Article
Proliferation and differentiation of mesenchymal stromal cells from osteoarthritic donors
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Authors
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H. Friedrich
- University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany
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C. Vater
- University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany
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J. Rauh
- University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany
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K.-P. Günther
- University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany
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C. Bünger
- University Hospital of Arhus, Dept. of Orthopaedic Surgery, Aarhus C, Denmark
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M. Stiehler
- University Hospital Carl Gustav Carus, Department of Orthopaedics, Dresden, Germany
| Published: | October 21, 2010 |
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Outline
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Objective: Osteoarthritis (OA) is one of the most frequent musculoskeletal disorders and represents the main indication for total joint arthroplasty http://www.jru.orthop.gu.se/. Multipo-tent mesenchymal stromal cells (MSCs) can be easily isolated and culture expanded from bone marrow aspirates and provide an excellent source of progenitor cells. [Stiehler et al. 2006]. To assess whether advanced-stage OA affects MSCs’ suitability for musculoskeletal regenerative therapy we compared proliferation and differentiation potential of MSCs from osteoarthritic versus healthy donors.
Methods: WOMAC, EQ-5D, Harris Hip Score, extended laboratory scan, and radiological classification of the affected hip joint (OA group only) were performed. Subjects on bone metabolism-affecting medication were excluded from the study. MSCs were isolated from bone marrow aspirates obtained from the pelvic compartments of n=14 advanced-stage osteoarthritic (Kellgren and Lawrence grade 3 or 4, mean age 67±6 years) and n=15 age-matched (61±4 years) healthy donors by gradient-separation and polystyrene adhesion method. MSCs were cultured in basic (DMEM, 10% fetal calf serum), osteogenic, adipogenic, or chondrogenic medium for up to 21 days. For all assays low-passage (<3) MSC populations were used. Proliferation was assessed by total DNA quantification, FACS analysis and CFU-F assay.
Differentiation was proofed by cell staining and by osteogenic (Runx-2, ALP, BSP2), chondrogenic (Sox9, ColIIa, ColX), and adipogenic (PPARγ, FABP4) qRT-PCR marker gene expression analysis. Additionally, osteogenic differentiation was evaluated by cell-specific alkaline phosphatase (ALP) activity assay.
Overall statistical significance was defined as p<0.05 (two-sided) based on all pairwise comparisons . The study was approved by the local ethics review board (protocol #EK203082008).
Results and conclusions: Algofunctional scores of osteoarthritic donors were significantly lower compared to healthy donors (p<0.0001). No significant differences between osteoarthritic and healthy donors concerning the proliferation potential, cell-specific ALP acitivity and differentiation marker gene expression were observed.
We therefore conclude that the regenerative potential of MSCs from osteoarthritic donors is compa-rable to MSCs from healthy donors. These data will help to facilitate the application of autologous cell-based strategies for musculoskeletal tissue regeneration.
