Human mesenchymal stem cells transduced with recombinant bone morphogenetic protein-9 adenovirus promote osteogenesis in rodents.
The present study was undertaken to determine whether ex vivo bone morphogenetic protein-9 (BMP-9) gene therapy using human mesenchymal stem cells (hMSCs) can induce endochondral bone formation in athymic nude rats. An in vitro study was initially performed on hMSCs to evaluate morphological changes and osteoblastic differentiation induced by replication-defective adenovirus type 5 with the cytomegalovirus promoter and either the BMP-9 (Ad-BMP-9) or beta-galactosidase (Ad-beta-gal) gene. In vivo, athymic nude rats received an injection (10(6) hMSCs transduced with recombinant adenovirus at 50 PFU/cell) into the anterior thigh musculature: Ad-BMP-9 on the left and Ad-beta-gal (control) on the right. Computed tomography scans and histological analysis were obtained 7, 14, 28, 42, 56, and 84 days postinjection. In vitro, human mesenchymal stem cells treated with Ad-BMP-9 (50 PFU/cell) showed signs of differentiation, whereas hMSCs treated with 250 and 1250 PFU/cell showed cytotoxicity. In vivo, computed tomography and histological analysis clearly demonstrated ectopic bone at hMSC/Ad-BMP-9 treatment sites, whereas the hMSC/Ad-beta-gal treatment sites showed no evidence of osteogenesis. None of the animals showed clinical evidence of toxicity. Ex vivo gene therapy with hMSC/BMP-9 may be efficacious for promoting bone formation for a variety of bone pathologies and certainly warrants further investigations.
Department of Neurological Surgery, University of Virginia, Charlottesville, Virginia 22908, USA.
Hayan
H
Dayoub
Randall J
RJ
Dumont
Jin Zhong
JZ
Li
Aaron S
AS
Dumont
Gerald R
GR
Hankins
David F
DF
Kallmes
Gregory A
GA
Helm
D000260:Adenoviruses, Human / Q000235:genetics; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000235:genetics / Q000502:physiology*; D002454:Cell Differentiation; D003587:Cytomegalovirus / Q000235:genetics; D003673:Defective Viruses / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D055427:Growth Differentiation Factor 2; D055412:Growth Differentiation Factors; D006801:Humans; D007527:Isoenzymes / Q000032:analysis; D007763:Lac Operon; D008648:Mesoderm / Q000166:cytology*; D010006:Osteoblasts / Q000166:cytology / Q000201:enzymology; D010012:Osteogenesis; D011401:Promoter Regions, Genetic; D051381:Rats; D011923:Rats, Nude; D011993:Recombinant Fusion Proteins / Q000502:physiology; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000502:physiology*; D014161:Transduction, Genetic; D014183:Transplantation, Heterologous
D019485:Bone Morphogenetic Proteins; C525887:GDF2 protein, human; D055427:Growth Differentiation Factor 2; D055412:Growth Differentiation Factors; D007527:Isoenzymes; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase