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Effect of reduced oxygen tension and long-term mechanical stimulation on chondrocyte-polymer constructs

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Abstract

We have investigated the influence of long-term confined dynamic compression and surface motion under low oxygen tension on tissue-engineered cell-scaffold constructs. Porous polyurethane scaffolds (8 mm × 4 mm) were seeded with bovine articular chondrocytes and cultured under normoxic (21% O2) or hypoxic (5% O2) conditions for up to 4 weeks. By means of our joint-simulating bioreactor, cyclic axial compression (10–20%; 0.5 Hz) was applied for 1 h daily with a ceramic ball, which simultaneously oscillated over the construct surface (±25°; 0.5 Hz). Culture under reduced oxygen tension resulted in an increase in mRNA levels of type II collagen and aggrecan, whereas the expression of type I collagen was down-regulated at early time points. A higher glycosaminoglycan content was found in hypoxic than in normoxic constructs. Immunohistochemical analysis showed more intense type II and weaker type I collagen staining in hypoxic than in normoxic cultures. Type II collagen gene expression was slightly elevated after short-term loading, whereas aggrecan mRNA levels were not influenced by the applied mechanical stimuli. Of importance, the combination of loading and low oxygen tension resulted in a further down-regulation of collagen type I mRNA expression, contributing to the stabilization of the chondrocytic phenotype. Histological results confirmed the beneficial effect of mechanical loading on chondrocyte matrix synthesis. Thus, mechanical stimulation combined with low oxygen tension is an effective tool for modulating the chondrocytic phenotype and should be considered when chondrocytes or mesenchymal stem cells are cultured and differentiated with the aim of generating cartilage-like tissue in vitro.

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Acknowledgement

We are grateful to Dr. Andreas Goessl, Baxter Biosurgery, Vienna, for providing the fibrin components and to Robert Peter for excellent technical assistance with cell culture and biochemical analysis.

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Author notes

  1. Ellen Wernike

    Present address: Department of Clinical Research, University of Bern, Bern, 3010, Switzerland

Authors and Affiliations

  1. Biomaterials and Tissue Engineering Program, AO Research Institute, Clavadelerstrasse 8, Davos Platz, 7270, Switzerland

    Ellen Wernike, Zhen Li, Mauro Alini & Sibylle Grad

  2. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, 310027, People’s Republic of China

    Zhen Li

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  1. Ellen Wernike
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  2. Zhen Li
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Correspondence to Sibylle Grad.

Additional information

This work was supported by the Swiss National Science Foundation (grant no. 3200B0-104083).

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Wernike, E., Li, Z., Alini, M. et al. Effect of reduced oxygen tension and long-term mechanical stimulation on chondrocyte-polymer constructs. Cell Tissue Res 331, 473–483 (2008). https://doi.org/10.1007/s00441-007-0500-9

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