Human osteochondritis dissecans fragment-derived chondrocyte characteristics ex vivo, after monolayer expansion-induced de-differentiation, and after re-differentiation in alginate bead culture
Background Autologous chondrocyte implantation (ACI) is a therapy for articular cartilage and osteochondral lesions that relies on notch- or trochlea-derived primary chondrocytes. An alternative cell source for ACI could be osteochondritis dissecans (OCD) fragment-derived chondrocyt...
| Main Authors: | , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
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BioMed Central
2018
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| Online Access: | http://hdl.handle.net/1721.1/115986 |
| _version_ | 1811095793210228736 |
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| author | Aurich, Matthias Gras, Florian Hofmann, Gunther O. Rolauffs, Bernd G |
| author2 | Massachusetts Institute of Technology. Center for Biomedical Engineering |
| author_facet | Massachusetts Institute of Technology. Center for Biomedical Engineering Aurich, Matthias Gras, Florian Hofmann, Gunther O. Rolauffs, Bernd G |
| author_sort | Aurich, Matthias |
| collection | MIT |
| description | Background
Autologous chondrocyte implantation (ACI) is a therapy for articular cartilage and osteochondral lesions that relies on notch- or trochlea-derived primary chondrocytes. An alternative cell source for ACI could be osteochondritis dissecans (OCD) fragment-derived chondrocytes. Assessing the potential of these cells, we investigated their characteristics ex vivo and after monolayer expansion, as monolayer expansion is an integral step of ACI. However, as monolayer expansion can induce de-differentiation, we asked whether monolayer-induced de-differentiation can be reverted through successive alginate bead culture.
Methods
Chondrocytes were isolated from the OCD fragments of 15 patient knees with ICRS grades 3–4 lesions for ex vivo analyses, primary alginate bead culture, monolayer expansion, and alginate bead culture following monolayer expansion for attempting re-differentiation. We determined yield, viability, and the mRNA expression of aggrecan and type I, II, and X collagen.
Results
OCD fragment-derived chondrocyte isolation yielded high numbers of viable cells with a low type I:II collagen expression ratio (< 1) and a relatively high aggrecan and type II and X collagen mRNA expression, indicating chondrogenic and hypertrophic characteristics. As expected, monolayer expansion induced de-differentiation. Alginate bead culture of monolayer-expanded cells significantly improved the expression profile of all genes investigated, being most successful in decreasing the hypertrophy marker type X collagen to 1.5% of its ex vivo value. However, the chondrogenic phenotype was not fully restored, as the collagen type I:II expression ratio decreased significantly but remained > 1.
Conclusion
OCD fragment derived human chondrocytes may hold not yet utilized clinical potential for cartilage repair. Keywords: Chondrocyte; Articular cartilage; De-differentiation Re-differentiation; Monolayer expansion; Alginate bead culture |
| first_indexed | 2024-09-23T16:28:04Z |
| format | Article |
| id | mit-1721.1/115986 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T16:28:04Z |
| publishDate | 2018 |
| publisher | BioMed Central |
| record_format | dspace |
| spelling | mit-1721.1/1159862022-09-29T19:57:37Z Human osteochondritis dissecans fragment-derived chondrocyte characteristics ex vivo, after monolayer expansion-induced de-differentiation, and after re-differentiation in alginate bead culture Aurich, Matthias Gras, Florian Hofmann, Gunther O. Rolauffs, Bernd G Massachusetts Institute of Technology. Center for Biomedical Engineering Rolauffs, Bernd G Background Autologous chondrocyte implantation (ACI) is a therapy for articular cartilage and osteochondral lesions that relies on notch- or trochlea-derived primary chondrocytes. An alternative cell source for ACI could be osteochondritis dissecans (OCD) fragment-derived chondrocytes. Assessing the potential of these cells, we investigated their characteristics ex vivo and after monolayer expansion, as monolayer expansion is an integral step of ACI. However, as monolayer expansion can induce de-differentiation, we asked whether monolayer-induced de-differentiation can be reverted through successive alginate bead culture. Methods Chondrocytes were isolated from the OCD fragments of 15 patient knees with ICRS grades 3–4 lesions for ex vivo analyses, primary alginate bead culture, monolayer expansion, and alginate bead culture following monolayer expansion for attempting re-differentiation. We determined yield, viability, and the mRNA expression of aggrecan and type I, II, and X collagen. Results OCD fragment-derived chondrocyte isolation yielded high numbers of viable cells with a low type I:II collagen expression ratio (< 1) and a relatively high aggrecan and type II and X collagen mRNA expression, indicating chondrogenic and hypertrophic characteristics. As expected, monolayer expansion induced de-differentiation. Alginate bead culture of monolayer-expanded cells significantly improved the expression profile of all genes investigated, being most successful in decreasing the hypertrophy marker type X collagen to 1.5% of its ex vivo value. However, the chondrogenic phenotype was not fully restored, as the collagen type I:II expression ratio decreased significantly but remained > 1. Conclusion OCD fragment derived human chondrocytes may hold not yet utilized clinical potential for cartilage repair. Keywords: Chondrocyte; Articular cartilage; De-differentiation Re-differentiation; Monolayer expansion; Alginate bead culture 2018-05-30T18:12:54Z 2018-05-30T18:12:54Z 2018-05 2017-12 2018-05-27T03:31:10Z Article http://purl.org/eprint/type/JournalArticle 1471-2474 http://hdl.handle.net/1721.1/115986 Aurich, Matthias et al. "Human osteochondritis dissecans fragment-derived chondrocyte characteristics ex vivo, after monolayer expansion-induced de-differentiation, and after re-differentiation in alginate bead culture." BMC Musculoskeletal Disorders 19 (May 2018): 168 © 2018 The Author(s) en https://doi.org/10.1186/s12891-018-2079-6 BMC Musculoskeletal Disorders Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ The Author(s). application/pdf BioMed Central BioMed Central |
| spellingShingle | Aurich, Matthias Gras, Florian Hofmann, Gunther O. Rolauffs, Bernd G Human osteochondritis dissecans fragment-derived chondrocyte characteristics ex vivo, after monolayer expansion-induced de-differentiation, and after re-differentiation in alginate bead culture |
| title | Human osteochondritis dissecans fragment-derived chondrocyte characteristics ex vivo, after monolayer expansion-induced de-differentiation, and after re-differentiation in alginate bead culture |
| title_full | Human osteochondritis dissecans fragment-derived chondrocyte characteristics ex vivo, after monolayer expansion-induced de-differentiation, and after re-differentiation in alginate bead culture |
| title_fullStr | Human osteochondritis dissecans fragment-derived chondrocyte characteristics ex vivo, after monolayer expansion-induced de-differentiation, and after re-differentiation in alginate bead culture |
| title_full_unstemmed | Human osteochondritis dissecans fragment-derived chondrocyte characteristics ex vivo, after monolayer expansion-induced de-differentiation, and after re-differentiation in alginate bead culture |
| title_short | Human osteochondritis dissecans fragment-derived chondrocyte characteristics ex vivo, after monolayer expansion-induced de-differentiation, and after re-differentiation in alginate bead culture |
| title_sort | human osteochondritis dissecans fragment derived chondrocyte characteristics ex vivo after monolayer expansion induced de differentiation and after re differentiation in alginate bead culture |
| url | http://hdl.handle.net/1721.1/115986 |
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