Max Planck Institute of Colloids and Interfaces, Department of Interfaces, Research Campus Golm, Potsdam, Germany.
Biomaterials. 2010 Sep;31(27):7167-74. doi: 10.1016/j.biomaterials.2010.06.002. Epub 2010 Jun 26.
A simple approach to the mechanical modulation of layer-by-layer (LbL) films is through manipulation of the film assembly. Here, we report results based on altering the salt concentration during film assembly and its effect on film rigidity. Based on changes in film rigidity, cell adhesion characteristics and transfection activity were investigated in vitro. LbL films consisting of reducible hyperbranched poly(amide amine) (RHB) have been implemented along with DNA for investigating fibroblast adhesion on RHB/DNA films with varying rigidities. The rigidity was varied by changing the ionic concentration of the deposition solution between 0.01 m NaCl and 1.0 m NaCl. Molecular force probe (MFP) measurements were performed to measure the apparent Young's modulus, E(APP), of the films in situ. Cell adhesion and stress-fiber characteristics were investigated using total internal reflectance microscopy (TIRF-M). The average cell peripheral area, fiber density and average fiber length during 5 days of cell growth on films with either low (below 2.0 MPa) or high (above 2.0 MPa) film elastic modulus were investigated. Transfection studies were performed using gfpDNA and SEAP-DNA to investigate if changes in cell adhesion affect transfection activity. Furthermore, cell proliferation and cytotoxicity studies were used to investigate cellular viability over a week. The results have shown that surface modification of bioreducible LbL films of controlled thickness and roughness promotes cellular adhesion, stress-fiber growth and increased transfection activity without the need for an additional adhesive protein pre-coating of the surface or chemical cross-linking of the film.
一种简单的方法是通过对膜组装的操作来实现对层层(LbL)膜的机械调制。在这里,我们报告了基于改变膜组装过程中盐浓度及其对膜刚性影响的结果。基于膜刚性的变化,研究了体外细胞黏附特性和转染活性。已经实施了由可还原超支化聚(酰胺-胺)(RHB)组成的 LbL 膜,并用 DNA 来研究在[RHB/DNA](n/2)膜上具有不同刚性的成纤维细胞黏附。通过将沉积溶液的离子浓度从 0.01 m NaCl 改变至 1.0 m NaCl 来改变刚性。使用分子力探针(MFP)原位测量来测量膜的表观杨氏模量,E(APP)。使用全内反射显微镜(TIRF-M)研究细胞黏附和应力纤维特征。研究了在具有低(低于 2.0 MPa)或高(高于 2.0 MPa)膜弹性模量的膜上细胞生长 5 天时的细胞外周区域、纤维密度和平均纤维长度的平均细胞周边区域、纤维密度和平均纤维长度。使用 GFP-DNA 和 SEAP-DNA 进行转染研究,以研究细胞黏附是否会影响转染活性。此外,还进行了细胞增殖和细胞毒性研究,以研究一周内的细胞活力。结果表明,具有受控厚度和粗糙度的生物可还原 LbL 膜的表面改性促进了细胞黏附、应力纤维生长和转染活性的增加,而无需对表面进行额外的粘合蛋白预涂层或对膜进行化学交联。