Canadian Institutes of Health Research Bioengineering of Skeletal Tissues Team, Department of Pathology and Laboratory Medicine, Mount Sinai Hospital and University of Toronto, Toronto, Canada.
Tissue Eng Part A. 2009 Dec;15(12):3789-98. doi: 10.1089/ten.TEA.2008.0530.
The inferior biomechanical properties of in vitro-formed tissue remain a significant obstacle in bioengineering articular cartilage tissue. We have previously shown that cyclic compression (30 minutes, 1 kPa, 1 Hz) of chondrocytes isolated from full-thickness cartilage can induce greater matrix synthesis, although articular cartilage is composed of different subpopulations of chondrocytes, and their individual contribution to enhanced tissue formation has not been fully characterized. This study examines the contribution of chondrocyte subpopulations to this response. Bovine articular chondrocytes were isolated from superficial to mid zones (SMZs) or deep zones (DZs), placed in three-dimensional culture, and subjected to cyclic compression. DZ chondrocytes on calcium polyphosphate substrates formed thicker tissue than those from SMZs. Compression increased matrix accumulation in SMZ chondrocytes while decreasing accumulation in DZ chondrocytes. The SMZ and DZ chondrocytes also differed in their type 1 membrane-bound matrix metalloproteinase (MMP) and MMP-13 expression, enzymes that play a crucial role in mediating the response to mechanical stimulation. In addition, the duration of the culture period was important in determining the DZ response, raising the possibility that matrix accumulation plays a role in the response to stimulation. Understanding the cellular response to mechanical stimulation during tissue formation will facilitate our understanding of tissue growth and allow for further optimization of cartilage tissue formation in vitro.
体外形成的组织的低生物力学性能仍然是生物工程关节软骨组织的一个重大障碍。我们之前已经表明,对从全层软骨分离的软骨细胞进行循环压缩(30 分钟,1 kPa,1 Hz)可以诱导更大的基质合成,尽管关节软骨由不同的软骨细胞亚群组成,但其对增强组织形成的个别贡献尚未得到充分表征。本研究检查了软骨细胞亚群对这种反应的贡献。从浅区到中区(SMZ)或深区(DZ)分离牛关节软骨细胞,置于三维培养中,并进行循环压缩。在磷酸钙聚集体上的 DZ 软骨细胞形成的组织比 SMZ 软骨细胞厚。压缩增加了 SMZ 软骨细胞中基质的积累,而减少了 DZ 软骨细胞中基质的积累。SMZ 和 DZ 软骨细胞在其 1 型膜结合基质金属蛋白酶(MMP)和 MMP-13 表达上也存在差异,这些酶在介导对机械刺激的反应中起着至关重要的作用。此外,培养期的持续时间对于确定 DZ 反应很重要,这表明基质积累在刺激反应中起作用。了解组织形成过程中细胞对机械刺激的反应将有助于我们理解组织生长,并允许进一步优化体外软骨组织形成。