Three-dimensional micropatterning of bioactive hydrogels via two-photon laser scanning photolithography for guided 3D cell migration
Three-dimensional micropatterning of bioactive hydrogels via two-photon laser scanning photolithography for guided 3D cell migration
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DOI：
10.1016/j.biomaterials.2008.04.004
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发表时间：
2008-07-01
期刊：
影响因子：
14
通讯作者：
West, Jennifer L.
中科院分区：
文献类型：
--
作者：
Lee, Soo-Hong;Moon, James J.;West, Jennifer L.
在微尺度上控制生物分子和细胞三维（3D）排列的微图案化技术将有助于开发由多种细胞类型组成且具有复杂结构的临床相关组织。尽管在调控各种材料中生物分子的空间和时间分布方面已经有了重大进展，但大多数微图案化技术仅适用于二维图案化。我们在此报道利用双光子激光扫描（TPLS）光刻技术在水凝胶中对细胞黏附配体（RGDS）进行微图案化，以引导细胞沿着预先定义的3D路径迁移。TPLS光刻技术在微尺度焦点体积内调控光反应过程，从而生成复杂的、无微观尺度缺陷的图案，并能控制水凝胶支架内生物分子的空间呈现和浓度。TPLS光刻技术被用于确定RGDS在胶原酶敏感的聚（乙二醇 - 共 - 肽）二丙烯酸酯水凝胶中的精确位置，并且使用荧光素标记的RGDS评估固定化RGDS的量。当在纤维蛋白簇中培养的人皮肤成纤维细胞被包封在微图案化的胶原酶敏感水凝胶中时，细胞仅在引导下向水凝胶的RGDS图案化区域进行3D迁移。这些结果展示了通过在高度明确的几何形状中提供适当的生物活性线索，在3D支架中微尺度引导组织再生的前景。（C）2008爱思唯尔有限公司。保留所有权利。
Micropatterning techniques that control three-dimensional (3D) arrangement of biomolecules and cells at the microscale will allow development of clinically relevant tissues composed of multiple cell types in complex architecture. Although there have been significant developments to regulate spatial and temporal distribution of biomolecules in various materials, most micropatterning techniques are applicable only to two-dimensional patterning. We report here the use of two-photon laser scanning (TPLS) photolithographic technique to micropattern cell adhesive ligand (RGDS) in hydrogels to guide cell migration along pre-defined 3D pathways, The TPLS photolithographic technique regulates photoreactive processes in microscale focal volumes to generate complex, free from microscale patterns with control over spatial presentation and concentration of biomolecules within hydrogel scaffolds. The TPLS photolithographic technique was used to dictate the precise location of RGDS in collagenase-sensitive poly(ethylene glycol-co-peptide) diacrylate hydrogels, and the amount of immobilized RGDS was evaluated using fluorescein-tagged RGDS. When human dermal fibroblasts cultured in fibrin clusters were encapsulated within the micropatterned collagenase-sensitive hydrogels, the cells underwent guided 3D migration only into the RGDS-patterned regions of the hydrogels. These results demonstrate the prospect of guiding tissue regeneration at the microscale in 3D scaffolds by providing appropriate bioactive cues in highly defined geometries. (C) 2008 Elsevier Ltd. All rights reserved.