不同刚度纳米粒配伍薄荷醇/莪术醇对bEnd.3细胞力学性质的影响研究

Effect of nanoparticles of different stiffness combined with menthol/curcumol on mechanical properties of bEnd.3 cells

探究不同刚度纳米粒PLGA-NPs和介孔二氧化硅纳米粒(MSNs)配伍薄荷醇或莪术醇前后对bEnd.3细胞力学性质的影响。采用马尔文粒度仪测定PLGA-NPs和MSNs的粒径分布情况,并通过原子力显微镜(AFM)对两者的刚度进行定量研究;体外培养bEnd.3细胞,以细胞表面形貌、粗糙程度、杨氏模量为考察指标,表征细胞表面的粗糙程度和刚度,采用AFM观察细胞力学性质的变化,并采用激光共聚焦显微镜成像观察细胞骨架肌动蛋白F-actin的结构和表达。结果显示,2种纳米粒均具有良好的分散性,PLGA-NPs的粒径为(98.77±2.04)nm,PDI为0.140±0.030,杨氏模量值为(104.717±8.475)MPa;MSNs的粒径为(97.47±3.92)nm,PDI为0.380±0.016,杨氏模量值为(306.019±8.822)MPa,PLGA-NPs刚度显著低于MSNs。PLGA-NPs和MSNs分别单独作用于bEnd.3细胞后,细胞表面均出现细微孔洞,粗糙程度增大,杨氏模量值减小,F-actin条带均出现轮廓模糊、断裂等现象,平均灰度值均显著降低;PLGA-NPs配伍薄荷醇或莪术醇后,与PLGA-NPs作用相比,bEnd.3细胞表面孔洞加深且密集分布,粗糙程度增加,杨氏模量值显著降低,F-actin条带断裂现象更加明显,平均灰度值显著降低。MSNs配伍薄荷醇后,与MSNs作用相比,bEnd.3细胞表面孔洞加深且密集分布,粗糙程度增加,杨氏模量值显著降低,F-actin条带断裂更加明显,平均灰度值显著降低;配伍莪术醇后,与单独作用相比,无显著差异。由此推测,芳香性成分能通过改变bEnd.3细胞的生物力学性质,增加纳米粒胞内摄取量和转运量。

This study aimed to investigate the effects of nanoparticles PLGA-NPs and mesoporous silicon nanoparticles(MSNs)of different stiffness before and after combination with menthol or curcumol on the mechanical properties of bEnd.3 cells.The particle size distributions of PLGA-NPs and MSNs were measured by Malvern particle size analyzer,and the stiffness of the two nanoparticles was quantified by atomic force microscopy(AFM).The bEnd.3 cells were cultured in vitro,and the cell surface morphology,roughness,and Young′s modulus were examined to characterize the roughness and stiffness of the cell surface.The changes in the mechanical properties of the cells were observed by AFM,and the structure and expression of cytoskeletal F-actin were observed by a laser-scanning confocal microscope.The results showed that both nanoparticles had good dispersion.The particle size of PLGA-NPs was(98.77±2.04)nm,the PDI was(0.140±0.030),and Young′s modulus value was(104.717±8.475)MPa.The particle size of MSNs was(97.47±3.92)nm,the PDI was(0.380±0.016),and Young′s modulus value was(306.019±8.822)MPa.The stiffness of PLGA-NPs was significantly lower than that of MSNs.After bEnd.3 cells were treated by PLGA-NPs and MSNs separately,the cells showed fine pores on the cell surface,increased roughness,decreased Young′s modulus,blurred and broken F-actin bands,and reduced mean gray value.Compared with PLGA-NPs alone,PLGA-NPs combined with menthol or curcumol could allow deepened and densely distributed surface pores of bEnd.3 cells,increase roughness,reduce Young′s modulus,aggravate F-actin band breakage,and diminish mean gray value.Compared with MSNs alone,MSNs combined with menthol could allow deepened and densely distributed surface pores of bEnd.3 cells,increase roughness,reduce Young′s modulus,aggravate F-actin band breakage,and diminish mean gray value,while no significant difference was observed in combination with curcumol.Therefore,it is inferred that the aromatic components can increase the intracellular uptake and transport of nanoparticles by altering the biomechanical properties of bEnd.3 cells.