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Tissue Engineering of Cartilage Using a Random Positioning Machine

Wehland, Markus and Steinwerth, Paul and Aleshcheva, Ganna and Sahana, Jayashree and Hemmersbach, Ruth and Lützenberg, Roland and Kopp, Sascha and Infanger, Manfred and Grimm, Daniela (2020) Tissue Engineering of Cartilage Using a Random Positioning Machine. International Journal of Molecular Sciences, 21 (24), p. 9596. Multidisciplinary Digital Publishing Institute (MDPI). doi: 10.3390/ijms21249596. ISSN 1661-6596.

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Official URL: http://dx.doi.org/10.3390/ijms21249596

Abstract

Articular cartilage is a skeletal tissue of avascular nature and limited self-repair capacity. Cartilage-degenerative diseases, such as osteoarthritis (OA), are difficult to treat and often necessitate joint replacement surgery. Cartilage is a tough but flexible material and relatively easy to damage. It is, therefore, of high interest to develop methods allowing chondrocytes to recolonize, to rebuild the cartilage and to restore joint functionality. Here we studied the in vitro production of cartilage-like tissue using human articular chondrocytes exposed to the Random Positioning Machine (RPM), a device to simulate certain aspects of microgravity on Earth. To screen early adoption reactions of chondrocytes exposed to the RPM, we performed quantitative real-time PCR analyses after 24 h on chondrocytes cultured in DMEM/F-12. A significant up-regulation in the gene expression of IL6, RUNX2, RUNX3, SPP1, SOX6, SOX9, and MMP1 was detected, while the levels of IL8, ACAN, PRG4, ITGB1, TGFB1, COL1A1, COL2A1, COL10A1, SOD3, SOX5, MMP1, and MMP2 mRNAs remained unchanged. The STRING (Search Tool for the Retrieval of Interacting Genes/Proteins) analysis demonstrated among others the importance of these differentially regulated genes for cartilage formation. Chondrocytes grown in DMEM/F-12 medium produced three-dimensional (3D) spheroids after five days without the addition of scaffolds. On day 28, the produced tissue constructs reached up to 2 mm in diameter. Using specific chondrocyte growth medium, similar results were achieved within 14 days. Spheroids from both types of culture media showed the typical cartilage morphology with aggrecan positivity. Intermediate filaments form clusters under RPM conditions as detected by vimentin staining after 7 d and 14 d. Larger meshes appear in the network in 28-day samples. Furthermore, they were able to form a confluent chondrocyte monolayer after being transferred back into cell culture flasks in 1 g conditions showing their suitability for transplantation into joints. Our results demonstrate that the cultivation medium has a direct influence on the velocity of tissue formation and tissue composition. The spheroids show properties that make them interesting candidates for cellular cartilage regeneration approaches in trauma and OA therapy.

Item URL in elib:https://elib.dlr.de/140211/
Document Type:Article
Title:Tissue Engineering of Cartilage Using a Random Positioning Machine
Authors:
AuthorsInstitution or Email of AuthorsAuthor's ORCID iDORCID Put Code
Wehland, MarkusOtto von Guericke University Magdeburg, Clinic for Plastic, Aesthetic and Hand Surgery, Department of Microgravity & Translational Regenerative Medicine, Magdeburg, Germanyhttps://orcid.org/0000-0002-8160-859XUNSPECIFIED
Steinwerth, PaulOtto von Guericke University, Clinic for Plastic, Aesthetic & Hand Surgery, Department of Microgravity & Translational Regenerative Medicine, Magdeburg, GermanyUNSPECIFIEDUNSPECIFIED
Aleshcheva, GannaOtto von Guericke University, Clinic for Plastic, Aesthetic & Hand Surgery, Department of Microgravity & Translational Regenerative Medicine, Magdeburg, GermanyUNSPECIFIEDUNSPECIFIED
Sahana, JayashreeUniversity Aarhus, Department of Biomedicine and Pharmacology, Aarhus, Denmarkhttps://orcid.org/0000-0002-6606-6315UNSPECIFIED
Hemmersbach, RuthGerman Aerospace Center (DLR), Institute of Aerospace Medicine, Gravitational Biology, Cologne, Germanyhttps://orcid.org/0000-0001-5308-6715UNSPECIFIED
Lützenberg, RolandOtto von Guericke University, Clinic for Plastic, Aesthetic & Hand Surgery, Department of Microgravity & Translational Regenerative Medicine, Magdeburg, GermanyUNSPECIFIEDUNSPECIFIED
Kopp, SaschaOtto von Guericke University, Clinic for Plastic, Aesthetic & Hand Surgery, Department of Microgravity & Translational Regenerative Medicine, Magdeburg, Germanyhttps://orcid.org/0000-0001-5829-6455UNSPECIFIED
Infanger, ManfredOtto von Guericke University, Clinic for Plastic, Aesthetic & Hand Surgery, Department of Microgravity & Translational Regenerative Medicine, Magdeburg, GermanyUNSPECIFIEDUNSPECIFIED
Grimm, DanielaUniversity Aarhus, Department of Biomedicine & Pharmacology, Aarhus, Denmark & Otto von Guericke University, Clinic for Plastic, Aesthetic & Hand Surgery, Department of Microgravity & Translational Regenerative Medicine, Magdeburg, Germanyhttps://orcid.org/0000-0002-5083-2812UNSPECIFIED
Date:20 December 2020
Journal or Publication Title:International Journal of Molecular Sciences
Refereed publication:Yes
Open Access:Yes
Gold Open Access:Yes
In SCOPUS:Yes
In ISI Web of Science:Yes
Volume:21
DOI:10.3390/ijms21249596
Page Range:p. 9596
Publisher:Multidisciplinary Digital Publishing Institute (MDPI)
ISSN:1661-6596
Status:Published
Keywords:Tissue engineering, Cartilage, Spheroids, Random positioning machine (RPM); Scaffold-free Cell Culturing
HGF - Research field:Aeronautics, Space and Transport
HGF - Program:Space
HGF - Program Themes:Research under Space Conditions
DLR - Research area:Raumfahrt
DLR - Program:R FR - Research under Space Conditions
DLR - Research theme (Project):R - Vorhaben Biowissenschaftliche Exp.-vorbereitung (old), R - Vorhaben Biowissenschaftliche Nutzerunterstützung (old)
Location: Köln-Porz
Institutes and Institutions:Institute of Aerospace Medicine > Gravitational Biology
Deposited By: Duwe, Helmut
Deposited On:11 Jan 2021 14:13
Last Modified:11 Jan 2021 14:13

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