Efficient gene transfer to periodontal ligament cells and human gingival fibroblasts by adeno-associated virus vectors

Kunze, Melanie; Huber, Anke; Krajewski, Anna; Lowden, Esther; Schuhmann, Natascha; Buening, Hildegard; Hallek, Michael; Noack, Michael; Perabo, Luca

doi:10.1016/j.jdent.2009.03.001

« Previous Next »Journal of Dentistry
Volume 37, Issue 7 , Pages 502-508, July 2009

Efficient gene transfer to periodontal ligament cells and human gingival fibroblasts by adeno-associated virus vectors☆

Melanie Kunze
- Center of Dental Medicine, Department of Operative Dentistry and Periodontology, University of Cologne, Germany
Anke Huber
- Clinic I for Internal Medicine, University of Cologne, Kerpener Str. 62, 50937 Cologne, Germany
Anna Krajewski
- Center of Dental Medicine, Department of Operative Dentistry and Periodontology, University of Cologne, Germany
Esther Lowden
- Center of Dental Medicine, Department of Operative Dentistry and Periodontology, University of Cologne, Germany
Natascha Schuhmann
- Clinic I for Internal Medicine, University of Cologne, Kerpener Str. 62, 50937 Cologne, Germany
Hildegard Buening
- Clinic I for Internal Medicine, University of Cologne, Kerpener Str. 62, 50937 Cologne, Germany
- Center for Molecular Medicine, University of Cologne, Germany
Michael Hallek
- Clinic I for Internal Medicine, University of Cologne, Kerpener Str. 62, 50937 Cologne, Germany
- Center for Molecular Medicine, University of Cologne, Germany
Michael Noack
- Center of Dental Medicine, Department of Operative Dentistry and Periodontology, University of Cologne, Germany
Luca Perabo
- Clinic I for Internal Medicine, University of Cologne, Kerpener Str. 62, 50937 Cologne, Germany
- Corresponding author. Tel.: +49 221 478 4469; fax: +49 221 478 87941.

Received 17 January 2009; accepted 8 March 2009.

Abstract

Objectives

We explored for the first time the possibility to deliver a reporter gene (Green Fluorescence Protein) to human primary periodontal ligament (PDL) cells and human gingival fibroblasts (HGF) using shuttle vectors derived from adeno-associated virus (AAV). Since AAV transduction rates on other human primary fibroblasts have been previously shown to depend on the particular cell lineage and on the employed viral serotype, we determined the most effective AAV variant for periodontal cells comparing different vector types.

Methods

AAV serotypes 1–5 encoding GFP in single stranded (ss) and self-complementary (sc) vector genome conformations were used to infect primary HGF and PDL cells. Two days post-infection, the percentage of GFP expressing cells was determined by flow cytometry.

Results

Highest transduction rates for both cell types were achieved with self-complementary vectors derived from AAV-2, resulting in GFP expression in up to 86% of PDL cells and 50% of HGF. Transgene expression could be observed by optical microscopy for 2 months after infection. Lower but detectable rates were obtained with serotypes 1, 3 and 5.

Conclusions

The efficacy demonstrated here and the safety and versatility of AAV technology indicated in previous studies clearly suggest the potential of AAV vectors as tools for gene transfer to periodontal tissues.

Keywords: Gene transfer, Adeno-associated virus, Human gingival fibroblasts, Periodontal ligament cells