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Interaktion zwischen humanen Chondrozyten und extrazellulärer Matrix in vitro

Ein Beitrag zur autologen Chondrozytentransplantation

Interaction between human chondrocytes and extracellular matrix in vitro

A contribution to autologous chondrocyte transplantation

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Zusammenfassung

Hintergrund

Die autologe Chondrozytentransplantation (ACT) hat als Therapiemethode bei kleineren Knorpeldefekten Erfolge gezeigt. Die Vermehrung der Chondrozyten für ACT gelingt nur in vitro und diese führt zur Dedifferenzierung der Zellen. Ziel dieser Arbeit war, die Züchtung von Chondrozyten zu optimieren und eine Dedifferenzierung der Zellen zu verhindern.

Material und Methode

Die humanen Chondrozyten wurden auf mit und ohne Typ-II-Kollagen beschichteten Oberflächen kultiviert. Die Zellen wurden morphologisch und mittels „Immunoblotting“ untersucht.

Ergebnisse

Auf Typ-II-Kollagen bleibt der chondrogene Phänotyp bis zum 20. Tag stabil und es werden mehr aktivierte intrazelluläre Proteine sowie das Adaptorprotein Shc (sarc homology collagen) nachgewiesen. Behandlung mit β1-Integrin-Antikörpern führte zu einer deutliche Abnahme (82%) der Zell-Adhäsion. Typ-II-Kollagen und Chondrozyteninteraktion führte zur Aktivierung der Integrine, die über das Shc-Protein den Ras-MAPK-Signalübertragungsweg aktivieren, welcher den chondrogenen Phänotyp stabilisiert.

Schlussfolgerung

Diese Kenntnisse könnten langfristig die In-vitro-Vermehrung autologer Chondrozyten verbessern und dem Einsatz dieser Technik bei größeren oder sogar arthrotischen Knorpeldefekten mehr Aussicht geben.

Abstract

Background

Autologous chondrocyte transplantation (ACT) has had reasonable success for repairing small articular cartilage defects. A limiting factor for ACT is, however, the in vitro cultivation of chondrocytes because it leads to dedifferentiation. Therefore, the goal of this work was to optimize the monolayer culture of chondrocytes in vitro.

Material and method

Human articular chondrocytes were plated on either collagen type II or untreated surfaces. The cells were evaluated morphologically and with immunoblotting.

Results

On collagen type II surfaces, a stable chondrogenic phenotype, expression of β1-integrin, and a significant activation of phosphorylated intracellular proteins and the adaptor protein Shc could be observed up to day 20 in culture. Treatment with β1 integrin antibody led to a loss of cell adhesion (82%). The results indicate that on collagen type II, β1-integrin receptors are activated. Through the activation of Shc, these stimulate the Ras-MAPK pathway, which stabilizes the chondrogenic phenotype.

Conclusion

Our results provide a practical and low-cost solution for improved long-term chondrocyte cultivation, thus providing a new perspective for using ACT on larger or arthrotic cartilage defects.

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Danksagung

Der besondere Dank der Autoren gilt der technischen Unterstützung von Fr. Karoline Fischer.

Interessenkonflikt

Der korrespondierende Autor gibt an, dass kein Interessenkonflikt besteht.

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Authors and Affiliations

  1. Forschungsgruppe Muskuloskelettales System, Institut für Anatomie, Medizinische Fakultät, Ludwig-Maximilians-Universität München, Pettenkoferstraße 11, 80336, München, Deutschland

    M. Shakibaei, C. Csaki & R. Putz

  2. Internationales Centrum für Gelenk- und Knochenchirurgie Berlin, Berlin, Deutschland

    M. Rahmanzadeh

Authors
  1. M. Shakibaei
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  2. C. Csaki
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  3. M. Rahmanzadeh
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  4. R. Putz
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Correspondence to M. Shakibaei.

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Shakibaei, M., Csaki, C., Rahmanzadeh, M. et al. Interaktion zwischen humanen Chondrozyten und extrazellulärer Matrix in vitro. Orthopäde 37, 440–447 (2008). https://doi.org/10.1007/s00132-008-1260-2

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  • DOI: https://doi.org/10.1007/s00132-008-1260-2

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