Kalinina Sviatlana, Gliemann Hartmut, López-García Mónica, Petershans Andre, Auernheimer Jörg, Schimmel Thomas, Bruns Michael, Schambony Alexandra, Kessler Horst, Wedlich Doris
Institut für Zoologie II (Entwicklungs- und Zellphysiologie), Universität Karlsruhe (TH), Karlsruhe, Germany.
Biomaterials. 2008 Jul;29(20):3004-13. doi: 10.1016/j.biomaterials.2008.04.003. Epub 2008 Apr 22.
With the advances made in surface patterning by micro- and nanotechnology, alternative methods to immobilize biomolecules for different purposes are highly desired. RGD peptides are commonly used to create cell-attractive surfaces for cell-biological and also medical applications. We have developed a fast, one-step method to bind RGD peptides covalently to surfaces by thiourea formation, which can be applied to structured and unstructured materials. RGD peptides were fused to an isothiocyanate anchor during synthesis and directly immobilized on amino-terminated surfaces. The spreading behavior of fibroblasts and the formation of focal contacts served to prove the applicability of the coupling method. Two different linear peptides and one cyclic peptide were compared. All the peptides induced spreading behavior and the formation of focal contacts in murine fibroblasts. Adhesion was specific as cells neither recognized the corresponding negative control peptides nor spread in the presence of soluble H-RGDS-OH peptide. We successfully applied our coupling method to functionalize surface patterns created by microcontact printing (microCP) and chemical etching. Cells recognize areas selectively coated with RGD-containing peptides, proliferate and maintain this preference during long-term cultivation. Our method significantly facilitates surface modification with any kind of peptide - even for the preparation of peptide-functionalized small surface areas.
随着微纳技术在表面图案化方面取得的进展，人们迫切需要用于不同目的固定生物分子的替代方法。RGD肽通常用于为细胞生物学及医学应用创建具有细胞吸引力的表面。我们开发了一种快速的一步法，通过硫脲形成将RGD肽共价结合到表面，该方法可应用于结构化和非结构化材料。在合成过程中，RGD肽与异硫氰酸酯锚定基团融合，并直接固定在氨基端表面。成纤维细胞的铺展行为和粘着斑的形成证明了偶联方法的适用性。比较了两种不同的线性肽和一种环肽。所有肽均诱导小鼠成纤维细胞的铺展行为和粘着斑的形成。细胞粘附具有特异性，因为细胞既不识别相应的阴性对照肽，也不在可溶性H-RGDS-OH肽存在下铺展。我们成功地将偶联方法应用于微接触印刷（microCP）和化学蚀刻创建的表面图案功能化。细胞选择性识别涂有含RGD肽的区域，在长期培养过程中增殖并保持这种偏好。我们的方法显著促进了用任何种类的肽进行表面修饰，甚至用于制备肽功能化的小表面积。
