Institute of Advanced Biomedical Engineering and Science Tokyo Women's Medical University, Tokyo, Japan.
Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Kanagawa, Japan.
J Tissue Eng Regen Med. 2017 Oct;11(10):2885-2894. doi: 10.1002/term.2192. Epub 2016 Jul 12.
Endeavouring to repair and regenerate articular cartilage using cell sheets, we have previously established a co-culture system of chondrocytes and synoviocytes, and have reported the successful and rapid production of chondrocyte sheets. In the present study, to examine the effects of oxygen concentration on the chondrocyte sheets, we co-cultured human articular chondrocytes and human synoviocytes in 2%, 5% and 21% oxygen, and measured chondrocyte metabolic activity and proliferation activities under each condition for 14 days in culture. Layered chondrocyte sheets were also created under each condition and the proteoglycan (PG) level was compared with the gene expression of type I collagen (COL1), COL2, COL27, tissue metallopeptidase inhibitor 1 (TIMP1), fibronectin-1 (FN1), SRY-related HMG Box 9 (SOX9), aggrecan-1 (ACAN), integrin-α10 (ITGα10), matrix metalloproteinase 3 (MMP3), MMP13 and a disintegrin and metalloproteinase with thrombospondin motif 5 (ADAMTS5). Compared with 5% and 21% oxygen, the 2% condition caused significantly greater cell metabolic activity and proliferation (p < 0.05). The 2% condition produced a 10% greater PG level compared with 21% oxygen (p < 0.05). All conditions increased the expression of chondrocyte-specific genes, such as COL2, and were associated with low expression levels of catabolic factors, such as MMP3 and MMP13. These observations indicated that the specificity of the chondrocyte sheets was maintained under all conditions. The culture times did not differ between the 5% and 21% conditions. Compared with 21% oxygen, layered chondrocyte sheets rich in extracellular matrix were created 2.85 days earlier in 2% oxygen, which is similar to the level found in deep cartilage. Copyright © 2016 John Wiley & Sons, Ltd.
使用细胞片努力修复和再生关节软骨，我们之前建立了软骨细胞和滑膜细胞的共培养体系，并报告了成功快速产生软骨细胞片。在本研究中，为了研究氧浓度对软骨细胞片的影响，我们在 2%、5%和 21%氧中共同培养了人关节软骨细胞和人滑膜细胞，并在每种条件下培养 14 天测量了软骨细胞代谢活性和增殖活性。在每种条件下也创建了分层的软骨细胞片，并将蛋白聚糖 (PG) 水平与 I 型胶原 (COL1)、COL2、COL27、组织金属蛋白酶抑制剂 1 (TIMP1)、纤维连接蛋白-1 (FN1)、Sry 相关高迁移率族框 9 (SOX9)、聚集蛋白聚糖-1 (ACAN)、整合素-α10 (ITGα10)、基质金属蛋白酶 3 (MMP3)、MMP13 和解整合素金属蛋白酶与凝血酶 5 (ADAMTS5) 的基因表达进行比较。与 5%和 21%氧相比，2%条件下细胞代谢活性和增殖显著增加 (p < 0.05)。与 21%氧相比，2%条件下 PG 水平增加了 10% (p < 0.05)。所有条件均增加了软骨细胞特异性基因的表达，如 COL2，并与分解代谢因子如 MMP3 和 MMP13 的低表达水平相关。这些观察结果表明，在所有条件下均保持了软骨细胞片的特异性。5%和 21%条件下的培养时间没有差异。与 21%氧相比，在 2%氧中，富含细胞外基质的分层软骨细胞片早 2.85 天形成，与深层软骨中的水平相似。版权所有 © 2016 约翰威立父子公司
