Photoreactive elastin-like proteins for use as versatile bioactive materials and surface coatings.
Photoreactive elastin-like proteins for use as versatile bioactive materials and surface coatings.
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光电反应性弹性蛋白样蛋白用作多功能生物活性材料和表面涂层。
DOI：
10.1039/c2jm31768k
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发表时间：
2012-10-07
影响因子：
--
通讯作者：
Heilshorn S
中科院分区：
文献类型：
--
作者：
Raphel J;Parisi-Amon A;Heilshorn S
可光交联的蛋白质工程生物材料联合收割机将快速、可控、细胞相容性交联方法与模块化设计策略相结合，以创建新的生物活性材料家族。这些材料具有广泛的生物医学应用，包括生物活性植入物涂层、药物递送载体和组织工程支架的开发。我们提出了成功的功能化的生物活性弹性蛋白样蛋白与光反应性二氮杂环丙烷部分。使用标准重组蛋白表达宿主，然后用异双功能N-羟基琥珀酰亚胺酯-二氮杂环丙烷交联剂对赖氨酸残基进行位点特异性修饰，实现可扩展的合成。所得到的生物材料被证明是可加工的旋涂，滴铸，软平版印刷图案，和模具铸造制造各种二维和三维的光交联的生物材料的长度尺度跨越纳米到毫米的范围。蛋白质薄膜被证明在三周内高度稳定。掺入工程蛋白材料的细胞粘附功能结构域显示出在光处理后保持活性。与对照基质相比，人类脂肪来源的干细胞在工程蛋白薄膜上实现了更快的细胞粘附速率和更大的铺展面积。材料生产的简易性和可扩展性、加工的多功能性和模块化生物活性功能性使这种重组工程蛋白成为开发新型生物材料涂层、薄膜和支架的理想候选物。
Photocrosslinkable, protein-engineered biomaterials combine a rapid, controllable, cytocompatible crosslinking method with a modular design strategy to create a new family of bioactive materials. These materials have a wide range of biomedical applications, including the development of bioactive implant coatings, drug delivery vehicles, and tissue engineering scaffolds. We present the successful functionalization of a bioactive elastin-like protein with photoreactive diazirine moieties. Scalable synthesis is achieved using a standard recombinant protein expression host followed by site-specific modification of lysine residues with a heterobifunctional N-hydroxysuccinimide ester-diazirine crosslinker. The resulting biomaterial is demonstrated to be processable by spin coating, drop casting, soft lithographic patterning, and mold casting to fabricate a variety of two- and three-dimensional photocrosslinked biomaterials with length scales spanning the nanometer to millimeter range. Protein thin films proved to be highly stable over a three-week period. Cell-adhesive functional domains incorporated into the engineered protein materials were shown to remain active post-photo-processing. Human adipose-derived stem cells achieved faster rates of cell adhesion and larger spread areas on thin films of the engineered protein compared to control substrates. The ease and scalability of material production, processing versatility, and modular bioactive functionality make this recombinantly engineered protein an ideal candidate for the development of novel biomaterial coatings, films, and scaffolds.