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Activation of Angiogenesis and Wound Healing in Diabetic Mice Using NO-Delivery Dinitrosyl Iron Complexes

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    Activation of Angiogenesis and Wound Healing in Diabetic Mice Using NO-Delivery Dinitrosyl Iron Complexes
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    • Yu-Jen Chen
      Yu-Jen Chen
      Department of Biological Science and Technology, Institute of Molecular Medicine and Bioengineering, Center For Intelligent Drug Systems and Smart Bio-devices (IDS2B), National Chiao Tung University, Hsinchu 300, Taiwan
      More by Yu-Jen Chen
    • Shou-Cheng Wu
      Shou-Cheng Wu
      Department of Biological Science and Technology, Institute of Molecular Medicine and Bioengineering, Center For Intelligent Drug Systems and Smart Bio-devices (IDS2B), National Chiao Tung University, Hsinchu 300, Taiwan
      More by Shou-Cheng Wu
    • Hsiang-Ching Wang
      Hsiang-Ching Wang
      Biomedical Technology and Device Research Laboratories, Industrial Technology Research Institute, Hsinchu 310, Taiwan
      More by Hsiang-Ching Wang
    • Tung-Ho Wu
      Tung-Ho Wu
      Division of Cardiovascular Surgery, Department of Surgery and Division of Surgical Critical Care, Department of Critical Care Medicine, Veterans General Hospital, Kaohsiung 813, Taiwan
      More by Tung-Ho Wu
    • Shyng-Shiou F. Yuan*
      Shyng-Shiou F. Yuan
      Translational Research Center and Department of Obstetrics and Gynecology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 807, Taiwan
      *E-mail: [email protected] (S.S.F.Y.).
      More by Shyng-Shiou F. Yuan
    • Tsai-Te Lu*
      Tsai-Te Lu
      Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan
      *E-mail: [email protected] (T.T.L.).
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      Orcidhttp://orcid.org/0000-0001-6977-2568
    • Wen-Feng Liaw*
      Wen-Feng Liaw
      Department of Chemistry, National Tsing Hua University, Hsinchu 300, Taiwan
      *E-mail: [email protected] (W.F.L.).
      More by Wen-Feng Liaw
      Orcidhttp://orcid.org/0000-0001-9949-8344
    • Yun-Ming Wang*
      Yun-Ming Wang
      Department of Biological Science and Technology, Institute of Molecular Medicine and Bioengineering, Center For Intelligent Drug Systems and Smart Bio-devices (IDS2B), National Chiao Tung University, Hsinchu 300, Taiwan
      Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung 807, Taiwan
      *E-mail: [email protected] (Y.M.W.).
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      Orcidhttp://orcid.org/0000-0001-7179-5179
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    Molecular Pharmaceutics

    Cite this: Mol. Pharmaceutics 2019, 16, 10, 4241–4251
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    https://doi.org/10.1021/acs.molpharmaceut.9b00586
    Published August 22, 2019
    Copyright © 2019 American Chemical Society
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    Abstract

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    In diabetes, abnormal angiogenesis due to hyperglycemia and endothelial dysfunction impairs wound healing and results in high risks of diabetic foot ulcers and mortality. Alternative therapeutic methods were attempted to prevent diabetic complications through the activation of endothelial nitric oxide synthase. In this study, direct application of nitric oxide using dinitrosyl iron complexes (DNICs) to promote angiogenesis and wound healing under physiological conditions and in diabetic mice is investigated. Based on in vitro and in vivo studies, DNIC [Fe2(μ-SCH2CH2OH)2(NO)4] (DNIC-1) with a sustainable NO-release reactivity (t1/2 = 27.4 ± 0.5 h at 25 °C and 16.8 ± 1.8 h at 37 °C) activates the NO-sGC-cGMP pathway and displays the best pro-angiogenesis activity overwhelming other NO donors and the vascular endothelial growth factor. Moreover, this pro-angiogenesis effect of DNIC-1 restores the impaired angiogenesis in the ischemic hind limb and accelerates the recovery rate of wound closure in diabetic mice. This study translates synthetic DNIC-1 into a novel therapeutic agent for the treatment of diabetes and highlights its sustainable •NO-release reactivity on the activation of angiogenesis and wound healing.

    Copyright © 2019 American Chemical Society

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    Supporting Information

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.molpharmaceut.9b00586.

    • Interplay among diabetes and associated hyperglycemia (red), chemical compounds reported for pro-angiogenesis (black), upstream signaling pathway for regulation of nitric oxide level (blue), nitric oxide and downstream signaling pathway for angiogenesis and wound healing (green), and the •NO-delivery DNIC used in this study; time-dependent decomposition of DNIC-1 and DNIC-2; Cell proliferation assay of the EA.hy926 cells treated with 20 μM PTIO followed by 7.8 μM DNIC-1, 7.8 μM DNIC-2, 7.8 μM Spermine NONOate, 7.8 μM MAHMA NONOate, and 0.1 μg/mL of VEGF, respectively, for 24 h (***p-value < 0.001 vs PTIO nontreated group); cell proliferation assay of the EA.hy926 cells treated with 7.8 μM Fe3+, 7.8 μM HS(CH2)2OH, 7.8 μM HS(CH2)2NH2, 7.8 μM HS(CH2)2OH/Fe3+, 7.8 μM HS(CH2)2NH2/Fe3+, 7.8 μM degraded DNIC-1, and 7.8 μM degraded DNIC-2, respectively, for 24 h; wound healing assay of the EA.hy926 cells treated with 20 μM PTIO followed by 7.8 μM DNIC-1, 7.8 μM DNIC-2, 7.8 μM Spermine NONOate, 7.8 μM MAHMA NONOate, and 0.1 μg/mL of VEGF, respectively, for 24 h; wound healing assay of the EA.hy926 cells treated with 7.8 μM Fe3+, 7.8 μM HS(CH2)2OH, 7.8 μM HS(CH2)2NH2, 7.8 μM HS(CH2)2OH/Fe3+, 7.8 μM HS(CH2)2NH2/Fe3+, 7.8 μM degraded DNIC-1, and 7.8 μM degraded DNIC-2, respectively, for 24 h; in vitro pro-angiogenesis effect of NO donors and VEGF investigated by tube formation assay; change of body weight of the diabetic mice (db+/db+) treated with DNIC-1 (0.18 mg/kg), VEGF (0.17 μg/kg), or PBS, respectively, via tail-vein injection every 2 days before the mice were sacrificed; representative images of the wound closure in normal mice and diabetic mice treated with PBS; quantitative analysis of wound closure (%); cell proliferation of endothelial cells upon treatment with alternative NO donors and VEGF at different concentrations for 24 h; increased percentage (%) of endothelial junctions, endothelial meshes, and total branching length upon treatment with alternative NO donors and VEGF for 24 h; vascular density index in chicken CAM assay upon treatment with alternative NO donors and VEGF for 48 h; relative SIV length in zebrafish embryos upon treatment with alternative NO donors and VEGF for 24 and 48 h (PDF)

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    Molecular Pharmaceutics

    Cite this: Mol. Pharmaceutics 2019, 16, 10, 4241–4251
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acs.molpharmaceut.9b00586
    Published August 22, 2019
    Copyright © 2019 American Chemical Society
    Request reuse permissions

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