Next Article in Journal
Role of Cyclosporine in Gingival Hyperplasia: An In Vitro Study on Gingival Fibroblasts
Previous Article in Journal
Inactivity and Skeletal Muscle Metabolism: A Vicious Cycle in Old Age
Previous Article in Special Issue
Supporting Cell-Based Tendon Therapy: Effect of PDGF-BB and Ascorbic Acid on Rabbit Achilles Tenocytes In Vitro
Open AccessArticle

SV40 Transfected Human Anterior Cruciate Ligament Derived Ligamentocytes—Suitable as a Human in Vitro Model for Ligament Reconstruction?

1
Institute of Anatomy and Cell Biology, Paracelsus Medical Private University, Salzburg and Nuremberg, 90419 Nuremberg, Germany
2
Institute of Anatomy, Christian Albrecht Universität, Otto-Hahn-Platz 8, 24118 Kiel, Germany
3
Department of Biosciences, Paris Lodron University Salzburg, 5020 Salzburg, Austria
4
Leibniz-Institut für Polymerforschung Dresden e. V., 01069 Dresden, Germany
5
Forschungsinstitut für Leder und Kunststoffbahnen (FILK) gGmbH, 09599 Freiberg, Germany
6
Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin and Berlin Institute of Health, Department of Traumatology and Reconstructive Surgery, Campus Benjamin Franklin, Hindenburgdamm 30, 12203 Berlin, Germany
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(2), 593; https://doi.org/10.3390/ijms21020593
Received: 25 December 2019 / Revised: 10 January 2020 / Accepted: 13 January 2020 / Published: 16 January 2020
(This article belongs to the Special Issue Tendon/Ligament Reconstruction by Tissue Engineering)
Cultured human primary cells have a limited lifespan undergoing dedifferentiation or senescence. Anterior cruciate ligaments (ACL) are hypocellular but tissue engineering (TE) requires high cell numbers. Simian virus (SV) 40 tumor (T) antigen expression could extend the lifespan of cells. This study aimed to identify cellular changes induced by SV40 expression in human ACL ligamentocytes by comparing them with non-transfected ligamentocytes and tissue of the same donor to assess their applicability as TE model. Human ACL ligamentocytes (40-year-old female donor after ACL rupture) were either transfected with a SV40 plasmid or remained non-transfected (control) before monitored for SV40 expression, survival, and DNA content. Protein expression of cultured ligamentocytes was compared with the donor tissue. Ligamentocyte spheroids were seeded on scaffolds embroidered either from polylactic acid (PLA) threads solely or combined PLA and poly (L-lactide-co-ε-caprolactone) (P(LA-CL)) threads. These scaffolds were further functionalized with fluorination and fibrillated collagen foam. Cell distribution and survival were monitored for up to five weeks. The transfected cells expressed the SV40 antigen throughout the entire observation time, but often exhibited random and incomplete cell divisions with significantly more dying cells, significantly more DNA and more numerous nucleoli than controls. The expression profile of non-transfected and SV40-positive ligamentocytes was similar. In contrast to controls, SV40-positive cells formed larger spheroids, produced less vimentin and focal adhesions and died on the scaffolds after 21 d. Functionalized scaffolds supported human ligamentocyte growth. SV40 antigen expressing ligamentocytes share many properties with their non-transfected counterparts suggesting them as a model, however, applicability for TE is limited. View Full-Text
Keywords: anterior cruciate ligament; tissue engineering; PLA; P(LA-CL); embroidered scaffold; ligamentocytes; SV40 anterior cruciate ligament; tissue engineering; PLA; P(LA-CL); embroidered scaffold; ligamentocytes; SV40
Show Figures

Graphical abstract

MDPI and ACS Style

Schulze-Tanzil, G.; Arnold, P.; Gögele, C.; Hahn, J.; Breier, A.; Meyer, M.; Kohl, B.; Schröpfer, M.; Schwarz, S. SV40 Transfected Human Anterior Cruciate Ligament Derived Ligamentocytes—Suitable as a Human in Vitro Model for Ligament Reconstruction? Int. J. Mol. Sci. 2020, 21, 593.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop