Morphologic analysis of BMP-9 gene therapy-induced osteogenesis.
The present study was performed to determine the histological, ultrastructural, and radiographic changes that occur over time at intramuscular BMP-9 gene therapy treatment sites. Several members of the bone morphogenetic protein (BMP) family have the potential to induce osteochondrogenesis when the protein is delivered to rodents, canines, rabbits, and nonhuman primates. Previous studies have also demonstrated that BMP gene therapy utilizing adenoviral vectors can also stimulate orthotopic and heterotopic bone formation in rodents and rabbits. Athymic nude and Sprague-Dawley rats were injected with Ad-BMP-9 or Ad-beta-Gal (3.75 x 10(9) particles) in their thigh musculature and light microscopic, electron microscopic, and computerized tomography analysis was performed 3, 6, 9, 12, 15, 18, 21, and 100 days later. To assess early mesenchymal cell proliferation, a bromodeoxyuridine (BrdU) immunohistochemical analysis was also performed 48, 60, and 72 hr postinjection in athymic nude rats. All animals demonstrated extensive endochondral bone formation at the Ad-BMP-9 treatment sites within 3 weeks. The Sprague-Dawley rats also exhibited a massive, acute inflammatory infiltrate during the first week. Proliferating mesenchymal stem cells were clearly evident as early as 2 days after treatment, which differentiated into small or hypertrophied chondrocytes during the next week. During the third week, the cartilaginous matrix mineralized and formed woven bone, which converted to lamellar bone by 3 months. No evidence of bone formation was demonstrated at the Ad-beta-Gal injection sites in the athymic nude or Sprague-Dawley rats. In addition, no cellular proliferation was seen at the Ad-beta-Gal treatment sites in the athymic nude animals as assessed by light microscopy and BrdU immunohistochemistry. The extensive bone formation induced by Ad-BMP-9 suggests that BMP gene therapy may have potential utility in the treatment of degenerative, rheumatic, or traumatic bone pathology.
Varady
P
P
Department of Neurosurgery, Health Sciences Center, Charlottesville, VA 22908, USA.
Li
J Z
JZ
Cunningham
M
M
Beres
E J
EJ
Das
S
S
Engh
J
J
Alden
T D
TD
Pittman
D D
DD
Kerns
K M
KM
Kallmes
D F
DF
Helm
G A
GA
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D019485:Bone Morphogenetic Proteins / Q000235:genetics*; D001842:Bone and Bones / Q000000981:diagnostic imaging / Q000648:ultrastructure*; D001973:Bromodeoxyuridine; D002460:Cell Line; D019902:Chondrocytes / Q000648:ultrastructure; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D055427:Growth Differentiation Factor 2; D007124:Immunoenzyme Techniques; D008854:Microscopy, Electron; D010012:Osteogenesis / Q000235:genetics* / Q000502:physiology*; D051381:Rats; D011923:Rats, Nude; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014057:Tomography, X-Ray Computed
D019485:Bone Morphogenetic Proteins; D017931:DNA Primers; D055427:Growth Differentiation Factor 2; D001973:Bromodeoxyuridine