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Updated Method for In Vitro Analysis of Nanoparticle Hemolytic Properties
Barry W Neun1, Anna N Ilinskaya1, Marina A Dobrovolskaia2
1Cancer Research Technology Program, Nanotechnology Characterization Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, P.O. Box B, Frederick, MD, 21702, USA.
Methods in Molecular Biology (Clifton, N.J.)
|October 18, 2017
Summary
This study revises an in vitro assay for nanoparticle-induced red blood cell (RBC) damage, measuring hemoglobin release. The updated protocol refines nanoparticle preparation and controls for accurate hemolysis analysis.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Hematology
Background:
- Hemolysis, or red blood cell (RBC) damage, releases hemoglobin into plasma.
- An existing in vitro assay analyzes nanoparticle hemolytic properties.
- A revised protocol is presented to improve RBC damage assessment.
Purpose of the Study:
- To provide an updated and refined in vitro protocol for evaluating nanoparticle-induced hemolysis.
- To enhance the accuracy and reliability of assessing nanoparticle interactions with RBCs.
Main Methods:
- Nanoparticles and controls are incubated with blood.
- Red blood cells (RBCs) are separated post-incubation via centrifugation.
- Released hemoglobin is quantified using Drabkin's reagent and spectrophotometry.
Main Results:
- The revised protocol includes updated nanoparticle sample preparation and concentration selection.
- Enhanced details on assay controls and potential nanoparticle interference are provided.
- The method quantifies nanoparticle-induced hemolysis as a percentage of total hemoglobin.
Conclusions:
- The revised in vitro hemolysis assay offers improved methodology for nanoparticle safety assessment.
- Accurate quantification of nanoparticle-induced RBC damage is crucial for material safety evaluations.
- This updated protocol facilitates reliable analysis of nanoparticle hemolytic activity.