Department of Clinical Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392, Japan.
Biomacromolecules. 2020 Sep 14;21(9):3765-3771. doi: 10.1021/acs.biomac.0c00871. Epub 2020 Aug 3.
Three-dimensional (3D) cell scaffolds are essential for tissue engineering. So far, various polymer hydrogels have been utilized to design 3D scaffolds as a biomaterial. In this study, we focused on a biocompatible polysaccharide, agarose, as a potential biomaterial candidate. We have previously established a laminin-derived cell adhesive peptide library, and these peptides are useful for incorporating cell adhesiveness into inert materials. We synthesized aldehyde-functionalized agarose (CHO-agarose) by (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO) oxidation of agarose and developed peptide-agarose scaffolds using two laminin peptides, CGG-A99 (CGGAGTFALRGDNPQG, binds to αvβ3 integrin) and CGG-AG73 (CGGRKRLQVQLSIRT, binds to syndecan). The peptides were effectively loaded onto the CHO-agarose gels via thiazolidine formation without coupling reagents. Two-dimensional (2D) cell culture assay using human dermal fibroblasts (HDFs) showed that the peptide-agarose gels have potent cell adhesion activity and promoted cell proliferation. Furthermore, we developed a simple preparation method of 3D cell culture scaffolds using peptide-agarose microgels. HDFs cultured in a 3D environment of the A99-agarose microgel scaffold exhibited cell spreading morphology and proliferated significantly. These results suggest that 3D cell culture systems using peptide-agarose microgel scaffolds are promising as a biomaterial for tissue engineering.
三维（3D）细胞支架对于组织工程至关重要。到目前为止，已经利用各种聚合物水凝胶来设计 3D 支架作为生物材料。在本研究中，我们专注于一种具有生物相容性的多糖琼脂糖，将其作为一种潜在的生物材料候选物。我们之前已经建立了层粘连蛋白衍生的细胞黏附肽文库，这些肽对于将细胞黏附性纳入惰性材料中非常有用。我们通过琼脂糖的（2,2,6,6-四甲基哌啶-1-基）氧自由基（TEMPO）氧化合成了醛基化琼脂糖（CHO-agarose），并使用两种层粘连蛋白肽 CGG-A99（CGGAGTFALRGDNPQG，与αvβ3 整合素结合）和 CGG-AG73（CGGRKRLQVQLSIRT，与 syndecan 结合）开发了肽-琼脂糖支架。通过噻唑烷形成，无需偶联试剂，肽就可以有效地负载到 CHO-琼脂糖凝胶上。用人真皮成纤维细胞（HDFs）进行的二维（2D）细胞培养试验表明，肽-琼脂糖凝胶具有很强的细胞黏附活性，并促进了细胞增殖。此外，我们开发了一种使用肽-琼脂糖微凝胶制备 3D 细胞培养支架的简单方法。在 A99-琼脂糖微凝胶支架的 3D 环境中培养的 HDFs 表现出细胞扩展形态并显著增殖。这些结果表明，使用肽-琼脂糖微凝胶支架的 3D 细胞培养系统有望成为组织工程的生物材料。
