Aoki T, Nagao Y, Sanui K, Ogata N, Kikuchi A, Sakurai Y, Kataoka K, Okano T
Department of Chemistry, Faculty of Science and Technology, Sophia University, Tokyo, Japan.
J Biomater Sci Polym Ed. 1997;9(1):1-14. doi: 10.1163/156856297x00227.
Copolymers (IABb) composed of N-isopropylacrylamide (I), N-(3-dimethylaminopropyl)-acrylamide (A), 3-acrylamidophenylboronic acid (B), and a hydrophobic comonomer, n-butyl methacrylate (b), were synthesized as cell culture substrata since we previously learned that bovine aortic endothelial cells (BAECs) cultured on the phenylboronic acid-containing copolymer (IAB) differentiated into capillary structures after 26 days culture. The synthesized IABb copo ymers contained higher boron and amine moieties than the IAB copolymer. The results of the dynamic contact angle measurement revealed that IABb copolymer-coated surfaces showed a relatively hydrophobic nature, changing to hydrophilic in response to the aqueous environment. BAECs cultured on the copolymer substrata developed into capillary networks after 7 days. This is probably due to the enrichment of boron and amine segments in the vicinity of the hydrophilic copolymer surface, enhancing more pronounced interaction of boronates with cell membrane glycocalyx. The introduction of n-butyl methacrylate into the polymers might enhance the diffusion of the hydrophobic segments to the bulk polymers and the concentration of relatively hydrophilic segments at the outermost polymer surfaces by contact with water. A copolymer (IAP) without boronic acid groups was also prepared using N-phenylacrylamide (P) as a comonomer instead of 3-acrylamidophenylboronic acid in the IAB copolymer to investigate the effect of boronic acid moieties on the capillary formation of the cultured cells. Although the endothelial cells seeded on the copolymer without boronic acid groups adhered during the early culture period, these cells showed neither proliferation nor differentiation and detached from the surface after 13 days. These results strongly support the opinion that the phenylboronic acid groups in the copolymers are responsible for the specific induction of tissue formation of BAECs through the interaction with glycoconjugates on the cell membranes.
合成了由N-异丙基丙烯酰胺（I）、N-（3-二甲基氨基丙基）-丙烯酰胺（A）、3-丙烯酰胺基苯硼酸（B）和疏水性共聚单体甲基丙烯酸正丁酯（b）组成的共聚物（IABb）作为细胞培养基质，因为我们之前了解到，在含苯硼酸的共聚物（IAB）上培养的牛主动脉内皮细胞（BAECs）在培养26天后会分化成毛细血管结构。合成的IABb共聚物比IAB共聚物含有更高的硼和胺部分。动态接触角测量结果表明，IABb共聚物涂层表面呈现相对疏水的性质，在水环境中会转变为亲水。在共聚物基质上培养的BAECs在7天后发育成毛细血管网络。这可能是由于亲水性共聚物表面附近硼和胺链段的富集，增强了硼酸盐与细胞膜糖萼更明显的相互作用。将甲基丙烯酸正丁酯引入聚合物中可能会增强疏水链段向本体聚合物的扩散以及通过与水接触使相对亲水链段在聚合物最外层表面的浓度增加。还制备了一种不含硼酸基团的共聚物（IAP），使用N-苯基丙烯酰胺（P）作为共聚单体代替IAB共聚物中的3-丙烯酰胺基苯硼酸，以研究硼酸部分对培养细胞毛细血管形成的影响。尽管接种在不含硼酸基团的共聚物上的内皮细胞在早期培养阶段附着，但这些细胞既不增殖也不分化，13天后从表面脱落。这些结果有力地支持了这样一种观点，即共聚物中的苯硼酸基团通过与细胞膜上的糖缀合物相互作用，对BAECs组织形成的特异性诱导起作用。
