van den Beucken Jeroen J J P, Vos Matthijn R J, Thüne Peter C, Hayakawa Tohru, Fukushima Tadao, Okahata Yoshio, Walboomers X Frank, Sommerdijk Nico A J M, Nolte Roeland J M, Jansen John A
Department of Periodontology and Biomaterials, College of Dental Science, Radboud University Nijmegen Medical Center, P.O. Box 9101, the Netherlands.
Biomaterials. 2006 Feb;27(5):691-701. doi: 10.1016/j.biomaterials.2005.06.015. Epub 2005 Aug 1.
This study describes the fabrication of two types of multilayered coatings onto titanium by electrostatic self-assembly (ESA), using deoxyribosenucleic acid (DNA) as the anionic polyelectrolyte and poly-d-lysine (PDL) or poly(allylamine hydrochloride) (PAH) as the cationic polyelectrolyte. Both coatings were characterized using UV-vis spectrophotometry, atomic force microscopy (AFM), X-ray photospectroscopy (XPS), contact angle measurements, Fourier transform infrared spectroscopy (FTIR), and for the amount of DNA immobilized. The mutagenicity of the constituents of the coatings was assessed. Titanium substrates with or without multilayered DNA-coatings were used in cell culture experiments to study cell proliferation, viability, and morphology. Results of UV-vis spectrophotometry, AFM, and contact angle measurements clearly indicated the progressive build-up of the multilayered coatings. Furthermore, AFM and XPS data showed a more uniform build-up and morphology of [PDL/DNA]-coatings compared to [PAH/DNA]-coatings. DNA-immobilization into both coatings was linear, and approximated 3microg/cm(2) into each double-layer. The surface morphology of both types of multilayered DNA-coatings showed elevations in the nanoscale range. No mutagenic effects of DNA, PDL, or PAH were detected, and cell viability and morphology were not affected by the presence of either type of multilayered DNA-coating. Still, the results of the proliferation assay revealed an increased proliferation of primary rat dermal fibroblasts on both types of multilayered DNA-coatings compared to non-coated controls. The biocompatibility and functionalization of the coatings produced here, will be assessed in subsequent cell culture and animal-implantation studies.
本研究描述了通过静电自组装（ESA）在钛表面制备两种类型多层涂层的方法，使用脱氧核糖核酸（DNA）作为阴离子聚电解质，聚-D-赖氨酸（PDL）或聚（烯丙胺盐酸盐）（PAH）作为阳离子聚电解质。使用紫外可见分光光度法、原子力显微镜（AFM）、X射线光电子能谱（XPS）、接触角测量、傅里叶变换红外光谱（FTIR）对两种涂层进行表征，并测定固定化DNA的量。评估了涂层成分的致突变性。在细胞培养实验中使用有无多层DNA涂层的钛基底来研究细胞增殖、活力和形态。紫外可见分光光度法、AFM和接触角测量结果清楚地表明了多层涂层的逐步形成。此外，AFM和XPS数据显示，与[PAH/DNA]涂层相比，[PDL/DNA]涂层的形成更加均匀，形态更好。两种涂层中DNA的固定化呈线性，每层双层中约为3μg/cm²。两种类型的多层DNA涂层的表面形态在纳米尺度范围内均有凸起。未检测到DNA、PDL或PAH的致突变作用，两种类型的多层DNA涂层的存在均未影响细胞活力和形态。然而，增殖试验结果显示，与未涂层对照相比，原代大鼠皮肤成纤维细胞在两种类型的多层DNA涂层上的增殖均增加。此处制备的涂层的生物相容性和功能化将在后续的细胞培养和动物植入研究中进行评估。
