Gene expression analysis of growth factor receptors in human chondrocytes in monolayer and 3D pellet cultures

Witt,Anika; Salamon,Achim; Boy,Diana; Hansmann,Doris; Büttner,Andreas; Wree,Andreas; Bader,Rainer; Jonitz‑Heincke,Anika

doi:10.3892/ijmm.2017.2994

Gene expression analysis of growth factor receptors in human chondrocytes in monolayer and 3D pellet cultures

Authors:
- Anika Witt
- Achim Salamon
- Diana Boy
- Doris Hansmann
- Andreas Büttner
- Andreas Wree
- Rainer Bader
- Anika Jonitz‑Heincke
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Affiliations: Department of Orthopaedics, Biomechanics and Implant Technology Research Laboratory, University Medical Centre Rostock, D‑18057 Rostock, Germany, Department of Cell Biology, University Medical Centre Rostock, D‑18057 Rostock, Germany, Institute of Forensic Medicine, University Medical Centre Rostock, D‑18057 Rostock, Germany, Institute of Anatomy, University Medical Centre Rostock, D‑18057 Rostock, Germany
Published online on: May 18, 2017 https://doi.org/10.3892/ijmm.2017.2994
Pages: 10-20
Copyright: © Witt et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The main goal of cartilage repair is to create functional tissue by enhancing the in vitro conditions to more physiological in vivo conditions. Chondrogenic growth factors play an important role in influencing cartilage homeostasis. Insulin‑like growth factor (IGF)‑1 and transforming growth factor (TGF)‑β1 affect the expression of collagen type II (Col2) and glycosaminoglycans (GAGs) and, therefore, the targeted use of growth factors could make chondrogenic redifferentiation more efficient. In the present study, human chondrocytes were postmortally isolated from healthy articular cartilage and cultivated as monolayer or 3D pellet cultures either under normoxia or hypoxia and stimulated with IGF‑1 and/or TGF‑β1 to compare the impact of the different growth factors. The mRNA levels of the specific receptors (IGF1R, TGFBR1, TGFBR2) were analyzed at different time points. Moreover, gene expression rates of collagen type 1 and 2 in pellet cultures were observed over a period of 5 weeks. Additionally, hyaline‑like Col2 protein and sulphated GAG (sGAG) levels were quantified. Stimulation with IGF‑1 resulted in an enhanced expression of IGF1R and TGFBR2 whereas TGF‑β1 stimulated TGFBR1 in the monolayer and pellet cultures. In monolayer, the differences reached levels of significance. This effect was more pronounced under hypoxic culture conditions. In pellet cultures, increased amounts of Col2 protein and sGAGs after incubation with TGF‑β1 and/or IGF‑1 were validated. In summary, constructing a gene expression profile regarding mRNA levels of specific growth factor receptors in monolayer cultures could be helpful for a targeted application of growth factors in cartilage tissue engineering.

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