Iranian Journal of Allergy, Asthma and Immunology 2017. 16(3):205-218.

Effects of Intraperitoneal Injection of Allogeneic Bone Marrow-derived Mesenchymal Stem Cells on Bronchiolitis Obliterans in Mice Model
Sakine Işık, Nevin Uzuner, Meral Karaman, Özkan Karaman, Müge Kıray, İlknur Kozanoğlu, Hüsnü Alper Bağrıyanık, Zeynep Arıkan-Ayyıldız, Melis Kartal Yandım, Yusuf Baran


Bone marrow-derived mesenchymal stem cells (BMSCs) can ameliorate a variety of lung diseases such as asthma, lung fibrosis, and acute lung injury by its anti-inflammatory and immunmodulatory effects. In this study, we developed a mouse model of bronchiolitis obliterans (BO) and evaluated the effects of the intraperitoneal administration of BMSCs on lung histopathology and cytokine levels. 25 BALB/c mice were divided into four groups; control group (Group I), BO developed and 1x10BMSCs-injected group (Group II), non-BO, 1x106 BMSCs-injected group (Group III), and BO developed and saline-injected group (Group IV). Histological and immunohistochemical findings of the lung tissue and the migration of BMSCs to the lung were evaluated using light and confocal microscopy techniques. Confocal microscopy evaluations showed that there was no noteworthy amount of BMSCs in the lung tissue of group III while significant amount of BMSCs was detected in group II. Wall thicknesses of terminal bronchiole and periterminal bronchiolar collagen deposition were significantly lower in group II compared to the group IV (p<0.05). Furthermore, according to the immunohistochemical staining results, CD3, CD4, CD8, CD20, CD68 and neutrophil elastase positive immune cells of group II were stained more positive than group IV cells (p<0.05). IFN-γ IL-2 and TNF-α levels in bronchoalveolar lavage fluid (BALF) were significantly lower in group II compared to group IV (p<0.05). The findings of this study indicate that intraperitoneally administered BMSCs have potent effects on histopatological changes of the lung tissue and cytokine levels in the murine model of BO.


Bronchiolitis obliterans; Intraperitoneal administration; Mesenchymal stem cells; Murine model

Full Text:



1. Ryu JH, Myers JL, Swensen SJ. Bronchiolar disorders. Am J Respir Crit Care Med 2003; 168(11):1277-92.

2. Chan PWK, Muridan R, Debruyne JA. Bronchiolitis obliterans in children: clinical profile and diagnosis. Respirology 2000; 5(4):369-75.

3. Kotloff RM, Ahya VN, Crawford SW. Pulmonary complications of solid organ and hematopietic stme cell transplantation. Am J Respir Crit Care Med 2004; 170(1):22-48.

4. Yalçın E, Doğru D, Haliloğlu M, Özçelik U, Kiper N, Göçmen A. Postinfectious bronchiolitis obliterans in children: clinical and radiological profile and prognostic factors. Respiration 2003; 70(4):371-75.

5. Sarahrudi K, Carretta A, Wisser W, Senbaklavaci O, Ploner M, Neuhauser P, et al. The value of switching from cyclosporine to tacrolimus in the treatment of refractory acute rejection and obliterative bronchiolitis after lung transplantation. Transplant Int 2002; 15(1):24-8.

6. Ding IB, Baumgartner RA, Schwaiblamir M, Behr J. Admistration of anti-interleukine 2Rα monoclonal antibody in bronchiolitis obliterans syndrome after lung transplantation. Transplantation 2003; 75(10):1767-9.

7. Dosanh A, Ikonen T, Wan B, Morris RE. Pirfenidone: a novel anti-fibrotic agent and progressive chronic allograft rejection. Pulm Pharmacol Ther 2001; 15(5):433-7.

8. Gerhardt SG, McDyer JF, Girgis RE, Conte JV, Yang SC, Orens JB. Maintenance azithromycin therapy for bronchiolitis obliterans syndrome. Am J Respir Crit Care Med 2003; 168(1):121-5.

9. Hardy KA, Schidlow DV, Zacri N. Obliterative bronchiolitis in children. Chest 1988; 93(3):460-6.

10. Delorme B, Charbord P. Culture and characterization of human bone marrow mesenchymal stem cells. Methods Mol Med 2007; 140:67-81.

11. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284(5411):143-7.

12. Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem 2006; 98(5):1076-84.

13. Loebinger MR, Aguilar S, Janes SM. Therapeutic potential of stem cells in lung disease: progress and pitfalls. Clin Sci (Lond) 2008; 114(2):99-108.

14. Mohammadian M, Boskabady MH, Kashani IR, Jahromi GP, Omidi A, Mejad AK, et al. Effect of bone marrow derived mesenchymal stem cells on lung pathology and inflammation in ovaalbumin-induced asthma in mouse. Iran J Basic Med Sci 2016; 19(1):55-63.

15. Ghorbani A, Feizpour A, Hashemzahi M, Gholami L, Hosseini M, Soukhtanloo M, et al. The effect of adipose derived stromal cells on oxidative stress level, lungemphysema and white blood cells of guinea pigs model of chronic obstructive pulmonary disease. Daru 2014; 22(1):26.

16. Matthay MA, Thompson BT, Read EJ, McKenna DH Jr, Liu KD, Calfee CS, et al. Therapeutic potential of mesenchymal stem cells for severe acute lung injury. Chest 2010; 138(4):965-72.

17. Horie S, Laffey JG. Recent insights: mesencyhmal stromal/stem cell tharapy for acute respiratory distress syndrome. F1000Res 2016; 28:5.

18. Garippo AL, Parra ER, Teodoro WR, Veloza AP, Yoshinari NH, Capelozzi VL. Immune cell infiltration and broncovascular remodeling after nitric acid nasal instillation in a mouse bronchiolitis obliterans model. Lung 2006; 184(4):229-38.

19. Firinci F, Karaman M, Baran Y, Bagriyanik A, Ayyildiz ZA, Kiray M, et al. Mesenchymal stem cells ameliorate the histopathological changes in a murine model of chronic asthma. Int Immunopharmacol 2011; 11(8):1120-6.

20. Sunay O, Can G, Cakir Z, Denek Z, Kozanoğlu İ, Erbil G, et al. Autologous Rabbit Adipose Tissue Derived Mesenchymal Stem Cells For The Treatment Of Bone Injuries With Distraction Osteogenesis. Cytotherapy 2013; 15(6):690-702.

21. Baran Y, Ural AU, Avcu F, Sarper M, Elçi P, Pekel A. Optimisation of Transfection of Green Flourescent Protein in Pursuing Mesenchymal Stem Cells, In Vivo. Turk J Haematol 2008; 25(4):172-5.

22. Loebinger MR, Aguilar S, Janes SM. Therapeutic potential of stem cells in lung disease: progress and pitfalls. Clin Sci (Lond) 2008; 114(2):99-108.

23. Aris RM, Walsh S, Chalermskulrat W, Hathwar V, Neuringer IP. Growth factor upregulation during obliterative bronchiolitis in the mouse model. Am J Respir Crit Care Med 2002; 166 (3):417-22.

24. Hertz MI, Henke CA, Nakhleh RE, Harmon KR, Marinelli WA, Fox JM, et al. Obliterative bronchliolitis after lung transplantation: a fibroobliterative disorder associated with platelet-derived growth factor. Proc Natl Acad Sci USA 1992; 89(21):10385-9.

25. Laohaburanakit P, Chan A, Allen RP. Bronchiolitis obliterans. Clin Rev Allergy Immunol 2003; 25(3):259-74.

26. Krause DS, Theise ND, Collector MI, Henegariu O, Hwang S, Gardner R, et al. Multiorgan,multi-lineage engraftment by a single bone marrow-derived stem cell. Cell 2001; 105(3):369–77.

27. Kotton DN, Ma BY, Cardoso WV, Sanderson EA, Summer RS, Williams MC, et al. Bone marrow derived cells as progenitors of lung alveolar epithelium. Development 2001; 128(24):5181–8.

28. Theise ND, Henegariu O, Grove J, Jagirdar J, Kao PN, Crawford JM, et al. Radiation pneumonitis in mice: a severe injury model for pneumocyte engraftment from bone marrow. Exp Hematol 2002; 30(11):1333–8.

29. Ortiz LA, Gambelli F, McBride C, Gaupp D, Baddoo M, Kaminski N, et al. Mesenchymal stem cell engraftment in lung is enhanced in response to bleomycin exposure and ameliorates its fibrotic effects. Proc Natl Acad Sci U S A 2003; 100(14):8407-11.

30. Ishizawa K, Kubo H, Yamada M, Kobayashi S, Suzuki T, Mizuno S, et al. Hepatocyte growth factor induces angiogenesis in injured lungs through mobilizing endothelial progenitor cells. Biochem Biophys Res Commun 2004; 324(1):276–80.

31. Mei SH, Haitsma J, Dos Santos CC, Deng Y, Lai PF, Slutsky AS, et al. Mesenchymal stem cells reduce inflammation while enhancing bacterial clearance and improving survival in sepsis. Am J Respir Crit Care Med 2010; 182(8):1047–57.

32. Krasnodembskaya A, Song Y, Fang X, Gupta N, Serikov V, Lee JW, et al. Antibacterial effect of human mesenchymal stem cells is mediated in part from secretion of the antimicrobial peptide LL-37. Stem Cells 2010; 28(12):2229–38.

33. Ribeiro-Paes JT, Bilaqui A, Greco OT, Ruiz MA, Marcelino MY, Stessuk T, et al. Unicentric study of cell therapy in chronic obstructive pulmonary disease/ pulmonary emphysema. Int J Chron Obstruct Pulmon Dis 2011; 6:63–71.

34. Chang YS, Ahn SY, Yoo HS, Sung SI, Choi SJ, Oh WI, et al. Mesenchymal stem cells for bronchopulmonary dysplasia: phase 1 dose-escalation clinical trial. J Pediatr 2014; 164(5):966-72.

35. Zhao Y, Gillen JR, Harris DA, Kron IL, Murphy MP, Lau CL. Treatment with placenta-derived mesencyhmal stem cells mitigates development of bronchiolitis obliterans in a murine model. J Thorac Cardiovasc Surg 2014;147(5):1668-1677.e5.doi:10.1016/j.jtcvs.2013.09.041.

36. Grove DA, Xu J, Joodi R, Torres-Gonzales E, Neujahr D, Mora AL, et al. Attenuation of early airway obstruction by mesencyhmal stem cells in a murine model of heterotopic tracheal transplantation. J Heart Lung Transplant 2011; 30(3):341-50.

37.Espinel Jde O, Uribe C, Meyer FS, Bringheti R, Kulczynskı J, Saueressig MG. Cell therapy in thetreatment of bronchiolitis obliterans in a murine model. Rev Col Bras Cir 2015; 42(3):181-8.

38. Lee SH, Jang AS, Kim YE, Cha JY, Kim TH, Jung S, et al. Modulation of cytokine and nitric oxide by mesenchymal stem cell transfer in lung injury/fibrosis.Respir Res 2010; 8:11-6.

39. Ortiz LA, Dutreil M, Fattman C, Pandey AC, Torres G, Go K, et al. Interleukin 1 receptor antagonist mediates the antiinflammatory and antifibrotic effect of mesenchymal stem cells during lung injury. Proc Natl Acad Sci USA 2007; 104(26):11002–07.

40. Moodley Y, Atienza D, Manuelpillai U, Samuel CS, Tchongue J, Ilancheran S, et al. Human umbilical cord mesenchymal stem cells reduce fibrosis of bleomycin-induced lung injury. Am J Pathol 2009; 175(1):303-13.

41. Salama M, Andrukhova O, Jaksch P, Taghavi S, Kelpetko W, Dekan G, et al. Endothelin-1 governs proliferation and migration of bronchoalveolar lavage-derived lung mesenchymal stem cells in bronchiolitis obliterans syndrome. Transplantation 2011; 92(2):155-62.

42. Walker N, Badri L, Wettlaufer S, Flint A, Sajjan U, Krebsbach PH, et al. Resident tissue-specific mesenchymal progenitor cells contribute to fibrogenesis in human lung allografts. Am J Pathol 2011; 178(6):2461-9.

43. Rojas M, Xu J, Woods CR, Mora AL, Spears W, Roman J, et al. Bone marrow-derived mesenchymal stem cells in repair of the injured lung. Am J Respir Cell Mol Biol 2005; 33(2):145–52.

44.Tian ZF, Du J, Fu XM, Wang B, Hong XY, Feng ZC. Influence of human bone marrow-derived mesenchymal stem cells on the lung of newborn rats damaged by hyperoxia. Zhonghua Er Ke Za Zhi 2008; 46(1):4-8.

45. Abreu SC, Antunes MA, Maron- Gutierrez T, Maron-Gutierrez T, Cruz FF, Carmo LG, et al. Effects of bone marrow-derived mononuclear cells on airway and lung parenchyma remodeling in a murine model of chronic allergic inflammation. Respir Physiol Neurobiol 2010; 175(1):153–63.

46. Mistchenko AS, Diez RA, Mariani AL, Robaldo J, Maffey AF, Bayley-Bustamante G, et al. Cytokines in adenoviraldisease in children: association of interleukine-6, interleukine-8 and tumor necrosis factor alpha levels with clinical outcome. J Pediatr 1994; 124(5):714-20.

47. Boehler A, Bai XH, Liu M, Cassivi S, Chamberlain D, Slutsky AS, et al. Upregulation of T-helper 1 cytokines and chemokine expression in post-transplant airway obliteration. Am J Respir Crit Care Med 1999; 159(6):1910-7.

48. Zhao Y, Gillen JR, Harris DA, Kron IL, Murphy MP, Lau CL. Treatment with placenta-derived mesenchymal stem cells mitigates development of bronchiolitis obliterans in a murine model. J Thorac Cardiovasc Surg 2014; 147(5):1668-77.


  • There are currently no refbacks.

Creative Commons Attribution-NonCommercial 3.0

This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.