Physikalische Charakterisierung dezellularisierter Knorpelmatrix zur Anwendung in der Rhinochirurgie

Zusammenfassung

Hintergrund: Der Einsatz von autologem Knorpel aus Ohrmuschel und Rippe zur Rekonstruktion von Knorpeldefekten und Deformitäten der Nase birgt eine Reihe von Problemen, die sowohl den Chirurgen als auch den Patienten betreffen. Daher ist die Entwicklung geeigneter Ersatzmaterialien Gegenstand intensiver Forschung. Dezellularisierter Knorpel hat sich in vorangegangenen Experimenten als vielversprechendes Rekonstruktionsmaterial herausgestellt.

Material/Methode: Zur genaueren Definition der Oberflächeneigenschaften und Untersuchung der zellulären Reaktion auf die Kollagenmatrix wurden in einer Langzeit-3D-Kultur die Materialgeometrie von dezellularisiertem Schweineseptumknorpel durch Mikrocomputertomografie und die Materialcharakteristika mittels Raster- bzw. Transmissionselektronenmikroskopie näher untersucht. Weiterhin wurde die Integrin-Expression humaner Chondrozyten nach Besiedelung und Migration in das Trägermaterial ermittelt.

Ergebnisse: Dabei zeigte sich nach Dezellularisierung des Knorpels eine deutliche Vergrößerung der Gesamtoberfläche. Eine intensive Interaktion der Chondrozyten mit der Kollagenmatrix konnte anhand der Veränderung des Integrin-Expressionprofils mit Hochregulierung von ITGB1 und ITGA1 gezeigt werden, die für eine zunehmende chondrozytäre Differenzierung sprechen. Darüber hinaus ließ sich eine aktive Migration in das Material beobachten.

Schlussfolgerung: Dezellularisierter porciner Knorpel bietet somit hinsichtlich der zur Verfügung stehenden Oberfläche und daraus resultierenden Zellintegration eine geeignete Mikrostruktur für humane Chondrozyten. Er besitzt folglich die notwendigen Vor­aussetzungen für die klinische Anwendung in der rekonstruktiven Chirurgie.

Abstract

Background: The use of autologous auricular and rib cartilage for the reconstruction of nasal defects and deformities is associated with a number of disadvantages. The development of alternative materials is therefore the focus of intensive research. Recent studies demonstrated that decellularized cartilage is a promising material for cartilage tissue engineering. Hence, the aim of this study was to characterize the materials surface and cellular reactions to the decellularized cartilage matrix in long term-3D-culture.

Material and Methods: Material geometry of decellularized cartilage was examined by microcomputed tomography as well as material characteristics by scanning and transmission electron microscopy. The expression of integrins on the surface of human chondrocytes was determined after seeding and migration into the scaffold.

Results: After decellularization an obvious enlargement of the matrix surface and an intensive interaction between the chondrocytes and the collagen matrix was observed. ITGA1 and ITGB1 were upregulated indicating chondrogenic differentiation.

Conclusion: Therefore, decellularized porcine cartilage provides an optimal microstructure for human chondrocytes with respect to cell integration and matrix production. Thus, it offers promising characteristics for clinical application in reconstructive surgery.

Schlüsselwörter

Knorpelersatz - ECM Matrices - Zelladhäsion - Integrine - Mikrocomputertomografie

Key words

cartilage reconstruction - ECM matrices - cell adhesion - integrins - micro-CT

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