Polyelectrolyte multilayers (PEMs) consisting of the polysaccharides hyaluronic acid (HA) as the polyanion and chitosan (Chi) as the polycation were prepared with layer-by-layer technique (LbL). The [Chi/HA]₅ multilayers were exposed to solutions of metal ions (Ca²⁺, Co²⁺, Cu²⁺ and Fe³⁺). Binding of metal ions to [Chi/HA]₅ multilayers by the formation of complexes with functional groups of polysaccharides modulates their physical properties and the bioactivity of PEMs with regard to the adhesion and function of multipotent murine C3H10T1/2 embryonic fibroblasts. Characterization of multilayer formation and surface properties using different analytical methods demonstrates changes in the wetting, surface potential and mechanical properties of multilayers depending on the concentration and type of metal ion. Most interestingly, it is observed that Fe³⁺ metal ions greatly promote adhesion and spreading of C3H10T1/2 cells on the low adhesive [Chi/HA]₅ PEM system. The application of intermediate concentrations of Cu²⁺, Ca²⁺ and Co²⁺ as well as low concentrations of Fe³⁺ to PEMs also results in increased cell spreading. Moreover, it can be shown that complex formation of PEMs with Cu²⁺ and Fe³⁺ ions leads to increased metabolic activity in cells after 24 h and induces cell differentiation towards adipocytes in the absence of any additional adipogenic media supplements. Overall, complex formation of [Chi/HA]₅ PEM with metal ions like Cu²⁺ and Fe³⁺ represents an interesting and cheap alternative to the use of growth factors for making cell-adhesive coatings and guiding stem cell differentiation on implants and scaffolds to regenerate connective-type of tissues.

中文翻译：

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金属离子对透明质酸和壳聚糖多层物理性质的影响，以及多能小鼠成纤维细胞的粘附、生长和成脂分化

采用逐层技术 (LbL) 制备由多糖透明质酸 (HA) 作为聚阴离子和壳聚糖 (Chi) 作为聚阳离子组成的聚电解质多层 (PEM)。[Chi/HA] ₅多层被暴露于金属离子（Ca ²⁺、Co ²⁺、Cu ²⁺和Fe ³⁺）的溶液中。金属离子与 [Chi/HA] _{5 的结合}通过与多糖官能团形成复合物，多层膜可调节 PEM 的物理特性和生物活性，与多能小鼠 C3H10T1/2 胚胎成纤维细胞的粘附和功能有关。使用不同分析方法表征多层膜的形成和表面性质表明多层膜的润湿性、表面电位和机械性质会根据金属离子的浓度和类型发生变化。最有趣的是，观察到 Fe ³⁺金属离子极大地促进了 C3H10T1/2 细胞在低粘附性 [Chi/HA] ₅ PEM 系统上的粘附和扩散。中等浓度Cu ²⁺、Ca ²⁺和Co ^{2+ 的应用}以及低浓度的 Fe ³⁺到 PEM 也会导致细胞扩散增加。此外，可以证明 PEM 与 Cu ²⁺和 Fe ³⁺离子的复合形成导致 24 小时后细胞中的代谢活性增加，并在没有任何额外的脂肪生成培养基补充剂的情况下诱导细胞向脂肪细胞分化。总体而言，[Chi/HA] ₅ PEM 与 Cu ²⁺和 Fe ³⁺等金属离子的复合形成代表了一种有趣且廉价的替代品，可以替代使用生长因子来制造细胞粘附涂层和引导植入物和支架上的干细胞分化再生结缔组织类型。