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bioRxiv · 10.64898/2025.12.19.695439

Fibroblast alignment is governed by stiffness interfaces in hydrogels.

Dijkstra, R.·Burgess, J. K.·Petridis, X.·Sharma, P. K.·Harmsen, M. C.
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Abstract

Transitions in mechanical properties between different tissue scaffolds, such as extracellular matrices (ECMs), are common in the human body and influence cellular orientation. For example, in teeth, the abrupt transition between soft ([~]5 kPa) dental pulp and stiff (>100 kPa) dentin directs the alignment of odontoblasts. In this study, hydrogels, with defined mechanical transitions were used to evaluate the phenotypic response of fibroblasts to either a soft/soft (3 kPa/3 kPa) or soft/stiff (3 kPa/200 kPa) interface. Near the soft/soft interface fibroblasts aligned in a more parallel fashion to the interface (11{degrees} angle to the interface), compared to the more perpendicular fashion (69{degrees} angle to the interface) observed at the soft-stiff interface. Notably, regions of alignment also showed an increased proportion of alpha smooth muscle actin (SMA)-positive cells, indicating myofibroblast differentiation in response to the mechanical transition. These findings demonstrate that modifying a biomaterial by introducing an interface alters the bioactivity of responding cells solely through mechanical mechanisms, without any change to the cells biochemical environment. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=147 HEIGHT=200 SRC="FIGDIR/small/695439v1_ufig1.gif" ALT="Figure 1000"> View larger version (42K): org.highwire.dtl.DTLVardef@d0ca04org.highwire.dtl.DTLVardef@17ee49forg.highwire.dtl.DTLVardef@1d389a5org.highwire.dtl.DTLVardef@f1b2a0_HPS_FORMAT_FIGEXP M_FIG C_FIG Schematic summary showing interface-induced fibroblast orientation and variety in phenotypic expression. On the bottom left the perpendicular orientation of fibroblast (skin tone) and myofibroblast (pink) on a soft/soft (3kPa/3kPa) GelMA hydrogel interface and on the right side the parallel orientation near a soft/stiff (3kPa/200kPa) GelMA hydrogel interface with greater collagen deposition, depicted by purple halos.

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Dijkstra, R., Burgess, J. K., Petridis, X., Sharma, P. K., Harmsen, M. C.. 2025-12-22. Fibroblast alignment is governed by stiffness interfaces in hydrogels.. https://doi.org/10.64898/2025.12.19.695439

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