Laminin‐1 peptide‐conjugated chitosan membranes as a novel approach for cell engineering
Laminin‐1 peptide‐conjugated chitosan membranes as a novel approach for cell engineering
复制标题
DOI：
10.1096/fj.02-0564fje
复制
发表时间：
2003-05
期刊：
影响因子：
--
通讯作者：
M. Mochizuki;Y. Kadoya;Y. Wakabayashi;Kozue Kato;I. Okazaki;M. Yamada;Taku Sato;N. Sakairi
中科院分区：
文献类型：
--
作者：
M. Mochizuki;Y. Kadoya;Y. Wakabayashi;Kozue Kato;I. Okazaki;M. Yamada;Taku Sato;N. Sakairi
层粘连蛋白是基底膜的主要成分，具有多种生物学活性。最近，我们通过使用大量合成肽鉴定了层粘连蛋白-1上的各种生物活性序列。壳聚糖是一种可生物降解的多糖，已被用作生物材料。在这里，我们将几种具有生物活性的层粘连蛋白肽缀合到壳聚糖膜上，并测量了肽缀合的壳聚糖膜与各种细胞类型的细胞附着活性。活性层粘连蛋白肽缀合的壳聚糖膜促进具有细胞类型特异性的细胞附着。A99（AGTFALRGDNPQG）-壳聚糖膜促进细胞与组织良好的肌动蛋白应力纤维的附着。这种粘附被EDTA抑制，但不被肝素抑制。AG 73（RKRLQVQLSIRT）-壳聚糖膜促进细胞附着并形成丝状伪足，这种粘附被肝素抑制，但不被EDTA抑制。这些数据表明，A99-壳聚糖膜与整合素细胞受体相互作用，并且AG 73-壳聚糖膜促进蛋白聚糖介导的细胞附着，如先前所报道的。此外，AG 73-壳聚糖和A99-壳聚糖膜均有效促进PC 12大鼠嗜铬细胞瘤细胞的神经突生长。我们的结论是，共轭壳聚糖膜是适用于定量测试的合成肽的生物活性，这些结构具有潜在的能力，作为生物粘附材料的组织再生和工程。
Laminin, a major component of the basement membrane, has diverse biological activities. Recently, we identified various biologically active sequences on laminin‐1 by using a large set of synthetic peptides. Chitosan, a polysaccharide, is biodegradable and has been used as a biomaterial. Here, we conjugated several biologically active laminin peptides onto chitosan membranes and measured the cell attachment activity of peptide‐conjugated chitosan membranes with various cell types. The active laminin peptide‐conjugated chitosan membranes promoted cell attachment with cell type specificity. A99 (AGTFALRGDNPQG)‐chitosan membrane promoted cell attachment with well‐organized actin stress fibers. This adhesion was inhibited by EDTA but not by heparin. AG73 (RKRLQVQLSIRT)‐chitosan membrane promoted cell attachment with filopodia formation, and this adhesion was inhibited by heparin but not by EDTA. These data suggest that the A99‐chitosan membrane interacted with an integrin cellular receptor and that the AG73‐chitosan membrane promoted proteoglycan‐mediated cell attachment, as previously reported. Furthermore, both AG73‐chitosan and A99‐chitosan membranes effectively promoted neurite outgrowth with PC12 rat pheochromocytoma cells. We conclude that conjugation on a chitosan membrane is applicable for testing quantitatively the biological activity of synthetic peptides and that these constructs have a potential ability to serve as bioadhesive materials for tissue regeneration and engineering.