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Z Orthop Ihre Grenzgeb 2005; 143(6): 669-676
DOI: 10.1055/s-2005-836744
Grundlagenforschung

© Georg Thieme Verlag Stuttgart · New York

Zelldifferenzierung einer humanen Knochenmarkzellkultur unter dem Einfluss von UHMW-PE Abriebpartikeln

Cell Differentiation of a Human Bone Marrow Cell Culture under the Influence of UHMW-PE DebrisS. Endres1 , 2 , I. Bartsch2 , M. Kratz2 , D. Jones2 , A. Wilke1 , 2
  • 1Klinik für Orthopädie und Rheumatologie der Philipps-Universität Marburg
  • 2Experimentelle Orthopädie und Biomechanik der Philipps-Universität Marburg
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Publication History

Publication Date:
28 December 2005 (online)

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Zusammenfassung

Studienziel: Ziel der vorliegenden Arbeit war es, den Einfluss von Partikeln des hochmolekulargewichtigen Polyethylens (UHMW-PE), wie es standardmäßig in der Endoprothetik verwendet wird, auf Veränderungen der zellulären Differenzierung in einem neuartigen in vitro Modell (direkter Zell-Partikel-Kontakt) zu untersuchen. Methode: UHMW-PE Partikel mit einer durchschnittlichen Größe von Ø ≤ 7,5 µm wurden in unterschiedlichen Konzentrationen (105, 106 und 107 Partikel pro Kulturschale) in einer Kollagen I-Suspension gelöst und in den Kulturschalen befestigt. Dadurch wurde ein direkter Kontakt der Zellen mit den UHMW-PE Abriebpartikeln gesichert. Im Anschluss wurden jeweils 3 × 106 humane Knochenmarkzellen pro Kulturschale auf die befestigten Partikel ausgesät und für 72 h kultiviert. Die Reaktion der Zellen auf die Partikel wurde lichtmikroskopisch, rasterelektronenmikroskopisch und anhand einer FACS-Analyse mit Kollagen befestigten Zellkulturen ohne Partikel verglichen. Ergebnisse: Die lichtmikroskopische Auswertung zeigte vor allem eine Einscheidung der Partikel, die sich zu großen Konglomeraten (Ø 7,5 µm) zusammen gelagert hatten und weniger phagozytierte Partikel, welches Rasterelektronenmikroskopisch bestätigt wurde. Die Ergebnisse der FACS-Analyse ergaben signifikante Unterschiede im Fall der CD3/CD4-positiven, CD14-positiven und CD19-positiven Zellen (p < 0,05). Eine signifikante Zunahme der CD3/CD4-positiven and CD14-positiven Zellen (p < 0,05) konnte nach 72 h Kulturdauer beobachtet werden, wohingegen eine signifikante Abnahme der CD19-positiven Zellen nachweisbar war. Schlussfolgerung: Die Ergebnisse der vorliegenden Studien belegen, dass die partikel-induzierte Abwehrreaktion durch UHMW-PE sich nicht nur auf den Partikel-Makrophagen-Kontakt beschränkt, sondern auch die Zelldifferenzierung im Knochenmark beeinflussen kann. Des Weiteren bekräftigen die Ergebnisse, dass die vorliegende Methode nützlich zur In-vitro-Analyse von UHMW-PE Abriebpartikeln mit direkten Partikel-Zell-Kontakt ist. Obwohl die Partikel hauptsächlich zu Konglomeraten zusammenlagert waren, konnte gezeigt werden, dass eine Änderung der immunkompetenten Zellen hervorgerufen wurde.

Abstract

Aim: The purpose of the present study was to evaluate the influence of ultra-high-molecular-weight polyethylene (UHMW-PE), which is the major constituent of the material debris formed as a result of orthopaedic implant wear, on the cellular differentiation in a modified in vitro model. Methods: UHMW-PE particles (Ø ≤ 7,5 µm) were suspended in soluble collagen type I and subsequently solidified in different concentrations (105, 106 and 107 particles per well) on the bottom of the wells. Human bone marrow cells in a concentration of 3 × 106 cells per well were seeded on the collagen-particle substrate and maintained for up to 72 h. The response of the cells to the particles was examined by light microscopy, scanning electron microscopy and FACS analysis compared to cells on control collagen surfaces without any particles. Results: Light and scanning microscopic evaluation revealed that the UHMW-PE particles, which had built large conglomerates (Ø 7.5 µm), were mainly surrounded by the cells and less phagocytosed. The results of the FACS analysis revealed significant differences in CD3/CD4 positive, CD14 positive and CD19 positive cells (p < 0.05). A significant elevation of CD3/CD4 positive and CD14 positive cells (p < 0.05) was observed after the period of culture (72 h) whereas a significant decrease could be detected in the case of CD19 positive cells. Conclusion: The results demonstrate that the particle-induced response by UHMW-PE limits itself not only to the particle macrophage contact but influences also the differentiation of the bone marrow. Moreover, the results confirm that the present method is useful to evaluate the in vitro effects of UHMW-PE wear particles with direct particle cell contact. Although the particles built large conglomerates, it could be shown that a change of the immune-competent cells also occurred.

Schlüsselwörter

UHMW-PE - Abrieb - Zelldifferenzierung - direkter Zellkontakt

Key words

UHMW-PE - wear debris - cell differentiation - direct cell contact

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Dr. med. S. Endres

Philipps-Universität Marburg

Baldingerstrasse

35033 Marburg

Deutschland

Phone: 00 49/64 21/2 86 36 91

Email: endres@med.uni-marburg.de