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Fluorescently Labeled Cellulose Nanofibers for Environmental Health and Safety Studies
Ilabahen Patel1, Jeremiah Woodcock2, Ryan Beams2
1Department of Chemistry, American University, Washington, DC 20016, USA.
Nanomaterials (Basel, Switzerland)
|April 30, 2021
Summary
A new fluorescent probe, mDTEB, effectively labels cellulose nanofibers (CNFs) for tracking in biological studies. This method confirmed CNFs are non-toxic in both in vitro and in vivo environmental health and safety assessments.
Area of Science:
- Nanomaterial science
- Environmental toxicology
- Biomedical imaging
Background:
- Cellulose nanofibers (CNFs) are increasingly used, necessitating methods to assess their environmental health and safety (EHS).
- Tracking CNFs in biological systems is essential for understanding their fate and potential toxicity.
- Existing methods for CNF tracking are limited, especially for in vitro and in vivo studies.
Purpose of the Study:
- To develop and validate a novel fluorescent probe for labeling CNFs.
- To confirm the covalent attachment, purity, and photoluminescence properties of labeled CNFs.
- To evaluate the in vitro and in vivo toxicity of CNFs using the developed labeling method.
Main Methods:
- Synthesis and application of a new boron-dipyrromethene (BODIPY) reactive fluorescent probe, meso-DichlorotriazineEthyl BODIPY (mDTEB).
- Time-correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) for confirming labeling and purity.
- Fluorescence spectroscopy to characterize photoluminescence properties (pH stability, quantum yield).
- In vitro triculture small intestinal epithelial model and in vivo zebrafish (Danio rerio) embryo studies for toxicity assessment.
Main Results:
- mDTEB successfully labeled CNFs with confirmed covalent attachment and high purity via FLIM.
- mDTEB-labeled CNFs exhibited excellent photoluminescence stability across a wide pH range (2-10) and high quantum yield for low-concentration detection.
- Lignin-like impurities on CNFs were found to quench mDTEB fluorescence, highlighting the impact of CNF composition.
- No cytotoxicity was observed for either unlabeled or mDTEB-labeled CNFs in both the in vitro triculture model and in vivo zebrafish embryos.
Conclusions:
- The mDTEB probe provides an optimal methodology for locating and tracking CNFs in biological systems.
- CNF purity, influenced by production methods, affects fluorescence detection and requires consideration in EHS studies.
- Cellulose nanofibers, labeled or unlabeled, demonstrate a lack of cytotoxicity, supporting their safe use in various applications.
