Osterix overexpression in mesenchymal stem cells stimulates healing of critical-sized defects in murine calvarial bone.

Qisheng Tu, Paloma Valverde, Shu Li, Jin Zhang, Pishan Yang, Jake Chen
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引用次数: 103

Abstract

Osterix (Osx) is a zinc-finger-containing transcription factor that is expressed in osteoblasts of all endochondral and membranous bones. In Osx null mice, osteoblast differentiation is impaired, and bone formation is absent. We hypothesized that overexpression of Osx in bone marrow-derived mesenchymal stem cells (BMSCs) would enhance osteogenic differentiation during bone regeneration in vivo. Overexpression of Osx in mouse BMSCs was achieved using retroviral infection together with a green fluorescent protein (GFP) vector to monitor transduction efficiency and determine the source of regenerative cells in implantation studies. Bone regeneration in vivo was evaluated by implanting BMSCs overexpressing Osx into 4-mm calvarial bone defects in adult mice using type I collagen sponge as a carrier. New bone formation in the defects was quantified using radiological and histological procedures 5 weeks after implantation. The results showed that implantation of Osx-transduced BMSCs resulted in 85% healing of calvarial bone defects as detected using radiological analyses. Histological examination of the implants demonstrated that the Osx-transduced group exhibited amounts of newly formed bone that was five times as high as in a group transduced with the empty vector. Immunohistochemistry for GFP showed positive immunoreaction localized to areas of newly engineered bone in the Osx-transduced group. Immunohistochemistry with antibodies against the extracellular matrix protein bone sialoprotein resulted in strong staining in areas of new bone formation. In addition, the clonal BMSCs showed an osteogenic potential similar to that of primary cultures of BMSCs, suggesting the usefulness of this model in bone tissue engineering. These results indicate that ex vivo gene therapy of Osx is a useful therapeutic approach in regenerating adult bone tissue.

间充质干细胞中Osterix的过度表达刺激小鼠颅骨临界尺寸缺陷的愈合。
Osterix (Osx)是一种含锌指的转录因子,在所有软骨内骨和膜性骨的成骨细胞中表达。在Osx缺失小鼠中,成骨细胞分化受损,骨形成缺失。我们假设Osx在骨髓间充质干细胞(BMSCs)中的过度表达会在体内骨再生过程中增强成骨分化。在移植研究中,利用逆转录病毒感染和绿色荧光蛋白(GFP)载体实现了Osx在小鼠骨髓间充质干细胞中的过表达,以监测转导效率并确定再生细胞的来源。以I型胶原海绵为载体,将过表达Osx的骨髓间充质干细胞植入成年小鼠4 mm颅骨骨缺损中,观察骨再生情况。植入后5周采用放射学和组织学方法量化缺损的新骨形成。结果显示osx转导的骨髓间充质干细胞植入后,颅骨骨缺损的愈合率为85%。植入物的组织学检查表明,osx转导组的新骨形成量是空载体转导组的五倍。在osx转导组中,GFP免疫组化显示新工程骨区域的阳性免疫反应。免疫组化抗体对抗细胞外基质蛋白骨唾液蛋白导致新骨形成区域的强染色。此外,克隆骨髓间充质干细胞显示出与原代培养的骨髓间充质干细胞相似的成骨潜能,表明该模型在骨组织工程中的实用性。这些结果表明Osx离体基因治疗是一种有效的成人骨组织再生治疗方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Tissue engineering
Tissue engineering CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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