Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage
Objective: To quantify the structural characteristics and nanomechanical properties of aggrecan produced by adult bone marrow stromal cells (BMSCs) in peptide hydrogel scaffolds and compare to aggrecan from adult articular cartilage. Design: Adult equine BMSCs were encapsulated in 3D-peptide...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Elsevier Ltd.
2011
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| Online Access: | http://hdl.handle.net/1721.1/67287 https://orcid.org/0000-0003-3511-5679 https://orcid.org/0000-0003-0026-6215 https://orcid.org/0000-0002-4942-3456 |
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| author | Lee, H.-Y. Kopesky, Paul Wayne Plaas, A. Sandy, J. Kisiday, John D. Frisbie, David D. Grodzinsky, Alan J. Ortiz, Christine |
| author2 | Massachusetts Institute of Technology. Center for Biomedical Engineering |
| author_facet | Massachusetts Institute of Technology. Center for Biomedical Engineering Lee, H.-Y. Kopesky, Paul Wayne Plaas, A. Sandy, J. Kisiday, John D. Frisbie, David D. Grodzinsky, Alan J. Ortiz, Christine |
| author_sort | Lee, H.-Y. |
| collection | MIT |
| description | Objective:
To quantify the structural characteristics and nanomechanical properties of aggrecan produced by adult bone marrow stromal cells (BMSCs) in peptide hydrogel scaffolds and compare to aggrecan from adult articular cartilage.
Design:
Adult equine BMSCs were encapsulated in 3D-peptide hydrogels and cultured for 21 days with TGF-β1 to induce chondrogenic differentiation. BMSC-aggrecan was extracted and compared with aggrecan from age-matched adult equine articular cartilage. Single molecules of aggrecan were visualized by atomic force microcopy-based imaging and aggrecan nanomechanical stiffness was quantified by high resolution force microscopy. Population-averaged measures of aggrecan hydrodynamic size, core protein structures and CS sulfation compositions were determined by size-exclusion chromatography, Western analysis, and fluorescence-assisted carbohydrate electrophoresis (FACE).
Results:
BMSC-aggrecan was primarily full-length while cartilage-aggrecan had many fragments. Single molecule measurements showed that core protein and GAG chains of BMSC-aggrecan were markedly longer than those of cartilage-aggrecan. Comparing full-length aggrecan of both species, BMSC-aggrecan had longer GAG chains, while the core protein trace lengths were similar. FACE analysis detected a ∼1:1 ratio of chondroitin-4-sulfate to chondroitin-6-sulfate in BMSC-GAG, a phenotype consistent with aggrecan from skeletally-immature cartilage. The nanomechanical stiffness of BMSC-aggrecan was demonstrably greater than that of cartilage-aggrecan at the same total sGAG (fixed charge) density.
Conclusions:
The higher proportion of full-length monomers, longer GAG chains and greater stiffness of the BMSC-aggrecan makes it biomechanically superior to adult cartilage-aggrecan. Aggrecan stiffness was not solely dependent on fixed charge density, but also on GAG molecular ultrastructure. These results support the use of adult BMSCs for cell-based cartilage repair. |
| first_indexed | 2024-09-23T11:01:47Z |
| format | Article |
| id | mit-1721.1/67287 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:01:47Z |
| publishDate | 2011 |
| publisher | Elsevier Ltd. |
| record_format | dspace |
| spelling | mit-1721.1/672872022-09-27T16:38:52Z Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage Lee, H.-Y. Kopesky, Paul Wayne Plaas, A. Sandy, J. Kisiday, John D. Frisbie, David D. Grodzinsky, Alan J. Ortiz, Christine Massachusetts Institute of Technology. Center for Biomedical Engineering Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Massachusetts Institute of Technology. Department of Materials Science and Engineering Massachusetts Institute of Technology. Department of Mechanical Engineering Grodzinsky, Alan J. Kopesky, Paul Wayne Lee, H.-Y. Grodzinsky, Alan J. Ortiz, Christine Kisiday, John D. Objective: To quantify the structural characteristics and nanomechanical properties of aggrecan produced by adult bone marrow stromal cells (BMSCs) in peptide hydrogel scaffolds and compare to aggrecan from adult articular cartilage. Design: Adult equine BMSCs were encapsulated in 3D-peptide hydrogels and cultured for 21 days with TGF-β1 to induce chondrogenic differentiation. BMSC-aggrecan was extracted and compared with aggrecan from age-matched adult equine articular cartilage. Single molecules of aggrecan were visualized by atomic force microcopy-based imaging and aggrecan nanomechanical stiffness was quantified by high resolution force microscopy. Population-averaged measures of aggrecan hydrodynamic size, core protein structures and CS sulfation compositions were determined by size-exclusion chromatography, Western analysis, and fluorescence-assisted carbohydrate electrophoresis (FACE). Results: BMSC-aggrecan was primarily full-length while cartilage-aggrecan had many fragments. Single molecule measurements showed that core protein and GAG chains of BMSC-aggrecan were markedly longer than those of cartilage-aggrecan. Comparing full-length aggrecan of both species, BMSC-aggrecan had longer GAG chains, while the core protein trace lengths were similar. FACE analysis detected a ∼1:1 ratio of chondroitin-4-sulfate to chondroitin-6-sulfate in BMSC-GAG, a phenotype consistent with aggrecan from skeletally-immature cartilage. The nanomechanical stiffness of BMSC-aggrecan was demonstrably greater than that of cartilage-aggrecan at the same total sGAG (fixed charge) density. Conclusions: The higher proportion of full-length monomers, longer GAG chains and greater stiffness of the BMSC-aggrecan makes it biomechanically superior to adult cartilage-aggrecan. Aggrecan stiffness was not solely dependent on fixed charge density, but also on GAG molecular ultrastructure. These results support the use of adult BMSCs for cell-based cartilage repair. National Institutes of Health (U.S.) (NIH grant EB003805) National Institutes of Health (U.S.) (Grant AR33236) National Science Foundation (U.S.) (NSF grant NIRT-0403903) National Science Foundation (U.S.) (CMMI-0758651) National Institutes of Health (U.S.) (NIH Molecular, Cell, and Tissue Biomechanics Training Grant) Massachusetts Institute of Technology (Whitaker Health Science Fund Fellowship) 2011-11-21T22:30:12Z 2011-11-21T22:30:12Z 2010-11 2010-01 Article http://purl.org/eprint/type/JournalArticle 1522-9653 1063-4584 http://hdl.handle.net/1721.1/67287 Lee, H.-Y. et al. “Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage.” Osteoarthritis and Cartilage 18 (2010): 1477-1486. https://orcid.org/0000-0003-3511-5679 https://orcid.org/0000-0003-0026-6215 https://orcid.org/0000-0002-4942-3456 en_US http://dx.doi.org/10.1016/j.joca.2010.07.015 Osteoarthritis and Cartilage Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf Elsevier Ltd. PubMed Central |
| spellingShingle | Lee, H.-Y. Kopesky, Paul Wayne Plaas, A. Sandy, J. Kisiday, John D. Frisbie, David D. Grodzinsky, Alan J. Ortiz, Christine Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage |
| title | Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage |
| title_full | Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage |
| title_fullStr | Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage |
| title_full_unstemmed | Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage |
| title_short | Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage |
| title_sort | adult bone marrow stromal cell based tissue engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage |
| url | http://hdl.handle.net/1721.1/67287 https://orcid.org/0000-0003-3511-5679 https://orcid.org/0000-0003-0026-6215 https://orcid.org/0000-0002-4942-3456 |
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