Hahn Mariah S, Taite Lakeshia J, Moon James J, Rowland Maude C, Ruffino Katie A, West Jennifer L
Department of Bioengineering, Rice University, 6100 Main Street/MS 142, Houston, TX 77005, USA.
Biomaterials. 2006 Apr;27(12):2519-24. doi: 10.1016/j.biomaterials.2005.11.045. Epub 2005 Dec 20.
A simple, inexpensive photolithographic method for surface patterning deformable, solvated substrates is demonstrated using photoactive poly(ethylene glycol) (PEG)-diacrylate hydrogels as model substrates. Photolithographic masks were prepared by printing the desired patterns onto transparencies using a laser jet printer. Precursor solutions containing monoacryloyl-PEG-peptide and photoinitiator were layered onto hydrogel surfaces. The acrylated moieties in the precursor solution were then conjugated in monolayers to specific hydrogel regions by exposure to UV light through the transparency mask. The effects of UV irradiation time and precursor solution concentration on the levels of immobilized peptide were characterized, demonstrating that bound peptide concentration can be controlled by tuning these parameters. Multiple peptides can be immobilized to a single hydrogel surface in distinct patterns by sequential application of this technique, opening up its potential use in co-cultures. In addition, 3D structures can be generated by incorporating PEG-diacrylate into the precursor solution. To evaluate the feasibility of using these patterned surfaces for guiding cell behavior, human dermal fibroblast adhesion on hydrogel surfaces patterned with acryloyl-PEG-RGDS was investigated. This patterning method may find use in tissue engineering, the elucidation of fundamental structure-function relationships, and the formation of immobilized cell and protein arrays for biotechnology.
利用光活性聚乙二醇(PEG)-二丙烯酸酯水凝胶作为模型底物,展示了一种用于对可变形、溶剂化底物进行表面图案化的简单、低成本光刻方法。通过使用激光打印机将所需图案打印到透明胶片上来制备光刻掩膜。将含有单丙烯酰基-PEG-肽和光引发剂的前体溶液铺展在水凝胶表面。然后,通过透过透明掩膜暴露于紫外光,使前体溶液中的丙烯酸化部分在单层中与特定的水凝胶区域共轭。对紫外照射时间和前体溶液浓度对固定化肽水平的影响进行了表征,表明可以通过调节这些参数来控制结合肽的浓度。通过依次应用该技术,可以将多种肽以不同图案固定在单个水凝胶表面,这为其在共培养中的潜在应用开辟了道路。此外,通过将PEG-二丙烯酸酯加入前体溶液中可以生成三维结构。为了评估使用这些图案化表面引导细胞行为的可行性,研究了人真皮成纤维细胞在以丙烯酰基-PEG-RGDS图案化的水凝胶表面上的粘附情况。这种图案化方法可能在组织工程、阐明基本结构-功能关系以及形成用于生物技术的固定化细胞和蛋白质阵列方面得到应用。