Yiming Sun, Ailan Huang, Zhi Zhao, Chen Song, Guihua Lai
{"title":"[Immunogenic and toxic effects of graphene oxide nanoparticles in mouse skeletal muscles and human red blood cells].","authors":"Yiming Sun, Ailan Huang, Zhi Zhao, Chen Song, Guihua Lai","doi":"10.12122/j.issn.1673-4254.2024.04.01","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To investigate immunogenic and toxic effects of graphene oxide (GO) nanoparticles in mouse skeletal muscles and in human blood <i>in vitro</i>.</p><p><strong>Methods: </strong>GO nanoparticles prepared using a probe sonicator were supended in deionized H<sub>2</sub>O or PBS, and particle size and surface charge of the nanoparticles were measured with dynamic light scattering (DLS). Different concentrations (0.5, 1.0 and 2.0 mg/mL) of GO suspension or PBS were injected at multiple sites in the gastrocnemius muscle (GN) of C57BL/6 mice, and inflammatory response and immune cell infiltrations were detected with HE and immunofluorescence staining. We also examined the effects of GO nanoparticles on human red blood cell (RBC) morphology, hemolysis and blood coagulation using scanning electron microscope (SEM), spectrophotometry, and thromboelastography (TEG).</p><p><strong>Results: </strong>GO nanoparticles suspended in PBS exhibited better colloidal dispersity, stability and surface charge effects than those in deionized H<sub>2</sub>O. In mouse GNs, injection of GO suspensions dose- and time-dependently resulted in sustained muscular inflammation and myofiber degeneration at the injection sites, which lasted till 8 weeks after the injection; immunofluorescence staining revealed obvious infiltration of monocytes, macrophages, dendritic cells and CD4<sup>+</sup> T cells around the injection sites in mouse GNs. In human RBCs, incubation with GO suspensions at 0.2, 2.0 and 20 mg/mL, but not at 0.002 or 0.02 mg/mL, caused significant alterations of cell morphology and hemolysis. TEG analysis showed significant abnormalities of blood coagulation parameters following treatment with high concentrations of GO.</p><p><strong>Conclusion: </strong>GO nanoparticles can induce sustained inflammatory and immunological responses in mouse GNs and cause RBC hemolysis and blood coagulation impairment, suggesting its muscular toxicity and hematotoxicity at high concentrations.</p>","PeriodicalId":18962,"journal":{"name":"Nan fang yi ke da xue xue bao = Journal of Southern Medical University","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11073950/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Nan fang yi ke da xue xue bao = Journal of Southern Medical University","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.12122/j.issn.1673-4254.2024.04.01","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 0
Abstract
Objective: To investigate immunogenic and toxic effects of graphene oxide (GO) nanoparticles in mouse skeletal muscles and in human blood in vitro.
Methods: GO nanoparticles prepared using a probe sonicator were supended in deionized H2O or PBS, and particle size and surface charge of the nanoparticles were measured with dynamic light scattering (DLS). Different concentrations (0.5, 1.0 and 2.0 mg/mL) of GO suspension or PBS were injected at multiple sites in the gastrocnemius muscle (GN) of C57BL/6 mice, and inflammatory response and immune cell infiltrations were detected with HE and immunofluorescence staining. We also examined the effects of GO nanoparticles on human red blood cell (RBC) morphology, hemolysis and blood coagulation using scanning electron microscope (SEM), spectrophotometry, and thromboelastography (TEG).
Results: GO nanoparticles suspended in PBS exhibited better colloidal dispersity, stability and surface charge effects than those in deionized H2O. In mouse GNs, injection of GO suspensions dose- and time-dependently resulted in sustained muscular inflammation and myofiber degeneration at the injection sites, which lasted till 8 weeks after the injection; immunofluorescence staining revealed obvious infiltration of monocytes, macrophages, dendritic cells and CD4+ T cells around the injection sites in mouse GNs. In human RBCs, incubation with GO suspensions at 0.2, 2.0 and 20 mg/mL, but not at 0.002 or 0.02 mg/mL, caused significant alterations of cell morphology and hemolysis. TEG analysis showed significant abnormalities of blood coagulation parameters following treatment with high concentrations of GO.
Conclusion: GO nanoparticles can induce sustained inflammatory and immunological responses in mouse GNs and cause RBC hemolysis and blood coagulation impairment, suggesting its muscular toxicity and hematotoxicity at high concentrations.
目的研究氧化石墨烯(GO)纳米颗粒在小鼠骨骼肌和人体血液中的体外免疫原性和毒性效应:用探针声波发生器制备 GO 纳米粒子,将其置于去离子水或 PBS 中,用动态光散射法(DLS)测量纳米粒子的粒径和表面电荷。在 C57BL/6 小鼠腓肠肌(GN)的多个部位注射不同浓度(0.5、1.0 和 2.0 mg/mL)的 GO 悬浮液或 PBS,用 HE 和免疫荧光染色法检测炎症反应和免疫细胞浸润。我们还使用扫描电子显微镜(SEM)、分光光度计和血栓弹力图(TEG)研究了 GO 纳米粒子对人类红细胞(RBC)形态、溶血和血液凝固的影响:结果:与去离子水相比,悬浮在 PBS 中的 GO 纳米粒子具有更好的胶体分散性、稳定性和表面电荷效应。在小鼠GNs中,注射GO悬浮液可导致注射部位持续的肌肉炎症和肌纤维变性,这种情况在注射后持续8周;免疫荧光染色显示小鼠GNs注射部位周围有明显的单核细胞、巨噬细胞、树突状细胞和CD4+T细胞浸润。在人红细胞中,与 0.2、2.0 和 20 毫克/毫升(而不是 0.002 或 0.02 毫克/毫升)的 GO 悬浮液一起孵育会导致细胞形态和溶血的显著改变。TEG分析表明,使用高浓度的GO处理后,血液凝固参数出现明显异常:结论:GO 纳米粒子可诱导小鼠 GN 产生持续的炎症和免疫反应,并导致 RBC 溶血和血液凝固障碍,这表明高浓度 GO 纳米粒子具有肌肉毒性和血液毒性。