Comparison of different chondrocytes for use in tissue engineering of cartilage model structures
Comparison of different chondrocytes for use in tissue engineering of cartilage model structures
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DOI：
10.1089/ten.2006.12.691
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发表时间：
2006-04-01
影响因子：
--
通讯作者：
Landis, William J.
中科院分区：
文献类型：
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作者：
Isogai, Noritaka;Kusuhara, Hirohisa;Landis, William J.
本研究比较了从四个不同的动物位置-鼻中隔，关节，肋骨，和耳廓-组织工程软骨建模收获的牛软骨细胞。虽然这项工作作为制造人耳模型的初步研究，但其结果对组织工程软骨具有重要意义。软骨细胞进行培养和检查，以确定相对细胞增殖率，II型胶原蛋白和聚集蛋白聚糖基因表达，和细胞外基质的生产。然后将相应的软骨细胞接种到可生物降解的聚（L-丙交酯-ε-己内酯）盘形支架上。培养细胞-共聚物构建体，随后将其植入无胸腺小鼠的皮下空间长达20周。通过分子和组织学方法评估收获结构中的新软骨发育。经过长达4周的细胞培养，显示组织来源的软骨细胞增殖不同，II型胶原蛋白和聚集蛋白聚糖基因表达水平也不同。对于这两个基因，最高的表达被发现肋软骨细胞，其次是鼻中隔，关节，耳廓细胞。从小鼠中取出20周的椎间盘，发现不同软骨细胞的结构尺寸发生变化。最大的光盘直径被发现的支架接种耳廓软骨细胞，其次是那些与肋，鼻中隔，关节细胞。最大的椎间盘厚度测量的支架含有肋软骨细胞，其次是与鼻中隔，耳廓和关节细胞。单独回收的共聚物的直径和厚度最小。只有耳支架在植入20周后形成弹性纤维。在植入20周的椎间盘的定量RT-PCR上检测到不同表达水平的II型胶原和聚集蛋白聚糖。这些数据表明，牛软骨细胞从不同的软骨网站在动物中可能会引起不同的反应，在他们各自的发展组织工程新软骨。因此，每种软骨细胞类型建立或保持其特定的发育特征，并且这种观察在任何组织工程化软骨模型的设计和制作中是至关重要的。
This study compares bovine chondrocytes harvested from four different animal locations-nasoseptal, articular, costal, and auricular-for tissue-engineered cartilage modeling. While the work serves as a preliminary investigation for fabricating a human ear model, the results are important to tissue-engineered cartilage in general. Chondrocytes were cultured and examined to determine relative cell proliferation rates, type II collagen and aggrecan gene expression, and extracellular matrix production. Respective chondrocytes were then seeded onto biodegradable poly(L-lactide-epsilon-caprolactone) disc-shaped scaffolds. Cell-copolymer constructs were cultured and subsequently implanted in the subcutaneous space of athymic mice for up to 20 weeks. Neocartilage development in harvested constructs was assessed by molecular and histological means. Cell culture followed over periods of up to 4 weeks showed chondrocyte proliferation from the tissue sources varied, as did levels of type II collagen and aggrecan gene expression. For both genes, highest expression was found for costal chondrocytes, followed by nasoseptal, articular, and auricular cells. Retrieval of 20-week discs from mice revealed changes in construct dimensions with different chondrocytes. Greatest disc diameter was found for scaffolds seeded with auricular chondrocytes, followed by those with costal, nasoseptal, and articular cells. Greatest disc thickness was measured for scaffolds containing costal chondrocytes, followed by those with nasoseptal, auricular, and articular cells. Retrieved copolymer alone was smallest in diameter and thickness. Only auricular scaffolds developed elastic fibers after 20 weeks of implantation. Type II collagen and aggrecan were detected with differing expression levels on quantitative RT-PCR of discs implanted for 20 weeks. These data demonstrate that bovine chondrocytes obtained from different cartilaginous sites in an animal may elicit distinct responses during their respective development of a tissue-engineered neocartilage. Thus, each chondrocyte type establishes or maintains its particular developmental characteristics, and this observation is critical in the design and elaboration of any tissue-engineered cartilage model.