open access

Vol 57, No 3 (2019)
ORIGINAL PAPERS
Published online: 2019-09-05
Submitted: 2019-03-28
Accepted: 2019-08-26
Get Citation

Stem cells and metformin synergistically promote healing in experimentally induced cutaneous wound injury in diabetic rats

Lamiaa M. Shawky, Eman A. El. Bana, Ahmed A. Morsi
DOI: 10.5603/FHC.a2019.0014
·
Pubmed: 31489604
·
Folia Histochem Cytobiol 2019;57(3):127-138.

open access

Vol 57, No 3 (2019)
ORIGINAL PAPERS
Published online: 2019-09-05
Submitted: 2019-03-28
Accepted: 2019-08-26

Abstract

Introduction. Diabetes mellitus (DM) is a serious, chronic metabolic disorder commonly complicated by diabetic foot ulcers with delayed healing. Metformin was found to have a wound healing effect through several mechanisms. The current study investigated the effect of both bone marrow-derived mesenchymal stem cells (BM-MSCs) and metformin, considered alone or combined, on the healing of an experimentally induced cutaneous wound injury in streptozotocin-induced diabetic rats.
Material and methods. Forty adult male albino rats were used. Diabetes was induced by single intravenous (IV) injection of streptozotocin (STZ). Next, two circular full thickness skin wounds were created on the back of the animals, then randomly assigned into 4 groups, ten rats each. BM-MSCs were isolated from albino rats, 8 weeks of age and labeled by PKH26 before intradermal injection into rats of Group III and IV. Groups I (diabetic positive control), II (metformin-treated, 250 mg/kg/d), III (treated with 2×106 BM-MSCs), and IV (wounded rats treated both with metformin and BM-MSCs cells). Healing was assessed 3, 7, 14, and 21 days post wound induction through frequent measuring of wound diameters. Skin biopsies were obtained at the end of the experiment.
Results. Gross evaluation of the physical healing of the wounds was done. Skin biopsies from the wound areas were processed for hematoxylin and eosin (H&E), Masson’s trichrome staining and immunohistochemical staining for CD31. The results showed better wound healing in the combined therapy group (IV) as compared to monotherapy groups.
Conclusions. Although both metformin and BM-MSCs were effective in the healing of experimentally induced skin wounds in diabetic rats, the combination of both agents appears to be a better synergistic option for the treatment of diabetic wound injuries.

Abstract

Introduction. Diabetes mellitus (DM) is a serious, chronic metabolic disorder commonly complicated by diabetic foot ulcers with delayed healing. Metformin was found to have a wound healing effect through several mechanisms. The current study investigated the effect of both bone marrow-derived mesenchymal stem cells (BM-MSCs) and metformin, considered alone or combined, on the healing of an experimentally induced cutaneous wound injury in streptozotocin-induced diabetic rats.
Material and methods. Forty adult male albino rats were used. Diabetes was induced by single intravenous (IV) injection of streptozotocin (STZ). Next, two circular full thickness skin wounds were created on the back of the animals, then randomly assigned into 4 groups, ten rats each. BM-MSCs were isolated from albino rats, 8 weeks of age and labeled by PKH26 before intradermal injection into rats of Group III and IV. Groups I (diabetic positive control), II (metformin-treated, 250 mg/kg/d), III (treated with 2×106 BM-MSCs), and IV (wounded rats treated both with metformin and BM-MSCs cells). Healing was assessed 3, 7, 14, and 21 days post wound induction through frequent measuring of wound diameters. Skin biopsies were obtained at the end of the experiment.
Results. Gross evaluation of the physical healing of the wounds was done. Skin biopsies from the wound areas were processed for hematoxylin and eosin (H&E), Masson’s trichrome staining and immunohistochemical staining for CD31. The results showed better wound healing in the combined therapy group (IV) as compared to monotherapy groups.
Conclusions. Although both metformin and BM-MSCs were effective in the healing of experimentally induced skin wounds in diabetic rats, the combination of both agents appears to be a better synergistic option for the treatment of diabetic wound injuries.

Get Citation

Keywords

rat; stem cells, BMSCs; STZ diabetes; metformin; skin wound; healing; angiogenesis; CD31

About this article
Title

Stem cells and metformin synergistically promote healing in experimentally induced cutaneous wound injury in diabetic rats

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 57, No 3 (2019)

Pages

127-138

Published online

2019-09-05

DOI

10.5603/FHC.a2019.0014

Pubmed

31489604

Bibliographic record

Folia Histochem Cytobiol 2019;57(3):127-138.

Keywords

rat
stem cells
BMSCs
STZ diabetes
metformin
skin wound
healing
angiogenesis
CD31

Authors

Lamiaa M. Shawky
Eman A. El. Bana
Ahmed A. Morsi

References (41)
  1. Guariguata L, Whiting DR, Hambleton I, et al. Global estimates of diabetes prevalence for 2013 and projections for 2035. Diabetes Res Clin Pract. 2014; 103(2): 137–149.
  2. Katsuda Y, Ohta T, Miyajima K, et al. Diabetic complications in obese type 2 diabetic rat models. Exp Anim. 2014; 63(2): 121–132.
  3. Anderson K, Hamm RL. Factors That Impair Wound Healing. J Am Coll Clin Wound Spec. 2012; 4(4): 84–91.
  4. Li DJ, Huang F, Lu WJ, et al. Metformin promotes irisin release from murine skeletal muscle independently of AMP-activated protein kinase activation. Acta Physiol (Oxf). 2015; 213(3): 711–721.
  5. Rena G, Hardie DG, Pearson ER. The mechanisms of action of metformin. Diabetologia. 2017; 60(9): 1577–1585.
  6. Otranto M, Nascimento AP, Monte-Alto-Costa A. Insulin resistance impairs cutaneous wound healing in mice. Wound Repair Regen. 2013; 21(3): 464–472.
  7. Rahmati M, Pennisi CP, Mobasheri A, et al. Bioengineered Scaffolds for Stem Cell Applications in Tissue Engineering and Regenerative Medicine. Adv Exp Med Biol. 2018; 1107: 73–89.
  8. Council NR. Guide for the care and use of laboratory animals. 8th ed. Wishington, DC: National Academies Press. ; 2010.
  9. Yu JW, Deng YP, Han X, et al. Metformin improves the angiogenic functions of endothelial progenitor cells via activating AMPK/eNOS pathway in diabetic mice. Cardiovasc Diabetol. 2016; 15: 88.
  10. Dunn L, Prosser HCG, Tan JTM, et al. Murine model of wound healing. J Vis Exp. 2013(75): e50265.
  11. Kumar V, Abbas AK, Aster JC. Inflammation and repair. In: Kumar V, Abbas AK, Aster JC, editors. Robbins Basic Pathology. 10th ed. Elsevier Health Sciences; 2017. p. 57.
  12. Furman BL. Streptozotocin-Induced Diabetic Models in Mice and Rats. Curr Protoc Pharmacol. 2015; 70: 5.47.1–5.4720.
  13. Lee CH, Hsieh MJ, Chang SH, et al. Enhancement of diabetic wound repair using biodegradable nanofibrous metformin-eluting membranes: in vitro and in vivo. ACS Appl Mater Interfaces. 2014; 6(6): 3979–3986.
  14. Basiouny HS, Salama NM, Maadawi ZM, et al. Effect of bone marrow derived mesenchymal stem cells on healing of induced full-thickness skin wounds in albino rat. Int J Stem Cells. 2013; 6(1): 12–25.
  15. Hu Y, Lou B, Wu X, et al. Comparative Study on Culture of Mouse Bone Marrow Mesenchymal Stem Cells. Stem Cells Int. 2018; 2018: 6704583.
  16. Baghaei K, Hashemi SM, Tokhanbigli S, et al. Isolation, differentiation, and characterization of mesenchymal stem cells from human bone marrow. Gastroenterol Hepatol Bed Bench. 2017; 10(3): 208–213.
  17. Strober W. Trypan Blue Exclusion Test of Cell Viability. Current Protocols in Immunology. 2015: A3.B.1–A3.B.3.
  18. Ghaneialvar H, Soltani L, Rahmani HR, et al. Characterization and Classification of Mesenchymal Stem Cells in Several Species Using Surface Markers for Cell Therapy Purposes. Indian J Clin Biochem. 2018; 33(1): 46–52.
  19. Bancroft JD, Lyton C. The Hematoxylins and Eosin. In: Suvarna SK, Bancroft JD, Lyton C, editors. Theory and practice of histological techniques. 8th ed, Ed. 2018. p. 126–38.
  20. Kiernan JA. Immunohistochemistry. In: Kiernan J, editor. Histological and histochemical methods Theory and practice. 4–th ed. Scion Publishing Ltd; 2015. p. 454–90.
  21. Reis RM, Reis-Filho JS, Longatto Filho A, et al. Differential Prox-1 and CD 31 expression in mucousae, cutaneous and soft tissue vascular lesions and tumors. Pathol Res Pract. 2005; 201(12): 771–776.
  22. Armstrong DG, Boulton AJM, Bus SA. Diabetic Foot Ulcers and Their Recurrence. N Engl J Med. 2017; 376(24): 2367–2375.
  23. Vileikyte L, Crews RT, Reeves ND. Psychological and Biomechanical Aspects of Patient Adaptation to Diabetic Neuropathy and Foot Ulceration. Curr Diab Rep. 2017; 17(11): 109.
  24. Ojeh N, Pastar I, Tomic-Canic M, et al. Stem Cells in Skin Regeneration, Wound Healing, and Their Clinical Applications. Int J Mol Sci. 2015; 16(10): 25476–25501.
  25. Zhao P, Sui BD, Liu Nu, et al. Anti-aging pharmacology in cutaneous wound healing: effects of metformin, resveratrol, and rapamycin by local application. Aging Cell. 2017; 16(5): 1083–1093.
  26. Čriepoková Z, Lenhardt L’, Gál P. Basic Roles of Sex Steroid Hormones in Wound Repair with Focus on Estrogens (A Review). Folia Veterinaria. 2016; 60(1): 41–46.
  27. Vig K, Chaudhari A, Tripathi S, et al. Advances in Skin Regeneration Using Tissue Engineering. Int J Mol Sci. 2017; 18(4).
  28. Wong SL, Demers M, Martinod K, et al. Diabetes primes neutrophils to undergo NETosis, which impairs wound healing. Nat Med. 2015; 21(7): 815–819.
  29. Li Y, Zhang J, Yue J, et al. Epidermal Stem Cells in Skin Wound Healing. Adv Wound Care (New Rochelle). 2017; 6(9): 297–307.
  30. Seo E, Lim JS, Jun JB, et al. Exendin-4 in combination with adipose-derived stem cells promotes angiogenesis and improves diabetic wound healing. J Transl Med. 2017; 15(1): 35.
  31. Kuo YR, Wang CT, Cheng JT, et al. Adipose-Derived Stem Cells Accelerate Diabetic Wound Healing Through the Induction of Autocrine and Paracrine Effects. Cell Transplant. 2016; 25(1): 71–81.
  32. Serra MB, Barroso WA, da Silva NN, et al. From Inflammation to Current and Alternative Therapies Involved in Wound Healing. Int J Inflam. 2017; 2017: 3406215.
  33. Miller EJ. Collagen types: structure, distribution, and functions. In: Collagen. CRC Press; 2018. p. 139–56.
  34. Elsharawy MA, Naim M, Greish S. Human CD34+ stem cells promote healing of diabetic foot ulcers in rats. Interact Cardiovasc Thorac Surg. 2012; 14(3): 288–293.
  35. Das SK, Yuan YiF, Li MQ. An Overview on Current Issues and Challenges of Endothelial Progenitor Cell-Based Neovascularization in Patients with Diabetic Foot Ulcer. Cell Reprogram. 2017; 19(2): 75–87.
  36. Pacelli S, Basu S, Whitlow J, et al. Strategies to develop endogenous stem cell-recruiting bioactive materials for tissue repair and regeneration. Adv Drug Deliv Rev. 2017; 120: 50–70.
  37. Hu MS, Borrelli MR, Lorenz HP, et al. Mesenchymal Stromal Cells and Cutaneous Wound Healing: A Comprehensive Review of the Background, Role, and Therapeutic Potential. Stem Cells Int. 2018; 2018: 6901983.
  38. Hernandez S, Gong J, Chen L, et al. Characterization of Circulating and Endothelial Progenitor Cells in Patients With Extreme-Duration Type 1 Diabetes. Diabetes Care. 2014; 37(8): 2193–2201.
  39. Ochoa-Gonzalez F, Cervantes-Villagrana AR, Fernandez-Ruiz JC, et al. Metformin Induces Cell Cycle Arrest, Reduced Proliferation, Wound Healing Impairment In Vivo and Is Associated to Clinical Outcomes in Diabetic Foot Ulcer Patients. PLoS One. 2016; 11(3): e0150900.
  40. Lian Z, Yin X, Li H, et al. Synergistic effect of bone marrow-derived mesenchymal stem cells and platelet-rich plasma in streptozotocin-induced diabetic rats. Ann Dermatol. 2014; 26(1): 1–10.

Important: This website uses cookies. More >>

The cookies allow us to identify your computer and find out details about your last visit. They remembering whether you've visited the site before, so that you remain logged in - or to help us work out how many new website visitors we get each month. Most internet browsers accept cookies automatically, but you can change the settings of your browser to erase cookies or prevent automatic acceptance if you prefer.

By "Via Medica sp. z o.o." sp.k., ul. Świętokrzyska 73, 80–180 Gdańsk

tel.:+48 58 320 94 94, faks:+48 58 320 94 60, e-mail:  viamedica@viamedica.pl