Chondrocyte phenotype in engineered fibrous matrix is regulated by fiber size
Chondrocyte phenotype in engineered fibrous matrix is regulated by fiber size
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DOI：
10.1089/ten.2006.12.1775
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发表时间：
2006-07-01
影响因子：
--
通讯作者：
Tuan, Rocky S.
中科院分区：
文献类型：
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作者：
Li, Wan-Ju;Jiang, Yi Jen;Tuan, Rocky S.
作为天然细胞外基质的功能替代物的生物材料支架为细胞容纳提供空间。在这项研究中，我们种植软骨细胞，分离自4至6个月大的小牛，在2种类型的聚（L-乳酸）支架，由微米和纳米纤维，并比较对细胞活性的影响。扫描电子显微镜显示了良好的传播形态的软骨细胞上生长的微纤维。相比之下，发现纳米纤维上的软骨细胞具有圆形形态，并且与在微纤维支架上良好分散的软骨细胞培养物中观察到的有组织的细胞骨架相比，显示出无序的肌动蛋白细胞骨架结构。两种支架都支持软骨细胞增殖，在纳米纤维支架中的培养物中观察到更高的速率。定量逆转录-聚合酶链反应分析表明，两种文化支持表达胶原蛋白I型和II型和聚集蛋白聚糖。生化分析表明，在培养的硫酸化糖胺聚糖的更高水平，证实了更强烈的阿尔新蓝组织染色。软骨培养物还显示出较高的免疫染色的II型和IX型胶原蛋白，聚集蛋白聚糖，和软骨蛋白聚糖连接蛋白。基于这些结果，我们得出结论，软骨细胞对不同直径的纤维支架有不同的反应，并且由纳米纤维生物材料制成的支架促进有效的基于细胞的软骨组织工程。
A biomaterial scaffold acting as a functional substitute for the native extracellular matrix provides space for cell accommodation. In this study, we seeded chondrocytes, isolated from 4- to 6-month-old calves, in 2 types of poly(L-lactide) scaffolds, composed of micro- and nanofibers, and compared the effects on cellular activities. Scanning electron microscopy revealed a well-spread morphology for chondrocytes grown on microfibers. In contrast, chondrocytes on the nanofibers were found to have a rounded morphology and displayed a disorganized actin cytoskeletal structure compared to the organized cytoskeleton seen in well-spread chondrocytes culture on the microfibrous scaffold. Both scaffolds supported chondrocyte proliferation, with a higher rate seen in cultures in nanofibrous scaffold. Quantitative reverse transcription-polymerase chain reaction analysis showed that both cultures supported expression of collagen types I and II and aggrecan. Biochemical analysis showed a higher level of sulfated glycosaminoglycan in the nanofiber culture, confirmed by more intense alcian blue histologic staining. The nanofiber cultures also showed higher immunostaining for collagen types II and IX, aggrecan, and cartilage proteoglycan link protein. Based on these results, we conclude that chondrocytes respond differently to fibrous scaffolds of varying diameters, and that the scaffolds made of nanofibrous biomaterial promote efficient cell-based cartilage tissue engineering.