Modulation of the Effect of Transforming Growth Factor-β3 by Low-Intensity Pulsed Ultrasound on Scaffold-Free Dedifferentiated Articular Bovine Chondrocyte Tissues.
Modulation of the Effect of Transforming Growth Factor-β3 by Low-Intensity Pulsed Ultrasound on Scaffold-Free Dedifferentiated Articular Bovine Chondrocyte Tissues.
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DOI：
10.1089/ten.tec.2014.0428
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发表时间：
2015-04
期刊：
影响因子：
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通讯作者：
Stephanie Yin Wai Ting;K. Montagne;Y. Nishimura;T. Ushida;K. Furukawa
中科院分区：
文献类型：
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作者：
Stephanie Yin Wai Ting;K. Montagne;Y. Nishimura;T. Ushida;K. Furukawa
本研究的目的是评估低强度脉冲超声（LIPUS）如何调节转化生长因子-β3（TGF-β3）对无支架去分化牛关节软骨细胞组织向软骨样表型分化的影响。具体而言，分析了这些刺激对无支架模型的肥大标志物I型胶原蛋白、X型胶原蛋白和软骨降解胶原酶基因表达的影响。将LIPUS直接从换能器通过硅凝胶施加到含有组织的六孔板的生物反应器允许进行简单、无菌和大规模的实验。在10天的时间内，在有或没有TGF-β3（10 ng/mL）的情况下，对组织进行LIPUS，功率为55 mW/cm（2），频率为200 μs，频率为1 MHz。暴露于TGF-β3的组织具有显著增加的糖胺聚糖和总胶原蛋白产生沿着上调的软骨特异性基因表达，导致具有较高杨氏模量的组织。然而，这些组织还上调了肥大标志物I型胶原、X型胶原、MMP-1、MMP-13、MMP-2的基因表达，并且还增加了p38的磷酸化。这些基质降解酶的表达通过肥大发育进行补救，并使去分化的牛关节软骨细胞向软骨形成谱系分化，使其成为软骨组织工程中有价值的工具。
The aim of this study was to evaluate how low-intensity pulsed ultrasound (LIPUS) modulates the effect of transforming growth factor-β3 (TGF-β3) on the differentiation of scaffold-free dedifferentiated bovine articular chondrocyte tissues toward a cartilage-like phenotype. Specifically, the effect of these stimuli on the expression of hypertrophic markers collagen type I, collagen type X, and cartilage-degrading collagenase gene expression for a scaffold-free model was analyzed. A bioreactor that applied LIPUS directly from the transducer through a silicone gel to a six-well plate containing the tissues allowed simple, sterile, and large-scale experiments. Tissues were subjected to LIPUS of 55 mW/cm(2) in a 200 μs burst sine wave of 1 MHz over a 10-day period with or without TGF-β3 (10 ng/mL). Tissues exposed to TGF-β3 had significantly increased glycosaminoglycan and total collagen protein production along with upregulated cartilage-specific gene expression, resulting in tissues with a higher Young's Modulus. However, these tissues had also upregulated gene expression for hypertrophic markers collagen type I, collagen type X, MMP-1, MMP-13, MMP-2, and also an increase in the phosphorylation of p38. The expression of these matrix-degrading enzymes was remediated by hypertrophic development and differentiate dedifferentiated bovine articular chondrocytes towards a chondrogenic lineage allowing it to be a valuable tool in cartilage tissue engineering.