Korean Circ J.  2012 Jun;42(6):390-396. 10.4070/kcj.2012.42.6.390.

Magnetic Bionanoparticle Enhances Homing of Endothelial Progenitor Cells in Mouse Hindlimb Ischemia

• Kang HJ
• Kim JY
• Lee HJ
• Kim KH
• Kim TY
• Lee CS
• Lee HC
• Park TH
• Kim HS
• Park YB

• KMID: 1433858
• DOI: http://doi.org/10.4070/kcj.2012.42.6.390

Abstract

BACKGROUND AND OBJECTIVES
Poor homing efficiency is one of the major limitations of current stem cell therapy. Magnetic bionanoparticles (MPs) obtained from Magnetospirillum sp. AMB-1 have a lipid bilayer membrane and ferromagnetic properties. We evaluated a novel priming strategy using MPs to enhance the homing of transplanted progenitor cells to target tissue.
MATERIALS AND METHODS
Effects of MP on proliferation, viability, and migration of late human endothelial progenitor cells (EPCs) were examined in vitro. Additionally, effects of MP on gene and protein expression related to survival and adhesion were evaluated. Homing and angiogenic efficiency of MP transferred late EPCs was evaluated in nude mouse hindlimb ischemia model.
RESULTS
Below threshold concentration, MP transfer did not influence proliferation or survival of late EPCs, but enhanced migration and trans-endothelial migration of late EPCs toward magnet. Below threshold concentration, MP transfer did not influence gene and protein expression related to survival. In the mouse hindlimb ischemia model, late EPCs treated with high dose MP (5 ug/mL) showed enhanced homing of injected late EPCs in the ischemic limb by magnet, compared to low dose MP (1 ug/mL) treated late EPCs. In addition, high dose MP transferred EPC showed significantly better improvement of perfusion in ischemic limb compared to untreated EPC.
CONCLUSION
MP transfer with magnet application can be a promising novel strategy to enhance homing efficacy and outcomes of current stem cell therapy.

Keyword

Nanoparticles; Stem cells; Ischemia

MeSH Terms

Animals
Extremities
Hindlimb
Humans
Ischemia
Lipid Bilayers
Magnetics
Magnetospirillum
Magnets
Membranes
Mice
Mice, Nude
Nanoparticles
Perfusion
Phosphorylcholine
Stem Cells
Transplants
Lipid Bilayers
Phosphorylcholine

Figure

• 

  Fig. 1 Transfection of magnetic bionanoparticle. A: electron microscopic image of endothelial progenitor cell (EPC) which was transfected with magnetic bionanoparticles (MPs). B: transfection efficacy to EPCs was proportionally increased with concentration of MP, which was assessed by Prussian blue staining. C: viability of EPCs was significantly decreased at above 10 ug/mL of MPs, which was assessed by tryphan blue exclusion assay. D: proliferation of EPC was not influenced by MP transfection below threshold concentration (10 ug/mL) in normoxic and hypoxia-reoxygenation condition. There were no significant differences among groups (n=3 respectively). Error bars represented standard deviation. *p<0.05 compared with no MP group (n=3 respectively).

• 

  Fig. 2 Transfection of MP did not influence the gene and protein expression of key signaling and surface molecules related to survival and angiogenesis in normoxia and hypoxia-reoxygenation condition. A: reverse transcript-polymerase chain reaction. B: western blot (each experiments were repeated twice respectively and representative figures were presented). M: marker, MP: magnetic bionanoparticle, ICAM: intercellular adhesion mdecule, VCAM: vascular cell adhesion molecule, SDF: stromal cell-derived factor.

• 

  Fig. 3 Migration of EPC was enhanced by MP transfection with magnet apply. A and C: scratch wound assay: MP was transfected with concentration of 1 ug/mL and magnet was set in cell migrating direction (n=3 respectively). B and D: transendothelial migration assay: magnetic force enhanced MP transfected EPC transmigration through human umbilical vein endothelial cell monolayer: transmigrated cells at bottom of the porous membrane in the vehicle and MPs group (B). EPC were labeled CFSE before co-incubation (n=5 respectively). Error bars represented standard deviation. EPC: endothelial progenitor cell, MP: magnetic bionanoparticle, CFSE: Carboxyfluoroscein Diacetate Succinimidyl Ester.

• 

  Fig. 4 Therapeutic efficacy of MP-transfected hEPCs. A and B: in an athymic mouse hindlimb ischemia model, EPCs transfected with MPs showed enhanced homing into ischemic limb in dose dependent manner (n=3 respectively). C: high dose MP (10 ug/mL) and magnet application significantly improved perfusion of ischemic limb compared to control group: MP untreated group without magnet application (n=5 respectively). Error bars represented standard deviation. MP: magnetic bionanoparticle, EPC: endothelial progenitor cell.

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