Sonic hedgehog promotes chondrogenesis of rabbit bone marrow stem cells in a rotary cell culture system
Sonic hedgehog promotes chondrogenesis of rabbit bone marrow stem cells in a rotary cell culture system
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声波刺猬促进旋转细胞培养系统中兔骨髓干细胞的软骨形成
DOI：
10.1186/s12861-019-0198-4
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发表时间：
2019-08-12
影响因子：
--
通讯作者：
Wu, Xing
中科院分区：
文献类型：
--
作者：
Chen, Liyang;Liu, Gejun;Wu, Xing
背景Sonic hedgehog（Shh）是一种重要的信号蛋白，参与软骨细胞早期分化的诱导.在此，我们证明，Shh显着诱导软骨形成的兔骨髓基质细胞（BMSCs）在微重力条件下，并促进软骨regeneration.ResultsIn旋转细胞培养系统（RCCS），软骨分化揭示了较强的甲苯胺蓝和II型胶原免疫组化染色的Shh转染组，和软骨诱导活性的Shh相当于TGF-β。Western blotting和qRT-PCR分析结果证实了微重力环境中Shh或TGF-β处理的兔BMSCs中Sox 9、聚集蛋白聚糖（ACAN）和胶原II的表达增强。在所有组中均检测到低水平的软骨形成肥大、成骨和脂肪形成相关因子。体内移植后，组织学分析显示Shh transfection group.ConclusionsThese结果提示，Shh信号在微重力条件下促进兔BMSCs软骨形成，相当于TGF-β，并改善软骨和软骨下缺损修复的早期阶段。此外，RCCS提供了一个动态的培养微环境，有利于细胞增殖，聚集和分化。
BackgroundSonic hedgehog (Shh) is an important signalling protein involved in the induction of early cartilaginous differentiation. Herein, we demonstrate that Shh markedly induces chondrogenesis of rabbit bone marrow stromal cells (BMSCs) under microgravity conditions, and promotes cartilage regeneration.ResultsIn the rotary cell culture system (RCCS), chondrogenic differentiation was revealed by stronger Toluidine Blue and collagen II immunohistochemical staining in the Shh transfection group, and chondroinductive activity of Shh was equivalent to that of TGF-β. Western blotting and qRT-PCR analysis results verified the stronger expression of Sox9, aggrecan (ACAN), and collagen II in rabbit BMSCs treated with Shh or TGF-β in a microgravity environment. Low levels of chondrogenic hypertrophy, osteogenesis, and adipogenesis-related factors were detected in all groups. After transplantation in vivo, histological analysis revealed a significant improvement in cartilage and subchondral repair in the Shh transfection group.ConclusionsThese results suggested that Shh signalling promoted chondrogenesis in rabbit BMSCs under microgravity conditions equivalent to TGF-β, and improved the early stages of the repair of cartilage and subchondral defects. Furthermore, RCCS provided a dynamic culture microenvironment conducive for cell proliferation, aggregation and differentiation.