Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus
Bioscaffolds derived from the extracellular matrix (ECM) have shown the capacity to promote regeneration by providing tissue-specific biological instructive cues that can enhance cell survival and direct lineage-specific differentiation. This study focused on the development and characterization of...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.937239/full |
| _version_ | 1811258381561757696 |
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| author | Marco A. Herrera Quijano Marco A. Herrera Quijano Nadia Sharma Nadia Sharma Pascal Morissette Martin Cheryle A. Séguin Cheryle A. Séguin Lauren E. Flynn Lauren E. Flynn Lauren E. Flynn Lauren E. Flynn |
| author_facet | Marco A. Herrera Quijano Marco A. Herrera Quijano Nadia Sharma Nadia Sharma Pascal Morissette Martin Cheryle A. Séguin Cheryle A. Séguin Lauren E. Flynn Lauren E. Flynn Lauren E. Flynn Lauren E. Flynn |
| author_sort | Marco A. Herrera Quijano |
| collection | DOAJ |
| description | Bioscaffolds derived from the extracellular matrix (ECM) have shown the capacity to promote regeneration by providing tissue-specific biological instructive cues that can enhance cell survival and direct lineage-specific differentiation. This study focused on the development and characterization of two-dimensional (2-D) and three-dimensional (3-D) cell culture platforms incorporating decellularized nucleus pulposus (DNP). First, a detergent-free protocol was developed for decellularizing bovine nucleus pulposus (NP) tissues that was effective at removing cellular content while preserving key ECM constituents including collagens, glycosaminoglycans, and the cell-adhesive glycoproteins laminin and fibronectin. Next, novel 2-D coatings were generated using the DNP or commercially-sourced bovine collagen type I (COL) as a non-tissue-specific control. In addition, cryo-milled DNP or COL particles were incorporated within methacrylated chondroitin sulphate (MCS) hydrogels as a 3-D cell culture platform for exploring the effects of ECM particle composition. Culture studies showed that the 2-D coatings derived from the DNP could support cell attachment and growth, but did not maintain or rescue the phenotype of primary bovine NP cells, which de-differentiated when serially passaged in monolayer culture. Similarly, while bovine NP cells remained highly viable following encapsulation and 14 days of culture within the hydrogel composites, the incorporation of DNP particles within the MCS hydrogels was insufficient to maintain or rescue changes in NP phenotype associated with extended in vitro culture based on gene expression patterns. Overall, DNP produced with our new decellularization protocol was successfully applied to generate both 2-D and 3-D bioscaffolds; however, further studies are required to assess if these platforms can be combined with additional components of the endogenous NP microenvironment to stimulate regeneration or lineage-specific cell differentiation. |
| first_indexed | 2024-04-12T18:13:28Z |
| format | Article |
| id | doaj.art-dcbd6cf11b2b482ca0dca4524b300500 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-12T18:13:28Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-dcbd6cf11b2b482ca0dca4524b3005002022-12-22T03:21:45ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-09-011010.3389/fbioe.2022.937239937239Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposusMarco A. Herrera Quijano0Marco A. Herrera Quijano1Nadia Sharma2Nadia Sharma3Pascal Morissette Martin4Cheryle A. Séguin5Cheryle A. Séguin6Lauren E. Flynn7Lauren E. Flynn8Lauren E. Flynn9Lauren E. Flynn10Department of Anatomy and Cell Biology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, CanadaBone and Joint Institute, The University of Western Ontario, London, ON, CanadaBone and Joint Institute, The University of Western Ontario, London, ON, CanadaSchool of Biomedical Engineering, The University of Western Ontario, London, ON, CanadaDepartment of Anatomy and Cell Biology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, CanadaBone and Joint Institute, The University of Western Ontario, London, ON, CanadaDepartment of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, CanadaDepartment of Anatomy and Cell Biology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, CanadaBone and Joint Institute, The University of Western Ontario, London, ON, CanadaSchool of Biomedical Engineering, The University of Western Ontario, London, ON, CanadaDepartment of Chemical and Biochemical Engineering, Faculty of Engineering, The University of Western Ontario, London, ON, CanadaBioscaffolds derived from the extracellular matrix (ECM) have shown the capacity to promote regeneration by providing tissue-specific biological instructive cues that can enhance cell survival and direct lineage-specific differentiation. This study focused on the development and characterization of two-dimensional (2-D) and three-dimensional (3-D) cell culture platforms incorporating decellularized nucleus pulposus (DNP). First, a detergent-free protocol was developed for decellularizing bovine nucleus pulposus (NP) tissues that was effective at removing cellular content while preserving key ECM constituents including collagens, glycosaminoglycans, and the cell-adhesive glycoproteins laminin and fibronectin. Next, novel 2-D coatings were generated using the DNP or commercially-sourced bovine collagen type I (COL) as a non-tissue-specific control. In addition, cryo-milled DNP or COL particles were incorporated within methacrylated chondroitin sulphate (MCS) hydrogels as a 3-D cell culture platform for exploring the effects of ECM particle composition. Culture studies showed that the 2-D coatings derived from the DNP could support cell attachment and growth, but did not maintain or rescue the phenotype of primary bovine NP cells, which de-differentiated when serially passaged in monolayer culture. Similarly, while bovine NP cells remained highly viable following encapsulation and 14 days of culture within the hydrogel composites, the incorporation of DNP particles within the MCS hydrogels was insufficient to maintain or rescue changes in NP phenotype associated with extended in vitro culture based on gene expression patterns. Overall, DNP produced with our new decellularization protocol was successfully applied to generate both 2-D and 3-D bioscaffolds; however, further studies are required to assess if these platforms can be combined with additional components of the endogenous NP microenvironment to stimulate regeneration or lineage-specific cell differentiation.https://www.frontiersin.org/articles/10.3389/fbioe.2022.937239/fullintervertebral disc (IVD) degenerationnucleus pulposusdecellularizationextracellar matrixcoatingshydrogel composite |
| spellingShingle | Marco A. Herrera Quijano Marco A. Herrera Quijano Nadia Sharma Nadia Sharma Pascal Morissette Martin Cheryle A. Séguin Cheryle A. Séguin Lauren E. Flynn Lauren E. Flynn Lauren E. Flynn Lauren E. Flynn Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus Frontiers in Bioengineering and Biotechnology intervertebral disc (IVD) degeneration nucleus pulposus decellularization extracellar matrix coatings hydrogel composite |
| title | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
| title_full | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
| title_fullStr | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
| title_full_unstemmed | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
| title_short | Development of 2-D and 3-D culture platforms derived from decellularized nucleus pulposus |
| title_sort | development of 2 d and 3 d culture platforms derived from decellularized nucleus pulposus |
| topic | intervertebral disc (IVD) degeneration nucleus pulposus decellularization extracellar matrix coatings hydrogel composite |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.937239/full |
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