Platelet-derived growth factor (PDGF) gene delivery for application in periodontal tissue engineering.
BACKGROUND
A challenge in the reconstruction of periodontal structures is the targeted delivery of growth-promoting molecules to the tooth root surface. Polypeptide growth factors such as platelet-derived growth factor (PDGF) stimulate both cementogenesis and osteogenesis. Recent advances in gene therapy offer the advantage of delivering recombinant proteins to tissues for extended periods of time in vivo.


METHODS
Recombinant adenoviral vectors encoding for the PDGF-A gene were constructed to allow delivery of PDGF transgenes to cells. The recombinant adenoviruses were assembled using the viral backbone of Ad2/CMV/EGFP and replacing GFP (reporter gene encoding green fluorescent protein driven by the cytomegalovirus promoter [CMV] within adenovirus type 2) with the PDGF-A gene. Root lining cells (cloned cementoblasts) were transduced with Ad2/PDGF-A and evaluated for gene expression, DNA synthesis, and cell proliferation. PDGF-inducible genes, c-myc and osteopontin, were also evaluated following gene delivery of Ad2/PDGF-A.


RESULTS
The results revealed high level transduction of cementoblasts by gene transfer for 7 days as evidenced by flow cytometry and Northern blotting. Cementoblast DNA synthesis and subsequent proliferation were stimulated by Ad2/PDGF-A at levels equal to or greater than continuous rhPDGF-AA application. Strong message for the PDGF-A gene and protein as evidenced by Northern blotting and immunocytochemistry was noted. Furthermore, the potent induction of c-myc and osteopontin mRNA was found after PDGF gene delivery to cementoblasts.


CONCLUSIONS
These findings demonstrate that gene delivery of platelet-derived growth factor stimulates cementoblast activity that is sustained above that of rhPDGF-AA application. The use of gene therapy as a mode of growth factor delivery offers a novel approach to periodontal tissue engineering.
Giannobile
W V
WV
Department of Periodontics/Prevention/Geriatrics & Center for Biorestoration of Oral Health, The University of Michigan, Ann Arbor 48109-1078, USA. wgiannob@umich.edu
Lee
C S
CS
Tomala
M P
MP
Tejeda
K M
KM
Zhu
Z
Z
D000256:Adenoviridae / Q000235:genetics; D000469:Alkaline Phosphatase / Q000235:genetics; D000818:Animals; D015152:Blotting, Northern; D002455:Cell Division / Q000235:genetics; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000096:biosynthesis; D003739:Dental Cementum / Q000166:cytology / Q000502:physiology; D005434:Flow Cytometry; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D016259:Genes, myc / Q000235:genetics; D005818:Genetic Engineering; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D010012:Osteogenesis / Q000235:genetics; D053495:Osteopontin; D010510:Periodontal Diseases / Q000628:therapy*; D010982:Platelet-Derived Growth Factor / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012795:Sialoglycoproteins / Q000235:genetics; D013223:Statistics as Topic; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics
D008164:Luminescent Proteins; D010982:Platelet-Derived Growth Factor; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; D053495:Osteopontin; D049452:Green Fluorescent Proteins; D004247:DNA; D000469:Alkaline Phosphatase