Gene transfer of the Runx2 transcription factor enhances osteogenic activity of bone marrow stromal cells in vitro and in vivo
Gene transfer of the Runx2 transcription factor enhances osteogenic activity of bone marrow stromal cells in vitro and in vivo
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DOI：
10.1016/j.ymthe.2005.03.009
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发表时间：
2005-08-01
影响因子：
12.4
通讯作者：
Franceschi, RT
中科院分区：
文献类型：
--
作者：
Zhao, ZR;Zhao, M;Franceschi, RT
骨髓基质细胞（MSC）具有分化成多种间充质细胞类型的潜力。为了利用间充质干细胞进行骨再生的能力，必须开发方法将其选择性地分化为成骨细胞谱系。本研究的目的是检验利用离体 Runx2 基因转移增强 MSC 成骨活性的可行性。用编码 β-半乳糖苷酶或 Runx2 的腺病毒载体转导从 C57BL6 小鼠中分离的原代 MSC。用Ad-Runx2转导的细胞表达Runx2蛋白并经历成骨细胞分化，通过碱性磷酸酶活性和矿化的增加来测量。时程研究表明，Runx2 蛋白在转导后 1 天达到最高，并在 15 天时降至检测限以下。成骨细胞标志物 mRNA 表达与 Runx2 水平平行。相比之下，Runx2 依赖性矿化在实验期间持续存在。为了评估体内成骨活性，将 Ad-Runx2 转导的 MSC 和对照 MSC 吸附到两种不同的载体支架上，并皮下植入 C57BL6 小鼠体内。在这两种情况下，表达 Runx2 的 MSC 形成的骨量明显多于用对照病毒转导的细胞。综上所述，这些研究表明 Runx2 基因转移可能是增强 MSC 成骨潜力的有效途径。
Marrow stromal cells (MSCs) have the potential to differentiate into multiple mesenchymal cell types. To harness the power of MSCs for bone regeneration, methods must be developed to direct their differentiation selectively to the osteoblast lineage. The objective of this study was to examine the feasibility of using ex vivo Runx2 gene transfer to enhance the osteogenic activity of MSCs. Primary MSCs isolated from C57BL6 mice were transduced with adenoviral vectors encoding beta-galactosidase or Runx2. Cells transduced with Ad-Runx2 expressed Runx2 protein and underwent osteoblast differentiation as measured by increases in alkaline phosphatase activity and mineralization. Time-course studies revealed that Runx2 protein was highest 1 day after transduction and declined below the limits of detection by 15 days. Osteoblast marker mRNA expression paralleled Runx2 levels. In contrast, Runx2-dependent mineralization persisted for the duration of the experiment. To assess in vivo osteogenic activity, Ad-Runx2-transduced and control MSCs were adsorbed to two different carrier scaffolds and subcutaneously implanted into C57BL6 mice. In both cases, MSCs expressing Runx2 formed substantially more bone than cells transduced with control virus. Taken together, these studies indicate that Runx2 gene transfer may be an effective route to enhance the osteogenic potential of MSCs.