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Characterization of human chondrocytes exposed to simulated microgravity

机译：暴露于模拟微重力下的人类软骨细胞的表征

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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 (μg). Results: When cultured in simulated μg, 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 μg seem to change their extracellular matrix production behavior, while they rearrange their cytoskeletal proteins prior to forming 3D aggregates.
机译：背景：组织工程学是软骨再生的一种策略，但是软骨细胞3D生长所需的支架仍然是一个问题。方法：寻找改善无支架软骨工程的可能性的方法，我们表征了在随机定位机（RPM）上孵育的人类软骨细胞以模拟微重力（μg）。结果：当以模拟微克培养时，人类软骨细胞会在5天内开始形成3D细胞装配体。 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，Annexin V流式细胞仪显示1g培养物中18％的凋亡软骨细胞，而RPM仅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，两种凋亡率均未改变。 24小时后，模拟微重力也显着降低了I型和X型胶原，但没有改变IV型和层粘连蛋白，而II型胶原，硫酸软骨素和聚集蛋白聚糖则比1g对照升高。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被修饰。结论：暴露于模拟微克的软骨细胞似乎会改变其细胞外基质的生产行为，同时会在形成3D聚集体之前重新排列其细胞骨架蛋白。

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