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Reduced hypertrophy in vitro after chondrogenic differentiation of adult human mesenchymal stem cells following adenoviral SOX9 gene delivery

Authors
  • Weissenberger, M.1
  • Weissenberger, M. H.1, 2
  • Gilbert, F.1, 3
  • Groll, J.4
  • Evans, C. H.5
  • Steinert, A. F.1, 6
  • 1 Julius-Maximilians-University, Brettreichstrasse 11, Würzburg, D-97074, Germany , Würzburg (Germany)
  • 2 Caritas-Hospital, Bad Mergentheim, Germany , Bad Mergentheim (Germany)
  • 3 University Hospital Würzburg, Würzburg, Germany , Würzburg (Germany)
  • 4 Julius-Maximilians-University, Würzburg, Germany , Würzburg (Germany)
  • 5 Mayo Clinic, Rochester, MN, USA , Rochester (United States)
  • 6 Rhön-Klinikum Campus Bad Neustadt, Bad Neustadt a.d. Saale, Germany , Bad Neustadt a.d. Saale (Germany)
Type
Published Article
Journal
BMC Musculoskeletal Disorders
Publisher
Springer (Biomed Central Ltd.)
Publication Date
Feb 17, 2020
Volume
21
Issue
1
Identifiers
DOI: 10.1186/s12891-020-3137-4
Source
Springer Nature
Keywords
License
Green

Abstract

BackgroundMesenchymal stem cell (MSC) based-treatments of cartilage injury are promising but impaired by high levels of hypertrophy after chondrogenic induction with several bone morphogenetic protein superfamily members (BMPs). As an alternative, this study investigates the chondrogenic induction of MSCs via adenoviral gene-delivery of the transcription factor SOX9 alone or in combination with other inducers, and comparatively explores the levels of hypertrophy and end stage differentiation in a pellet culture system in vitro.MethodsFirst generation adenoviral vectors encoding SOX9, TGFB1 or IGF1 were used alone or in combination to transduce human bone marrow-derived MSCs at 5 × 102 infectious particles/cell. Thereafter cells were placed in aggregates and maintained for three weeks in chondrogenic medium. Transgene expression was determined at the protein level (ELISA/Western blot), and aggregates were analysed histologically, immunohistochemically, biochemically and by RT-PCR for chondrogenesis and hypertrophy.ResultsSOX9 cDNA was superior to that encoding TGFB1, the typical gold standard, as an inducer of chondrogenesis in primary MSCs as evidenced by improved lacuna formation, proteoglycan and collagen type II staining, increased levels of GAG synthesis, and expression of mRNAs associated with chondrogenesis. Moreover, SOX9 modified aggregates showed a markedly lower tendency to progress towards hypertrophy, as judged by expression of the hypertrophy markers alkaline phosphatase, and collagen type X at the mRNA and protein levels.ConclusionAdenoviral SOX9 gene transfer induces chondrogenic differentiation of human primary MSCs in pellet culture more effectively than TGFB1 gene transfer with lower levels of chondrocyte hypertrophy after 3 weeks of in vitro culture. Such technology might enable the formation of more stable hyaline cartilage repair tissues in vivo.

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