Korean Circ J.  2008 Oct;38(10):536-543. 10.4070/kcj.2008.38.10.536.

Human Mesenchymal Stem Cell Transplantation Induces Sympathetic Nerve Sprouting and Reduces the Gap Junction With Potential Proarrhythmias in Dogs

• Kim SK
• Pak HN
• Kim GI
• Park JH
• Fang YF
• Lim HE
• Kim BS
• Hwang C
• Kim YH

• KMID: 2225718
• DOI: http://doi.org/10.4070/kcj.2008.38.10.536

Abstract

BACKGROUND AND OBJECTIVES
Although human mesenchymal stem cell (hMSC) transplantation has been known to improve ventricular function, the potential proarrhythmic effects have not yet been studied. MATERIALS AND METHODS: We monitored the heart rhythm of 6 dogs for 4 weeks after transplantation of hMSC (1x10(7), epicardial injection) (hMSC group) and in 5 Sham dogs after the injection of the vehicle alone. Cardiac sympathetic nerve sprouting {nerve growth factor (NGF)-beta; tyrosine hydroxylase (TH)} and gap junction expression {connexin (Cx) 43} were evaluated in 10 dogs (5 hMSC and 5 Sham) that survived longer than 4 weeks. RESULTS: The hMSC group expressed higher levels of NGF-beta messenger ribonucleic acid (mRNA) (56.0+/-66.8 fold; p<0.01) with TH+ sympathetic nerves (0.51+/-0.40 vs. 0.15+/-0.13% area; p<0.03) than the Sham control. In contrast, the hMSC group expressed lower levels of Cx43 mRNA (0.59+/-0.29 fold, p<0.001) and Cx43+ (1.64+/-1.79 vs. 2.12+/-1.07% area, p<0.001) than the Sham control. The incidences of ventricular fibrillation were 33.3% and 0% in the hMSC group and Sham control, respectively. One of the dogs with ventricular fibrillation (VF) in the hMSC group died suddenly. CONCLUSION: hMSC transplantation may be proarrhythmic since NGF-beta expression increased with cardiac sympathetic hyperinnervation and the expression of Cx43 and the gap junction decreased.

Keyword

Mesenchymal stem cells; Nerve growth factor; Gap Junctions; Ventricular tachycardia

MeSH Terms

Animals
Connexin 43
Dogs
Gap Junctions
Heart
Humans
Incidence
Mesenchymal Stem Cell Transplantation
Mesenchymal Stromal Cells
Nerve Growth Factor
RNA
RNA, Messenger
Salicylamides
Tachycardia, Ventricular
Transplants
Tyrosine 3-Monooxygenase
Ventricular Fibrillation
Ventricular Function
Connexin 43
Nerve Growth Factor
RNA
RNA, Messenger
Salicylamides
Tyrosine 3-Monooxygenase

Figure

• 

  Fig. 1 Experimental procedures. A: the ICD lead was positioned using the left internal jugular venous approach, and the position of the ICD coil electrode was confirmed by fluoroscopic imaging. B: hMSC injection sites were marked with suture material, and the hMSCs were transplanted by direct epicardial injection. C: the ICD electrogram revealed an episode of spontaneously induced non-sustained VF. This dog survived for 4 weeks. ICD: implantable defibrillator, hMSC: human mesenchymal stem cell, VF: ventricular fibrillation.

• 

  Fig. 2 Human nucleoline (hNC) immunostaining of ventricular tissue close to the area of hMSC injection. A: the brown colored nucleus represents hNC positive human cells (hMSC) at 200× magnification. B: the 400× magnification high power field shows no evidence of immune rejection (no lymphocytic infiltration, but polymorphonuclear cells due to surgical inflammation).

• 

  Fig. 3 A: mRNA expression levels of NGF-β was significantly higher in the hMSC group compared with Sham control group. B: calculated % area of TH positive (sympathetic) nerves was significantly higher in the hMSC transplanted tissues compared with the Sham group. C and D: tyrosine hydroxylase (TH) immunostaining of ventricular tissue close to the area of hMSC injection at 200× magnification. The hMSC group shows significant sympathetic hyperinnervation after 4 weeks of survival. By contrast, no significant increase in sympathetic nerves was observed in the Sham group. qRT-PCR: quantitative Reverse Transcription Polymerase chain Reaction, NGF-β: nerve growth factor-β, hMSC: human mesenchymal stem cell.

• 

  Fig. 4 mRNA expression levels. A: Cx43 was significantly lower in the hMSC group compared with Sham control group. B: calculated % area of Cx43 positive gap junction was significantly lower in the hMSC group compared with the Sham group. C and D: Cx43 immunostainings of ventricular tissue close to the area of hMSC injection at 200× magnification. The density of Cx43 positive gap junction is lower in hMSC transplanted tissue compared with the Sham after 4 weeks of survival. qRT-PCR: quantitative Reverse Transcription Polymerase chain Reaction, hMSC: human mesenchymal stem cell.

• 

  Fig. 5 Isochronal map of LV endocardium during high right atrial pacing (cycle length 500 ms) in right anterior oblique view and anterior posterior cranial view in animals that were transplanted with hMSC. As showed in the color scale bar, the impulse conducts from the white color to the purple color, and the earliest activation site is a septum. However, there is a significant conduction delay at the LV anterior wall (white dotted circle) where the hMSC was injected and Cx43 expression was reduced. LV: left ventricle, hMSC: human mesenchymal stem cell.

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