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生物活性分子和碳颗粒接枝的聚乙烯提高了血管平滑肌细胞的黏附、生长和成熟。

Improved adhesion, growth and maturation of vascular smooth muscle cells on polyethylene grafted with bioactive molecules and carbon particles.

机构信息

Centre for Cardiovascular Research, Institute of Physiology, Academy of Sciences of the Czech Republic / Videnska 1083, CZ-14220 Prague 4, Czech Republic.

Department of Solid State Engineering, Institute of Chemical Technology / Technicka 5, 16628 Prague 6, Czech Republic.

出版信息

Int J Mol Sci. 2009 Nov 20;10(10):4352-4374. doi: 10.3390/ijms10104352.

Abstract

High-density polyethylene (PE) foils were modified by an Ar(+) plasma discharge and subsequent grafting with biomolecules, namely glycine (Gly), polyethylene glycol (PEG), bovine serum albumin (BSA), colloidal carbon particles (C) or BSA and C (BSA + C). As revealed by atomic force microscopy (AFM), goniometry and Rutherford Backscattering Spectroscopy (RBS), the surface chemical structure and surface morphology of PE changed dramatically after plasma treatment. The contact angle decreased for the samples treated by plasma, mainly in relation to the formation of oxygen structures during plasma irradiation. A further decrease in the contact angle was obvious after glycine and PEG grafting. The increase in oxygen concentration after glycine and PEG grafting proved that the two molecules were chemically linked to the plasma-activated surface. Plasma treatment led to ablation of the PE surface layer, thus the surface morphology was changed and the surface roughness was increased. The materials were then seeded with vascular smooth muscle cells (VSMC) derived from rat aorta and incubated in a DMEM medium with fetal bovine serum. Generally, the cells adhered and grew better on modified rather than on unmodified PE samples. Immunofluorescence showed that focal adhesion plaques containing talin, vinculin and paxillin were most apparent in cells on PE grafted with PEG or BSA + C, and the fibres containing alpha-actin, beta-actin or SM1 and SM2 myosins were thicker, more numerous and more brightly stained in the cells on all modified PE samples than on pristine PE. An enzyme-linked immunosorbent assay (ELISA) revealed increased concentrations of focal adhesion proteins talin and vinculin and also a cytoskeletal protein beta-actin in cells on PE modified with BSA + C. A contractile protein alpha-actin was increased in cells on PE grafted with PEG or Gly. These results showed that PE activated with plasma and subsequently grafted with bioactive molecules and colloidal C particles, especially with PEG and BSA + C, promotes the adhesion, proliferation and phenotypic maturation of VSMC.

摘要

高密度聚乙烯(PE)箔经 Ar(+)等离子体放电处理和随后接枝生物分子,即甘氨酸(Gly)、聚乙二醇(PEG)、牛血清白蛋白(BSA)、胶体碳颗粒(C)或 BSA 和 C(BSA+C),进行了改性。原子力显微镜(AFM)、接触角测量和卢瑟福背散射光谱(RBS)表明,等离子体处理后 PE 的表面化学结构和形貌发生了剧烈变化。经等离子体处理的样品的接触角减小,主要与等离子体辐照过程中形成的氧结构有关。接枝甘氨酸和 PEG 后,接触角进一步降低。接枝甘氨酸和 PEG 后氧浓度的增加证明了这两种分子与等离子体激活表面发生了化学结合。等离子体处理导致 PE 表面层的烧蚀,从而改变了表面形貌并增加了表面粗糙度。然后,将材料接种来源于大鼠主动脉的血管平滑肌细胞(VSMC),并在含有胎牛血清的 DMEM 培养基中孵育。通常,经改性的 PE 样品上的细胞比未经改性的 PE 样品更易于附着和生长。免疫荧光显示,在接枝有 PEG 或 BSA+C 的 PE 上的细胞中,含有 talin、vinculin 和 paxillin 的焦点粘附斑最为明显,并且在所有改性的 PE 样品上的细胞中的含有 alpha-actin、beta-actin 或 SM1 和 SM2 肌球蛋白的纤维更厚、更多且更亮。酶联免疫吸附测定(ELISA)显示,在经 BSA+C 改性的 PE 上的细胞中,焦点粘附蛋白 talin 和 vinculin 以及细胞骨架蛋白 beta-actin 的浓度增加。在接枝有 PEG 或 Gly 的 PE 上的细胞中,收缩蛋白 alpha-actin 增加。这些结果表明,经等离子体激活并随后接枝生物活性分子和胶体 C 颗粒的 PE,特别是接枝有 PEG 和 BSA+C 的 PE,促进了 VSMC 的粘附、增殖和表型成熟。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/245c/2790113/515445d2fd3c/ijms-10-04352f1.jpg

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