Conditioned medium obtained from human amniotic membrane-derived mesenchymal stem cell attenuates heart failure injury in rats

Document Type : Original Article


1 Physiology Research Center and Department of Physiology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran

2 Department of Physiology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran

3 Department of Histology and Neuroscience, Anti-microbial Resistance Research Center, School of Medicine, Iran University of Medical Sciences, Tehran, Iran


Objective(s): Heart failure (HF) is one of the leading causes of death worldwide. Due to beneficial effects of stem cells, paracrine secretion of them has recently been used by researchers. The purpose of this study was to investigate the effects of intravenous injection (IV) of conditioned medium (CM) of human amniotic membrane-derived mesenchymal stem cell (MSC-CM) on HF.
Materials and Methods: Male Wistar rats (n=35, 180 g) were randomly divided into five groups: sham, HF, HF+MSC-CM, HF+culture medium and HF+phosphate-buffered saline (PBS). To induce HF, isoproterenol (170 mg/kg/d) was injected subcutaneously for 4 consecutive days. After 28 days, induction of HF was evaluated by echocardiography. A day after echocardiography, 50 μg culture medium/5 ml PBS in HF+culture medium group, 50 μg MSC-CM/5 ml PBS in HF+MSC-CM group and 5 ml PBS in HF+PBS group were injected  two times for 4 successive days. The echocardiography was performed 4 weeks after the last injection of isoproterenol. To evaluate the fibrosis, morphology, and cardiac function, Trichrome Masson’s staining, Hematoxylin and Eosin staining and echocardiography were performed, respectively.
Results: CM significantly increased fractional shortening and ejection fraction, and also significantly decreased apoptotic nuclear condensation. Moreover, significant decreased level of fibrosis and increased level of angiogenesis was observed in the treatment group (P<0.05).
Conclusion: Our results indicated that IV injection of CM has therapeutic effects on HF by reducing fibrosis and preventing the progression of failure due to its paracrine effects.


Main Subjects

1. Cowie M, Wood D, Coats A, Thompson S, Suresh V, Poole-Wilson P, et al. Survival of patients with a new diagnosis of heart failure: a population based study. Heart 2000; 83:505-510.
2. Gheorghiade M, Bonow RO. Chronic heart failure in the United States: a manifestation of coronary artery disease. Circulation 1998; 97:282-289.
3. Bhat P, Tang WW. Biomarkers to Assess and Guide the Management of Heart Failure.  Biomarkers in Cardiovascular Disease: Elsevier 2019: 97-108.
4. Amani H, Habibey R, Hajmiresmail S, Latifi S, Pazoki-Toroudi H, Akhavan O. Antioxidant nanomaterials in advanced diagnoses and treatments of ischemia reperfusion injuries. J Mater Chem B  2017; 5:9452-9476.
5. Amani H, Ajami M, Maleki SN, Pazoki-Toroudi H, Daglia M, Sokeng AJT, et al. Targeting signal transducers and activators of transcription (STAT) in human cancer by dietary polyphenolic antioxidants. Biochimie 2017; 142:63-79.
6. Ajami M, Davoodi SH, Habibey R, Namazi N, Soleimani M, Pazoki‐Toroudi H, et al. Effect of DHA+ EPA on oxidative stress and apoptosis induced by ischemia‐reperfusion in rat kidneys.  Fundam Clin Pharmacol 2013; 27:593-602.
7. Habibey R, Ajami M, Ebrahimi SA, Hesami A, Babakoohi S, Pazoki-Toroudi H, et al. Nitric oxide and renal protection in morphine-dependent rats. Free Radic Biol Med 2010; 49:1109-1118.
8. Passier R, van Laake LW, Mummery CL. Stem-cell-based therapy and lessons from the heart. Nature 2008; 453:322-329.
9. Amani H, Mostafavi E, Arzaghi H, Davaran S, Akbarzadeh A, Akhavan O, et al. Three-Dimensional Graphene Foams: Synthesis, Properties, Biocompatibility, Biodegradability, and Applications in Tissue Engineering. ACS Biomater Sci Eng  2019; 5:193-214.
10. Teerlink JR, Metra M, Filippatos GS, Davison BA, Bartunek J, Terzic A, et al. Benefit of cardiopoietic mesenchymal stem cell therapy on left ventricular remodelling: results from the Congestive Heart Failure Cardiopoietic Regenerative Therapy (CHART‐1) study.  Eur J Heart Fail 2017; 19:1520-1529.
11. Bartolucci J, Verdugo FJ, González PL, Larrea RE, Abarzua E, Goset C, et al. Safety and efficacy of the intravenous infusion of umbilical cord mesenchymal stem cells in patients with heart failure: a phase 1/2 randomized controlled trial (RIMECARD trial [randomized clinical trial of intravenous infusion umbilical cord mesenchymal stem cells on cardiopathy]).  Circ Res 2017; 121:1192-1204.
12. Halkos ME, Zhao Z-Q, Kerendi F, Wang N-P, Jiang R, Schmarkey LS, et al. Intravenous infusion of mesenchymal stem cells enhances regional perfusion and improves ventricular function in a porcine model of myocardial infarction. Basic Res  Cardiol 2008; 103:525-536.
13. Patel AN, Silva F, Winters AA. Stem cell therapy for heart failure. Heart Fail Clin. 2015; 11:275-286.
14. Tse HF, Siu CW, Zhu SG, Songyan L, Zhang QY, Lai WH, et al. Paracrine effects of direct intramyocardial implantation of bone marrow derived cells to enhance neovascularization in chronic ischaemic myocardium. Eur J Heart Fail 2007; 9:747-753.
15. Vulliet PR, Greeley M, Halloran SM, MacDonald KA, Kittleson MD. Intra-coronary arterial injection of mesenchymal stromal cells and microinfarction in dogs. Lancet 2004; 363:783-784.
16. Trounson A, McDonald C. Stem cell therapies in clinical trials: progress and challenges. Cell Stem Cell 2015; 17:11-22.
17. Gnecchi MJSctm. Cell therapy for heart regeneration: Learning from the past to build a brighter future. Stem Cells Translational Medicine 2018;7:702–704.
18. Arsalan M, Dhein S, Aupperle H, Rastan AJ, Barten MJ, Walther T, et al. The reverse remodeling effect of mesenchymal stem cells is independent from the site of epimyocardial cell transplantation. Innovations: Technology and Techniques in Cardiothoracic and Vascular Surgery. Innovations (Phila) 2013; 8:433-439.
19. Bing W, Pang X, Qu Q, Bai X, Yang W, Bi Y, et al. Simvastatin improves the homing of BMSC s via the PI 3K/AKT/miR‐9 pathway. J Cell Mol Med 2016; 20:949-961.
20. Baglio SR, Pegtel DM, Baldini N. Mesenchymal stem cell secreted vesicles provide novel opportunities in (stem) cell-free therapy. Front Physiol 2012; 3:359.
21. Gnecchi M, Danieli P, Malpasso G, Ciuffreda MC. Paracrine mechanisms of mesenchymal stem cells in tissue repair. Methods Mol Biol 2016; 1416:123-146.
22. Aboutaleb N, Faezi M, Maleki SN, Nazarinia D, Tousi SMTR, Hashemirad NJJoCN. Conditioned medium obtained from mesenchymal stem cells attenuates focal cerebral ischemia reperfusion injury through activation of ERK1/ERK2-BDNF signaling pathway.  J Chem Neuroanat 2019; 97:87-98.
23. Cunningham CJ, Redondo-Castro E, Allan SMJJoCBF, Metabolism. The therapeutic potential of the mesenchymal stem cell secretome in ischaemic stroke. J Cereb Blood Flow Metab 2018; 38:1276-1292.
24. Rehman J, Traktuev D, Li J, Merfeld-Clauss S, Temm-Grove CJ, Bovenkerk JE, et al. Secretion of angiogenic and antiapoptotic factors by human adipose stromal cells. Circulation 2004; 109:1292-1298.
25. Barile L, Biemmi V, Ciullo A, Torre T, Demerzis S, Mauri P, et al. P4224 Superior exosome-mediated paracrine effects of cardiac progenitor cells compared to bone marrow mesenchymal stem cells derived from the same patient for cardiac repair. Eur Heart J 2018;  39:ehy563.P4224.
26. Kervadec A, Bellamy V, El Harane N, Arakélian L, Vanneaux V, Cacciapuoti I, et al. Cardiovascular progenitor–derived extracellular vesicles recapitulate the beneficial effects of their parent cells in the treatment of chronic heart failure.  J Heart Lung Transplant 2016; 35:795-807.
27. Golle L, Gerth HU, Beul K, Heitplatz B, Barth P, Fobker M, et al. Bone marrow-derived cells and their conditioned medium induce microvascular repair in uremic rats by stimulation of endogenous repair mechanisms.  Sci Rep 2017;7:9444.
28. Faezi M, Maleki SN, Aboutaleb N, Nikougoftar. The membrane mesenchymal stem cell derived conditioned medium exerts neuroprotection against focal cerebral ischemia by targeting apoptosis.  J Chem Neuroanat 2018; 94:21-31.
29. Liu Y-H, Lu M, Xie Z-Z, Hua F, Xie L, Gao JH, et al. Hydrogen sulfide prevents heart failure development via inhibition of renin release from mast cells in isoproterenol-treated rats.  Antioxid Redox Signal 2014; 20:759-769.
30. Pazoki-Toroudi H, Amani H, Ajami M, Nabavi SF, Braidy N, Kasi PD, et al. Targeting mTOR signaling by polyphenols: a new therapeutic target for ageing.  Ageing Res Rev 2016; 31:55-66.
31. Mehrjerdi FZ, Aboutaleb N, Pazoki-Toroudi H, Soleimani M, Ajami M, Khaksari M, et al. The protective effect of remote renal preconditioning against hippocampal ischemia reperfusion injury: role of KATP channels.  J Mol Neurosci 2015; 57:554-560.
32. Pazoki‐Toroudi HR, Ajami M, Habibey R. Pre‐medication and renal pre‐conditioning: a role for alprazolam, atropine, morphine and promethazine. Fundamental Clin Pharmacol 2010; 24:189-198.
33. Habibey R, Pazoki‐Toroudi HJC, Pharmacology E, Physiology. Morphine dependence protects rat kidney against ischaemia–reperfusion injury. Clin Exp Pharmacol Physiol 2008; 35:1209-1214.
34. Ajami M, Pazoki-Toroudi H, Amani H, Nabavi SF, Braidy N, Vacca RA, et al. Therapeutic role of sirtuins in neurodegenerative disease and their modulation by polyphenols. Neurosci Biobehav Rev, 2017; 73:39-47.
35. Höglund M. Stem cell transplantation. HemaSphere 2018; 2:159.
36. Naderi‐Meshkin H, Bahrami AR, Bidkhori HR, Mirahmadi M, Ahmadiankia N. Strategies to improve homing of mesenchymal stem cells for greater efficacy in stem cell therapy. Cell Biol Int 2015; 39:23-34
37. Luger D, Lipinski MJ, Westman PC, Glover DK, Frias JC, Albelda M, et al. Intravenously-delivered mesenchymal stem cells: systemic anti-inflammatory effects improve left ventricular dysfunction in acute myocardial infarction and ischemic cardiomyopathy. Circ Res 2017; 120:1598-1613.
38. Kim W-S, Park B-S, Sung J-H. The wound-healing and antioxidant effects of adipose-derived stem cells. Expert Opin Biol Ther 2009; 9:879-887.
39. Wang CY, Yang HB, Hsu HS, Chen LL, Tsai CC, Tsai KS, et al. Mesenchymal stem cell‐conditioned medium facilitates angiogenesis and fracture healing in diabetic rats. J Tissue Eng Regen Med 2012; 7:559-569.
40. Li Z, Guo J, Chang Q, Zhang A. Paracrine role for mesenchymal stem cells in acute myocardial infarction. Biol Pharm Bull 2009; 32:1343-1346.
41.Timmers L, Lim SK, Hoefer IE, Arslan F, Lai RC, van Oorschot AA, et al. Human mesenchymal stem cell-conditioned medium improves cardiac function following myocardial infarction. Stem Cell Res. (Amst.), 2011; 6:206-214.