Optimization of Methods for Articular Cartilage Surface Tissue Engineering: Cell Density and Transforming Growth Factor Beta Are Critical for Self-Assembly and Lubricin Secretion.
Optimization of Methods for Articular Cartilage Surface Tissue Engineering: Cell Density and Transforming Growth Factor Beta Are Critical for Self-Assembly and Lubricin Secretion.
复制标题
关节软骨表面组织工程方法的优化：细胞密度和转化生长因子β对于自组装和润滑素分泌至关重要。
DOI：
10.1089/ten.tec.2017.0121
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Reddi,AHari
中科院分区：
文献类型：
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作者：
Iwasa,Kenjiro;Reddi,AHari
目的：润滑素/浅表区蛋白(SZP)/蛋白多糖4(PRG4)在关节软骨边界润滑中起重要作用。滑膜表层软骨细胞和滑膜细胞分泌润滑素。本研究的具体目的是优化关节软骨表面组织工程的方法。本研究的目的是探讨细胞密度对浅层软骨细胞自组装和润滑素分泌的影响，作为功能评估。设计：浅层软骨细胞在低、中、高密度下作为单层培养。3种不同密度的软骨细胞经转化生长因子-β1处理后，每周或每天2次，免疫印迹法检测细胞培养上清液中的润滑量，酶联免疫吸附测定法测定细胞密度。结果：转化生长因子-β1能增加低密度和中密度软骨细胞的数量，而高密度细胞的细胞数量在每周2次的转化生长因子-β-1作用下明显减少，而高密度软骨细胞的细胞数量在每天2次的转化生长因子-β-1作用下保持不变。免疫印迹和EL ISA定量分析表明，转化生长因子-β1刺激不同密度的浅层软骨细胞分泌润滑素，每周2次转化生长因子-β处理。值得注意的是，与每周两次处理相比，每天处理转化生长因子-β1对润滑剂的增加要高得多。结论：在单层培养密度较高的情况下，每天处理对转化生长因子-β1的反应是最佳的。这些发现为关节软骨表面区软骨细胞的自组装在关节软骨表面组织工程中的应用奠定了基础。
Objective:Lubricin/superficial zone protein (SZP)/proteoglycan4 (PRG4) plays an important role in boundary lubrication in articular cartilage. Lubricin is secreted by superficial zone chondrocytes and synoviocytes of the synovium. The specific objective of this investigation is to optimize the methods for tissue engineering of articular cartilage surface. The aim of this study is to investigate the effect of cell density on the self-assembly of superficial zone chondrocytes and lubricin secretion as a functional assessment.Design:Superficial zone chondrocytes were cultivated as a monolayer at low, medium, and high densities. Chondrocytes at the three different densities were treated with transforming growth factor beta (TGF-β)1 twice a week or daily, and the accumulated lubricin in the culture medium was analyzed by immunoblots and quantitated by enzyme-linked immunosorbent assay (ELISA).Results:Cell numbers in low and medium densities were increased by TGF-β1; whereas cell numbers in high-density cell cultures were decreased by twice-a-week treatment of TGF-β1. On the other hand, the cell numbers were maintained by daily TGF-β treatment. Immunoblots and quantitation of lubricin by ELISA analysis indicated that TGF-β1 stimulated lubricin secretion by superficial zone chondrocytes at all densities with twice-a-week TGF-β treatment. It is noteworthy that the daily treatment of TGF-β1 increased lubricin much higher compared with twice-a-week treatment.Conclusions:These data demonstrate that daily treatment is optimal for the TGF-β1 response in a higher density of monolayer cultures. These findings have implications for self-assembly of surface zone chondrocytes of articular cartilage for application in tissue engineering of articular cartilage surface.