Guo Likun, Kawazoe Naoki, Fan Yujiang, Ito Yoshihiro, Tanaka Junzo, Tateishi Tetsuya, Zhang Xingdong, Chen Guoping
Biomaterials Center, National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan.
Biomaterials. 2008 Jan;29(1):23-32. doi: 10.1016/j.biomaterials.2007.08.043.
Human mesenchymal stem cells (MSCs) were cultured on polystyrene surfaces modified with photoreactive azidophenyl-derivatives of three different chargeable polymers, poly(acrylic acid) (PAAc), polyallylamine (PAAm), and poly(ethylene glycol) (PEG). The MSCs adhered and spread both on a PAAm-modified surface and on PAAc-modified and polystyrene (control) surfaces. However, the cells adhered more easily to the PAAm-modified surface. The MSCs did not attach to the PEG-modified surface and aggregated to form pellets immediately after cell seeding. The cells proliferated on the PAAc-, PAAm-modified and control surfaces with culture time, formed a monolayer, and aggregated to form pellets. The cells in the pellets that formed on the PAAm- and PEG-modified surfaces after 2 weeks culture had a round morphology and the extracellular matrices were positively stained by safranin O and toluidine blue, while those that formed on the PAAc-modified and control surfaces had a spindle, fibroblast-like morphology and were not positively stained by safranin O and toluidine blue. The pellets that formed on the PAAm- and PEG-modified surfaces contained significantly higher levels of sulfated glycosaminoglycans than did those that formed on the PAAc-modified and control surfaces. Type II collagen and cartilage proteoglycan were immunohistologically detected in the pellets that formed on PAAm- and PEG-modified surfaces, but not those that formed on the PAAc-modified and control surfaces. The MSCs cultured on the PAAm- and PEG-modified surfaces expressed a high level of cartilaginous genes encoding type II collagen and aggrecan, while the MSCs cultured on the PAAc-modified and control surfaces did not express these genes. These results suggest that the PAAm-modified surface supported cell adhesion and proliferation and also promoted chondrogenic differentiation of the MSCs. The PAAc-modified and polystyrene surfaces supported cell adhesion and proliferation, but not chondrogenic differentiation. The PEG-modified surfaces did not support cell adhesion, but did promote chondrogenic differentiation. The adhesion, proliferation, and differentiation of the MSCs could be controlled by surface chemistry.
将人间充质干细胞(MSCs)培养于用三种不同可充电聚合物的光反应性叠氮苯基衍生物修饰的聚苯乙烯表面,这三种聚合物分别是聚丙烯酸(PAAc)、聚烯丙胺(PAAm)和聚乙二醇(PEG)。MSCs能在PAAm修饰的表面以及PAAc修饰的和聚苯乙烯(对照)表面上黏附并铺展。然而,细胞更容易黏附于PAAm修饰的表面。MSCs不附着于PEG修饰的表面,且在细胞接种后立即聚集形成小球。随着培养时间的推移,细胞在PAAc、PAAm修饰的和对照表面上增殖,形成单层,并聚集形成小球。培养2周后在PAAm和PEG修饰表面上形成的小球中的细胞呈圆形形态,细胞外基质经番红O和甲苯胺蓝阳性染色,而在PAAc修饰的和对照表面上形成的小球中的细胞呈纺锤形、成纤维细胞样形态,且未被番红O和甲苯胺蓝阳性染色。在PAAm和PEG修饰表面上形成的小球中硫酸化糖胺聚糖的含量显著高于在PAAc修饰的和对照表面上形成的小球。在PAAm和PEG修饰表面上形成的小球中通过免疫组织化学检测到II型胶原蛋白和软骨蛋白聚糖,但在PAAc修饰的和对照表面上形成的小球中未检测到。在PAAm和PEG修饰表面上培养的MSCs表达高水平的编码II型胶原蛋白和聚集蛋白聚糖的软骨基因,而在PAAc修饰的和对照表面上培养的MSCs不表达这些基因。这些结果表明,PAAm修饰的表面支持细胞黏附与增殖,还促进了MSCs的软骨形成分化。PAAc修饰的和聚苯乙烯表面支持细胞黏附与增殖,但不支持软骨形成分化。PEG修饰的表面不支持细胞黏附,但促进软骨形成分化。MSCs的黏附、增殖和分化可通过表面化学来控制。