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Ahn, Min, Kim, Min, Yang, Kim, and Ha: Effect of Nanoparticle with VEGF in Mouse Ischemic Hindlimb Model
Original Article
Journal of the Korean Surgical Society

Journal of the Korean Surgical Society 2010; 79(4): 294-299.

Published online: 31 October 2010

DOI: https://doi.org/10.4174/jkss.2010.79.4.294

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., Jongwon Ha, M.D.

Department of Surgery, Seoul National University College of Medicine, Seoul, Korea.

Corresponding author (jwhamd@snu.ac.kr)

Received 26 May 2010       Accepted 2 July 2010

Copyright © 2010 The Korean Surgical Society

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 and Tables

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.
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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.
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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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