Recombinant AAV-mediated VEGF gene therapy induces mandibular condylar growth.
Craniofacial anomalies resulting from impaired growth of mandibular condyles require multidisciplinary interventions, which impose a substantial burden on patients and their families. So far, correcting such deformities with an alternative strategy - gene therapy - is still an uncharted territory. Here, we established an effective in vivo gene delivery system with recombinant adeno-associated virus (rAAV)-mediated vascular endothelial growth factor (VEGF) to enhance mandibular condylar growth. With in situ hybridization, RT-PCR, immunostaining and Western blot, transgene expression was clearly detected in the mandibular condyles during the whole experiment periods. At defined time points, specific osteogenetic markers (alkaline phosphatase and osteocalcin) and chondrogenetic markers (collagen type II and collagen type X) were assessed by means of biochemical analysis and their expression significantly changed from day 30. Proliferation index by proliferating cell nuclear antigen staining showed also a significant increase in cell proliferation. Morphological measurement identified that the size of mandibular condyle significantly increased from day 30. Taken together, rAAV-VEGF was successfully established as an efficient delivery system to induce mandibular condylar growth, which provides the basis for future gene therapy to treat patients with craniofacial deformities.
Rabie
A B M
AB
Biomedical and Tissue Engineering Group, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China. rabie@hkusua.hku.hk
Dai
J
J
Xu
R
R
D000818:Animals; D015153:Blotting, Western; D049109:Cell Proliferation; D019902:Chondrocytes / Q000166:cytology; D000229:Dependovirus / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008335:Mandibular Condyle / Q000254:growth & development* / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology; D008856:Microscopy, Fluorescence; D010010:Osteoclasts / Q000166:cytology; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis / Q000235:genetics*
D042461:Vascular Endothelial Growth Factor A; D049452:Green Fluorescent Proteins