Enzyme-assisted photolithography for spatial functionalization of hydrogels
Enzyme-assisted photolithography for spatial functionalization of hydrogels
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DOI：
10.1039/c001335h
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
Tang, Yi
中科院分区：
文献类型：
--
作者：
Gu, Zhen;Tang, Yi
形成功能性聚合物模式结构的能力对细胞生物学、组织工程和医学诊断的研究具有重要意义。我们开发了一种新的酶辅助光刻（EAPL）方法，通过高通量的方式实现水凝胶的空间功能化。在水凝胶聚合过程中使用双丙烯酸化肽交联剂，它含有蛋白酶可切割的氨基酸序列，并被光不稳定的片段笼住。采用简单的两步过程，包括紫外线暴露在所需区域破坏肽交联剂和蛋白酶开发，仅在紫外线处理区域特异性消化凝胶。重要的是，肽键的蛋白质水解在图案区域产生可进一步功能化的自由亲核胺基团。利用这种策略和caspase-3作为酶的开发物，我们展示了在聚乙二醇（PEG）水凝胶中同时生成地形和功能模式。我们发现，用含精氨酸-甘氨酸-天冬氨酸（RGD）肽功能化的20毫米宽的线阵列可以用于产生具有单个细胞分辨率的细胞图案。我们还制作了直径20毫米的阵列，用B淋巴细胞特异性抗cd19修饰腔，用于从0.1%的起始B细胞混合物中获得600倍的B细胞富集。简单的制造工艺、直接的化学反应以及全水基生物相容性和环境友好的方法使EAPL成为一个通用的平台，可以为芯片实验室系统和组织工程构建生物响应的2D模式或3D支架。
The ability to form functional polymeric patterning structures has important implications for the studies of cell biology, tissue engineering, and medical diagnostics. We have developed a novel enzyme-assisted photolithography (EAPL) method for spatial functionalization of hydrogels via a high throughput fashion. A bisacrylated peptide crosslinker, containing a protease cleavable amino acid sequence and caged by a photolabile moiety, is used during hydrogel polymerization. A facile two-step process is employed, including UV exposure to decage the peptide crosslinker at a desired area and protease development to specifically digest gels at UV treated regions only. Importantly, proteolysis of the peptide bonds generates free nucleophilic amine groups at the patterned area that can be further functionalized. Using this strategy and caspase-3 as the enzyme developer, we demonstrate the simultaneous generation of topographical and functional patterns into poly(ethylene glycol) (PEG) hydrogels. We show that 20 mm-wide line arrays functionalized with arginine-glycine-aspartic acid (RGD)-containing peptides can be used to generate cell patterns with individual cell resolution. We also fabricated arrays 20 mm diameter cavities decorated with B lymphocyte specific anti-CD19, which was used to achieve a 600-fold enrichment of B-cells from a 0.1% starting B-cell mixture. The simple fabrication process, straightforward chemistry and an all-aqueous based biocompatible and environmentally friendly approach render EAPL a versatile platform to construct biologically responsive 2D patterns or 3D scaffolds for lab-on-a-chip systems and tissue engineering.