Hemocompatibility of dextran-graft-polyacrylamide/zinc oxide nanosystems: hemolysis or eryptosis?

Anatolii I Onishchenko; Volodymyr Yu Prokopiuk; Vasyl A Chumachenko; Pavlo A Virych; Liliya Y Tryfonyuk; Nataliya V Kutsevol; Anton S Tkachenko

doi:10.1088/1361-6528/ad02a3

Hemocompatibility of dextran-graft-polyacrylamide/zinc oxide nanosystems: hemolysis or eryptosis?

Nanotechnology. 2023 Oct 30;35(3). doi: 10.1088/1361-6528/ad02a3.

Authors

Anatolii I Onishchenko¹, Volodymyr Yu Prokopiuk^{1

2}, Vasyl A Chumachenko³, Pavlo A Virych³, Liliya Y Tryfonyuk⁴, Nataliya V Kutsevol³, Anton S Tkachenko¹

Affiliations

¹ Research Institute of Experimental and Clinical Medicine, Kharkiv National Medical University, 4 Nauky ave., 61022 Kharkiv, Ukraine.
² Department of Cryobiochemistry, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, 23 Pereyaslavskaya st., 61015 Kharkiv, Ukraine.
³ Department of Chemistry, Taras Shevchenko National University of Kyiv, 60 Volodymyrska st., 01601 Kyiv, Ukraine.
⁴ Institute of Health, National University of Water and Environmental Engineering, 11 Sobornast, 33000 Rivne, Ukraine.

PMID: 37827140
DOI: 10.1088/1361-6528/ad02a3

Abstract

Aim. In this study, blood compatibility of ZnO nanoparticles-polymer nanocomplex (D-PAA/ZnONPs(SO42-)) synthesizedin situinto dextran-graft-polyacrylamide (D-PAA) using zinc sulphate as a precursor was tested using hemolysis, osmotic fragility and eryptosis assays.Materials and methods. Dose-dependent ability to induce eryptosis was assessed following 24 h incubation at concentrations of 0-800 mg l^-1analyzing hallmarks of eryptosis (cell shrinkage and phosphatidylserine externalization), as well as reactive oxygen species generation. Hemolysis was detected spectrophotometrically based on hemoglobin release following exposure to the D-PAA/ZnONPs(SO42-) nanocomplex. Osmotic fragility test (OFT) involved detection of hemolysis of red blood cells exposed to 0.2% saline solution following incubation with the D-PAA/ZnONPs(SO42-) nanocomplex. Additional incubation of the nanocomplex in the presence or absence of either ascorbic acid or EGTA was used to reveal the implication of oxidative stress- or Ca²⁺-mediated mechanisms in D-PAA/ZnONPs(SO42-) nanocomplex-induced erythrotoxicity.Results. Hemocompatibility assessment of the D-PAA/ZnONPs(SO42-) nanocomplex revealed that it induced hemolysis and reduced resistance of erythrocytes to osmotic stress at concentrations of above 400 and 200 mg l^-1, respectively. Oxidative stress- or Ca²⁺-mediated mechanisms were not involved in D-PAA/ZnONPs(SO42-) nanocomplex-induced hemolysis. Strikingly, the D-PAA/ZnONPs(SO42-) nanocomplex did not promote cell membrane scrambling, cell shrinkage and oxidative stress in red blood cells following the direct exposure for 24 h. Thus, the D-PAA/ZnONPs(SO42-) nanocomplex did not induce eryptosisin vitro. Eryptosis is generally considered to occur earlier than hemolysis in response to stress in order to prevent hemolytic cell death. Counterintuitively, our data suggest that hemolysis can be triggered by nanomaterials prior to eryptosis indicating that eryptosis and hemolysis assays should be used in combination for testing blood compatibility of nanomaterials.Conclusions. The D-PAA/ZnONPs(SO42-) nanocomplex has a good hemocompatibility profile at low concentrations. Hemocompatibility testing in nanotoxicology should include both eryptosis and hemolysis assays.

Keywords: cell death; nanoparticles; nanotechnology; reactive oxygen species; regulated cell death.

MeSH terms

Calcium
Cell Death
Dextrans
Eryptosis*
Erythrocytes
Hemolysis
Humans
Oxidative Stress
Reactive Oxygen Species / metabolism
Zinc Oxide* / toxicity

Substances

Zinc Oxide
polyacrylamide
Dextrans
Reactive Oxygen Species
Calcium