Heller Martin, Kämmerer Peer W, Al-Nawas Bilal, Luszpinski Marie-Anne, Förch Renate, Brieger Jürgen
Department of Otorhinolaryngolgy of the Johannes Gutenberg University Mainz, 55131, Mainz, Germany; Department of Oral and Maxillofacial Surgery of the Johannes Gutenberg University Mainz, 55131, Mainz, Germany; Max Planck Institute for Polymer Research, 55128, Mainz, Germany.
J Biomed Mater Res A. 2015 Jun;103(6):2035-44. doi: 10.1002/jbm.a.35340. Epub 2014 Oct 14.
Biomimetic surface modifications are regarded as promising approach to stimulate cellular behavior at the interface of implant materials. Aim of the study was an evaluation of the cellular response of human umbilical cord cells (HUVECS) and human osteoblasts (HOBS) on titanium covalently coated with the extracellular matrix (ECM) proteins fibrinogen, collagen, laminin, and osteopontin. For the surface modification, titanium discs were first amino-functionalized by plasma polymerization of allylamine. The ECM protein conjugation was performed using the linker molecule α, ω-bis-N-hydroxysuccinimide polyethylene glycol (Di-NHS linker). For surface characterization, infrared spectroscopy and fluorescein isothiocyanate staining (FITC) were used to evaluate the presence and distribution of primary amines in the plasma polymer film. Real-time analyses of the respective protein conjugation processes were performed via surface plasmon resonance kinetic measurements. All ECM proteins were immobilized successfully. Furthermore, the biological functionality of the conjugated factors fibronectin and collagen could be proven as they led to a distinct stimulation of cell adhesion of HUVECS and HOBS when compared to the control group. The highest cell coverage of HUVECS was observed on fibronectin-modified surfaces with approximately 35% and on collagen with 33% after 24 h (PT: 9.4%). For laminin, no additional effect was observed, and for osteopontin, only a slight enhancement of cell adhesion was found. A similar, cell-stimulating tendency of fibronectin and collagen was seen as well after 3 and 7 days. Biomimetic surface modification via plasma polymerization is a powerful method for biomolecule conjugation with a high retention of biological functionality and offer promising clinical perspectives.
仿生表面修饰被认为是在植入材料界面刺激细胞行为的一种有前景的方法。本研究的目的是评估人脐静脉内皮细胞（HUVECS）和人成骨细胞（HOBS）在共价包被有细胞外基质（ECM）蛋白纤维蛋白原、胶原蛋白、层粘连蛋白和骨桥蛋白的钛表面上的细胞反应。对于表面修饰，钛盘首先通过烯丙胺的等离子体聚合进行氨基功能化。使用连接分子α,ω-双-N-羟基琥珀酰亚胺聚乙二醇（双-NHS连接子）进行ECM蛋白偶联。为了进行表面表征，使用红外光谱和异硫氰酸荧光素染色（FITC）来评估等离子体聚合物膜中伯胺的存在和分布。通过表面等离子体共振动力学测量对各个蛋白质偶联过程进行实时分析。所有ECM蛋白均成功固定。此外，偶联因子纤连蛋白和胶原蛋白的生物学功能得到了证实，因为与对照组相比，它们导致HUVECS和HOBS的细胞粘附受到明显刺激。在纤连蛋白修饰的表面上，24小时后观察到HUVECS的最高细胞覆盖率约为35%，在胶原蛋白修饰的表面上为33%（PT：9.4%）。对于层粘连蛋白，未观察到额外的作用，对于骨桥蛋白，仅发现细胞粘附略有增强。在3天和7天后也观察到纤连蛋白和胶原蛋白有类似的细胞刺激趋势。通过等离子体聚合进行的仿生表面修饰是一种用于生物分子偶联的强大方法，具有高生物功能保留率，并提供了有前景的临床前景。
