Hubei-MOST KLOS & KLOBM, School and Hospital of Stomatology, Wuhan University, Wuhan 430079, People's Republic of China.
Biomed Mater. 2012 Oct;7(5):055009. doi: 10.1088/1748-6041/7/5/055009. Epub 2012 Sep 4.
To improve cell attachment and to understand the effects of positive charge on the behavior of osteoblasts, 2-(methacryloyloxy)ethyl-trimethylammonium chloride (MAETAC), a positively charged monomer, was incorporated into poly(ethylene glycol)-diacrylate (PEGDA) hydrogel. The physicochemical properties of the resultant polymers, including the degree of acrylation of PEGDA, pKa of MAETAC, swelling ratio, zeta potential, and protein adsorption were investigated. Meanwhile, osteoblast-like MC3T3-E1 cells were seeded on the hydrogel to evaluate the effect of the positive charge on the behavior of the cells, including attachment, proliferation, and differentiation. The results revealed that PEGDA was synthesized with 90 percent of acrylation and MAETAC had been successfully incorporated into PEGDA. The pKa value of MAETAC was 9.4 determined by acid-based titration. The electrically charged nature of modified hydrogels was confirmed by zeta potential. With increasing concentration of MAETAC, the swelling ratio of the hydrogel in deionized water increased, while the swelling ratio stayed constant in phosphate buffer solution. The protein adsorption of the hydrogel also increased with increasing concentration of MAETAC. The modification of positive charge not only enhanced the attachment and proliferation of osteoblast-like MC3T3-E1 cells on the hydrogel, but also up-regulated alkaline phosphatase activity in the cells as well as gene expression of focal adhesion kinase and osteogenic markers, including collagen I, osteopontin, and osteocalcin. These findings indicate that the positively charged PEGDA may not only be a promising scaffold candidate for bone tissue engineering, but also a good platform to study the effect of positive charge on cell behavior due to the controllable charge density.
为了改善细胞黏附性,并深入了解正电荷对成骨细胞行为的影响,将带正电荷的单体 2-(甲基丙烯酰氧基)乙基三甲基氯化铵(MAETAC)掺入聚乙二醇二丙烯酸酯(PEGDA)水凝胶中。研究了所得聚合物的物理化学性质,包括 PEGDA 的丙烯酰化程度、MAETAC 的 pKa、溶胀比、Zeta 电位和蛋白质吸附。同时,将成骨样细胞 MC3T3-E1 接种到水凝胶上,以评估正电荷对细胞行为(包括黏附、增殖和分化)的影响。结果表明,PEGDA 的丙烯酰化程度为 90%,并且成功地将 MAETAC 掺入到 PEGDA 中。通过酸滴定法确定 MAETAC 的 pKa 值为 9.4。通过 Zeta 电位证实了改性水凝胶的带电性质。随着 MAETAC 浓度的增加,水凝胶在去离子水中的溶胀比增加,而在磷酸盐缓冲溶液中的溶胀比保持不变。水凝胶的蛋白质吸附也随 MAETAC 浓度的增加而增加。正电荷的修饰不仅增强了成骨样细胞 MC3T3-E1 在水凝胶上的黏附和增殖,而且还上调了细胞中的碱性磷酸酶活性以及粘着斑激酶和骨生成标志物(包括胶原 I、骨桥蛋白和骨钙素)的基因表达。这些发现表明,带正电荷的 PEGDA 不仅可能是骨组织工程有前途的支架候选物,而且由于其可控的电荷密度,还是研究正电荷对细胞行为影响的良好平台。