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Cartilage tissue engineering of nasal septal chondrocyte-macroaggregates in human demineralized bone matrix

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Abstract

Tissue Engineering is an important method for generating cartilage tissue with isolated autologous cells and the support of biomaterials. In contrast to various gel-like biomaterials, human demineralized bone matrix (DBM) guarantees some biomechanical stability for an application in biomechanically loaded regions. The present study combined for the first time the method of seeding chondrocyte-macroaggregates in DBM for the purpose of cartilage tissue engineering. After isolating human nasal chondrocytes and creating a three-dimensional macroaggregate arrangement, the DBM was cultivated in vitro with the macroaggregates. The interaction of the cells within the DBM was analyzed with respect to cell differentiation and the inhibitory effects of chondrocyte proliferation. In contrast to chondrocyte-macroaggregates in the cell-DBM constructs, morphologically modified cells expressing type I collagen dominated. The redifferentiation of chondrocytes, characterized by the expression of type II collagen, was only found in low amounts in the cell-DBM constructs. Furthermore, caspase 3, a marker for apoptosis, was detected in the chondrocyte-DBM constructs. In another experimental setting, the vitality of chondrocytes as related to culture time and the amount of DBM was analyzed with the BrdU assay. Higher amounts of DBM tended to result in significantly higher proliferation rates of the cells within the first 48 h. After 96 h, the vitality decreased in a dose-dependent fashion. In conclusion, this study provides the proof of concept of chondrocyte-macroaggregates with DBM as an interesting method for the tissue engineering of cartilage. The as-yet insufficient redifferentiation of the chondrocytes and the sporadic initiation of apoptosis will require further investigations.

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Acknowledgments

The authors thank Mr. Schurig and Mr. Schweiger for their technical support and for making the DBM available.

Conflict of interest

No competing financial interests exist.

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Authors and Affiliations

  1. Department of General and Visceral Surgery, Johann Wolfgang Goethe University, Theodor-Stern-Kai 7, 60590, Frankfurt, Germany

    Juliane Liese

  2. Biotechnology, Berlin, Germany

    Ulrike Marzahn & Karym El Sayed

  3. Tissue Bank, Institute of Transfusion Medicine, Charité—Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany

    Axel Pruss

  4. Otolaryngology Center Berlin, Ferdinandstr. 34, 12203, Berlin, Germany

    Andreas Haisch

  5. Department of Otorhinolaryngology, Head and Neck Surgery, Charité—Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany

    Katharina Stoelzel

Authors
  1. Juliane Liese
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  2. Ulrike Marzahn
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  3. Karym El Sayed
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  4. Axel Pruss
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  5. Andreas Haisch
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  6. Katharina Stoelzel
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Corresponding author

Correspondence to Juliane Liese.

Additional information

Andreas Haisch and Katharina Stoelzel are joint senior authors.

The work was performed in the Department of Otorhinolaryngology, Head and Neck Surgery Charité—Universitätsmedizin Berlin, Germany.

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Liese, J., Marzahn, U., El Sayed, K. et al. Cartilage tissue engineering of nasal septal chondrocyte-macroaggregates in human demineralized bone matrix. Cell Tissue Bank 14, 255–266 (2013). https://doi.org/10.1007/s10561-012-9322-4

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  • DOI: https://doi.org/10.1007/s10561-012-9322-4

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