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. 2017 Sep 20;7:12022. doi: 10.1038/s41598-017-12363-x

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Temporal evolution of average cell-generated compressive and tensile stresses within growing 3D aggregates of tooth mesenchymal cells. (A) Average cellular anisotropic stresses for aggregates at 24, 48 and 60 hours (top), measured with droplets coated with RGD peptides (blue circles) or mPEG (orange squares) to quantify compressive and tensile stresses (see panel B). Confocal sections of RGD-coated droplets (red; surface label) embedded between cells (green) of the aggregate at different stages are shown; the increase in the magnitude of droplet deformations (i.e., stresses) over time is directly visible (bottom). Scale bars 20 μm. For 24, 48 and 60 hours, N = 30, 30, and 24, respectively, for droplets coated with RGD peptides, and N = 6, 5, and 5, respectively, for droplets coated with mPEG (N = number of aggregates). (B) Schematic representation of adhesive and non-adhesive droplets. Adhesive droplets, coated with ligands targeting cell adhesion receptors (RGD peptide in this case), enable the transmission of pulling cellular forces (tensile stresses), in addition to compressive stresses. Non-adhesive droplets are coated with PEG, preventing cell-droplet adhesion, and only allowing the measurement of (non-specific) compressive stresses.