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用于冠状动脉支架的等离子体纳米涂层的体外生物反应。

In vitro biological responses of plasma nanocoatings for coronary stent applications.

机构信息

Department of Mechanical and Aerospace Engineering, University of Missouri, Columbia, Missouri, USA.

Nanova, Inc., Columbia, Missouri, USA.

出版信息

J Biomed Mater Res A. 2023 Nov;111(11):1768-1780. doi: 10.1002/jbm.a.37587. Epub 2023 Jul 19.

Abstract

In-stent restenosis and thrombosis remain to be long-term challenges in coronary stenting procedures. The objective of this study was to evaluate the in vitro biological responses of trimethylsilane (TMS) plasma nanocoatings modified with NH /O (2:1 molar ratio) plasma post-treatment (TMS + NH /O nanocoatings) on cobalt chromium (CoCr) alloy L605 coupons, L605 stents, and 316L stainless steel (SS) stents. Surface properties of the plasma nanocoatings with up to 2-year aging time were characterized by wettability assessment and x-ray photoelectron spectroscopy (XPS). It was found that TMS + NH /O nanocoatings had a surface composition of 41.21 ± 1.06 at% oxygen, 31.90 ± 1.08 at% silicon, and 24.12 ± 1.7 at% carbon, and very small but essential amount of 2.77 ± 0.18 at% nitrogen. Surface chemical stability of the plasma coatings was noted with persistent O/Si atomic ratio of 1.292-1.413 and N/Si atomic ratio of ~0.087 through 2 years. The in vitro biological responses of plasma nanocoatings were studied by evaluating the cell proliferation and migration of porcine coronary artery endothelial cells (PCAECs) and smooth muscle cells (PCASMCs). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay results revealed that, after 7-day incubation, TMS + NH /O nanocoatings maintained a similar level of PCAEC proliferation while showing a decrease in the viability of PCASMCs by 73 ± 19% as compared with uncoated L605 surfaces. Cell co-culture of PCAECs and PCASMCs results showed that, the cell ratio of PCAEC/PCASMC on TMS + NH /O nanocoating surfaces was 1.5-fold higher than that on uncoated L605 surfaces, indicating enhanced selectivity for promoting PCAEC growth. Migration test showed comparable PCAEC migration distance for uncoated L605 and TMS + NH /O nanocoatings. In contrast, PCASMC migration distance was reduced nearly 8.5-fold on TMS + NH /O nanocoating surfaces as compared to the uncoated L605 surfaces. Platelet adhesion test using porcine whole blood showed lower adhered platelets distribution (by 70 ± 16%), reduced clotting attachment (by 54 ± 12%), and less platelet activation on TMS + NH /O nanocoating surfaces as compared with the uncoated L605 controls. It was further found that, under shear stress conditions of simulated blood flow, TMS + NH /O nanocoating significantly inhibited platelet adhesion compared to the uncoated 316L SS stents and TMS nanocoated 316L SS stents. These results indicate that TMS + NH /O nanocoatings are very promising in preventing both restenosis and thrombosis for coronary stent applications.

摘要

支架内再狭窄和血栓仍然是冠状动脉支架置入术的长期挑战。本研究旨在评估经 trimethylsilane(TMS)等离子体纳米涂层修饰并经 NH /O(2:1 摩尔比)等离子体后处理(TMS + NH /O 纳米涂层)后对钴铬(CoCr)合金 L605 小夹板、L605 支架和 316L 不锈钢(SS)支架的体外生物反应。通过润湿性评估和 X 射线光电子能谱(XPS)对等离子体纳米涂层的表面特性进行了长达 2 年老化时间的表征。结果发现,TMS + NH /O 纳米涂层的表面组成分别为 41.21 ± 1.06 at%氧、31.90 ± 1.08 at%硅和 24.12 ± 1.7 at%碳,以及非常小但必不可少的 2.77 ± 0.18 at%氮。通过 2 年的时间,观察到涂层表面化学稳定性,O/Si 原子比持续保持在 1.292-1.413,N/Si 原子比约为 0.087。通过评估猪冠状动脉内皮细胞(PCAECs)和平滑肌细胞(PCASMCs)的增殖和迁移来研究等离子体纳米涂层的体外生物反应。3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑(MTT)测定结果表明,经过 7 天孵育,TMS + NH /O 纳米涂层保持了 PCAEC 增殖的相似水平,而 PCASMCs 的活力降低了 73 ± 19%,与未涂覆的 L605 表面相比。PCAEC 和 PCASMC 细胞共培养的结果表明,TMS + NH /O 纳米涂层表面的 PCAEC/PCASMC 细胞比例是未涂覆 L605 表面的 1.5 倍,表明其对促进 PCAEC 生长具有更高的选择性。迁移试验表明,未涂覆的 L605 和 TMS + NH /O 纳米涂层的 PCAEC 迁移距离相当。相比之下,PCASMC 的迁移距离在 TMS + NH /O 纳米涂层表面减少了近 8.5 倍,而在未涂覆的 L605 表面。使用猪全血进行血小板黏附试验表明,TMS + NH /O 纳米涂层表面的黏附血小板分布减少了 70 ± 16%,血栓附着减少了 54 ± 12%,血小板激活减少了。进一步发现,在模拟血流剪切应力条件下,与未涂覆的 316L SS 支架和涂覆 TMS 的 316L SS 支架相比,TMS + NH /O 纳米涂层显著抑制了血小板黏附。这些结果表明,TMS + NH /O 纳米涂层在预防冠状动脉支架应用中的再狭窄和血栓形成方面非常有前景。

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