Atomic force microscopy study of red blood cell membrane nanostructure during oxidation-reduction processes

Area of Science:

• Biophysics
• Cell Biology
• Hematology

Background:

• Red blood cell (RBC) morphology and function depend on the spectrin network.
• Oxidation processes, induced by factors like UV irradiation, can damage this network.
• Measuring these oxidative changes in RBCs is challenging.

Purpose of the Study:

• To investigate RBC spectrin matrix and morphology changes during UV-induced oxidation using AFM.
• To evaluate the protective effect of Cytoflavin® against UV-induced RBC damage.
• To correlate morphological changes with biochemical markers of oxidation.

Main Methods:

• Atomic Force Microscopy (AFM) for imaging RBC ghosts and spectrin networks.
• Quantitative analysis of spectrin network section sizes.
• Measurement of methaemoglobin levels to assess oxidation intensity.
• Microscopic observation of RBC morphology (discocyte percentage).

Main Results:

• UV irradiation drastically reduced discocytes from 98% to 12%.
• AFM showed significant alterations in the spectrin network structure (section size increased from ~80-200 nm to 600-1000 nm).
• Cytoflavin® treatment post-irradiation maintained discocyte levels at 89% and preserved spectrin network integrity.
• Methaemoglobin levels increased from 1% to 70% during oxidation.

Conclusions:

• AFM is a powerful tool for direct, quantitative observation of RBC spectrin network damage during oxidation.
• UV irradiation causes substantial disruption of RBC morphology and spectrin structure.
• Cytoflavin® demonstrates a significant protective effect on RBCs against oxidative stress.