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. 2023 Mar 16;8(12):11447-11456.
doi: 10.1021/acsomega.3c00275. eCollection 2023 Mar 28.

Hydroxyethyl Starch-Bovine Hemoglobin Conjugate as an Effective Oxygen Carrier with the Ability to Expand Plasma

Wenying Yan  1   2 , Ming Sheng  3 , Weili Yu  1 , Lijuan Shen  1 , Jinming Qi  1 , Hong Zhou  3 , Tao Hu  1 , Lian Zhao  3
Affiliations

Hydroxyethyl Starch-Bovine Hemoglobin Conjugate as an Effective Oxygen Carrier with the Ability to Expand Plasma

Wenying Yan et al. ACS Omega. .

Abstract

Hemorrhagic shock leads to intravasal volume deficiency, tissue hypoxia, and cellular anaerobic metabolism. Hemoglobin (Hb) could deliver oxygen for hypoxic tissues but is unable to expand plasma. Hydroxyethyl starch (HES) could compensate for the intravasal volume deficiency but cannot deliver oxygen. Thus, bovine Hb (bHb) was conjugated with HES (130 kDa and 200 kDa) to develop an oxygen carrier with the ability to expand plasma. Conjugation with HES increased the hydrodynamic volume, colloidal osmotic pressure, and viscosity of bHb. It slightly perturbed the quaternary structure and heme environment of bHb. The partial oxygen pressures at 50% saturation (P 50) of the two conjugates (bHb-HES130 and bHb-HES200) were 15.1 and 13.9 mmHg, respectively. The two conjugates showed no apparent side effects on the morphology and rigidity, hemolysis, and platelet aggregation of red blood cells of Wistar rats. Thus, bHb-HES130 and bHb-HES200 were expected to function as an effective oxygen carrier with the ability to expand plasma.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Analysis of bHb-HES130 and bHb-HES200. The conjugates were analyzed by SDS-PAGE (a). Lanes 1 and 5 - marker, Lane 2 - bHb, Lane 3 - bHb-HES130, and Lane 4 - bHb-HES200. The conjugates were analyzed by dynamic light scattering (b) and a Superose 6 column (c). Values represent the mean value ± SD from three repeated measurements. The thiol reactivities of the conjugates (d) were estimated by measuring the conversion of 4-PDS to 4-thiopyridone at 324 nm as a function of time.
Figure 2
Figure 2
Structural characterization bHb-HES130 and bHb-HES200. The circular dichroism spectra (a), extrinsic fluorescence spectra (b), UV–vis spectra (c), and FT-IR spectra (d) of the conjugates were recorded.
Figure 3
Figure 3
Colloidal osmotic pressure and viscosity of bHb-HES130 and bHb-HES200. The colloidal osmotic pressure (a) was measured using Osmocoll reference standards. The viscosity (b) was measured at a shear rate of 100 s–1. Values represent the mean value ± SD from three repeated measurements.
Figure 4
Figure 4
Morphology of RBCs in the presence of different samples. The RBCs were incubated with normal saline (a), bHb (b), HES130 (c), HES200 (d), bHb-HES130 (e), and bHb-HES200 (f), respectively.
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