Cell-Laden Agarose-Collagen Composite Hydrogels for Mechanotransduction Studies
The increasing investigation of cellular mechanotransduction mechanisms requires biomaterials combining biofunctionality and suitable mechanical properties. Agarose is a standard biomaterial for cartilage and intervertebral disc mechanobiology studies, but lacks adhesion motifs and the necessary cel...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-04-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fbioe.2020.00346/full |
| _version_ | 1829105055998935040 |
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| author | Elena Cambria Silvio Brunner Sally Heusser Philipp Fisch Wolfgang Hitzl Wolfgang Hitzl Wolfgang Hitzl Stephen J. Ferguson Karin Wuertz-Kozak Karin Wuertz-Kozak Karin Wuertz-Kozak |
| author_facet | Elena Cambria Silvio Brunner Sally Heusser Philipp Fisch Wolfgang Hitzl Wolfgang Hitzl Wolfgang Hitzl Stephen J. Ferguson Karin Wuertz-Kozak Karin Wuertz-Kozak Karin Wuertz-Kozak |
| author_sort | Elena Cambria |
| collection | DOAJ |
| description | The increasing investigation of cellular mechanotransduction mechanisms requires biomaterials combining biofunctionality and suitable mechanical properties. Agarose is a standard biomaterial for cartilage and intervertebral disc mechanobiology studies, but lacks adhesion motifs and the necessary cell-matrix interaction for mechanotransduction. Here, collagen type I was blended at two concentrations (2 and 4.5 mg/mL) with agarose 2% wt/vol. The composite hydrogels were characterized in terms of structural homogeneity, rheological properties and size stability. Nucleus pulposus (NP) cell viability, proliferation, morphology, gene expression, GAG production, adhesion and mechanotransduction ability were further tested. Blended hydrogels presented a homogenous network of the two polymers. While the addition of 4.5 mg/mL collagen significantly decreased the storage modulus and increased the loss modulus of the gels, blended gels containing 2 mg/mL collagen displayed similar mechanical properties to agarose. Hydrogel size was conserved over 21 days for all agarose-based gels. Embedded cells were viable (>80%) and presented reduced proliferation and a round morphology typical of NP cells in vivo. Gene expression of collagen types I and II and aggrecan significantly increased in blended hydrogels from day 1 to 7, further resulting in a significantly superior GAG/DNA ratio compared to agarose gels at day 7. Agarose-collagen hydrogels not only promoted cell adhesion, contrary to agarose gels, but also showed a 5.36-fold higher focal adhesion kinase phosphorylation (pFAK/β-tubulin) when not compressed, and increased pFAK/FAK values 10 min after compression. Agarose-collagen thus outperforms agarose, mimics native tissues constituted of non-fibrillar matrix and collagens, and allows exploring complex loading in a highly reproducible system. |
| first_indexed | 2024-12-12T05:59:39Z |
| format | Article |
| id | doaj.art-537c3d3d76fb4878bc66c3819c9dbe16 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-12T05:59:39Z |
| publishDate | 2020-04-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-537c3d3d76fb4878bc66c3819c9dbe162022-12-22T00:35:28ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852020-04-01810.3389/fbioe.2020.00346525819Cell-Laden Agarose-Collagen Composite Hydrogels for Mechanotransduction StudiesElena Cambria0Silvio Brunner1Sally Heusser2Philipp Fisch3Wolfgang Hitzl4Wolfgang Hitzl5Wolfgang Hitzl6Stephen J. Ferguson7Karin Wuertz-Kozak8Karin Wuertz-Kozak9Karin Wuertz-Kozak10Institute for Biomechanics, ETH Zurich, Zurich, SwitzerlandInstitute for Biomechanics, ETH Zurich, Zurich, SwitzerlandInstitute for Biomechanics, ETH Zurich, Zurich, SwitzerlandInstitute for Biomechanics, ETH Zurich, Zurich, SwitzerlandResearch Office (Biostatistics), Paracelsus Medical University, Salzburg, AustriaDepartment of Ophthalmology and Optometry, Paracelsus Medical University, Salzburg, AustriaResearch Program Experimental Ophthalmology and Glaucoma Research, Paracelsus Medical University, Salzburg, AustriaInstitute for Biomechanics, ETH Zurich, Zurich, SwitzerlandInstitute for Biomechanics, ETH Zurich, Zurich, SwitzerlandDepartment of Biomedical Engineering, Rochester Institute of Technology, Rochester, NY, United StatesSpine Center, Schön Klinik München Harlaching, Academic Teaching Hospital and Spine Research Institute of the Paracelsus Private Medical University Salzburg (Austria), Munich, GermanyThe increasing investigation of cellular mechanotransduction mechanisms requires biomaterials combining biofunctionality and suitable mechanical properties. Agarose is a standard biomaterial for cartilage and intervertebral disc mechanobiology studies, but lacks adhesion motifs and the necessary cell-matrix interaction for mechanotransduction. Here, collagen type I was blended at two concentrations (2 and 4.5 mg/mL) with agarose 2% wt/vol. The composite hydrogels were characterized in terms of structural homogeneity, rheological properties and size stability. Nucleus pulposus (NP) cell viability, proliferation, morphology, gene expression, GAG production, adhesion and mechanotransduction ability were further tested. Blended hydrogels presented a homogenous network of the two polymers. While the addition of 4.5 mg/mL collagen significantly decreased the storage modulus and increased the loss modulus of the gels, blended gels containing 2 mg/mL collagen displayed similar mechanical properties to agarose. Hydrogel size was conserved over 21 days for all agarose-based gels. Embedded cells were viable (>80%) and presented reduced proliferation and a round morphology typical of NP cells in vivo. Gene expression of collagen types I and II and aggrecan significantly increased in blended hydrogels from day 1 to 7, further resulting in a significantly superior GAG/DNA ratio compared to agarose gels at day 7. Agarose-collagen hydrogels not only promoted cell adhesion, contrary to agarose gels, but also showed a 5.36-fold higher focal adhesion kinase phosphorylation (pFAK/β-tubulin) when not compressed, and increased pFAK/FAK values 10 min after compression. Agarose-collagen thus outperforms agarose, mimics native tissues constituted of non-fibrillar matrix and collagens, and allows exploring complex loading in a highly reproducible system.https://www.frontiersin.org/article/10.3389/fbioe.2020.00346/fullblended hydrogelsagarosecollagenmechanobiologyextracellular matrixdynamic compression |
| spellingShingle | Elena Cambria Silvio Brunner Sally Heusser Philipp Fisch Wolfgang Hitzl Wolfgang Hitzl Wolfgang Hitzl Stephen J. Ferguson Karin Wuertz-Kozak Karin Wuertz-Kozak Karin Wuertz-Kozak Cell-Laden Agarose-Collagen Composite Hydrogels for Mechanotransduction Studies Frontiers in Bioengineering and Biotechnology blended hydrogels agarose collagen mechanobiology extracellular matrix dynamic compression |
| title | Cell-Laden Agarose-Collagen Composite Hydrogels for Mechanotransduction Studies |
| title_full | Cell-Laden Agarose-Collagen Composite Hydrogels for Mechanotransduction Studies |
| title_fullStr | Cell-Laden Agarose-Collagen Composite Hydrogels for Mechanotransduction Studies |
| title_full_unstemmed | Cell-Laden Agarose-Collagen Composite Hydrogels for Mechanotransduction Studies |
| title_short | Cell-Laden Agarose-Collagen Composite Hydrogels for Mechanotransduction Studies |
| title_sort | cell laden agarose collagen composite hydrogels for mechanotransduction studies |
| topic | blended hydrogels agarose collagen mechanobiology extracellular matrix dynamic compression |
| url | https://www.frontiersin.org/article/10.3389/fbioe.2020.00346/full |
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