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细胞黏附受体在生物功能化聚己内酯表面培养的人成骨细胞中的表达

Expression of cell adhesion receptors in human osteoblasts cultured on biofunctionalized poly-(epsilon-caprolactone) surfaces.

作者信息

Amato Ilaria, Ciapetti Gabriela, Pagani Stefania, Marletta Giovanni, Satriano Cristina, Baldini Nicola, Granchi Donatella

机构信息

Laboratory for Pathophysiology, Istituti Ortopedici Rizzoli, Bologna, Italy.

出版信息

Biomaterials. 2007 Sep;28(25):3668-78. doi: 10.1016/j.biomaterials.2007.04.038. Epub 2007 May 3.

Abstract

This study was aimed to investigate whether the activation of poly-(epsilon-caprolactone) (PCL) surface by low-energy irradiation and/or the biofunctionalization by absorption of arginine-glycine-aspartic sequences (RGD), can modify the expression of integrins closely related to the osteoblast activity. For this purpose, we analysed the physicochemical changes induced by irradiation and RGD immobilization, the consequences on cell adhesion and spreading, and the effects on integrin expression. PCL irradiated with 5 x 10(15)He(+)/cm(2) (10 keV energy) (irr-PCL) showed an altered surface layer with a partial loss of carboxyl species and the formation of carbonyl groups. Moreover, irr-PCL showed a small smoothening effect and a less polar character in comparison to the pristine ones. The RGD immobilization was observed only on irr-PCL (surface coverage: 7.0 pmol/cm(2)). Human osteoblasts (hOB) were cultured on untreated PCL (ut-PCL), ut-PCL+RGD, irr-PCL, and irr-PCL+RGD. After 24h, ut-PCL hindered the cell adhesion, while a discrete layer of hOB with a good cytoskeleton organization was detected on irr-PCL and irr-PCL+RGD. Before seeding, the single hOB suspension expressed alpha1, alpha2, alpha3, alpha5, beta1, and alphaVbeta3; after 24h, cells cultured on tissue-plastic expressed high levels of beta1 and alphaVbeta3, while alpha1 showed a low intensity and alpha2, alpha3, and alpha5 were negative. beta1 and alphaVbeta3 were selected to evaluate the interaction between cells and PCL samples. The beta1 expression was higher in hOB cultured on irr-PCL than on the other samples. A significant increase in alphaVbeta3 expression was observed only in irr-PCL+RGD, and confirmed by the gene expression analysis. In conclusion, ion irradiation and RGD adsorption on PCL surfaces modulate the expression of integrin involved in hOB growth and function, indicating the effectiveness of biomimetic surfaces in promoting cell adhesion. Ultimately, the study of integrin expression may suggest proper changes to the surface structure in order to improve the osteoconductivity of selected materials.

摘要

本研究旨在探究低能辐照对聚(ε-己内酯)(PCL)表面的活化作用和/或通过吸收精氨酸-甘氨酸-天冬氨酸序列(RGD)进行生物功能化是否能够改变与成骨细胞活性密切相关的整合素的表达。为此,我们分析了辐照和RGD固定化诱导的物理化学变化、对细胞黏附和铺展的影响以及对整合素表达的作用。用5×10¹⁵He⁺/cm²(10 keV能量)辐照的PCL(irr-PCL)显示其表面层发生改变,羧基部分损失并形成羰基。此外,与原始PCL相比,irr-PCL表现出轻微的平滑效应和较低的极性。仅在irr-PCL上观察到RGD的固定化(表面覆盖率:7.0 pmol/cm²)。将人成骨细胞(hOB)培养在未处理的PCL(ut-PCL)、ut-PCL + RGD、irr-PCL和irr-PCL + RGD上。24小时后,ut-PCL阻碍细胞黏附,而在irr-PCL和irr-PCL + RGD上检测到具有良好细胞骨架组织的离散hOB层。在接种前,单个hOB悬液表达α1、α2、α3、α5、β1和αVβ3;24小时后,培养在组织塑料上的细胞表达高水平的β1和αVβ3,而α1表达强度较低,α2、α3和α5呈阴性。选择β1和αVβ3来评估细胞与PCL样品之间的相互作用。在irr-PCL上培养的hOB中β1表达高于其他样品。仅在irr-PCL + RGD中观察到αVβ3表达显著增加,基因表达分析证实了这一点。总之,PCL表面的离子辐照和RGD吸附调节了参与hOB生长和功能的整合素的表达,表明仿生表面在促进细胞黏附方面的有效性。最终,对整合素表达的研究可能会为表面结构提出适当的改变建议,以提高所选材料的骨传导性。

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