Fischell Department of Bioengineering, University of Maryland, College Park, MD 20742, USA.
J Tissue Eng Regen Med. 2012 Mar;6(3):179-92. doi: 10.1002/term.411. Epub 2011 Feb 24.
The delivery of growth factors to aid in cartilage engineering has received considerable attention. However, phenotypical differences between chondrocyte cell populations and their distinct responses to growth factors are not fully understood. To address this issue, we have investigated the gene expression of chondrocytes isolated from the superficial, middle, and deep zones of bovine articular cartilage. A three-dimensional (3D) alginate bead model was used to encapsulate zonal chondrocytes and culture with or without exogenous insulin-like growth factor-1(IFG-1) delivery. Following culture, mRNA expression of type I collagen, type II collagen, aggregan, IGF-1 and IGF-1 binding protein (IGF-BP3) were analysed at 1, 4 and 8 days. To the best of our knowledge, this is the first study to investigate gene expression of IGF-1 and IGF-BP3 by zone, and among the first studies to investigate growth factor delivery to chondrocytes in a 3D culture environment. Histological images and cell count data confirm the isolation of chondrocyte subpopulations, and gene expression data show distinct profiles for each zone, both with and without IGF-1 delivery. The data also show similar gene expression for the middle and deep zone cells, while the superficial zone group displays unique activity. Deep zone cells appear the most robust in their phenotype retention and most responsive to IGF-1 delivery. The results highlight differences in metabolic activity and varying responses to delivered growth factors between zonal chondrocyte populations.
生长因子的递送在软骨工程中受到了广泛关注。然而,软骨细胞群体之间表型的差异及其对生长因子的不同反应还不完全清楚。为了解决这个问题,我们研究了从牛关节软骨的浅层、中层和深层分离的软骨细胞的基因表达。使用三维(3D)藻酸盐珠模型包封区带软骨细胞,并在有或没有外源性胰岛素样生长因子-1(IGF-1)递送的情况下进行培养。培养后,在第 1、4 和 8 天分析 I 型胶原、II 型胶原、聚集蛋白、IGF-1 和 IGF-1 结合蛋白(IGF-BP3)的 mRNA 表达。据我们所知,这是第一项按区带研究 IGF-1 和 IGF-BP3 基因表达的研究,也是第一项在 3D 培养环境中研究生长因子向软骨细胞递送的研究之一。组织学图像和细胞计数数据证实了软骨细胞亚群的分离,并且基因表达数据显示每个区带都有独特的表达谱,无论是有还是没有 IGF-1 递送。数据还表明,中层和深层细胞的基因表达相似,而浅层区组表现出独特的活性。深层区细胞的表型保留最为稳健,对 IGF-1 的递送反应最为敏感。研究结果强调了区带软骨细胞群体之间代谢活性和对递送到的生长因子的不同反应的差异。