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bioRxiv · 10.64898/2026.01.20.700570

A Versatile, Biocompatible Additive for Spatiotemporal Volumetric Photografting of Biochemical and Mechanical Cues in Diverse Hydrogel Matrices

Falandt, M.·Bonhomme, C.·Florczak, S.·Vermonden, T.·Bernal, P. N.·Levato, R.
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Abstract

Engineering functional tissue constructs requires not only replicating their 3D architecture but also capturing their dynamic biochemical and mechanical environments. While 3D bioprinting technologies enable spatial control over cell and biomaterial deposition, post-fabrication modulation of material properties remains limited. Photografting approaches allow for spatiotemporal functionalization of certain 3D matrices by chemically binding bioactive factors onto spatially determined regions of a material, but current methods often rely on specialized chemistries with narrow material compatibility. Here, we introduce AddGraft, a biocompatible, off-the-shelf additive designed for semi-orthogonal thiol-ene photografting in vinyl-functionalized hydrogels. AddGraft, a heterobifunctional polyethylene glycol, carries an acrylate moiety for network incorporation during photocrosslinking and a norbornene group for post-crosslinking functionalization. AddGraft integrates into the polymer network during gel crosslinking without altering bulk mechanics, enabling precise modification at any time post-fabrication. We demonstrate compatibility with multiple acrylated biomaterial platforms and light-based volumetric photopatterning technology. Photopatterning achieves high spatial resolution and gradient formation in 3D, while grafting of multi-thiolated crosslinkers allows localized stiffening of hydrogels. Encapsulated human mesenchymal stromal cells exhibit high viability and undergo morphological changes in response to the dynamic tuning of their microenvironment. By decoupling structural and functional roles, AddGraft enables on-demand spatial and temporal control over hydrogel properties. This approach expands the biofabrication toolkit for engineering cell-instructive, 4D tissue environments with translational relevance in regenerative medicine.

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Falandt, M., Bonhomme, C., Florczak, S., Vermonden, T., Bernal, P. N., Levato, R.. 2026-01-23. A Versatile, Biocompatible Additive for Spatiotemporal Volumetric Photografting of Biochemical and Mechanical Cues in Diverse Hydrogel Matrices. https://doi.org/10.64898/2026.01.20.700570

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