Characterization of the Natural History of Extracellular Matrix Production in Tissue-Engineered Vascular Grafts during Neovessel Formation
Characterization of the Natural History of Extracellular Matrix Production in Tissue-Engineered Vascular Grafts during Neovessel Formation
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DOI：
10.1159/000331405
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发表时间：
2012-01-01
影响因子：
2.7
通讯作者：
Breuer, Christopher K.
中科院分区：
文献类型：
--
作者：
Naito, Yuji;Williams-Fritze, Misty;Breuer, Christopher K.
背景：细胞外基质（ECM）是新血管完整性的关键决定因素。材料与方法：将36个（聚乙醇酸+聚己内酯和聚乳酸）组织工程血管移植物与同基因骨髓单个核细胞一起植入C57 BL/6小鼠下腔静脉作为间置移植物。除了基质金属蛋白酶（MMP）外，还使用免疫组织化学染色和qPCR对标本进行表征。测定总胶原、弹性蛋白和糖胺聚糖（GAG）含量。采用酶谱法测定MMP活性。结果如下：组织学上的胶原蛋白产生在1周时表现出III型的初始增加，随后在2周时I型产生，在4周时IV型产生。I型和III型的基因表达在2周时达到峰值，而IV型在4周内继续增加。组织学显示，在1周时，E1蛋白-1沉积，随后在4周时产生弹性蛋白。弹性蛋白基因表达在4周时显著增加，而β-内酰胺酶-1在4周时降低。在组织学上，每个时间点的GAG均显示大量产生。核心蛋白聚糖的基因表达在4周时显著增加，而多功能蛋白聚糖随时间推移而降低。生化分析表明，总胶原蛋白的生产是最大的在2周，有一个显着增加弹性蛋白和GAG生产在4周。MMPs的组织学特征显示在每个时间点都有大量的MMP-2产生，而MMP-9在4周内下降。MMP-2的基因表达在4周时显著增加，而MMP-9在4周时显著降低。结论：新生血管形成过程中ECM的产生以早期ECM沉积和随后的广泛重塑为特征。版权所有（C）2011 S. Karger AG，巴塞尔
Background: The extracellular matrix (ECM) is a critical determinant of neovessel integrity. Materials and Methods: Thirty-six (polyglycolic acid + polycaprolactone and poly lactic acid) tissue-engineered vascular grafts seeded with syngeneic bone marrow mononuclear cells were implanted as inferior vena cava interposition grafts in C57BL/6 mice. Specimens were characterized using immunohistochemical staining and qPCR for representative ECM components in addition to matrix metalloproteinases (MMPs). Total collagen, elastin, and glycosaminoglycan (GAG) contents were determined. MMP activity was measured using zymography. Results: Collagen production on histology demonstrated an initial increase in type III at 1 week followed by type I production at 2 weeks and type IV at 4 weeks. Gene expression of both type I and type III peaked at 2 weeks, whereas type IV continued to increase over the 4-week period. Histology demonstrated fibrillin-1 deposition at 1 week followed by elastin production at 4 weeks. Elastin gene expression significantly increased at 4 weeks, whereas fibrillin-1 decreased at 4 weeks. GAG demonstrated abundant production at each time point on histology. Gene expression of decorin significantly increased at 4 weeks, whereas versican decreased over time. Biochemical analysis showed that total collagen production was greatest at 2 weeks, and there was a significant increase in elastin and GAG production at 4 weeks. Histological characterization of MMPs showed abundant production of MMP-2 at each time point, while MMP-9 decreased over the 4-week period. Gene expression of MMP-2 significantly increased at 4 weeks, whereas MMP-9 significantly decreased at 4 weeks. Conclusions: ECM production during neovessel formation is characterized by early ECM deposition followed by extensive remodeling. Copyright (C) 2011 S. Karger AG, Basel