Ruijter Nienke, Zanoni Ilaria, Persson Daniel, Arts Josje, Carriere Marie, Guiot Arnaud, Persson Michael, Katsumiti Alberto, Marshall Jessica, Boyles Matthew, Cassee Flemming R, Braakhuis Hedwig
National Institute for Public Health & The Environment (RIVM), Bilthoven, 3721 MA, The Netherlands.
Institute of Science, Technology and Sustainability for Ceramics, CNR-ISSMC, National Research Council of Italy, Faenza, 48018, Italy.
Part Fibre Toxicol. 2025 May 26;22(1):15. doi: 10.1186/s12989-025-00629-6.
The Safe and Sustainable by Design (SSbD) concept facilitates the design of safer and more sustainable chemicals and materials and is a crucial approach towards reaching the goals set out in the European Green Deal. It is critical that suitable guidance is provided on how to use new approach methodologies (NAMs) to fill hazard data gaps for nanomaterials (NMs) to facilitate SSbD decisions. Here, we showcase a nano-specific in vitro SSbD case study. The five colloidal silica nanoforms (SiO-NFs) under investigation in this study are surface modified with varying amounts of glycerolpropyl-organosilane groups. In this study, we use a simple yet comprehensive in vitro test battery along with thorough particle characterization to investigate the effect of surface silanization on in vitro toxicity to inform SSbD decisions.
Cytotoxic, pro-inflammatory and oxidative stress responses in A549, dTHP-1, and BEAS-2B cells after exposure to SiO-NFs submerged and at the air-liquid interface (ALI) decreased with increasing silane surface modification. None of the SiO-NFs showed surface reactivity or haemolytic potential. Deposition assessment using inductively coupled plasma - optical emission spectrometry (ICP-OES) revealed that increasing silane surface modification decreased particle settling. The two SiO-NFs with the highest amount of surface silanization did not reach the cells in a submerged exposure setting, and they were therefore only tested at the ALI. Identical dose-response curves were observed for both the submerged testing and testing at the ALI for the SiO-NFs without and with low/intermediate surface functionalization, again showing a decrease in effects with increasing surface functionalization.
We show that in vitro toxicity assays provide valuable information for SSbD decision making. In vitro cytotoxic, pro-inflammatory and oxidative stress responses can be reduced with increasing surface silane functionalization. The reduced deposition efficiency with increasing silane functionalization, however, highlights that thorough characterization of particle behaviour in cell culture medium should always be performed for SSbD hazard testing. The amount of silane required to reduce toxicity is important information for the future production of safer SiO-NFs and nano-enabled products. Exposure, functionality, and sustainability remain to be investigated to draw full SSbD conclusions.
“设计中确保安全与可持续性”（SSbD）概念有助于设计更安全、更可持续的化学品和材料，是实现欧洲绿色协议所设定目标的关键途径。至关重要的是，要提供关于如何使用新方法学（NAMs）来填补纳米材料（NMs）危害数据空白以促进SSbD决策的适当指导。在此，我们展示一个针对纳米材料的体外SSbD案例研究。本研究中所研究的五种胶体二氧化硅纳米形态（SiO-NFs）用不同量的甘油丙基有机硅烷基团进行了表面改性。在本研究中，我们使用一个简单但全面的体外测试组以及全面的颗粒表征来研究表面硅烷化对体外毒性的影响，以为SSbD决策提供信息。
暴露于浸没状态及气液界面（ALI）的SiO-NFs后，A549、dTHP-1和BEAS-2B细胞中的细胞毒性、促炎和氧化应激反应随着硅烷表面改性的增加而降低。没有一种SiO-NFs表现表面具有表面具有反应性或溶血潜力。使用电感耦合等离子体发射光谱法（ICP-OES）进行的沉积评估表明，硅烷表面改性增加会降低颗粒沉降。表面硅烷化程度最高的两种SiO-NFs在浸没暴露环境中未到达细胞，因此仅在ALI条件下进行了测试。对于未进行表面功能化以及具有低/中等表面功能化的SiO-NFs，浸没测试和ALI测试观察到相同的剂量反应曲线，再次表明随着表面功能化增加效应降低。
我们表明体外毒性测定为SSbD决策提供了有价值的信息。随着表面硅烷功能化增加，体外细胞毒性、促炎和氧化应激反应会降低。然而，随着硅烷功能化增加沉积效率降低，这突出表明在进行SSbD危害测试时，应始终对细胞培养基中颗粒行为进行全面表征。降低毒性所需的硅烷量是未来生产更安全的SiO-NFs和含纳米材料产品的重要信息。要得出完整的SSbD结论，暴露、功能和可持续性仍有待研究。
