Kawazoe Naoki, Guo Likun, Wozniak Michal J, Imaizumi Yumie, Tateishi Tetsuya, Zhang Xingdong, Chen Guoping
Biomaterials Center National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan.
J Nanosci Nanotechnol. 2009 Jan;9(1):230-9. doi: 10.1166/jnn.2009.j003.
Three kinds of photoreactive polyelectrolytes of polyallylamine (PAAm), poly(acrylic acid) (PAAc), and poly(vinyl alcohol) (PVA) were synthesized by the introduction of azidophenyl groups in the respective polymers. The photoreactive PAAm, PAAc, and PVA were micropatterned on polystyrene surfaces by photolithography. Observation with optical microscopy and scanning probe microscopy demonstrated the formation of a striped pattern of polyelectrolytes with a width of 200 microm. The micropatterned polyelectrolytes swelled in water. The micropatterned surfaces were used for cell culture of mesenchymal stem cells (MSCs) and their effects on adipogenic differentiation were investigated. The MSCs adhered to and proliferated evenly on the PAAm- and PAAc-patterned surfaces while they formed a cell pattern on the PVA-patterned surface. The PAAm-, PAAc-grafted, and polystyrene surfaces supported cell adhesion while the PVA-grafted surface did not. When cultured in adipogenic differentiation medium, the adipogenic differentiation of MSCs on the polyelectrolyte-patterned surfaces was demonstrated by the formation of lipid vacuoles and gene expression analysis. Oil Red-O-positive cells showed an even distribution on the PAAm- and PAAc-patterned surfaces, while they showed a pattern on the PVA-patterned surface. The fraction of Oil RedO-positive cells increased with culture time. The MSCs cultured on the PAAm-, PAAc-grafted, and polystyrene surfaces in adipogenic differentiation medium expressed the adipogenesis marker genes of peroxisome proliferator-activated receptor gamma2 (PPARgamma2), lipoprotein lipase (LPL), and fatty acid binding protein 4 (FABP4). These results indicate that the PAAm-, and PAAc-grafted, and polystyrene surfaces supported the adipogenesis of MSCs while a PVA-grafted surface did not.
通过在各自聚合物中引入叠氮苯基,合成了三种光反应性聚电解质,分别是聚烯丙胺(PAAm)、聚丙烯酸(PAAc)和聚乙烯醇(PVA)。通过光刻技术将光反应性PAAm、PAAc和PVA微图案化在聚苯乙烯表面。光学显微镜和扫描探针显微镜观察表明形成了宽度为200微米的聚电解质条纹图案。微图案化的聚电解质在水中膨胀。将微图案化表面用于间充质干细胞(MSC)的细胞培养,并研究其对成脂分化的影响。MSC在PAAm和PAAc图案化表面上均匀粘附并增殖,而在PVA图案化表面上形成细胞图案。PAAm、PAAc接枝表面和聚苯乙烯表面支持细胞粘附,而PVA接枝表面则不支持。当在成脂分化培养基中培养时,通过脂质空泡的形成和基因表达分析证明了MSC在聚电解质图案化表面上的成脂分化。油红O阳性细胞在PAAm和PAAc图案化表面上分布均匀,而在PVA图案化表面上呈现图案化。油红O阳性细胞的比例随培养时间增加。在成脂分化培养基中培养在PAAm、PAAc接枝表面和聚苯乙烯表面上的MSC表达过氧化物酶体增殖物激活受体γ2(PPARγ2)、脂蛋白脂肪酶(LPL)和脂肪酸结合蛋白4(FABP4)的成脂标记基因。这些结果表明,PAAm、PAAc接枝表面和聚苯乙烯表面支持MSC的成脂作用,而PVA接枝表面则不支持。