Hemoglobin-based oxygen carriers (HBOCs) are being developed to overcome limitations associated with transfusion of donated red blood cells (RBCs) such as potential transmission of blood-borne pathogens and limited ex vivo storage shelf-life. Annelid erythrocruorin (Ec) derived from the worm Lumbricus terrestris (Lt) is an acellular mega-hemoglobin that has shown promise as a potential HBOC due to the large size of its oligomeric structure, thus overcoming limitations of unmodified circulating cell-free hemoglobin (Hb). With a large molecular weight of 3.6 MDa compared to 64.5 kDa for human Hb (hHb) and 144 oxygen-binding globin subunits compared to the 4 globin subunits of hHb, LtEc does not extravasate from the circulation to the same extent as hHb. LtEc is stable in the circulation without RBC membrane encapsulation and has a lower rate of auto-oxidation compared to acellular hHb, which allows the protein to remain functional for longer periods of time in the circulation compared to HBOCs derived from mammalian Hbs. Surface coatings, such as poly(ethylene glycol) (PEG) and oxidized dextran (Odex), have been investigated to potentially reduce the immune response and improve the circulation time of LtEc in vivo. Polydopamine (PDA) is a hydrophilic, biocompatible, bioinspired polymer coating used for biomedical nanoparticle assemblies and coatings and has previously been investigated for the surface coating of hHb. PDA is typically synthesized via the self-polymerization of dopamine (DA) under alkaline (pH > 8.0) conditions. However, at pH > 8.0, the oligomeric structure of LtEc begins to dissociate. Therefore, in this study, we investigated a photocatalytic method of PDA polymerization on the surface of LtEc using 9-mesityl-10-methylacridinium tetrafluoroborate (Acr-Mes) to drive PDA polymerization under physiological conditions (pH 7.4, 25 °C) over 2, 5, and 16 h in order to preserve the size and structure of LtEc. The resulting structural, biophysical, and antioxidant properties of PDA surface-coated LtEc (PDA-LtEc) was characterized using various techniques. PDA-LtEc showed an increase in measured particle size, molecular weight, and surface ζ-potential with increasing reaction time from t = 2 to 16 h compared to unmodified LtEc. PDA-LtEc reacted for 16 h was found to have reduced oxygen-binding cooperativity and slower deoxygenation kinetics compared to PDA-LtEc with lower levels of polymerization (t = 2 h), but there was no statistically significant difference in oxygen affinity. The thickness of the PDA coating can be controlled and in turn the biophysical properties can be tuned by changing various reaction conditions. PDA-LtEc was shown to demonstrate an increased level of antioxidant capacity (ferric iron reduction and free-radical scavenging) when synthesized at a reaction time of t = 16 h compared to LtEc. These antioxidant properties may prove beneficial for oxidative protection of PDA-LtEc during its time in the circulation. Hence, we believe that PDA-LtEc is a promising oxygen therapeutic for potential use in transfusion medicine applications.

	Year	Citations

About