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

Biofabrication, vol. 16, pp. 035011

Abstract

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.

Authors 9

1. Ignasi Jorba corresponding

   Universitat de Barcelona · Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, The Netherlands

   Institute for Complex Molecular Systems (ICMS), PO Box 513, 5600 MB Eindhoven, The Netherlands

   Unitat de Biofísica i Bioenginyeria, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, 08036 Barcelona, Spain

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, Eindhoven, Noord-Brabant, 5600 MB, NETHERLANDS

2. Sil Gussenhoven

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, The Netherlands

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, Eindhoven, Noord-Brabant, 5600 MB, NETHERLANDS

3. Atze van der Pol

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, The Netherlands

   Institute for Complex Molecular Systems (ICMS), PO Box 513, 5600 MB Eindhoven, The Netherlands

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, Eindhoven, Noord-Brabant, 5600 MB, NETHERLANDS

4. Bart G. W. Groenen

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, The Netherlands

   Institute for Complex Molecular Systems (ICMS), PO Box 513, 5600 MB Eindhoven, The Netherlands

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, Eindhoven, Noord-Brabant, 5600 MB, NETHERLANDS

5. Maarten van Zon

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, The Netherlands

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, Eindhoven, Noord-Brabant, 5600 MB, NETHERLANDS

6. Marie‐José Goumans Aachen Department of Cell and Chemical Biology Department of Cell and Chemical Biology

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Department of Cell and Chemical Biology, Leiden University Medical Centre, Leiden, The Netherlands

   Department of Cell and Chemical Biology, Leiden University Medical Center, Einthovenweg 20, Leiden, Zuid-Holland, 2300 RC, NETHERLANDS

7. Nicholas A. Kurniawan

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, The Netherlands

   Institute for Complex Molecular Systems (ICMS), PO Box 513, 5600 MB Eindhoven, The Netherlands

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, Eindhoven, Noord-Brabant, 5600 MB, NETHERLANDS

8. Tommaso Ristori

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, The Netherlands

   Institute for Complex Molecular Systems (ICMS), PO Box 513, 5600 MB Eindhoven, The Netherlands

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, Eindhoven, Noord-Brabant, 5600 MB, NETHERLANDS

9. Carlijn V. C. Bouten

   Eindhoven University of Technology

   Affiliation as printed

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, The Netherlands

   Institute for Complex Molecular Systems (ICMS), PO Box 513, 5600 MB Eindhoven, The Netherlands

   Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, Eindhoven, Noord-Brabant, 5600 MB, NETHERLANDS