Mouw J K, Case N D, Guldberg R E, Plaas A H K, Levenston M E
George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA 30332-0405, USA.
Osteoarthritis Cartilage. 2005 Sep;13(9):828-36. doi: 10.1016/j.joca.2005.04.020.
To compare matrix composition and glycosaminoglycan (GAG) fine structure among five scaffolds commonly used for in vitro chondrocyte culture and cartilage tissue engineering.
Bovine articular chondrocytes were seeded into agarose, alginate, collagen I, fibrin and polyglycolic acid (PGA) constructs and cultured for 20 or 40 days. In addition to construct DNA and sulfated GAG (sGAG) contents, the delta-disaccharide compositions of the chondroitin/dermatan sulfate GAGs were determined for each scaffold group via fluorophore-assisted carbohydrate electrophoresis (FACE).
Significant differences were found in cell proliferation and extracellular matrix accumulation among the five scaffold groups. Significant cell proliferation was observed for all scaffold types but occurred later (20-40 days) in PGA constructs compared to the other groups (0-20 days). By 40 days, agarose constructs had the highest sGAG to DNA ratio, while alginate and collagen I had the lowest levels. Quantitative differences in the Delta-disaccharide composition of the GAGs accumulated in the different scaffolds were also found, with the most striking variations in unsulfated and disulfated delta-disaccharides. Agarose constructs had the highest fraction of disulfated residues and the lowest fraction of unsulfated residues, with a 6-sulfated/4-sulfated disaccharide ratio most similar to that of native articular cartilage.
The similarities and differences among scaffolds in proteoglycan accumulation and GAG composition suggest that the scaffold material directly or indirectly influences chondrocyte proteoglycan metabolism and may have an influence on the quality of tissue engineered cartilage.
比较常用于体外软骨细胞培养和软骨组织工程的五种支架的基质组成和糖胺聚糖（GAG）精细结构。
将牛关节软骨细胞接种到琼脂糖、藻酸盐、I型胶原、纤维蛋白和聚乙醇酸（PGA）构建体中，培养20或40天。除了测定构建体的DNA和硫酸化GAG（sGAG）含量外，还通过荧光辅助碳水化合物电泳（FACE）测定每个支架组中软骨素/硫酸皮肤素GAG的δ-二糖组成。
在五个支架组之间发现细胞增殖和细胞外基质积累存在显著差异。所有支架类型均观察到显著的细胞增殖，但与其他组（0 - 20天）相比，PGA构建体中的细胞增殖发生较晚（20 - 40天）。到40天时，琼脂糖构建体的sGAG与DNA比率最高，而藻酸盐和I型胶原的水平最低。还发现不同支架中积累的GAG的δ-二糖组成存在定量差异，其中未硫酸化和双硫酸化δ-二糖的变化最为显著。琼脂糖构建体中双硫酸化残基的比例最高，未硫酸化残基的比例最低，其6-硫酸化/4-硫酸化二糖比率与天然关节软骨最为相似。
支架在蛋白聚糖积累和GAG组成方面的异同表明，支架材料直接或间接影响软骨细胞蛋白聚糖代谢，并可能对组织工程软骨的质量产生影响。
