Cbfa1/osf2 Transduced Bone Marrow Stromal Cells Facilitate Bone Formation In Vitro and In Vivo
Cbfa1/osf2 Transduced Bone Marrow Stromal Cells Facilitate Bone Formation In Vitro and In Vivo
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DOI：
10.1007/s00223-003-0004-x
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发表时间：
2004-02
影响因子：
4.2
通讯作者：
H. Zheng;Zhao-qing Guo;Qingjun Ma;Hongti Jia;Gengting Dang
中科院分区：
文献类型：
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作者：
H. Zheng;Zhao-qing Guo;Qingjun Ma;Hongti Jia;Gengting Dang
核心结合因子a-1/成骨细胞特异性因子2（cbfa1/osf2）是成骨细胞分化和功能的关键调节因子，但其是否能在体外和体内诱导骨形成尚不清楚。为研究cbfa1/osf2对骨形成的影响，我们用携带小鼠cbfa1/osf2基因的重组腺病毒载体转染原代培养的BALB/c小鼠骨髓基质细胞（MSCs）。我们发现Ad-cbfa1/osf2转染的MSCs产生cbfa1/osf2蛋白并向成骨样细胞分化。体外实验结果表明，转染后的MSCs碱性磷酸酶活性增强，骨钙素、骨桥蛋白和骨唾液酸蛋白表达增加，基质矿化增加。将Ad-cbfa1/osf2基因转染的MSCs移植于BALB/c小鼠颅骨缺损处，以I型胶原为支架材料，观察其体内成骨诱导作用。治疗组和对照组的缺损愈合情况在4周时进行大体和组织学检查。用Ad-cbfa1/osf2转导的MSC移植的颅骨缺损在四周时平均有85%的骨闭合。对照组中，缺损未处理（第1组），仅用胶原蛋白处理（第2组），或用胶原蛋白和未转导的MSC处理（第3组）显示很少或没有骨愈合。这些研究表明，cbfa1/osf2基因在体外和体内均能诱导成骨细胞分化和骨形成，提示cbfa1/osf2基因转导的MSCs可用于治疗骨缺损。
It has been well established that core binding factor a-1/osteoblast-specific factor-2 (cbfa1/osf2) is a key regulator of osteoblast differentiation and function, however, it is not known whether it can induce bone formationin vitroandin vivo. To investigate the effect of cbfa1/osf2 on bone formation, we used a recombinant adenoviral vector carrying the mouse cbfa1/osf2 gene to transduce primary cultured bone marrow stromal cells (MSCs) of BALB/c mice. We found that Ad-cbfa1/osf2-transduced MSCs produced cbfa1/osf2 protein and differentiated into osteoblast-like cells. The transduced MSCs had increased alkaline phosphatase activity, increased expression of osteocalcin, osteopontin and bone sialoprotein, and increased matrix mineralizationin vitro. To observe the induction of bone formationin vivo, MSCs transduced with Ad-cbfa1/osf2 were transplanted into a 5 mm diameter critical-sized skull defect in BALB/c mice, with type I collagen as scaffolding material. Healing of the defect in treatment and control groups was examined grossly and histologically at four weeks. Skull defects transplanted with Ad-cbfa1/osf2-transduced MSCs had an average of 85% osseous closure at four weeks. Control groups in which the defects were not treated (group 1), treated with collagen only (group 2), or treated with collagen and nontransduced MSCs (group 3) showed little or no osseous healing. These studies indicate that cbfa1/osf2 can induce osteoblast differentiation and bone formation bothin vitroandin vivo, suggesting that MSCs transduced with the cbfa1/osf2 gene may be useful in treating bone defects.