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Experimentelle Untersuchung zur Oberflächenaktivierung von Implantaten durch liposomale Vektoren—eine Pilotstudie

Experimental pilot study on surface activation of implants with liposomal vectors

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Zusammenfassung

Hintergrund

Die Beschichtung von Titanimplantaten mit Mitogenen oder Morphogenen kann eine zeitlich verkürzte Einheilung und ein größeres Knochen-Implantat-Interface bedeuten [1, 5, 15]. Die Sicherheit des Einsatzes von Liposomen als DNA-Träger ist in der Literatur vielfach beschrieben [4, 13, 17]. Ziel der vorgestellten Pilotstudie war es, im Tiermodell Möglichkeiten der Oberflächenaktivierung von Implantaten und zur Behandlung periimplantärer Defekte durch Liposome zu evaluieren, die für BMP-2 („bone morphogenetic protein“) kodierende DNA enthielten.

Material und Methoden

Hierfür wurden neun 10×10 mm große Defekte im Os frontale des Hausschweins (n=3) angelegt und in diesen insgesamt 27 Implantate (3,5×14 mm) zentral so inseriert, dass der apikale Implantatbereich im direkten Knochen-Implantat-Kontakt stand. Somit war die Primärstabilität gewährleistet, während der zervikale Implantatbereich (obere 10 mm) den periimplantären Defekt simulierte. Zur Implantatlagerkonditionierung wurden Liposome angewandt, die BMP-2-DNA oder GFP-DNA („green fluorescence protein“) enthielten. Im ersten Versuchsarm wurden Liposomen mit GFP-DNA auf einer Kollagenmatrix direkt in den periimplantären Defekt appliziert. Im zweiten Versuchsarm erfolgte die Insertion von Implantaten, deren Oberfläche direkt mit BMP-2-DNA-haltigen Liposomen beschichtet war. Danach wurden die gewonnenen Proben der immunhistochemischen Aufarbeitung zum Nachweis von GFP und BMP-2 im Defektbereich und am Knochen-Implantat-Interface zugeführt.

Ergebnisse

Immunhistochemisch ließ sich zum Entnahmezeitpunkt am 3. postoperativen Tag im periimplantären Defekt eine erhöhte GFP-Expression erkennen, womit die Wirksamkeit des liposomalen Vektors für das gewählte Tiermodell nachgewiesen werden konnte. Am Knochen-Implantat-Interface der mit BMP-DNA beschichteten Implantate zeigte sich eine erhöhte BMP-2-Aktivität. So war eine In-vivo-Validierung des liposomalen Vektors auch für den Transfer von BMP-2-DNA im gewählten Versuchsmodell möglich. Daneben konnte ein System etabliert werden, mit dessen Hilfe BMP-2 kontinuierlich über einen größeren Zeitraum im Bereich der Implantatoberfläche freigesetzt werden kann.

Schlussfolgerung

Mithilfe dieser Pilotstudie gelang es, sowohl bezüglich des Einflusses der Implantatoberflächenaktivierung auf das Knochen-Implantat-Interface als auch zur Fragestellung der Behandlung periimplantärer Defekte Aussagen zu machen, die es ermöglichen, in einem laufenden Langzeitversuch entsprechend signifikante Daten zu gewinnen.

Abstract

Background

Surface coating with mitogenic or morphogenic proteins can improve the healing of bone adjacent to implants and increase the bone-implant interface [1, 5, 15]. Clinical surveys have shown liposome-mediated gene transfer to be a promising and safe new therapeutic method [4, 13, 17]. The aim of our study was to evaluate an experimental model of new approaches for topical treatment of the implant surface and of periimplant defects by using DNA liposomes encoding for BMP-2 (bone morphogenetic protein).

Material and methods

A total of 27 implants (3.5×14 mm) were placed in critically sized defects of the frontal skull bone of adult pigs (n=3). The bottom of the implant was placed in the base of the defect which guaranteed primary stability, whereas the superior part of the implant (10 mm) represented an implant in a defect area. Liposomes containing DNA encoding for BMP-2 and GFP (green fluorescence protein) were used. In a first trial GFP-DNA liposomes on a collagen matrix were directly applied to the periimplant defect. In a second stage, the surface of the implants was encoded with BMP-2 DNA liposomes. Subsequently, these implants were inserted in the manner described. The resulting bone samples were prepared for immunohistochemical staining. Staining for GFP was performed in the area of the defect and for BMP-2 on the bone-implant interface.

Results

Immunohistochemical staining on day 3 postoperatively revealed an increased GFP expression in the periimplant defect. Therefore, the effectiveness of the liposomal vector was verified for the chosen animal model. On the surface of the implants encoded with BMP-2 DNA liposomes an increased BMP-2 expression was found. Thus, the liposomal vector system was validated also for BMP-2 DNA transfer in the chosen animal model. Further, the established system allows a sustainable and delayed release of BMP-2 in the area of the bone-implant interface.

Conclusions

As a result of the study we were able to collect data concerning the influence of implant surface conditioning on the bone-implant interface and on therapeutically relevant options for the treatment of periimplant defects. These approaches are currently being evaluated in a long-term study.

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Authors and Affiliations

  1. Klinik und Poliklinik für Mund-, Kiefer-, Gesichtschirurgie, Friedrich-Alexander-Universität Erlangen-Nürnberg

    M. Thorwarth, K. A. Schlegel, J. Wiltfang, S. Rupprecht & J. H. Park

  2. Klinik und Poliklinik für Mund-, Kiefer-, Gesichtschirurgie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Glückstraße 11, 91054, Erlangen

    M. Thorwarth

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  1. M. Thorwarth
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Thorwarth, M., Schlegel, K.A., Wiltfang, J. et al. Experimentelle Untersuchung zur Oberflächenaktivierung von Implantaten durch liposomale Vektoren—eine Pilotstudie. Mund Kiefer GesichtsChir 8, 250–255 (2004). https://doi.org/10.1007/s10006-004-0536-x

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