Augmented neovascularization with magnetized endothelial progenitor cells in rats with hind-limb ischemia

Koiwaya, Hiroshi; Sasaki, Ken-ichiro; Ueno, Takayoshi; Yokoyama, Shinji; Toyama, Yasuyuki; Ohtsuka, Masanori; Nakayoshi, Takaharu; Mitsutake, Yoshiaki; Imaizumi, Tsutomu

doi:10.1016/j.yjmcc.2011.03.012

Cited by 15 publications

(19 citation statements)

References 28 publications

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“…This study conformed to the principles outlined in the Declaration of Helsinki and was approved by the Committees on the Ethics Review Board of Kurume University School of Medicine. We generated EPCs from peripheral blood-derived mononuclear cells (PB-MNCs), as we and others previously reported [9], [19]. In brief, PB-MNCs were isolated by density gradient centrifugation with Ficoll (Ficoll-Paque PLUS, GE Healthcare Bio-Sciences AB) and seeded onto a plate coated with fibronectin (Sigma) and cultured in endothelial basal medium with supplements (EBM-2 BulletKit, Clonetics) and 20% fatal bovine serum (FBS) for 4 days at 37°C, 5% CO 2 , in a humidified incubator.…”

Section: Methodsmentioning

confidence: 99%

“…The siRNA transfection of EPCs with either negative control siRNA (sequence: sense 5′-UAACGACGCGACGACGUAAtt, antisense UUACGUCGUCGCGUCGUUAtt-3′) or FOXO4 siRNA (sequence: sense 5′-CCGCGAUCAUAGACCUAGAtt, antisense UCUAGGUCUAUGAUCGCGGca-3′) (Silencer Select siRNA, Applied Biosystems) was performed using the magnetofection method [19], [20].…”

Section: Methodsmentioning

confidence: 99%

“…After detaching of EPCs from the culture plate, we incubated the cells with antibodies to the following cell surface antigens: CD133 (PE-labeled, Miltenyi Biotec GmbH), CD34, CD31 (PE-labeled, BD Biosciences), and KDR (Biotin conjugated, Sigma; SAv-FITC conjugated, BD Biosciences). The expressions of the antigens were analyzed by a flow cytometer (FACSCanto II, Becton Dickinson) [9], [19].…”

Section: Methodsmentioning

confidence: 99%

“…We assessed oxidative stress and neovascularization of ischemic limbs using a unilateral hindlimb ischemia animal model with 8-to-12-week-old athymic nude rats (F344/N-rnu/rnu, CLEA Japan Inc.) [19]. The proximal portion of the femoral artery and the distal portion of the saphenous artery were occluded using an electrical coagulator (Vetroson V-10 bi-polar electrosurgical unit, Summit Hill Laboratories), and the overlying skin was closed using nylon sutures.…”

Section: Methodsmentioning

confidence: 99%

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FOXO4-Knockdown Suppresses Oxidative Stress-Induced Apoptosis of Early Pro-Angiogenic Cells and Augments Their Neovascularization Capacities in Ischemic Limbs

et al. 2014

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The effects of therapeutic angiogenesis by intramuscular injection of early pro-angiogenic cells (EPCs) to ischemic limbs are unsatisfactory. Oxidative stress in the ischemic limbs may accelerate apoptosis of injected EPCs, leading to less neovascularization. Forkhead transcription factor 4 (FOXO4) was reported to play a pivotal role in apoptosis signaling of EPCs in response to oxidative stress. Accordingly, we assessed whether FOXO4-knockdown EPCs (FOXO4KD-EPCs) could suppress the oxidative stress-induced apoptosis and augment the neovascularization capacity in ischemic limbs. We transfected small interfering RNA targeted against FOXO4 of human EPCs to generate FOXO4KD-EPCs and confirmed a successful knockdown. FOXO4KD-EPCs gained resistance to apoptosis in response to hydrogen peroxide in vitro. Oxidative stress stained by dihydroethidium was stronger for the immunodeficient rat ischemic limb tissue than for the rat non-ischemic one. Although the number of apoptotic EPCs injected into the rat ischemic limb was greater than that of apoptotic EPCs injected into the rat non-ischemic limb, FOXO4KD-EPCs injected into the rat ischemic limb brought less apoptosis and more neovascularization than EPCs. Taken together, the use of FOXO4KD-EPCs with resistance to oxidative stress-induced apoptosis may be a new strategy to augment the effects of therapeutic angiogenesis by intramuscular injection of EPCs.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

FOXO4-Knockdown Suppresses Oxidative Stress-Induced Apoptosis of Early Pro-Angiogenic Cells and Augments Their Neovascularization Capacities in Ischemic Limbs

et al. 2014

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“…In vitro, EPCs can differentiated into endothelial lineage cells, and in animal models of ischemia, EPCs are shown to incorporate into sites of active neovascularization [10,11,12]. Endothelial nitric oxide synthase (eNOS) and its bioactive product nitric oxide (NO) are well-established proangiogenic molecules [13].…”

Section: Introductionmentioning

confidence: 99%

Deterioration of Cardiac Function After Acute Myocardial Infarction is Prevented by Transplantation of Modified Endothelial Progenitor Cells Overexpressing Endothelial NO Synthases

Chen

Zhao

et al. 2013

Cell Physiol Biochem

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Background/Aims: Stem cell transplantation and gene therapies have been shown to attenuate myocardial dysfunction after myocardial infarction (AMI) in different acute and chronic animal models. The aim of this study was to assess the potential therapeutic efficacy of endothelial NO synthases (eNOS)-expressing endothelial progenitor cells (EPCs) on infarcted hearts. Methods: Lentiviral eNOS-infected EPCs were injected after 1 h of ligation of the left anterior descending artery (LAD). The pro-inflammatory cytokines TNF-α and IL-1β levels in cardiac tissue were measured by ELISA at 3 days after transplantation. 28 days post AMI (before sacrifice), Left ventricular function of each group was determined by echocardiography and pressure-volume system. Cardiac tissues were analyzed with hematoxylin and eosin (H&E) staining and immunohistochemisty. eNOS expression in cardiac tissues was detected by western blot, and NO production of cardiac tissues was determined using an NO assay kit. Results: TNF-α and IL-1β levels in cardiac tissue were decreased significantly at 3 days after transplantation of lentiviral with eNOS infected EPCs compared to medium control, eNOS lentiviral vector and normal EPCs. At the 28 day after AMI, echocardiography and hemodynamic measurements, and isolated heart studies showed great therapeutic efficacy in improvement of cardiac function, reduction of infarcted size and improvement of vascular densities in the peri-infarct region after intramyocardial application of lentiviral eNOS-infected EPCs compared to medium control, eNOS lentiviral vector and normal EPCs. The eNOS over-expression in cardiac tissue was observed in the eNOS-EPCs and eNOS lentiviral vector group, and NO levels were increased significantly in the eNOS-EPCs and eNOS lentiviral vector group compared to the other three groups (sham operated group, transplantation of medium or EPCs group). Conclusion: EPCs can be an attractive vehicle for the exogenous eNOS expression into heart after infarction, which is beneficial to prevent deterioration and promote restoration of cardiac function after AMI by improving angiogenesis.

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Silica‐coated magnetic nanoparticles labeled endothelial progenitor cells alleviate ischemic myocardial injury and improve long‐term cardiac function with magnetic field guidance in rats with myocardial infarction

Zhang

Jiang

Chen

et al. 2019

Journal Cellular Physiology

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Low retention of endothelial progenitor cells (EPCs) in the infarct area has been suggested to be responsible for the poor clinical efficacy of EPC therapy for myocardial infarction (MI). This study aimed to evaluate whether magnetized EPCs guided through an external magnetic field could augment the aggregation of EPCs in an ischemia area, thereby enhancing therapeutic efficacy. EPCs from male rats were isolated and labeled with silica‐coated magnetic iron oxide nanoparticles to form magnetized EPCs. Then, the proliferation, migration, vascularization, and cytophenotypic markers of magnetized EPCs were analyzed. Afterward, the magnetized EPCs (1 × 10 6 ) were transplanted into a female rat model of MI via the tail vein at 7 days after MI with or without the guidance of an external magnet above the infarct area. Cardiac function, myocardial fibrosis, and the apoptosis of cardiomyocytes were observed at 4 weeks after treatment. In addition, EPC retention and the angiogenesis of ischemic myocardium were evaluated. Labeling with magnetic nanoparticles exhibited minimal influence to the biological functions of EPCs. The transplantation of magnetized EPCs guided by an external magnet significantly improved the cardiac function, decreased infarction size, and reduced myocardial apoptosis in MI rats. Moreover, enhanced aggregations of magnetized EPCs in the infarcted border zone were observed in rats with external magnet‐guided transplantation, accompanied by the significantly increased density of microvessels and upregulated the expression of proangiogenic factors, when compared with non‐external‐magnet‐guided rats. The magnetic field‐guided transplantation of magnetized EPCs was associated with the enhanced aggregation of EPCs in the infarcted border zone, thereby improving the therapeutic efficacy of MI.

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Augmented neovascularization with magnetized endothelial progenitor cells in rats with hind-limb ischemia

Cited by 15 publications

References 28 publications

FOXO4-Knockdown Suppresses Oxidative Stress-Induced Apoptosis of Early Pro-Angiogenic Cells and Augments Their Neovascularization Capacities in Ischemic Limbs

FOXO4-Knockdown Suppresses Oxidative Stress-Induced Apoptosis of Early Pro-Angiogenic Cells and Augments Their Neovascularization Capacities in Ischemic Limbs

Deterioration of Cardiac Function After Acute Myocardial Infarction is Prevented by Transplantation of Modified Endothelial Progenitor Cells Overexpressing Endothelial NO Synthases

Silica‐coated magnetic nanoparticles labeled endothelial progenitor cells alleviate ischemic myocardial injury and improve long‐term cardiac function with magnetic field guidance in rats with myocardial infarction

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