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Neovaskularisation und freier mikrochirurgischer Transfer von in vitro gezüchteten Knorpelkonstrukten

Neovascularisation and free microsurgical transfer of cartilage-engineered constructs

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Zusammenfassung

Hintergrund

Um im Bereich des Tissue-Engineerings das Überleben, die Form und die Funktion des gezüchteten Gewebes sicherzustellen, müssen in vitro gezüchtete Zellkonstrukte neovaskularisiert werden. Dieser Artikel präsentiert eine effektive Methode der Neovaskularisation und des anschließenden freien mikrochirurgischen Transfers von Knorpelzellkonstrukten.

Material und Methoden

Zwölf weibliche Chinchilla-Bastard-Kaninchen wurden operiert. Für die Herstellung der Knorpelkonstrukte wurden Chondrozyten aus einer Ohrknorpelbiopsie in Monolayerkultur amplifiziert und anschließend in poröse Konstrukte aus Polycaprolacton gesiedelt. In jeden präfabrizierten Hautlappen wurden 3 Knorpelkonstrukte (2×2×0,5 cm) und ein Konstrukt ohne Zellen (als Kontrolle) implantiert. Der randomisierte Hautlappen wurde mit einer Dimension von 8×15 cm geplant und mittels Implantation eines a.-v.-Gefäßstiels neovaskularisiert. Nach 6 Wochen wurde der 3-D-Hautlappen komplett entfernt und – basierend auf dem implantierten Gefäßstiel – mikrochirurgisch frei replantiert. Die Auswertung der Konstruktvitalität, Neovaskularisation und Bildung von neuem Knorpel erfolgte makroskopisch, mikroangiographisch, histologisch und immunhistologisch.

Ergebnisse

Alle neovaskularisierten Hautlappen mit integrierten gezüchteten Knorpelkonstrukten konnten erfolgreich frei transplantiert werden. Die Knorpelkonstrukte waren gut im Lappen integriert und geschützt, gut neovaskularisiert und histologisch in Bezug auf Größe und Form stabil. Immunhistologisch zeigte sich knorpelähnliches Gewebe mit einer Neusynthese von extrazellulärer Matrix.

Fazit

Diese experimentelle Studie zeigte eine verlässliche Methode der Neovaskularisation und freien mikrochirurgischen Transplantation von gezüchteten Knorpelzellkonstrukten im präfabrizierten Lappen. In Bezug auf eine mögliche klinische Anwendung ist sie aufgrund des guten ästhetischen Ergebnisses mit minimalem Hebedefekt vielversprechend.

Abstract

Background

Clinical imperatives for new cartilage to replace or restore the function of traumatized or missing tissue as a consequence of trauma, inherent malformations or disease has led to the need for therapies or procedures to generate cartilage for clinical applications. To ensure shape, function, and survival, in vitro cartilage-engineered constructs need to be revascularized. This study presents a viable method for neovascularization and free microsurgical transfer of these in vitro constructs.

Material and methods

Twelve female Chinchilla Bastard rabbits were operated. Cartilage-engineered constructs were created by isolating chondrocytes from auricular biopsies, amplifying in monolayer culture, and then seeding them onto polycaprolactone scaffolds. In each prefabricated skin flap, three in vitro cartilage-engineered constructs measuring 2×2×0.5 cm and one construct without cells, which served as the control, were implanted beneath an 8×15-cm random-pattern skin flap, neovascularized by implantation of an arteriovenous vascular pedicle with maximal blood flow. After 6 weeks, the neovascularized flaps with embedded cartilage-engineered constructs were completely removed based on the newly implanted vascular pedicle, and then freely retransferred into position using microsurgery. Macroscopic observation, histology, selective microangiography, and immunohistochemistry were performed to determine the construct vitality, neovascularization, and new cartilage formation.

Results

All neovascularized skin flaps with embedded tissue-engineered cartilage constructs were effectively free-transferred as free flaps. The implanted constructs were protected and well integrated within the flap. All constructs were well neovascularized and showed histologically stability in both form and size. Immunohistology showed the existence of cartilage-like tissue with extracellular matrix neosynthesis.

Conclusion

Our experimental study revealed the reliable ability of neovascularization and free microsurgical transplantation of cartilage-engineered constructs using prefabricated flaps. With respect to effective clinical application, engineered cartilage composed of a patient’s own cells can become a feasible option for the reconstruction of large cartilage defects or auricular reconstruction using this method. The procedure also represents a promising alternative for clinical practice due to minimal donor site morbidity and favorable aesthetic outcomes.

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Interessenkonflikt

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Authors and Affiliations

  1. Klinik für Hals-, Nasen- und Ohrenheilkunde, Universität Ulm, Frauensteige 12, 89075, Ulm, Deutschland

    A. Feucht

  2. Klinik für Hals-, Nasen- und Ohrenheilkunde, Klinikum rechts der Isar, Technische Universität, München, Deutschland

    C. Hoehnke, P.T. Hien, V. Mandlik, K. Storck & R. Staudenmaier

  3. Klinik für Orthopädie und Traumatologie, Abteilung für Hand- und Mikrochirurgie, Zentrales Universitätsklinikum 108, Hanoi, Vietnam

    N.T. Hoang

Authors
  1. A. Feucht
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  2. N.T. Hoang
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  3. C. Hoehnke
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  4. P.T. Hien
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  5. V. Mandlik
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  6. K. Storck
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  7. R. Staudenmaier
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Corresponding author

Correspondence to A. Feucht.

Additional information

Unsere mit dem APKO-Preis der Arbeitsgemeinschaft Plastische, rekonstruktive und ästhetische Kopf-Hals-Chirurgie ausgezeichnete Originalarbeit zu dieser Übersichtsarbeit und die meisten Abbildungen wurden bereits mit dem Titel „Neovascularisation and free mircosurgical transfer of cartilage engineered constructs“ in der Zeitschrift Microsurgery veröffentlicht: [7]. Der Abdruck erfolgt mit freundl. Genehmigung von John Wiley & Sons, Inc.

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Feucht, A., Hoang, N., Hoehnke, C. et al. Neovaskularisation und freier mikrochirurgischer Transfer von in vitro gezüchteten Knorpelkonstrukten. HNO 59, 239–247 (2011). https://doi.org/10.1007/s00106-011-2270-7

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  • DOI: https://doi.org/10.1007/s00106-011-2270-7

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