Chollet C, Labrugère C, Durrieu M C
National Institute on Health and Medical Research, U577, Bordeaux, France.
Annu Int Conf IEEE Eng Med Biol Soc. 2007;2007:5123-6. doi: 10.1109/IEMBS.2007.4353493.
The aim of this study was to evaluate the impact of different densities on MC3T3 cells attachment onto Poly (ethylene terephthalate) (PET) film surfaces. Biomimetic modifications were performed by means of a three-step reaction procedure: creation of COOH functions onto PET surface, coupling agent grafting and finally immobilization of peptides. The originality of this work consist, in one hand on quantifying RGD peptides densities grafted onto PET, and on the other hand on studying MC3T3 cells responses after seeding on such biomimetic surfaces. After each functionalization step, modifications were validated by several physico-chemical techniques: X-Ray Photoelectron Spectroscopy allowed to prove the grafting and high-resolution micro-imager coupled with use of radiolabelled amino acids enabled the evaluation of peptides densities. Moreover, this last technique permit us to ensure stability of binding between peptides and polymer. The efficiency of this new route for biomimetic modification of PET surface was demonstrated by measuring the adhesion at 15h of osteoblast like cells. Study of cellular comportement was realised by means of focal contact proteins (vinculin, actin) immunostaining.
本研究的目的是评估不同密度对MC3T3细胞附着在聚对苯二甲酸乙二酯（PET）薄膜表面的影响。通过三步反应程序进行仿生修饰：在PET表面形成羧基官能团、接枝偶联剂以及最后固定肽段。这项工作的独到之处在于，一方面对接枝到PET上的RGD肽段密度进行定量，另一方面研究在这种仿生表面接种后MC3T3细胞的反应。在每个功能化步骤之后，通过几种物理化学技术对修饰进行验证：X射线光电子能谱用于证明接枝情况，结合使用放射性标记氨基酸的高分辨率显微成像仪能够评估肽段密度。此外，最后这项技术使我们能够确保肽段与聚合物之间结合的稳定性。通过测量成骨样细胞在15小时时的粘附情况，证明了这种PET表面仿生修饰新途径的有效性。通过粘着斑蛋白（纽蛋白、肌动蛋白）免疫染色来研究细胞行为。
