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体外评估多聚 N-乙烯基吡咯烷酮/丙烯酸纳米粒子在微血管内皮细胞模型中的生物相容性。

In Vitro Assessment of Poly-N-Vinylpyrrolidone/Acrylic Acid Nanoparticles Biocompatibility in a Microvascular Endothelium Model.

机构信息

Laboratory of Histology-Embryology, School of Medicine, University of Crete, 71003 Heraklion, Greece.

Department of Technology of Chemical Pharmaceutical and Cosmetic Substances, D. Mendeleev University of Chemical Technology of Russia, 125047 Moscow, Russia.

出版信息

Int J Mol Sci. 2022 Oct 18;23(20):12446. doi: 10.3390/ijms232012446.

Abstract

An amphiphilic copolymer of N-vinyl-2-pyrrolidone and acrylic acid-namely, p(VP-AA)-OD6000 (p(VP-AA))-was synthesized to prepare p(VP-AA) nanoparticles (NPs). Furthermore, the copolymer was linked with CFSE, and the so-prepared nanoparticles were loaded with the DiI dye to form D nanoparticles (DNPs). In this study, as demonstrated by immunofluorescence microscopy, immunofluorescence, and confocal microscopy, DNPs were readily taken up by human microvascular endothelial cells (HMEC-1) cells in a concentration-dependent manner. Upon uptake, both the CFSE dye (green stain) and the DiI dye (red stain) were localized to the cytoplasm of treated cells. Treatment with p(VP-AA) did not affect the viability of normal and challenged with LPS, HMEC-1 cells at 0.010 mg/mL and induced a dose-dependent decrease of these cells' viability at the higher concentrations of 0.033 and 0.066 mg/mL ( ≤ 0.01; ≤ 0.001, respectively). Furthermore, we focused on the potential immunological activation of HMEC-1 endothelial cells upon p(VP-AA) NPs treatment by assessing the expression of adhesion molecules (E-Selectin, ICAM-1, and V-CAM). NPs treatments at concentrations utilized ( = NS) did not affect individual adhesion molecules' expression. p(VP-AA) NPs do not activate the endothelium and do not affect its viability at pharmacologically relevant concentrations.

摘要

合成了 N-乙烯基-2-吡咯烷酮和丙烯酸的两亲性共聚物,即 p(VP-AA)-OD6000(p(VP-AA)),用于制备 p(VP-AA)纳米颗粒(NPs)。此外,该共聚物与 CFSE 相连,所制备的纳米颗粒负载 DiI 染料形成 D 纳米颗粒(DNPs)。在这项研究中,如免疫荧光显微镜、免疫荧光和共聚焦显微镜所示,DNPs 以浓度依赖的方式被人微血管内皮细胞(HMEC-1)细胞摄取。摄取后,CFSE 染料(绿色染色)和 DiI 染料(红色染色)均定位于处理细胞的细胞质中。p(VP-AA)处理不会影响 0.010 mg/mL 浓度下正常和 LPS 挑战的 HMEC-1 细胞的活力,并在较高浓度 0.033 和 0.066 mg/mL 时诱导这些细胞活力呈剂量依赖性下降(≤0.01;≤0.001,分别)。此外,我们专注于 p(VP-AA) NPs 处理对 HMEC-1 内皮细胞潜在免疫激活的影响,通过评估粘附分子(E-选择素、ICAM-1 和 V-CAM)的表达来评估。在使用的浓度下(= NS), NPs 处理不会影响单个粘附分子的表达。在药理学相关浓度下,p(VP-AA) NPs 不会激活内皮细胞,也不会影响其活力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b9a9/9604021/41b264cd90ff/ijms-23-12446-g001.jpg

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