Simvastatin combined with bone marrow mesenchymal stromal cells (BMSCs) improve burn wound healing by ameliorating angiogenesis through SDF-1α/CXCR4 pathway

Document Type : Original Article

Authors

1 Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

2 Physiology research center and department of physiology, faculty of medicine, Iran University of Medical Sciences, Tehran, Iran

3 Department of Anatomy, faculty of medicine, Alborz University of Medical Sciences, Karaj, Iran

4 Dietary Supplements and Probiotics Research Center, Alborz University of Medical Sciences, Karaj, Iran

5 Deparment of medical biotechnology, faculty of allied medicine, Iran University of Medical Sciences, Tehran, Iran

6 Burn research center, Iran University of Medical Sciences, Tehran, Iran

7 Anti-Microbial Resistance Research Center, Iran University of Medical Sciences, Tehran, Iran

Abstract

Objective(s): Chemokines are wound mediators that promote angiogenesis during wound healing. We hypothesized that Simvastatin in combination with the bone marrow mesenchymal stromal cells (BMSCs) improve burn wound healing by ameliorating angiogenesis via SDF-1α/CXCR4 pathway.
Materials and Methods: Under general anesthesia, deep partial-thickness burns were created on the inter-scapular area of 48 male rats. Study groups were administrated with petroleum jelly (Simvastatin Vehicle), a single dose of intradermal BMSCs (1×106), topical Simvastatin (0.5 mg/kg) daily and combination of BMSCs and Simvastatin for 14 days. In this study, we used MTT assay, in vivo and in vitro wound closure, H&E and Trichorome staining, immunohistochemistry (IHC), real- time PCR, Western blot and tube formation assay.
Results: A significant improvement in wound closure percentage, epithelial thickness, collagen remodeling, and up-regulation of stromal cell-derived factor 1 alpha (SDF1α), C-X-C chemokine receptor type 4 (CXCR4), protein kinase B (AKT), and phosphatidylinositol 3- kinase (PI3K), as well as CD31 and vascular endothelial growth factor (VEGF) expression were observed after treatment with simvastatin, BMSCs and combination of them compared to the vehicle group. However, the co-treatment group revealed considerable superiority in examined factors. BMSCs treated with Simvastatin showed the highest viability in the concentration of 0.5 and 1 Nanomolar (nM). Increment in proliferation and capillary vessels formation of BMSCs was observed in the 0.5 nM and 1 nM concentrations of Simvastatin in vitro.
Conclusion: Treatment of deep partial-thickness of burns with co-treatment of BMSCs and Simvastatin resulted in improved burn wound healing through up-regulating of SDF-1α/CXCR4 pathway.

Keywords

References

1. Hunt TK. The physiology of wound healing. Ann Emerg Med 1988; 17:1265-1273.
2. Sheridan RL. Burns: A Practical Approach to Immediate Treatment and Long Term Care: CRC Press; 2011.
3. Branski LK, Gauglitz GG, Herndon DN, Jeschke MG. A review of gene and stem cell therapy in cutaneous wound healing. Burns 2009; 35:171-180.
4. Gowda S, Weinstein DA, Blalock TD, Gandhi K, Mast BA, Chin G, et al. Topical application of recombinant platelet-derived growth factor increases the rate of healing and the level of proteins that regulate this response. Int Wound J 2015; 12:564-571.
5. Ghieh F, Jurjus R, Ibrahim A, Geagea AG, Daouk H, El Baba B, et al. The use of stem cells in burn wound healing: a review. Biomed Res Int 2015; 2015.
6. Zhang CP, Fu XB. Therapeutic potential of stem cells in skin repair and regeneration. Chin J Traumatol 2008; 11:209-221.
7. Dennis JE, Merriam A, Awadallah A, Yoo JU, Johnstone B, Caplan AI. A quadripotential mesenchymal progenitor cell isolated from the marrow of an adult mouse. J Bone Miner Res 1999; 14:700-709.
8. Jackson WM, Nesti LJ, Tuan RS. Mesenchymal stem cell therapy for attenuation of scar formation during wound healing. Stem Cell Res Ther 2012; 3:20.
9. Oswald J, Boxberger S, Jorgensen B, Feldmann S, Ehninger G, Bornhauser M, et al. Mesenchymal stem cells can be differentiated into endothelial cells in vitro. Stem cells 2004; 22:377-384.
10. Natesan S, Stone II R, Chan RK, Christy RJ. Mesenchymal Stem Cell–Based Therapies for Repair and Regeneration of Skin Wounds.  A Roadmap to Non-Hematopoietic Stem Cell-based Therapeutics: Elsevier; 2019. p. 173-222.
11. Ridiandries A, Tan JTM, Bursill CA. The Role of Chemokines in Wound Healing. Int J Mol Sci 2018; 19.
12. De La Luz Sierra M, Yang F, Narazaki M, Salvucci O, Davis D, Yarchoan R, et al. Differential processing of stromal-derived factor-1alpha and stromal-derived factor-1beta explains functional diversity. Blood 2004; 103:2452-2459.
13. Yu H, Feng Y. The potential of statin and stromal cell-derived factor-1 to promote angiogenesis. Cell adhesion & migration 2008; 2:254-257.
14. Claes F, Vandevelde W, Moons L, Tjwa M. Another angiogenesis-independent role for VEGF: SDF1-dependent cardiac repair via cardiac stem cells. Cardiovasc Res 2011; 91:369-370.
15. Zheng H, Dai T, Zhou B, Zhu J, Huang H, Wang M, et al. SDF-1alpha/CXCR4 decreases endothelial progenitor cells apoptosis under serum deprivation by PI3K/Akt/eNOS pathway. Atherosclerosis 2008; 201:36-42.
16. Hwang JH, Shim SS, Seok OS, Lee HY, Woo SK, Kim BH, et al. Comparison of cytokine expression in mesenchymal stem cells from human placenta, cord blood, and bone marrow. J Korean Med Sci 2009; 24:547-554.
17. Stancu C, Sima A. Statins: mechanism of action and effects. J Cell Mol Med 2001; 5:378-387.
18. Butterick TA, Igbavboa U, Eckert GP, Sun GY, Weisman GA, Muller WE, et al. Simvastatin stimulates production of the antiapoptotic protein Bcl-2 via endothelin-1 and NFATc3 in SH-SY5Y cells. Mol Neurobiol 2010; 41:384-391.
19. Lyngdoh T, Vollenweider P, Waeber G, Marques-Vidal P. Association of statins with inflammatory cytokines: a population-based Colaus study. Atherosclerosis 2011; 219:253-258.
20. Cui X, Chopp M, Zacharek A, Roberts C, Lu M, Savant-Bhonsale S, et al. Chemokine, vascular and therapeutic effects of combination Simvastatin and BMSC treatment of stroke. Neurobiol Dis 2009; 36:35-41.
21. Asai J, Takenaka H, Hirakawa S, Sakabe J, Hagura A, Kishimoto S, et al. Topical simvastatin accelerates wound healing in diabetes by enhancing angiogenesis and lymphangiogenesis. Am J Pathol 2012; 181:2217-2224.
22. BAGHBAN EM, Nazarian H, TAGHIYAR L. Mesenchymal stem cell isolation from the removed medium of rat’s bone marrow primary culture and their differentiation into skeletal cell lineages.  2008.
23. Xie D, Ju D, Speyer C, Gorski D, Kosir MA. Strategic Endothelial Cell Tube Formation Assay: Comparing Extracellular Matrix and Growth Factor Reduced Extracellular Matrix. J Vis Exp 2016.
24. Kwon DS, Gao X, Liu YB, Dulchavsky DS, Danyluk AL, Bansal M, et al. Treatment with bone marrow-derived stromal cells accelerates wound healing in diabetic rats. Int Wound J 2008; 5:453-463.
25. Veith AP, Henderson K, Spencer A, Sligar AD, Baker AB. Therapeutic strategies for enhancing angiogenesis in wound healing. Adv Drug Deliv Rev 2019; 146:97-125.
26. Geesala R, Bar N, Dhoke NR, Basak P, Das A. Porous polymer scaffold for on-site delivery of stem cells--Protects from oxidative stress and potentiates wound tissue repair. Biomaterials 2016; 77:1-13.
27. Campeau L, Ding J, Tredget E. A potential role of SDF-1/CXCR4 chemotactic pathway in wound healing and hypertrophic scar formation. Receptors & Clinical Investigation 2015; 2.
28. Heidari F, Yari A, Rasoolijazi H, Soleimani M, Dehpoor A, Sajedi N, et al. Bulge Hair Follicle Stem Cells Accelerate Cutaneous Wound Healing in Rats. Wounds 2016; 28:132-141.
29. Singer AJ, Choi Y, Rashel M, Toussaint J, McClain SA. The effects of topical nitric oxide on healing of partial thickness porcine burns. Burns 2018; 44:423-428.
30. Suvik A, Effendy A. The use of modified Masson’s trichrome staining in collagen evaluation in wound healing study. Mal J Vet Res 2012; 3:39-47.
31. Chithra P, Sajithlal G, Chandrakasan G. Influence of Aloe vera on the healing of dermal wounds in diabetic rats. Journal of ethnopharmacology 1998; 59:195-201.
32. Bitto A, Minutoli L, Altavilla D, Polito F, Fiumara T, Marini H, et al. Simvastatin enhances VEGF production and ameliorates impaired wound healing in experimental diabetes. Pharmacol Res 2008; 57:159-169.
33. Relja B, Meder F, Wilhelm K, Henrich D, Marzi I, Lehnert M. Simvastatin inhibits cell growth and induces apoptosis and G0/G1 cell cycle arrest in hepatic cancer cells. Int J Mol Med 2010; 26:735-741.


Iranian Journal of Basic Medical Sciences
Volume 23, Issue 6
June 2020
Pages 751-759

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