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