Characterization of Human Chondrocytes Exposed to Simulated Microgravity
Characterization of Human Chondrocytes Exposed to Simulated Microgravity
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DOI：
10.1159/000303059
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发表时间：
2010-01-01
影响因子：
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通讯作者：
Grimm, Daniela
中科院分区：
文献类型：
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作者：
Ulbrich, Claudia;Westphal, Kriss;Grimm, Daniela
背景：组织工程是一种软骨再生策略，但软骨细胞 3D 生长所需的支架仍然是一个问题。方法：为了寻找改进软骨无支架工程的可能性，我们对在随机定位机（RPM）上孵育以模拟微重力（μg）的人类软骨细胞进行了表征。结果：在模拟 μg 中培养时，人类软骨细胞在 5 天内开始形成 3D 细胞组件。 24小时后，我们无法在这些细胞中检测到caspase-3、Fas、p53或Bcl-2蛋白，然而，Annexin V流式细胞术显示1g培养物中有18%的软骨细胞凋亡，但在RPM上只有10%。当添加血管内皮生长因子（VEGF）或碱性成纤维细胞生长因子（bFGF）时，两种细胞凋亡率都没有改变。 24小时，模拟微重力也显着降低了I型和X型胶原蛋白，但没有改变IV型胶原蛋白和层粘连蛋白，而II型胶原蛋白、硫酸软骨素和聚集蛋白聚糖与1g对照相比有所升高。当应用外部 bFGF 或 VEGF 时，II/X 型胶原蛋白、硫酸软骨素和聚集蛋白聚糖的产生会发生变化。结论：暴露于模拟 μg 的软骨细胞似乎改变了它们的细胞外基质生产行为，同时它们在形成 3D 聚集体之前重新排列了它们的细胞骨架蛋白。版权所有 (C) 2010 S. Karger AG，巴塞尔
Background: Tissue engineering is a strategy of cartilage regeneration, but scaffolds, required for 3D growth of chondrocytes, are still a problem. Methods: Searching for possibilities to improve scaffold-free engineering of cartilage, we characterized human chondrocytes incubated on a random positioning machine (RPM) to simulate microgravity (mu g). Results: When cultured in simulated mu g, human chondrocytes start forming 3D cell assemblies within 5 days. After 24h, we could not detect caspase-3, Fas, p53 or Bcl-2 proteins in these cells, Annexin V flow cytometry, however, revealed 18% of apoptotic chondrocytes in 1g cultures but only 10% on the RPM. Both rates of apoptosis were not changed, when vascular endothelial growth factor (VEGF) or basic fibroblast growth factor (bFGF) was added. 24 h, simulated microgravity also had significantly decreased collagen type I and X, but did not change collagen type IV and laminin, while collagen type II, chondroitin sulfate and aggrecan were elevated as compared with 1g controls. The production of collagen type II/X, chondroitin sulfate and aggrecan was modified, when external bFGF or VEGF had been applied. Conclusion: Chondrocytes exposed to simulated mu g seem to change their extracellular matrix production behavior, while they rearrange their cytoskeletal proteins prior to forming 3D aggregates. Copyright (C) 2010 S. Karger AG, Basel