Role of bovine bone morphogenetic proteins in bone matrix protein and osteoblast-related gene expression during rat bone marrow stromal cell differentiation
Role of bovine bone morphogenetic proteins in bone matrix protein and osteoblast-related gene expression during rat bone marrow stromal cell differentiation
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DOI：
10.1097/01.scs.0000170449.72040.ee
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发表时间：
2005-11-01
影响因子：
0.9
通讯作者：
Clokie, CML
中科院分区：
文献类型：
--
作者：
Hu, ZM;Peel, SAF;Clokie, CML
已知骨形态发生蛋白（BMP）促进骨生成，目前正在进行临床试验，评估某些BMP促进骨移植和骨折愈合的能力。为探讨BMP诱导成骨的机制，将100 μ g牛BMP分别作用于大鼠骨髓基质细胞（MSCs）和C2 C12细胞系，诱导成骨分化14和28天。我们检测碱性磷酸酶（ALP）的测定，骨基质蛋白的免疫组化研究，骨基质蛋白的mRNA表达和成骨细胞相关的分析，通过逆转录聚合酶链反应。牛骨形态发生蛋白可使MSC培养液中ALP活性提高2 ~ 5倍（P < 0.05-0.001）。DNA和蛋白质含量在14天内增加。在成骨细胞分化过程中，BMP显著增加I型胶原、ALP、osterix、骨钙素、骨桥蛋白、血管内皮生长因子（VEGF）、血小板衍生生长因子（PDGF）-A和甲状旁腺激素受体的mRNA表达，并呈时间依赖性。骨涎蛋白（BSP）和3-磷酸甘油醛脱氢酶mRNA表达与对照组相比无明显增强。免疫组化结果还显示BMP在C2 C12分化过程中增加I型胶原、骨钙素、骨连接素、骨桥蛋白和BSP的免疫反应阳性。这些数据表明，BMP增强了我们刺激成骨样细胞分化的能力，并在体外成骨祖细胞分化过程中增加骨诱导性、骨基质蛋白形成和矿化、血管生成和软骨形成，而软骨形成的作用较弱。
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 ug 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.