. By measuring the emission intensity of the scaffold, the local oxygen level can be monitored. The hydrogel scaffolds are transparent, and thus 3D fluorescence intensity can be mapped by confocal microscopy. Human bone marrow stromal cells HS-5 were successfully cultured on the oxygen sensing scaffold, and the observed Ru(dpp)(3) emission intensity from the scaffold was stronger in cell rich area, which indicates a lower oxygen level due to the consumption of the cells.