Tissue Engineering of Cartilage Using a Random Positioning Machine
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 damag...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/24/9596 |
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| author | Markus Wehland Paul Steinwerth Ganna Aleshcheva Jayashree Sahana Ruth Hemmersbach Ronald Lützenberg Sascha Kopp Manfred Infanger Daniela Grimm |
| author_facet | Markus Wehland Paul Steinwerth Ganna Aleshcheva Jayashree Sahana Ruth Hemmersbach Ronald Lützenberg Sascha Kopp Manfred Infanger Daniela Grimm |
| author_sort | Markus Wehland |
| collection | DOAJ |
| description | 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 <i>IL6</i>, <i>RUNX2</i>, <i>RUNX3</i>, <i>SPP1</i>, <i>SOX6</i>, <i>SOX9</i>, and <i>MMP13</i> was detected, while the levels of <i>IL8</i>, <i>ACAN</i>, <i>PRG4</i>, <i>ITGB1</i>, <i>TGFB1</i>, <i>COL1A1</i>, <i>COL2A1</i>, <i>COL10A1</i>, <i>SOD3</i>, <i>SOX5</i>, <i>MMP1</i>, and <i>MMP2</i> 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 <i>g</i> 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. |
| first_indexed | 2024-03-10T14:01:02Z |
| format | Article |
| id | doaj.art-682d9bda5011405784be5057345e9d8b |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T14:01:02Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-682d9bda5011405784be5057345e9d8b2023-11-21T01:08:12ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-12-012124959610.3390/ijms21249596Tissue Engineering of Cartilage Using a Random Positioning MachineMarkus Wehland0Paul Steinwerth1Ganna Aleshcheva2Jayashree Sahana3Ruth Hemmersbach4Ronald Lützenberg5Sascha Kopp6Manfred Infanger7Daniela Grimm8Department of Microgravity and Translational Regenerative Medicine, Clinic for Plastic, Aesthetic and Hand Surgery, Otto von Guericke University, 39106 Magdeburg, GermanyDepartment of Microgravity and Translational Regenerative Medicine, Clinic for Plastic, Aesthetic and Hand Surgery, Otto von Guericke University, 39106 Magdeburg, GermanyDepartment of Microgravity and Translational Regenerative Medicine, Clinic for Plastic, Aesthetic and Hand Surgery, Otto von Guericke University, 39106 Magdeburg, GermanyDepartment for Biomedicine, Aarhus University, 8000 Aarhus, DenmarkGravitational Biology, Institute of Aerospace Medicine, German Aerospace Center, 51147 Cologne, GermanyDepartment of Microgravity and Translational Regenerative Medicine, Clinic for Plastic, Aesthetic and Hand Surgery, Otto von Guericke University, 39106 Magdeburg, GermanyDepartment of Microgravity and Translational Regenerative Medicine, Clinic for Plastic, Aesthetic and Hand Surgery, Otto von Guericke University, 39106 Magdeburg, GermanyDepartment of Microgravity and Translational Regenerative Medicine, Clinic for Plastic, Aesthetic and Hand Surgery, Otto von Guericke University, 39106 Magdeburg, GermanyDepartment of Microgravity and Translational Regenerative Medicine, Clinic for Plastic, Aesthetic and Hand Surgery, Otto von Guericke University, 39106 Magdeburg, GermanyArticular 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 <i>IL6</i>, <i>RUNX2</i>, <i>RUNX3</i>, <i>SPP1</i>, <i>SOX6</i>, <i>SOX9</i>, and <i>MMP13</i> was detected, while the levels of <i>IL8</i>, <i>ACAN</i>, <i>PRG4</i>, <i>ITGB1</i>, <i>TGFB1</i>, <i>COL1A1</i>, <i>COL2A1</i>, <i>COL10A1</i>, <i>SOD3</i>, <i>SOX5</i>, <i>MMP1</i>, and <i>MMP2</i> 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 <i>g</i> 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.https://www.mdpi.com/1422-0067/21/24/9596tissue engineeringcartilagespheroidsrandom positioning machinescaffold-free |
| spellingShingle | Markus Wehland Paul Steinwerth Ganna Aleshcheva Jayashree Sahana Ruth Hemmersbach Ronald Lützenberg Sascha Kopp Manfred Infanger Daniela Grimm Tissue Engineering of Cartilage Using a Random Positioning Machine International Journal of Molecular Sciences tissue engineering cartilage spheroids random positioning machine scaffold-free |
| title | Tissue Engineering of Cartilage Using a Random Positioning Machine |
| title_full | Tissue Engineering of Cartilage Using a Random Positioning Machine |
| title_fullStr | Tissue Engineering of Cartilage Using a Random Positioning Machine |
| title_full_unstemmed | Tissue Engineering of Cartilage Using a Random Positioning Machine |
| title_short | Tissue Engineering of Cartilage Using a Random Positioning Machine |
| title_sort | tissue engineering of cartilage using a random positioning machine |
| topic | tissue engineering cartilage spheroids random positioning machine scaffold-free |
| url | https://www.mdpi.com/1422-0067/21/24/9596 |
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