Steering cell orientation through light-based spatiotemporal modulation of the mechanical environment

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Abstract

The anisotropic organization of cells and the extracellular matrix (ECM) is essential for the physiological function of numerous biological tissues, including the myocardium. This organization changes gradually in space and time, during disease progression such as myocardial infarction. The role of mechanical stimuli has been demonstrated to be essential in obtaining, maintaining and de-railing this organization, but the underlying mechanisms are scarcely known. To enable the study of the mechanobiological mechanisms involved, in vitro techniques able to spatiotemporally control the multiscale tissue mechanical environment are thus necessary. Here, by using light-sensitive materials combined with light-illumination techniques, we fabricated 2D and 3D in vitro model systems exposing cells to multiscale, spatiotemporally resolved stiffness anisotropies. Specifically, spatial stiffness anisotropies spanning from micron-sized (cellular) to millimeter-sized (tissue) were achieved. Moreover, the light-sensitive materials allowed to introduce the stiffness anisotropies at defined timepoints (hours) after cell seeding, facilitating the study of their temporal effects on cell and tissue orientation. The systems were tested using cardiac fibroblasts (cFBs), which are known to be crucial for the remodeling of anisotropic cardiac tissue. We observed that 2D stiffness micropatterns induced cFBs anisotropic alignment, independent of the stimulus timing, but dependent on the micropattern spacing. cFBs exhibited organized alignment also in response to 3D stiffness macropatterns, dependent on the stimulus timing and temporally followed by (slower) ECM co-alignment. In conclusion, the developed model systems allow improved fundamental understanding of the underlying mechanobiological factors that steer cell and ECM orientation, such as stiffness guidance and boundary constraints.

Original languageEnglish
Article number035011
Number of pages19
JournalBiofabrication
Volume16
Issue number3
DOIs
Publication statusPublished - 1 Jul 2024

Funding

The authors acknowledge Dr Koen Pieterse from the ICMS Animation Studio (TU/e) for his valuable inputs in the figure design. Moreover, the authors thank Mirko D\u2019Urso, Dylan Mostert and Dr Jasper Foolen from the TU/e Biomedical Engineering Department for the insightful discussions. I J, A v d P, N A K and C V C B acknowledge financial support by the Gravitation Program \u2018Materials Driven Regeneration\u2019, funded by the Netherlands Organization for Scientific Research (024.003.013). I J acknowledges financial support from the Netherlands Organization for Scientific Research (OCENW.XS21.4.146). N A K acknowledges financial support the European Research Council (ERC) (CoEvolve, Grant No. 851960).

FundersFunder number
Case Western Reserve University
Nederlandse Organisatie voor Wetenschappelijk Onderzoek024.003.013, OCENW.XS21.4.146
European Research Council851960

    Keywords

    • cardiac fibroblasts
    • cellular orientation
    • extracellular matrix
    • in vitro models
    • mechanobiology
    • tissue engineering

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