Sun Min, Zhou Ping, Pan Luan-Feng, Liu Shui, Yang Hua-Xiao
The Key Laboratory of Molecular Engineering of Polymers, Department of Macromolecular Science, Fudan University, Shanghai, PR China.
J Mater Sci Mater Med. 2009 Aug;20(8):1743-51. doi: 10.1007/s10856-009-3739-8. Epub 2009 Mar 31.
Cell affinity is one of the important issues required for developing tissue engineering materials. Although the poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) has been attractive for its controllable mechanical properties recent years, its cell affinity is still necessary to be improved for the requirements. For this purpose, the regenerated silk fibroin (SF) was coated on the PHBHHx films and its porous scaffolds. The mechanical test showed that SF-modified PHBHHx (SF/PHBHHx) film has a maximum tensile strength of 11.5 +/- 0.5 MPa and elongation at break of 175 +/- 5%. ATR-FTIR spectroscopy demonstrated that SF firmly attached on the scaffold by the hydrogen bonding interaction between SF and PHBHHx even flushed for 21 days in the phosphate-buffer saline (PBS) solution (pH = 7.4). In order to characterize the cell affinity of the SF-modified material, endothelial-like cell line ECV304 cells were seeded on the SF/PHBHHx films and its porous scaffolds. The histochemical analyses of cells stained by the hematoxylin and eosin (HE) as well as cell nuclei stained by the 4',6-diamindine-2'-phenylindole (DAPI) demonstrated that cell attached and reached nearly 100% confluence on the SF/PHBHHx films when cultured for 4 days, which was much faster than that on the pure PHBHHx film. Moreover, the assay of cell activity by the 3-(4, 5-dimethyl thiazol -2-yl)-2, 5-diphenyl terazolium bromide (MTT) showed quantitatively that the number of cells on the SF/PHBHHx porous scaffolds was significant more than that on the unmodified ones after 4, 8, and 14 days culture, respectively. Scanning electron microscopy (SEM) revealed the similar results. Therefore, the SF-modified PHBHHx material is maybe a potential material applicable in the cardiovascular tissue engineering.
细胞亲和力是开发组织工程材料所需解决的重要问题之一。尽管聚（3-羟基丁酸酯-co-3-羟基己酸酯）（PHBHHx）近年来因其可控的机械性能而备受关注，但根据需求其细胞亲和力仍有待提高。为此，将再生丝素蛋白（SF）涂覆在PHBHHx薄膜及其多孔支架上。力学测试表明，SF改性的PHBHHx（SF/PHBHHx）薄膜的最大拉伸强度为11.5±0.5MPa，断裂伸长率为175±5%。衰减全反射傅里叶变换红外光谱（ATR-FTIR）表明，即使在磷酸盐缓冲盐水（PBS）溶液（pH = 7.4）中冲洗21天，SF也通过与PHBHHx之间的氢键相互作用牢固地附着在支架上。为了表征SF改性材料的细胞亲和力，将内皮样细胞系ECV304细胞接种在SF/PHBHHx薄膜及其多孔支架上。苏木精和伊红（HE）染色的细胞以及4',6-二脒基-2'-苯基吲哚（DAPI）染色的细胞核的组织化学分析表明，培养4天时，细胞附着在SF/PHBHHx薄膜上并达到近100%汇合，这比在纯PHBHHx薄膜上快得多。此外，3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑（MTT）细胞活性测定定量显示，分别在培养4天、8天和14天后，SF/PHBHHx多孔支架上的细胞数量明显多于未改性的支架。扫描电子显微镜（SEM）显示了类似的结果。因此，SF改性的PHBHHx材料可能是一种适用于心血管组织工程的潜在材料。
