Fibrinmatrix erhöht Adhärenz von regenerativen peripheren Nervenzellen

Zusammenfassung

Zur Schaffung eines artifiziellen Nervenersatzes ist es optimal, ein Konduit aus resorbierbarem Material zu verwenden, welches mit axonstimulierenden Zellen wie Schwann-Zellen oder modifizierbaren Stammzellen besiedelt ist. Idealerweise beinhaltet die Beschichtung eines Poly-3-hydroxybutyrat (PHB)-Konduits mit Zellen zur Aktivierung der Axonsprossung 1. eine homogene Verteilung der transplantierten Zellen, 2. ein Milieu, das das Überleben der Zellen garantiert, sowie 3. eine Matrix, die sich rasch resorbiert, um die Axonregeneration nicht zu blockieren. Die vorliegende In-vitro-Arbeit zeigt eine neue Methode zur Beschichtung von Konduitmaterial in Form von Matten mit Stamm- und Schwann-Zellen mit verdünntem, resorbierbaren Fibrinkleber Tisseel®. Anhand eines Quantifikationsmodells und Immunohistochemie (S100, DAPI) wird gezeigt, dass undifferenzierte (uMSC) und differenzierte (dMSC) mesenchymale Stammzellen (Schwann-Zell-Analogas) sowie Schwann-Zellen (SC) der Sprague-Dawley^©-Ratte in verdünnter Fibrinmatrix signifikant besser applizierbar sind und in Position gehalten werden als die Vergleichsgruppe ohne Matrixapplikation. Die Analyse der Gruppen mit der Fibrinmatrix ergab hoch signifikant bessere Werte (p < 0,001) für die Zelladhärenz auf den Matten sowie signifikant bessere Werte (p < 0,05) für den Zellverlust.

Abstract

Optimal seeding of a nerve conduit with cells is a core problem in tissue engineering of constructing an artificial nerve substitute to gap lesions in the peripheral nerve system. An ideal nerve gap substitute would have to present an equally distributed number of cells that can activate the regrowing axons. This work shows a new in vitro technique of two-step seeding of cells inside a conduit and on layered mats that allows a valuable targeting of the cells and a proven survival in the environment of poly-3-hydroxybutyrate (PHB) conduits. The technique uses two components of diluted fibrin glue Tisseel®. Initially, the chosen area on the mat was coated with thrombin followed from the seeding of a fibrinogen-cell compound. Using Sprague Dawley® rat cells, we could demonstrate with immunohistochemistry (S100, DAPI) techniques that undifferentiated (uMSC) and Schwann cells (SC) mimicking differentiated mesenchymal stem cells (dMSC) as well as SC can be suspended and targeted significantly better in dissolvable diluted fibrin glue than in growth medium. Analysis showed significantly better values for adherence (p < 0.001) and drop off (p < 0.05) from seeded cells. Using this two-step application allows the seeding of the cells to be more precise and simplifies the handling of cell transplantation.

Schlüsselwörter

Chirurgie der peripheren Nerven - Nervenregeneration - Tissue Engineering - Transplantation experimentell

Key words

peripheral nerve surgery - fibrin glue - Tisseel® - PHB conduit

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