Long-Term Comparison between Human Normal Conchal and Microtia Chondrocytes Regenerated by Tissue Engineering on Nanofiber Polyglycolic Acid Scaffolds
Long-Term Comparison between Human Normal Conchal and Microtia Chondrocytes Regenerated by Tissue Engineering on Nanofiber Polyglycolic Acid Scaffolds
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DOI：
10.1097/prs.0000000000003201
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发表时间：
2017-04-01
影响因子：
3.6
通讯作者：
Landis, William J.
中科院分区：
文献类型：
--
作者：
Nakao, Hitomi;Jacquet, Robin D.;Landis, William J.
背景资料：以前通过组织工程进行的耳廓软骨再生研究没有解决的问题是，将人耳廓软骨细胞接种到聚合物支架上的软骨细胞表型是否能长期保留，以及小耳畸形残余物是否可以作为耳廓重建的活细胞来源。从人耳（正常耳甲或小耳软骨残余物）分离软骨细胞，体外扩增，并接种到纳米级直径的聚乙醇酸片上。这些组织工程构建体被植入无胸腺小鼠体内长达40周。在收获时间的5，10，20，和40周，样品记录的大体形态，组织学，和逆转录定量聚合酶链反应analysis.Results：从两种类型的手术组织产生的新软骨是相似的外观，其细胞外基质和阳性染色的弹性蛋白和蛋白多糖。在5- 40周的时间间隔，有一个增加的趋势，在基因表达的II型胶原蛋白，弹性蛋白，和性别决定区Y盒5，重要的正常软骨表型，和一个下降的趋势，在基因表达的III型胶原蛋白，成纤维细胞和去分化标志物。经过40周的植入，原始纳米级直径的聚乙醇酸支架尺寸（1cm × 1cm × 80 μ m）的组织工程软骨的长度和宽度基本保持不变，而厚度在统计学上显著增加。耳软骨可以在长期内再生（40周）从手术残留物的组织工程技术纳入纳米级直径聚乙醇酸支架。根据目前的分析，小耳新生软骨非常类似于从正常耳甲软骨分离的软骨细胞再生的组织工程软骨。
Background: Previous regeneration studies of auricle-shaped cartilage by tissue engineering leave unresolved whether the chondrocyte phenotype from human auricular chondrocytes seeded onto polymeric scaffolds is retained over the long term and whether microtia remnants may be a viable cell source for auricular reconstruction.Methods: Chondrocytes were isolated from human ears, either normal conchal ear or microtia cartilage remnants, expanded in vitro, and seeded onto nanoscale-diameter polyglycolic acid sheets. These tissue-engineered constructs were implanted into athymic mice for up to 40 weeks. At harvest times of 5, 10, 20, and 40 weeks, samples were documented by gross morphology, histology, and reverse transcription-quantitative polymerase chain reaction analysis.Results: Neocartilages generated from the two types of surgical tissues were similar in appearance of their extracellular matrices and positive staining for elastin and proteoglycans. In the 5- to 40-week time interval, there was an increasing trend in gene expression for type II collagen, elastin, and sex determining region Y box 5, important to normal cartilage phenotype, and a decreasing trend in gene expression for type III collagen, a fibroblast and dedifferentiation marker. Over 40 weeks of implantation, the original nanoscale-diameter polyglycolic acid scaffold dimensions (1 cm x 1 cm x 80 mu m) were generally maintained in tissue-engineered cartilage length and width, and thickness was statistically significantly increased.Conclusions: Auricular cartilage can be regenerated over the long term (40 weeks) from surgical remnants by tissue-engineering techniques incorporating nanoscale-diameter polyglycolic acid scaffolds. Based on the present assays, microtia neocartilage very closely resembles tissue-engineered cartilage regenerated from chondrocytes isolated from normal conchal cartilage.