Role of bovine bone morphogenetic proteins in bone matrix protein and osteoblast-related gene expression during rat bone marrow stromal cell differentiation.
Bone morphogenetic proteins (BMPs) are known to promote osteogenesis, and clinical trials are currently underway evaluating the ability of certain BMPs to promote bone graft and fracture healing. To observe the mechanism of osteoinductive and bone formation, 100 microg of bovine BMP was tested during osteogenic differentiation of rat bone marrow stromal cells (MSCs) and C2C12 line culture for 14 and 28 days. We examined alkaline phosphatase (ALP) by assay, immunohistochemical studies for bone matrix proteins, and mRNA expression of bone matrix proteins and osteoblast-related analysis by reverse-transcription polymerase chain reaction. ALP activity in MSC cultures was elevated by bovine BMP by two to fivefold (P < 0.05-0.001). DNA and protein content increased over 14 days. BMP significantly increased the mRNA expression of type I collagen, ALP, osterix, osteocalcin, osteopontin, vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF)-A, and parathyroid hormone receptor time dependently during the osteoblastic differentiation. There was no markedly enhanced mRNA expression of bone sialoprotein (BSP) and glyceraldehyde-3-phosphate dehydrogenase compared with that of control. Immunohistochemical results also showed BMP increased immunoreactive positivity of type I collagen, osteocalcin, osteonectin, osteopontin, and BSP during the C2C12 differentiation. These data indicated that BMP enhances our ability to stimulate the differentiation of osteoblast-like cells and increases osteoinductivity, bone matrix protein formation and mineralization, angiogenesis, and chondrogenesis during osteoblast progenitor cell differentiation in vitro and that the role of chondrogenic is weak.
Hu
ZhenMing
Z
Orthobiologics Laboratory, Discipline of Oral and Maxillofacial Surgery, Faculty of Dentistry, University of Toronto, Toronto, ON, Canada. zminghu@hotmail.com
Peel
Sean A F
SA
Ho
Stephen K C
SK
Sándor
George K B
GK
Clokie
Cameron M L
CM
D000469:Alkaline Phosphatase / Q000187:drug effects; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects*; D001857:Bone Matrix / Q000737:chemistry*; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002478:Cells, Cultured; D024042:Collagen Type I / Q000187:drug effects; D004247:DNA / Q000187:drug effects; D008297:Male; D010006:Osteoblasts / Q000378:metabolism*; D015675:Osteocalcin / Q000187:drug effects; D010012:Osteogenesis / Q000187:drug effects; D053495:Osteopontin; D010750:Phosphoproteins / Q000187:drug effects; D010982:Platelet-Derived Growth Factor / Q000187:drug effects; D051381:Rats; D011917:Rats, Inbred Lew; D012795:Sialoglycoproteins / Q000187:drug effects; D017154:Stromal Cells / Q000187:drug effects; D014157:Transcription Factors / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000187:drug effects
D019485:Bone Morphogenetic Proteins; D024042:Collagen Type I; D010750:Phosphoproteins; D010982:Platelet-Derived Growth Factor; D012795:Sialoglycoproteins; C495181:Sp7 protein, rat; C506897:Spp1 protein, rat; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; C075278:platelet-derived growth factor A; D015675:Osteocalcin; D053495:Osteopontin; D004247:DNA; D000469:Alkaline Phosphatase