Protista Biotechnology AB, IDEON, SE-22370, Lund, Sweden.
Colloids Surf B Biointerfaces. 2010 Feb 1;75(2):510-9. doi: 10.1016/j.colsurfb.2009.09.028. Epub 2009 Sep 26.
Boronate-containing thin polyacrylamide gels (B-Gel), polymer brushes (B-Brush) and chemisorbed organosilane layers (B-COSL) were prepared on the surface of glass slides and studied as substrates for carbohydrate-mediated cell adhesion. B-COSL- and B-Brush-modified glass samples exhibited multiple submicron structures densely and irregularly distributed on the glass surface, as found by scanning electron microscopy and atomic force microscopy. B-Gel was ca. 0.1 mm thick and contained pores with effective size of 1-2 microm in the middle and of 5-20 microm on the edges of the gel sample as found by confocal laser scanning microscopy. Evidence for the presence of phenylboronic acid in the samples was given by time-of-flight secondary ion mass-spectrometry (ToF SIMS), contact angle measurements performed in the presence of fructose, and staining with Alizarin Red S dye capable of formation specific, fluorescent complexes with boronic acids. A comparative study of adhesion and cultivation of animal cells on the above substrates was carried out using murine hybridoma M2139 cell line as a model. M2139 cells adhered to the substrates in the culture medium without glucose or sodium pyruvate at pH 8.0, and then were cultivated in the same medium at pH 7.2 for 4 days. It was found that the substrates of B-Brush type were superior both regarding cell adhesion and viability of the adhered cells, among the substrates studied. MTT assay confirmed proliferation of M2139 cells on B-Brush substrates. Some cell adhesion was also registered in the macropores of B-Gel substrate. The effects of surface microstructure of the boronate-containing polymers on cell adhesion are discussed. Transparent glass substrates grafted with boronate-containing copolymers offer good prospects for cell adhesion studies and development of cell-based assays.
硼酸酯功能化的薄聚丙烯酰胺凝胶（B-Gel）、聚合物刷（B-Brush）和化学吸附的有机硅烷层（B-COSL）被制备在载玻片表面，并作为糖介导的细胞黏附的基底进行了研究。扫描电子显微镜和原子力显微镜发现，B-COSL 和 B-Brush 修饰的玻璃样品在玻璃表面上呈现出密集且不规则分布的多个亚微米结构。共聚焦激光扫描显微镜发现，B-Gel 的厚度约为 0.1mm，凝胶样品边缘的有效孔径为 5-20μm，中间的有效孔径为 1-2μm。飞行时间二次离子质谱（ToF SIMS）、在存在果糖时进行的接触角测量以及能够与硼酸形成特异性荧光复合物的茜素红 S 染料的染色为样品中存在苯硼酸提供了证据。使用鼠杂交瘤 M2139 细胞系作为模型，对上述基底上的动物细胞黏附和培养进行了比较研究。在 pH 值为 8.0、不含葡萄糖或丙酮酸钠的培养基中，M2139 细胞黏附在基底上，然后在 pH 值为 7.2 的相同培养基中培养 4 天。研究发现，在所研究的基底中，B-Brush 型基底在细胞黏附性和黏附细胞的活力方面均具有优势。MTT 测定法证实了 M2139 细胞在 B-Brush 基底上的增殖。在 B-Gel 基底的大孔中也观察到一些细胞黏附。讨论了硼酸酯聚合物的表面微观结构对细胞黏附的影响。接枝有硼酸酯共聚物的透明玻璃基底为细胞黏附研究和基于细胞的测定方法的发展提供了良好的前景。
