Tumor necrosis factor-alpha pretreated umbilical cord blood mesenchymal stem cell transplantation for treatment of myocardial infarction

doi:10.3969/j.issn.2095-4344.2016.32.018

Abstract

Abstract:

BACKGROUND: A large amount of inflammatory mediators from myocardial cells are secreted in response to myocardial injury after myocardial ischemia. Moreover, inflammatory cytokines in infarction and ischemia regions contribute to myocardial tissue repair and adaptation.
OBJECTIVE: To investigate the effect of tumor necrosis factor-α (TNF-α) pretreatment on cardiac function of myocardial infarction rabbits undergoing umbilical cord blood mesenchymal stem cell transplantation.
METHODS: Thirty-six white rabbits were equally randomized into sham, model, non-TNF-α, and TNF-α groups. Animal models of myocardial infarction were made in the latter three groups. Twenty-four hours after modeling, PBS, umbilical cord blood mesenchymal stem cells pretreated with or without TNF-α were injected at infarct center and border, respectively, in the model, TNF-α and non-TNF-α groups.
RESULTS AND CONCLUSION: Model rabbits in the TNF-α and non-TNF-α groups showed better cardiac function and lower size of myocardial infarction and fibrosis than those in the model group. Compared with the non-TNF-α group, moreover, the TNF-α group showed better outcomes in these indicators. These findings indicate that TNF-α pretreatment can markedly improve the therapeutic efficacy of umbilical cord blood mesenchymal stem cell transplantation on myocardial infarction.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Myocardial Ischemia, Cord Blood Stem Cell Transplantation, Tissue Engineering

CLC Number:

R394.2

Wang Wei, Li Xiao-fu, Li Zhong-jian. Tumor necrosis factor-alpha pretreated umbilical cord blood mesenchymal stem cell transplantation for treatment of myocardial infarction[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(32): 4831-4837.

Figures/Tables 1

2.1 脐血间充质干细胞的形态胎儿脐血间充质该细胞在培养48 h时开始出现贴壁生长，3 d时有细胞集落生成，细胞呈纺锤形或者梭形，培养7 d时开始形成大的细胞集落，细胞呈成纤维样，形态比较均一，排列有一定的方向性(图1A)。传代培养4 h细胞便出现完全贴壁生长，生长速度比较快，细胞培养至第3代时，细胞生长比较旺盛，增殖能力非常强，细胞呈梭形，向外延伸，方向比较一致(图1B)。 2.2 胎儿脐血间充质干细胞的鉴定结果胎儿脐血间充质干细胞的流式细胞仪鉴定结果显示，细胞呈CD24、CD44、CD45及CD69阳性，CD105和CD166阴性。符合胎儿脐血间充质干细胞的细胞免疫表型特点，表明分离的细胞为胎儿脐血间充质干细胞。 2.3 实验动物数量分析 36只兔均未出现死亡，均进入实验分析。 2.4 心肌梗死大白兔模型构建结果造模后第1天，假手术组兔心电图变化不明显。心肌梗死模型兔的心电图出现ST段抬高，表现为心肌梗死心电图波形，表明心肌梗死大白兔模型建造成功(图2)。 2.5 心脏功能与假手术组相比，模型组大白兔左室舒张末期前后径明显增大(P < 0.05)，且左室射血分数降低(P < 0.05)。肿瘤坏死因子α组和无肿瘤坏死因子α组大白兔的左室舒张末期前后径明显小于模型组(P < 0.05)，左室射血分数大于模型组(P < 0.05)。肿瘤坏死因子α组大白兔的左室舒张末期前后径明显小于无肿瘤坏死因子α预处理移植(P < 0.05)，左室射血分数大于无肿瘤坏死因子α预处理移植(P < 0.05；表1)。 2.6 心肌梗死形态苏木精-伊红染色结果显示，移植后4周，模型组、肿瘤坏死因子α组和无肿瘤坏死因子α组大白兔左心室均可见心肌细胞的变性坏死，心肌梗死区域周围有岛状分布的正常心肌细胞，其中在模型组大白兔的左心室间脂肪大量增生。肿瘤坏死因子α组和无肿瘤坏死因子α预处理移植大白兔的心肌梗死面积明显低于模型组(P < 0.05)，且肿瘤坏死因子α组大白兔的心肌梗死面积明显低于无肿瘤坏死因子α组(P < 0.05；图3，表2)。 2.7 心肌纤维化面积 Masson's染色结果显示，移植后4周，模型组、肿瘤坏死因子α组和无肿瘤坏死因子α组大白兔左心室均可见纤维瘢痕。肿瘤坏死因子α组和无肿瘤坏死因子α预处理移植大白兔的心肌纤维化面积明显低于模型组(P < 0.05)，且肿瘤坏死因子α组大白兔的心肌纤维化面积明显低于无肿瘤坏死因子α预处理移植的心肌梗死面积(P < 0.05；图4，表3)。"

References

[1] Guo Y, Lip GY, Banerjee A. Heart failure in East Asia. Curr Cardiol Rev. 2013;9(2):112-122.
[2] Go AS, Mozaffarian D, Roger VL, et al. Heart disease and stroke statistics--2013 update: a report from the American Heart Association. Circulation. 2013;127(1): e6-e245.
[3] Laflamme MA, Murry CE. Heart regeneration. Nature. 2011;473(7347):326-335.
[4] Assmus B, Zeiher AM. Early cardiac retention of administered stem cells determines clinical efficacy of cell therapy in patients with dilated cardiomyopathy. Circ Res. 2013;112(1):6-8.
[5] Harris DT. Non-haematological uses of cord blood stem cells. Br J Haematol. 2009;147(2):177-184.
[6] Rehman J. Empowering self-renewal and differentiation: the role of mitochondria in stem cells. J Mol Med (Berl). 2010;88(10):981-986.
[7] Wagers AJ. The stem cell niche in regenerative medicine. Cell Stem Cell. 2012;10(4):362-369.
[8] Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem. 2006;98(5): 1076-1084.
[9] Zhang M, Mal N, Kiedrowski M, et al. SDF-1 expression by mesenchymal stem cells results in trophic support of cardiac myocytes after myocardial infarction. FASEB J. 2007;21(12):3197-3207.
[10] Govindasamy V, Ronald VS, Abdullah AN, et al. Differentiation of dental pulp stem cells into islet-like aggregates. J Dent Res. 2011;90(5):646-652.
[11] Sasaki M, Abe R, Fujita Y, et al. Mesenchymal stem cells are recruited into wounded skin and contribute to wound repair by transdifferentiation into multiple skin cell type. J Immunol. 2008;180(4):2581-2587.
[12] Toma C, Pittenger MF, Cahill KS, et al. Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation. 2002;105(1):93-98.
[13] Martens TP, See F, Schuster MD, et al. Mesenchymal lineage precursor cells induce vascular network formation in ischemic myocardium. Nat Clin Pract Cardiovasc Med. 2006;3 Suppl 1:S18-22.
[14] Aggarwal S, Pittenger MF. Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood. 2005;105(4):1815-1822.
[15] Chen X, Armstrong MA, Li G. Mesenchymal stem cells in immunoregulation. Immunol Cell Biol. 2006;84(5): 413-421.
[16] Nauta AJ, Fibbe WE. Immunomodulatory properties of mesenchymal stromal cells. Blood. 2007;110(10): 3499-3506.
[17] Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8(4):315-317.
[18] Yu YS, Shen ZY, Ye WX, et al. AKT-modified autologous intracoronary mesenchymal stem cells prevent remodeling and repair in swine infarcted myocardium. Chin Med J (Engl). 2010;123(13): 1702-1708.
[19] Zhuang Y, Chen X, Xu M, et al. Chemokine stromal cell-derived factor 1/CXCL12 increases homing of mesenchymal stem cells to injured myocardium and neovascularization following myocardial infarction. Chin Med J (Engl). 2009;122(2):183-187.
[20] Gavira JJ, Perez-Ilzarbe M, Abizanda G, et al. A comparison between percutaneous and surgical transplantation of autologous skeletal myoblasts in a swine model of chronic myocardial infarction. Cardiovasc Res. 2006;71(4):744-753.
[21] Ding J, Ghali O, Lencel P, et al. TNF-alpha and IL-1beta inhibit RUNX2 and collagen expression but increase alkaline phosphatase activity and mineralization in human mesenchymal stem cells. Life Sci. 2009;84(15-16):499-504.
[22] 樊志刚,刘芳.TNF-α预处理脐血间充质干细胞移植心肌梗死大鼠心功能影响[J].现代仪器与医疗,2015(4): 4-7,18.
[23] Carlin R, Davis D, Weiss M, et al. Expression of early transcription factors Oct-4, Sox-2 and Nanog by porcine umbilical cord (PUC) matrix cells. Reprod Biol Endocrinol. 2006;4:8.
[24] Williams AR, Hare JM. Mesenchymal stem cells: biology, pathophysiology, translational findings, and therapeutic implications for cardiac disease. Circ Res. 2011;109(8):923-940.
[25] Gnecchi M, Zhang Z, Ni A, et al. Paracrine mechanisms in adult stem cell signaling and therapy. Circ Res. 2008;103(11):1204-1219.
[26] Malliaras K, Zhang Y, Seinfeld J, et al. Cardiomyocyte proliferation and progenitor cell recruitment underlie therapeutic regeneration after myocardial infarction in the adult mouse heart. EMBO Mol Med. 2013;5(2): 191-209.
[27] Xiong Q, Ye L, Zhang P, et al. Functional consequences of human induced pluripotent stem cell therapy: myocardial ATP turnover rate in the in vivo swine heart with postinfarction remodeling. Circulation. 2013;127(9):997-1008.
[28] Li T, Ma Q, Ning M, et al. Cotransplantation of human umbilical cord-derived mesenchymal stem cells and umbilical cord blood-derived CD34? cells in a rabbit model of myocardial infarction. Mol Cell Biochem. 2014;387(1-2):91-100.
[29] Guo J, Lin GS, Bao CY, et al. Anti-inflammation role for mesenchymal stem cells transplantation in myocardial infarction. Inflammation. 2007;30(3-4):97-104.
[30] Egea V, von Baumgarten L, Schichor C, et al. TNF-α respecifies human mesenchymal stem cells to a neural fate and promotes migration toward experimental glioma. Cell Death Differ. 2011;18(5):853-863.
[31] Böcker W, Docheva D, Prall WC, et al. IKK-2 is required for TNF-alpha-induced invasion and proliferation of human mesenchymal stem cells. J Mol Med (Berl). 2008;86(10):1183-1192.
[32] Perng DW, Yang DM, Hsiao YH, et al. miRNA-146a expression positively regulates tumor necrosis factor-α-induced interleukin-8 production in mesenchymal stem cells and differentiated lung epithelial-like cells. Tissue Eng Part A. 2012;18(21-22): 2259-2267.
[33] Du YY, Zhou SH, Zhou T, et al. Immuno-inflammatory regulation effect of mesenchymal stem cell transplantation in a rat model of myocardial infarction. Cytotherapy. 2008;10(5):469-478.
[34] Qi CM, Ma GS, Liu NF, et al. Transplantation of magnetically labeled mesenchymal stem cells improves cardiac function in a swine myocardial infarction model. Chin Med J (Engl). 2008;121(6): 544-550.
[35] Lencel P, Delplace S, Pilet P, et al. Cell-specific effects of TNF-α and IL-1β on alkaline phosphatase: implication for syndesmophyte formation and vascular calcification. Lab Invest. 2011;91(10):1434-1442.