Influence of Bone Morphogenetic Protein-2 on the Extracellular Matrix, Material Properties, and Gene Expression of Long-Term Articular Chondrocyte Cultures: Loss of Chondrocyte Stability
Influence of Bone Morphogenetic Protein-2 on the Extracellular Matrix, Material Properties, and Gene Expression of Long-Term Articular Chondrocyte Cultures: Loss of Chondrocyte Stability
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DOI：
10.1089/ten.tea.2008.0249
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发表时间：
2009-06-01
影响因子：
4.1
通讯作者：
Lewis, Jack L.
中科院分区：
文献类型：
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作者：
Krawczak, David A.;Westendorf, Jennifer J.;Lewis, Jack L.
本研究的目的是确定骨形态发生蛋白-2（BMP-2）对体外培养8周的单层关节软骨细胞组织的影响。从新西兰白色兔中分离关节软骨细胞，并接种在单层培养物中。培养物补充100 ng/mL的BMP-2长达8周，并分析细胞外基质（ECM）组成、材料特性和信使RNA（mRNA）表达。软骨特异性基因、II型胶原和聚集蛋白聚糖的mRNA表达表明，BMP-2增强软骨细胞稳定性长达3周。培养3周后，BMP-2处理的5周和8周培养物中I型胶原表达显著增加，骨桥蛋白和run-related转录因子2表达显著增加。此外，在用BMP-2处理的5周和8周培养物中，基质金属蛋白酶-13和ADAMTS-5（具有血小板反应蛋白5的解整合素样和金属蛋白酶）上调，与ECM密度、胶原和蛋白多糖的损失一致。与对照组相比，用BMP-2刺激的8周组织更脆弱，从培养皿中取出时更容易撕裂，这表明BMP-2在单层系统中的有效性的时间限制，也许是其他模型，以增强软骨样组织的产生用于组织工程的目的。
The aim of this study was to determine the effects of bone morphogenetic protein-2 (BMP-2) on articular chondrocyte tissues grown as monolayers in vitro for up to 8 weeks. Articular chondrocytes were isolated from New Zealand White rabbits and plated in monolayer cultures. The cultures were supplemented with 100 ng/mL of BMP-2 for up to 8 weeks and the extracellular matrix (ECM) composition, material properties, and messenger RNA (mRNA) expression were analyzed. mRNA expression of cartilage-specific genes, type II collagen, and aggrecan showed that BMP-2 enhanced chondrocyte stability for up to 3 weeks. After 3 weeks in culture, there was substantially more type I collagen expression and more osteopontin and runt-related transcription factor 2 expression in 5- and 8-week cultures treated with BMP-2 than in controls. Additionally, matrix metalloproteinase-13 and ADAMTS-5 (A disintegrin-like and metalloproteinase with thrombospondin 5) were upregulated in 5-and 8-week cultures treated with BMP-2, coinciding with a loss of ECM density, collagen, and proteoglycan. Eight-week tissue stimulated with BMP-2 was more fragile and tore more easily when removed from the culture dish as compared to controls, suggesting temporal limitations to the effectiveness of BMP-2 in monolayer systems and perhaps other models to enhance the generation of a cartilage-like tissue for tissue engineering purposes.