在含血清/无血清培养条件下,评估L929成纤维细胞在固定有精氨酸-甘氨酸-天冬氨酸-丝氨酸(RGDS)的壳聚糖上的附着和增殖情况。
Evaluation of L929 fibroblast attachment and proliferation on Arg-Gly-Asp-Ser (RGDS)-immobilized chitosan in serum-containing/serum-free cultures.
作者信息
Karakecili Ayşe Gönen, Demirtas Tugrul Tolga, Satriano Cristina, Gümüsderelioglu Menemşe, Marletta Giovanni
机构信息
Hacettepe University, Chemical Engineering and Bioengineering Departments, Beytepe, Ankara, Turkey.
出版信息
J Biosci Bioeng. 2007 Jul;104(1):69-77. doi: 10.1263/jbb.104.69.
In this study, chitosan membranes prepared by the solvent casting method were modified with the Arg-Gly-Asp-Ser (RGDS) sequence of fibronectin using the photochemical immobilization technique. The results obtained from attenuated total reflection-Fourier transform infrared spectra and X-ray photoelectron spectroscopy studies confirmed the successful immobilization of RGDS on chitosan membranes. The immobilized peptide concentration was determined by ninhydrin analysis on the order of 10(-7) mol/cm(2). In vitro cell culture studies were performed with L929 mouse fibroblasts to investigate the effect of biomodification on fibroblast cell behaviour in serum-free and 10% serum-containing media. The results obtained from cell culture studies pointed out the specific interactions between biosignal RGDS molecules and fibroblast cells. A triggered cell attachment and proliferation were observed on RGDS-modified chitosan membranes that were more distinguishable in serum-free medium. In addition, the photochemical immobilization technique was realized in the presence of a photomask that was used to immobilize the RGDS molecules in a defined micropattern. L929 mouse fibroblasts attached on the RGDS-micropatterned areas indicating integrin-mediated interactions.
在本研究中,采用溶剂浇铸法制备的壳聚糖膜通过光化学固定技术用纤连蛋白的精氨酸-甘氨酸-天冬氨酸-丝氨酸(RGDS)序列进行了修饰。衰减全反射-傅里叶变换红外光谱和X射线光电子能谱研究所得结果证实了RGDS成功固定在壳聚糖膜上。通过茚三酮分析确定固定化肽的浓度约为10(-7) mol/cm(2)。用L929小鼠成纤维细胞进行体外细胞培养研究,以考察生物修饰对无血清和含10%血清培养基中成纤维细胞行为的影响。细胞培养研究所得结果指出了生物信号RGDS分子与成纤维细胞之间的特异性相互作用。在RGDS修饰的壳聚糖膜上观察到了触发的细胞附着和增殖,在无血清培养基中这种现象更明显。此外,光化学固定技术是在光掩模存在的情况下实现的,该光掩模用于将RGDS分子固定在确定的微图案中。L929小鼠成纤维细胞附着在RGDS微图案区域,表明存在整合素介导的相互作用。