Patel Ilabahen, Woodcock Jeremiah, Beams Ryan, Stranick Stephan J, Nieuwendaal Ryan, Gilman Jeffrey W, Mulenos Marina R, Sayes Christie M, Salari Maryam, DeLoid Glen, Demokritou Philip, Harper Bryan, Harper Stacey, Ong Kimberly J, Shatkin Jo Anne, Fox Douglas M
Department of Chemistry, American University, Washington, DC 20016, USA.
Materials Science and Engineering Division, Materials Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.
Nanomaterials (Basel). 2021 Apr 15;11(4):1015. doi: 10.3390/nano11041015.
An optimal methodology for locating and tracking cellulose nanofibers (CNFs) in vitro and in vivo is crucial to evaluate the environmental health and safety properties of these nanomaterials. Here, we report the use of a new boron-dipyrromethene (BODIPY) reactive fluorescent probe, meso-DichlorotriazineEthyl BODIPY (mDTEB), tailor-made for labeling CNFs used in simulated or in vivo ingestion exposure studies. Time-correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) was used to confirm covalent attachment and purity of mDTEB-labeled CNFs. The photoluminescence properties of mDTEB-labeled CNFs, characterized using fluorescence spectroscopy, include excellent stability over a wide pH range (pH2 to pH10) and high quantum yield, which provides detection at low (μM) concentrations. FLIM analysis also showed that lignin-like impurities present on the CNF reduce the fluorescence of the mDTEB-labeled CNF, via quenching. Therefore, the chemical composition and the methods of CNF production affect subsequent studies. An in vitro triculture, small intestinal, epithelial model was used to assess the toxicity of ingested mDTEB-labeled CNFs. Zebrafish () were used to assess in vivo environmental toxicity studies. No cytotoxicity was observed for CNFs, or mDTEB-labeled CNFs, either in the triculture cells or in the zebrafish embryos.
一种用于在体外和体内定位与追踪纤维素纳米纤维（CNFs）的最佳方法，对于评估这些纳米材料的环境健康与安全特性至关重要。在此，我们报告了一种新型硼二吡咯亚甲基（BODIPY）反应性荧光探针，即中-二氯三嗪乙基BODIPY（mDTEB）的使用情况，该探针是为标记用于模拟或体内摄入暴露研究的CNFs而量身定制的。采用时间相关单光子计数（TCSPC）荧光寿命成像显微镜（FLIM）来确认mDTEB标记的CNFs的共价连接和纯度。使用荧光光谱对mDTEB标记的CNFs的光致发光特性进行表征，结果表明其在较宽的pH范围（pH2至pH10）内具有出色的稳定性和高量子产率，这使得能够在低（μM）浓度下进行检测。FLIM分析还表明，CNF上存在的类木质素杂质通过淬灭作用降低了mDTEB标记的CNF的荧光。因此，CNF的化学成分和生产方法会影响后续研究。使用体外三培养小肠上皮模型来评估摄入的mDTEB标记的CNFs的毒性。利用斑马鱼来评估体内环境毒性研究。在三培养细胞或斑马鱼胚胎中，均未观察到CNFs或mDTEB标记的CNFs具有细胞毒性。
