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Structural elucidation of cell membrane-derived nanoparticles using molecular probes.
Zhiyuan Fan1, Hao Zhou, Peter Y Li
1Department of Materials Science and Engineering, Drexel University, LeBow 332B, 3141 Chestnut Street, Philadelphia, PA 19104, USA. hcheng@drexel.edu.
Journal of Materials Chemistry. B
|April 9, 2020
Summary
Researchers developed a new method to quantitatively assess cell membrane-derived nanoparticle (CNP) orientation. This technique revealed red blood cell membrane-derived nanoparticles (RBC-NPs) exhibit 84% correct orientation and high permeability, crucial for medical applications.
Area of Science:
- Biomaterials Science
- Nanotechnology
- Cell Biology
Background:
- Cell membrane-derived nanoparticles (CNPs) offer advantages over synthetic nanomaterials due to their biological origin.
- Understanding the fundamental structure, particularly membrane orientation, of CNPs is crucial for optimizing their pharmacokinetic properties.
- Previous characterizations of CNP membrane orientation have been largely qualitative.
Purpose of the Study:
- To develop and validate a quantitative method for assessing the membrane orientation of CNPs.
- To determine the membrane orientation and permeability of red blood cell membrane-derived nanoparticles (RBC-NPs).
Main Methods:
- Utilized a Förster Resonance Energy Transfer (FRET) based approach employing a 6-FAM ssDNA probe and a BHQ1 ssDNA quencher.
- Conjugated ssDNA probes to cell membranes to specifically mark the outer leaflet.
- Quantified CNP orientation by measuring fluorescence changes upon quencher addition and assessed RBC-NP permeability using NBD quenching with dithionite ions.
Main Results:
- Fabricated RBC-NPs demonstrated a high correct outside-out (right-side-out) membrane orientation of 84%.
- RBC-NPs exhibited significantly higher permeability to dithionite ions compared to live cells and liposomes.
- The FRET-based method provides a quantitative and broadly applicable tool for CNP characterization.
Conclusions:
- The developed FRET method enables precise quantitative analysis of CNP membrane orientation.
- RBC-NPs possess favorable structural and permeability characteristics for potential biomedical applications.
- This approach facilitates direct comparisons and optimization of CNPs derived from various cell sources.
