Synthesis and physicochemical characterization of a series of hemoglobin-based oxygen carriers: Objective comparison between cellular and acellular types

Hiromi Sakai, Minako Yuasa, Hiroto Onuma, Shinji Takeoka, Eishun Tsuchida

    Research output: Contribution to journalArticle

    70 Citations (Scopus)

    Abstract

    A series of hemoglobin (Hb)-based O2 carriers, acellular and cellular types, were synthesized and their physicochemical characteristics were compared. The acellular type includes intramolecularly cross-linked Hb (XLHb), polyoxyethylene (POE)-conjugated pyridoxalated Hb (POE-PLP-Hb), hydroxyethylstarch-conjugated Hb (HES-XLHb), and glutaraldehyde-polymerized XLHb (Poly-XLHb). The cellular type is Hb-vesicles (HbV) of which the surface is modified with POE (POE-HbV). Their particle diameters are 7 ± 2, 22 ± 2, 47 ± 17, 68 ± 24, and 224 ± 76 nm, respectively, thus all the materials penetrate across membrane filters with 0.4 μm pore size, though only the POE-HbV cannot penetrate across the filter with 0.2 μm pore size. These characteristics of permeability are important to consider an optimal particle size in microcirculation in vivo. POE-PLP-Hb ([Hb] = 5 g/dL) showed viscosity of 6.1 cP at 332 s-1 and colloid osmotic pressure (COP) of 70.2 Torr, which are beyond the physiological conditions (human blood, viscosity = 3-4 cP, COP = ca. 25 Torr). XLHb and Poly-XLHb showed viscosities of 1.0 and 1.5 cp, respectively, which are significantly lower than that of blood. COP of POE-HbV is regulated to 20 Torr in 5% human serum albumin (HSA). HES-XLHb and POE-HbV/HSA showed comparable viscosity with human blood. Microscopic observation of human red blood cells (RBC) after mixing blood with POE-PLP-Hb or HES-XLHb disclosed aggregates of RBC, a kind of sludge, indicating a strong interaction with RBC, which is anticipated to modify peripheral blood flow in vivo. On the other hand, XLHb and POE-HbV showed no rouleaux or aggregates of RBC. The acellular Hbs (P50 = 14-32 Torr) have their specific O2 affinities determined by their structures, while that of the cellular POE-HbV is regulated by coencapsulating an appropriate amount of an allosteric effector (e.g., P50 = 18, 32 Torr). These differences in physicochemical characteristics between the acellular and cellular types indicate the advantages of the cellular type from the physiological points of view.

    Original languageEnglish
    Pages (from-to)56-64
    Number of pages9
    JournalBioconjugate Chemistry
    Volume11
    Issue number1
    DOIs
    Publication statusPublished - 2000 Jan

    Fingerprint

    Hemoglobin
    Polyethylene glycols
    Hemoglobins
    Oxygen
    Blood
    Osmotic Pressure
    Colloids
    Erythrocytes
    Viscosity
    Serum Albumin
    Pore size
    Microcirculation
    Blood Viscosity
    Glutaral
    Cellular Structures
    Sewage
    Particle Size
    Permeability
    Particle size
    Cells

    ASJC Scopus subject areas

    • Chemistry(all)
    • Organic Chemistry
    • Clinical Biochemistry
    • Biochemistry, Genetics and Molecular Biology(all)
    • Biochemistry

    Cite this

    Synthesis and physicochemical characterization of a series of hemoglobin-based oxygen carriers : Objective comparison between cellular and acellular types. / Sakai, Hiromi; Yuasa, Minako; Onuma, Hiroto; Takeoka, Shinji; Tsuchida, Eishun.

    In: Bioconjugate Chemistry, Vol. 11, No. 1, 01.2000, p. 56-64.

    Research output: Contribution to journalArticle

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    T1 - Synthesis and physicochemical characterization of a series of hemoglobin-based oxygen carriers

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    AU - Takeoka, Shinji

    AU - Tsuchida, Eishun

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    N2 - A series of hemoglobin (Hb)-based O2 carriers, acellular and cellular types, were synthesized and their physicochemical characteristics were compared. The acellular type includes intramolecularly cross-linked Hb (XLHb), polyoxyethylene (POE)-conjugated pyridoxalated Hb (POE-PLP-Hb), hydroxyethylstarch-conjugated Hb (HES-XLHb), and glutaraldehyde-polymerized XLHb (Poly-XLHb). The cellular type is Hb-vesicles (HbV) of which the surface is modified with POE (POE-HbV). Their particle diameters are 7 ± 2, 22 ± 2, 47 ± 17, 68 ± 24, and 224 ± 76 nm, respectively, thus all the materials penetrate across membrane filters with 0.4 μm pore size, though only the POE-HbV cannot penetrate across the filter with 0.2 μm pore size. These characteristics of permeability are important to consider an optimal particle size in microcirculation in vivo. POE-PLP-Hb ([Hb] = 5 g/dL) showed viscosity of 6.1 cP at 332 s-1 and colloid osmotic pressure (COP) of 70.2 Torr, which are beyond the physiological conditions (human blood, viscosity = 3-4 cP, COP = ca. 25 Torr). XLHb and Poly-XLHb showed viscosities of 1.0 and 1.5 cp, respectively, which are significantly lower than that of blood. COP of POE-HbV is regulated to 20 Torr in 5% human serum albumin (HSA). HES-XLHb and POE-HbV/HSA showed comparable viscosity with human blood. Microscopic observation of human red blood cells (RBC) after mixing blood with POE-PLP-Hb or HES-XLHb disclosed aggregates of RBC, a kind of sludge, indicating a strong interaction with RBC, which is anticipated to modify peripheral blood flow in vivo. On the other hand, XLHb and POE-HbV showed no rouleaux or aggregates of RBC. The acellular Hbs (P50 = 14-32 Torr) have their specific O2 affinities determined by their structures, while that of the cellular POE-HbV is regulated by coencapsulating an appropriate amount of an allosteric effector (e.g., P50 = 18, 32 Torr). These differences in physicochemical characteristics between the acellular and cellular types indicate the advantages of the cellular type from the physiological points of view.

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