Journal Article

Impact of surface functional group modification on cellular internalization and cytotoxicity of silica nanoparticles

Wakahara, Takatsugu, Sato, Akira, Tran, Lang, Watanabe, Eri, Vranic, Sandra, Terashima, Yuya, Takeuchi, Sakie, Matsushima, Kouji, Boland, Sonja, Wu, Wenting, Ichihara, Sahoko, Zong, Cai, Suzuki, Toshihiro, Abe, Ryo, Osada, Yurika, Ikegami, Akihiko, Hara, Yasushi, Ichihara, Gaku, Yamazaki, Kyoka, Sakurai, Toshihiro, Miyakawa, Kayoko

2025

Overview

Background Silica nanoparticles (SiO 2 NPs) are widely used in industrial products. Surface modification of SiO 2 NPs is one of the promising strategies to develop safer nanomaterials by design. The present study was designed to determine the effects of amino or carboxyl functionalization of rhodamine-labeled SiO 2 NPs on cellular uptake and cytotoxicity. Methods In the in vivo arm of the study, male mice were randomly divided into seven groups (n = 6, each) and exposed to either amino (NH 2 )- or carboxyl (COOH)-functionalized, or non-functionalized (OH)-rhodamine-labeled SiO 2 NPs at 2 or 10 mg/kg bw, or endotoxin-free water as a control, by pharyngeal aspiration. At 24 h after administration, the mice were euthanized and bronchoalveolar lavage fluid (BALF) was collected for differential cell count and assessment of silica nanoparticle uptake using confocal microscopy. In the in vitro arm of the study, murine RAW264.7 macrophages were exposed to NH 2 -or COOH-functionalized or OH- rhodamine-labeled SiO 2 NPs. Nonspecific caspase inhibitor, necroptosis inhibitor, pyroptosis inhibitor and autophagy inhibitor were used to determine the roles of cell death signaling in cytotoxicity. Results The in vivo studies demonstrated significant increase in lung weight at 2 and 10 mg/kg bw by OH-SiO 2 NPs but not the other two SiO 2 NPs. At 10 mg/kg bw, COOH-SiO2NPs induced a significant increase in BALF macrophages, whereas OH- SiO 2 NPs significantly decreased macrophages. OH-SiO 2 NPs at 2 mg/kg bw and NH 2 - and COOH-SiO 2 NPs at 10 mg/kg bw significantly increased BALF neutrophiles. The in vitro studies showed greater NH 2 -SiO 2 NPs internalization into RAW264.7 macrophages than OH-SiO 2 NPs, while OH-SiO 2 NPs induced cytotoxicity and upregulation of IL-1β and TNF-α to greater extent than the other two types. Co-treatment with pan-caspase inhibitor and necroptosis inhibitor attenuated (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) cytotoxicity of OH-SiO 2 NPs. Conclusion NH 2 - or COOH-functionalization reduced the harmful changes observed with OH- SiO 2 NPs, which included increase in lung weight and BALF neutrophils at low dose in mice as well as decrease in cell viability and upregulation of proinflammatory cytokines in RAW264.7 macrophages. The results suggested that OH-SiO 2 NPs-induced cytotoxicity against macrophages was mediated at least in part through apoptotic/necroptotic signaling but was not related to internalization of particles. The results imply possible development of safer silica nanoparticles by amino- or carboxyl-functionalization of their silanols.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Animals / Apoptosis / Autophagy / Autophagy - drug effects / Biocompatibility / Biomedical and Life Sciences / Biomedicine / Bronchoalveolar Lavage Fluid - cytology / Bronchus / Caspase / Caspase inhibitors / Cell death / Cell Survival - drug effects / Cell viability / Confocal microscopy / Cytotoxicity / Endotoxins / Environmental Health / Functional groups / Health aspects / In vivo methods and tests / Industrial products / Inflammation / Internalization / Lavage / Leukocytes (neutrophilic) / Lung - drug effects / Lung - metabolism / Lung - pathology / Lungs / Macrophages / Macrophages - drug effects / Macrophages - metabolism / Male / Medical research / Medicine, Experimental / Mice / Nanomaterials / Nanoparticles / Nanoparticles - chemistry / Nanoparticles - toxicity / Nanotechnology / Necroptosis / Neutrophils / Pharmacology/Toxicology / Pneumology/Respiratory System / Polyethylene glycol / Public Health / Pyroptosis / RAW 264.7 Cells / Rhodamine / Rhodamines - chemistry / Rhodamines - toxicity / Silica / Silicon dioxide / Silicon Dioxide - chemistry / Silicon Dioxide - metabolism / Silicon Dioxide - toxicity / Surface Properties / Toxicity / Tumor necrosis factor-α / Up-regulation / Weight