Open Access, Peer-reviewed
pISSN 2288-6575
eISSN 2288-6796
    J Korean Surg Soc. 2010 Oct;79(4):294-299. Korean.
    Published online Oct 31, 2010.
    https://doi.org/10.4174/jkss.2010.79.4.294
    Copyright © 2010 The Korean Surgical Society
    Original Article

    Effect of Nanoparticle with VEGF in Mouse Ischemic Hindlimb Model

    Sang Hyun Ahn, M.D., Sang-Il Min, M.D., Seong Yup Kim, M.D., Seung-Kee Min, M.D., Han-Kwang Yang, M.D., Sang Joon Kim, M.D. and Jongwon Ha, M.D.
      • Department of Surgery, Seoul National University College of Medicine, Seoul, Korea.
    • Corresponding author (Email: jwhamd@snu.ac.kr )
    Received May 26, 2010; Accepted July 02, 2010.

    Abstract

    Purpose

    Vascular endothelial growth factor (VEGF) is one of the factors regulating angiogenesis. For angiogenesis, the local concentration of VEGF has to be maintained. Because of its short half-life, VEGF has been conjugated with nanoparticles. Some nanoparticles, such as poly (lactic-co-glycolic acid (PLGA)) or polyethylenimine (PEI) are commonly used in this field, but have weak points such as faster release than expected and cell toxicity. We investigated the effect of core/shell nanoparticles including lecithin lipid cores in the ischemic hindlimb model.

    Methods

    Mice were anesthetized and a region of the common femoral artery and vein was ligated and excised. Hindlimb ischemic mice (n=28) were divided randomly into four groups: Control group (normal saline, n=7), mouse VEGF group (mVEGF, n=7), nanoparticle including mVEGF group (N-mVEGF, n=7), and nanoparticle/hydrogel mouse VEGF group (NH-mVEGF, n=7). The drug was injected postoperatively into the thigh muscle of the ischemic limb. Perfusion, capillary number and H&E stain were assessed 28 d after treatment.

    Results

    The capillary number increased in N-mVEGF and mVEGF group (P=0.026). Improvements of ischemic limb perfusion were inferior in N-mVEGF, NH-mVEGF groups (P=0.006) compared to other groups. Mice received N-mVEGF, NH-mVEGF treatment showed significant inflammation in the H&E staining.

    Conclusion

    Sustained VEGF delivery via core/shell nanoparticle with lecithin core did not show improved perfusion rate despite an increase in capillary number. Furthermore, vacuolization and induction of inflammation requiring a different composition of nanoparticle should be tested.

    Keywords
    Hindlimb ischemia; Vascular endothelial growth factor (VEGF); Nanoparticle

    Figures

    Fig. 1
    H&E staining of the muscles on week 4 (A: ×400), and percent of vacuolar degeneration (B). N/S = normal saline; mVEGF = mouse vascular endothelial growth factor; N-mVEGF = nanoparticle mouse vascular endothelial growth factor; NH-mVEGF = nanoparticle/hydrogel mouse vascular endothelial growth factor.

    Fig. 2
    Laser Doppler blood perfusion (A) and blood perfusion (B) in ischemic hind limbs is measured before, just after, on day 3, and at weeks 1, 2, 3 and 4 after right femoral artery ligation.

    Fig. 3
    (A) Cryosections of muscle obtained from the mice at day 28 were stained for alkaline phosphatase. Representative microscopic photographs of capillaries identified by staining of alkaline phosphatase. (B) Quantification of capillary density on the tissue sections. The ratio of the number of capillaries to the number of muscle fiber was measured. N/S = normal saline; mVEGF = mouse vascular endothelial growth factor; N-mVEGF = nanoparticle mouse vascular endothelial growth factor; NH-mVEGF = nanoparticle/hydrogel mouse vascular endothelial growth factor.

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    MeSH Terms
    Animals
    Capillaries
    Extremities
    Femoral Artery
    Half-Life
    Hindlimb
    Inflammation
    Lecithins
    Mice
    Muscles
    Nanoparticles
    Perfusion
    Polyethyleneimine
    Thigh
    Vascular Endothelial Growth Factor A
    Veins
    Metrics
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