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Cell-Dense Bioink Design for Xolography: Coupling Refractive Index-Matching with Increased Photoreactivity

Created on 10 Jun 2026

Authors

Balciunaite, A., Inacker, S., Badolato, A., Brauer, E., Konig, N. F., Lima, L. V., Humphreys, G. R., Polinari, C., Palato, S., Hernandez, P. P., Filippi, M., Hecht, S., Katzschmann, R.

Abstract

Bioxolography enables high-resolution fabrication of geometrically complex, cell-laden constructs for tissue engineering. However, tissue-relevant cell densities conflict with the optical transparency required for efficient dual-color volumetric printing. In this work, we extend the Bioxolography toolbox to include refractive index (RI) matching for cell-laden bioresins using iodixanol (IDX). Remarkably, IDX enhances optical transparency and boosts reactivity - a phenomenon unique to Xolography. Yet, excessive IDX compromises dual-color efficiency through increased absorption and undesired UV-only curing, underscoring a central trade-off between optical clarity and photochemical performance. Systematic tuning of resin compositions along an iso-refractive index line demonstrated the versatility of Bioxolography, with IDX enhancing polymerization and 4-Hydroxy-TEMPO providing biocompatible inhibition. Optimizing composition and printing parameters yielded GelMA hydrogels with cell densities up to 5x10<6> cells/mL. Cell-laden prints achieved sub-100 m resolution and complex geometries such as channels and gyroids. Using skeletal muscle tissue as a model, we validated RI matched Bioxolography as a promising strategy for tissue engineering by demonstrating cell alignment along printed grooves and formation of mature muscle fibers characterized by MyoHC+ staining and fusion index. By integrating physical, chemical, and biological perspectives, this work advances Xolography toward biomaterials development and reinforces its position as an emerging volumetric (bio)printing technology.

Preprint server: bioRxiv
The authors list and abstract were imported from bioRxiv on 10 Jun 2026.

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