Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Apr 4:6:35.
doi: 10.3389/fbioe.2018.00035. eCollection 2018.

Matrix Metalloproteinase-2 Impairs Homing of Intracoronary Delivered Mesenchymal Stem Cells in a Porcine Reperfused Myocardial Infarction: Comparison With Intramyocardial Cell Delivery

Katrin Zlabinger  1 , Dominika Lukovic  1 , Rayyan Hemetsberger  1 , Alfred Gugerell  1 , Johannes Winkler  1 , Ljubica Mandic  1 , Denise Traxler  1 , Andreas Spannbauer  1 , Susanne Wolbank  2 , Gerald Zanoni  2 , Christoph Kaun  1 , Aniko Posa  3 , Andrea Gyenes  3 , Zsolt Petrasi  4 , Örs Petnehazy  4 , Imre Repa  4 , Renate Hofer-Warbinek  5 , Rainer de Martin  5 , Florian Gruber  6 , Silvia Charwat  1 , Kurt Huber  7 , Noemi Pavo  1 , Imre J Pavo  1 , Noemi Nyolczas  1 , Dara L Kraitchman  8 , Mariann Gyöngyösi  1
Affiliations

Matrix Metalloproteinase-2 Impairs Homing of Intracoronary Delivered Mesenchymal Stem Cells in a Porcine Reperfused Myocardial Infarction: Comparison With Intramyocardial Cell Delivery

Katrin Zlabinger et al. Front Bioeng Biotechnol. .

Abstract

Background: Intracoronary (IC) injection of mesenchymal stem cells (MSCs) results in a prompt decrease of absolute myocardial blood flow (AMF) with late and incomplete recovery of myocardial tissue perfusion. Here, we investigated the effect of decreased AMF on oxidative stress marker matrix metalloproteinase-2 (MMP-2) and its influence on the fate and homing and paracrine character of MSCs after IC or intramyocardial cell delivery in a closed-chest reperfused myocardial infarction (MI) model in pigs.

Methods: Porcine MSCs were transiently transfected with Ad-Luc and Ad-green fluorescent protein (GFP). One week after MI, the GFP-Luc-MSCs were injected either IC (group IC, 11.00 ± 1.07 × 106) or intramyocardially (group IM, 9.88 ± 1.44 × 106). AMF was measured before, immediately after, and 24 h post GFP-Luc-MSC delivery. In vitro bioluminescence signal was used to identify tissue samples containing GFP-Luc-MSCs. Myocardial tissue MMP-2 and CXCR4 receptor expression (index of homing signal) were measured in bioluminescence positive and negative infarcted and border, and non-ischemic myocardial areas 1-day post cell transfer. At 7-day follow-up, myocardial homing (cadherin, CXCR4, and stromal derived factor-1alpha) and angiogenic [fibroblast growth factor 2 (FGF2) and VEGF] were quantified by ELISA of homogenized myocardial tissues from the bioluminescence positive and negative infarcted and border, and non-ischemic myocardium. Biodistribution of the implanted cells was quantified by using Luciferase assay and confirmed by fluorescence immunochemistry. Global left ventricular ejection fraction (LVEF) was measured at baseline and 1-month post cell therapy using magnet resonance image.

Results: AMF decreased immediately after IC cell delivery, while no change in tissue perfusion was found in the IM group (42.6 ± 11.7 vs. 56.9 ± 16.7 ml/min, p = 0.018). IC delivery led to a significant increase in myocardial MMP-2 64 kD expression (448 ± 88 vs. 315 ± 54 intensity × mm2, p = 0.021), and decreased expression of CXCR4 (592 ± 50 vs. 714 ± 54 pg/tissue/ml, p = 0.006), with significant exponential decay between MMP-2 and CXCR4 (r = 0.679, p < 0.001). FGF2 and VEGF of the bioluminescence infarcted and border zone of homogenized tissues were significantly elevated in the IM goups as compared to IC group. LVEF increase was significantly higher in IM group (0.8 ± 8.4 vs 5.3 ± 5.2%, p = 0.046) at the 1-month follow up.

Conclusion: Intracoronary stem cell delivery decreased AMF, with consequent increase in myocardial expression of MMP-2 and reduced CXCR4 expression with lower level of myocardial homing and angiogenic factor release as compared to IM cell delivery.

Keywords: cell delivery; homing; intracoronary; intramyocardial; ischemic injured heart tissue; mesenchymal stem cells; oxidative stress; translational research.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Study design of the experiment. MSC, mesenchymal stem cell; AMF, absolute myocardial blood flow; FUP, follow-up; MMP-2, matrix metalloproteinase; GFP, green fluorescent protein; micro CT, micro computed tomography; MRI, magnet resonance image.
Figure 2
Figure 2
In vitro bioluminescence of pig hearts with intracoronary (IC) or intramyocardial delivery of green fluorescent protein (GFP)-Luc-mesenchymal stem cells (MSCs). In vitro bioluminescence images of Luc-transfected porcine MSC show epicardial perivascular location of cells after IC injections (upper right) in contrast to intramyocardial delivery (upper left). Highly positive endocardial punctual signals 24 h after intramyocardial GFP-Luc-MSCs delivery (bottom left). Weak confluent signal of GFP-Luc-MSCs on the endocardial surface of the heart after IC delivery (bottom right).
Figure 3
Figure 3
Oxidative stress and homing signals of the myocardium 24 h post acute myocardial infarction. Zymography (A) and statistical results (B,C) of matrix metalloproteinase-2 (MMP-2), 72 kD and its active form 64 kD expression of the myocardium 1 day after intramyocardial or IC green fluorescent protein (GFP)-Luc-mesenchymal stem cell (MSCs) cell delivery in different location. IM, intramyocardial delivery; IC, intracoronary delivery; NIM, non-ischemic myocardium (remote posterior wall), border, border zone of infarction; MI, infarcted area; (D) CXCR4 expression in the myocardial tissues 24 h after cell treatment. (E) Exponential decay between MMP-2 and CXCR4. (F) Logarithmic correlation between MMP-2 64 kD and luciferase activity (index of number of GFP-Luc-MSCs).
Figure 4
Figure 4
Expression of homing and angiogenic signals of the myocardium 7 days after cardiac transfer of green fluorescent protein (GFP)-Luc-mesenchymal stem cell (MSCs). Fluorescent immunohistochemistry of the bioluminescence positive myocardial areas 7 days after intramyocardial [left panel (A,C,E,G)] or intracoronary [right panel (B,D,F,H)] GFP-Luc-MSCs delivery shows increased expression of homing signals cadherin (A,B), and angiogenic factors fibroblast growth factor 2 (FGF2) (C,D) and vascular endothelial growth factor (VEGF) (E,F) in group IM. Infarct area border zone (G,H) exhibited higher number of myocardial cells and higher level of VEGF expression in group IM (G). Hoechst staining of the nuclei, 40× magnification. Expression of homing signals cadherin (I), stromal-derived factor-1alpha (J), and angiogenic factors FGF2 (K) and VEGF (L).
Figure 5
Figure 5
Time-dependent biodistribution of the intramyocardial and intracoronary (IC) delivered green fluorescent protein (GFP)-Luc-mesenchymal stem cell (MSCs). GFP+ positive cells in the infarcted heart tissue (A,B), lung (C,D), mediastinal lymph node (E,F), liver (G,H), and spleen (I,J) 7 days after intramyocardial (left panel, group IM) or IC (right panel, group IC) delivery of GFP-Luc-MSCs. Time response of luciferase activity is shown in the graphs on the right side.
Figure 6
Figure 6
MicroCT of the infarcted and green fluorescent protein-Luc-mesenchymal stem cell (MSCs)-treated hearts. Microvascularization in the infarcted area (yellow arrow) 1 month after intramyocardial (group IM) or intracoronary (group IC) delivery of MSCs. Repeated micro CT image focusing on the anterior apical area of infarction (arrow).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Aarnoudse W., Van’t Veer M., Pijls N. H., Ter Woorst J., Vercauteren S., Tonino P., et al. (2007). Direct volumetric blood flow measurement in coronary arteries by thermodilution. J. Am. Coll. Cardiol. 50, 2294–2304.10.1016/j.jacc.2007.08.047 - DOI - PubMed
    1. Bassiouny H. S., Song R. H., Hong X. F., Singh A., Kocharyan H., Glagov S. (1998). Flow regulation of 72-kD collagenase IV (MMP-2) after experimental arterial injury. Circulation 98, 157–163.10.1161/01.CIR.98.2.157 - DOI - PubMed
    1. Ben-Haim S. A., Osadchy D., Schuster I., Gepstein L., Hayam G., Josephson M. E. (1996). Nonfluoroscopic, in vivo navigation and mapping technology. Nat. Med. 2, 1393–1395.10.1038/nm1296-1393 - DOI - PubMed
    1. Bussche L., Van de Walle G. R. (2014). Peripheral blood-derived mesenchymal stromal cells promote angiogenesis via paracrine stimulation of vascular endothelial growth factor secretion in the equine model. Stem Cells Transl. Med. 3, 1514–1525.10.5966/sctm.2014-0138 - DOI - PMC - PubMed
    1. Chavakis E., Urbich C., Dimmeler S. (2008). Homing and engraftment of progenitor cells: a prerequisite for cell therapy. J. Mol. Cell. Cardiol. 45, 514–522.10.1016/j.yjmcc.2008.01.004 - DOI - PubMed

LinkOut - more resources