Cyclically stretched ACL fibroblasts emigrating from spheroids adapt their cytoskeleton and ligament-related expression profile
Cell and Tissue Research, vol. 384, pp. 675–690
Abstract
Mechanical stress of ligaments varies; hence, ligament fibroblasts must adapt their expression profile to novel mechanomilieus to ensure tissue resilience. Activation of the mechanoreceptors leads to a specific signal transduction, the so-called mechanotransduction. However, with regard to their natural three-dimensional (3D) microenvironment cell reaction to mechanical stimuli during emigrating from a 3D spheroid culture is still unclear. This study aims to provide a deeper understanding of the reaction profile of anterior cruciate ligament (ACL)-derived fibroblasts exposed to cyclic uniaxial strain in two-dimensional (2D) monolayer culture and during emigration from 3D spheroids with respect to cell survival, cell and cytoskeletal orientation, distribution, and expression profile. Monolayers and spheroids were cultured in crosslinked polydimethyl siloxane (PDMS) elastomeric chambers and uniaxially stretched (14% at 0.3 Hz) for 48 h. Cell vitality, their distribution, nuclear shape, stress fiber orientation, focal adhesions, proliferation, expression of ECM components such as sulfated glycosaminoglycans, collagen type I, decorin, tenascin C and cell-cell communication-related gap junctional connexin (CXN) 43, tendon-related markers Mohawk and tenomodulin (myodulin) were analyzed. In contrast to unstretched cells, stretched fibroblasts showed elongation of stress fibers, cell and cytoskeletal alignment perpendicular to strain direction, less rounded cell nuclei, increased numbers of focal adhesions, proliferation, amplified CXN43, and main ECM component expression in both cultures. The applied cyclic stretch protocol evoked an anabolic response and enhanced tendon-related marker expression in ACL-derived fibroblasts emigrating from 3D spheroids and seems also promising to support in future tissue formation in ACL scaffolds seeded in vitro with spheroids.
Authors 7
-
University of Salzburg · Paracelsus Medical University · Paracelsus Medizinische Privatuniversität
Affiliation as printed
Department of Biosciences, Paris Lodron University Salzburg, Hellbrunnerstr. 34, 5020, Salzburg, Austria
Institute of Anatomy and Cell Biology, Paracelsus Medical University, Prof.-Ernst-Nathan Str. 1, 90419, Nuremberg and Salzburg, Nuremberg, Germany
-
Affiliation as printed
Institute of Biological Information Processing: IBI-2, Forschungszentrum Jülich, 52425, Jülich, Germany
-
Affiliation as printed
Institute of Biological Information Processing: IBI-2, Forschungszentrum Jülich, 52425, Jülich, Germany
-
Paracelsus Medical University · Paracelsus Medizinische Privatuniversität
Affiliation as printed
Institute of Anatomy and Cell Biology, Paracelsus Medical University, Prof.-Ernst-Nathan Str. 1, 90419, Nuremberg and Salzburg, Nuremberg, Germany
-
Affiliation as printed
Department of Anatomy and Cell Biology, RWTH Aachen University, Wendlingweg 2, 52074, Aachen, Germany
-
Affiliation as printed
Institute of Biological Information Processing: IBI-2, Forschungszentrum Jülich, 52425, Jülich, Germany
-
Gundula Gesine Schulze-Tanzil corresponding
Paracelsus Medical University · Paracelsus Medizinische Privatuniversität
Affiliation as printed
Institute of Anatomy and Cell Biology, Paracelsus Medical University, Prof.-Ernst-Nathan Str. 1, 90419, Nuremberg and Salzburg, Nuremberg, Germany. gundula.schulze@pmu.ac.at
Institute of Anatomy and Cell Biology, Paracelsus Medical University, Prof.-Ernst-Nathan Str. 1, 90419, Nuremberg and Salzburg, Nuremberg, Germany
Cited by 11 stored of 11
11 results
No patents citing this paper on Lens.org (checked 2026-10-06).