MMSL 2024, 93(1):100-111 | DOI: 10.31482/mmsl.2023.011

HISTOLOGICAL AND HISTOMORPHOMETRIC EVALUATION OF THE SYSTEMIC METFORMIN ADMINISTRATION ON BONE HEALING IN RABBITSOriginal article

Raad M. Hussein1*, Ghada A. Taqa ORCID...2
1 Ministry of Health, Kirkuk Health Directorate, Kirkuk, Iraq
2 Department of Dental Basic Sciences, College of Dentistry, University of Mosul, Mosul, Iraq

Background: After injuries, infections, or tumor removal, endogenous healing depends on bone repair. Disorders of bone healing are difficult to treat in clinical settings. There are numerous induced methods for correcting bone abnormalities, such as the induced membrane technique, allogenic bone grafting, synthetic bone grafting, artificial joint replacement, and autologous bone grafting. However, the delivery of the bone graft and bone filling materials necessitates surgical implantation at the fracture site, which could cause edema, infection, and the development of heterotopic bone locally. Therefore, systemically administered osteogenic drugs will provide an excellent method for bone lesion healing. Aim of the study: to evaluate the systemic effect of metformin on bone healing after surgical induction of bony defect and to determine the amount of newly formed bone using histological, histomorphometric analysis, and the surface area measurement of newly formed bone. Also to study the safety of metformin administration at the administered dose for this purpose.

Materials and methods: Twenty mature male New Zealand rabbits were separated into two groups, each including ten rabbits for the study. The same surgical procedure was performed on all rabbits. Two holes were made at the femur (3 mm in diameter and 3 mm in depth) and left empty. Metformin tablets were ground into a fine powder and the resultant powder was dissolved in 10ml of water to prepare a liquid dosage containing 50 mg /1ml of metformin. Metformin is administered orally to the rabbits through a feeding tube at a dose of 50 mg/kg body weight. Animals were euthanized at two-time intervals, 14 and 28 days. The femur was separated, sectioned preserved, and sent for histological analysis and histomor-phometry.

Results: The results revealed that there is an increase in new bone formation and bone-forming cells in the metformin-treated group.

Conclusion: Metformin increases bone healing by increasing the number of bone-forming cells and the surface area of newly formed bone tissues and causes less inflammatory response at the site of a bone lesion. So it possesses an osteogenic effect.

Keywords: Metformin; systemic; osteoblast; osteoclast; bone healing

Received: November 11, 2022; Revised: January 23, 2023; Accepted: February 22, 2023; Prepublished online: March 13, 2023; Published: March 1, 2024  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Hussein, R.M., & Taqa, G.A. (2024). HISTOLOGICAL AND HISTOMORPHOMETRIC EVALUATION OF THE SYSTEMIC METFORMIN ADMINISTRATION ON BONE HEALING IN RABBITS. MMSL, 93(1), 100-111. doi: 10.31482/mmsl.2023.011
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Pajarinen J, Lin T, Gibon E, et al. Mesenchymal stem cell-macrophage crosstalk and bone healing. Biomaterials. 2019;196:80-9. https://doi.org/10.1016/j.biomaterials.2017.12.025 Go to original source... Go to PubMed...
    2. Schell H, Duda GN, Peters A, et al. The hematoma and its role in bone healing. Journal of experimental orthopedics. 2017;4(1):1-1. https://doi.org/10.1186/s40634-017-0079-3 Go to original source... Go to PubMed...
    3. Bell A, Templeman D, Weinlein JC. Nonunion of the femur and tibia: an update. Orthopedic Clinics. 2016;47(2):365-375. https://doi.org/10.1016/j.ocl.2015.09.010 Go to original source... Go to PubMed...
    4. Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature. 2003 May;423(6937):337-342. https://doi.org/10.1038/nature01658 Go to original source... Go to PubMed...
    5. Medhat D, Rodríguez CI, Infante A. Immunomodulatory effects of MSCs in bone healing. International Journal of Molecular Sciences. 2019;20(21):5467. https://doi.org/10.3390/ijms20215467. Go to original source... Go to PubMed...
    6. Aoki K, Saito N. Biodegradable polymers as drug delivery systems for bone regeneration. Pharmaceutics. 2020;12(2):95. https://doi.org/10.3390/pharmaceutics12020095. Go to original source... Go to PubMed...
    7. Ratko TA, Belinson SE, Samson DJ, et al. Bone Morphogenetic Protein: The State of the Evidence of On-Label and Off-Label Use (Internet). https://europepmc.org/article/nbk/nbk284865
    8. Wang P, Ma T, Guo D, et al. Metformin induces osteoblastic differentiation of human induced pluripotent stem cell-derived mesenchymal stem cells. Journal of tissue engineering and regenerative medicine. 2018;12(2):437-446. https://doi.org/10.1002/term.2470 Go to original source... Go to PubMed...
    9. Rothe R, Schulze S, Neuber C, et al. Adjuvant drug-assisted bone healing: Part III-Further strategies for local and systemic modulation. Clinical Hemorheology and Microcirculation. 2019;73(3):439-488. https://doi.org/10.3233/CH-199104 Go to original source... Go to PubMed...
    10. Kobayashi M, Yamazaki K, Hirao K, et al. The status of diabetes control and antidiabetic drug therapy in Japan-a cross-sectional survey of 17,000 patients with diabetes mellitus (JDDM 1). Diabetes research and clinical practice. 2006;73(2):198-204. https://doi.org/10.1016/j.diabres.2006.01.013 Go to original source... Go to PubMed...
    11. Andrews M, Soto N, Arredondo M. Effect of metformin on the expression of tumor necrosis factor-α, Toll-like receptors 2/4 and C reactive protein in obese type-2 diabetic patients. Revista medica de Chile. 2012;140(11):1377-1382. https://doi.org/10.4067/s0034-98872012001100001 Go to original source... Go to PubMed...
    12. Koh SJ, Kim JM, Kim IK, et al. Anti-inflammatory mechanism of metformin and its effects in intestinal inflammation and colitis-associated colon cancer. Journal of gastroenterology and hepatology. 2014;29(3):502-510. https://doi.org/10.1111/jgh.12435 Go to original source... Go to PubMed...
    13. Ren C, Hao X, Wang L, et al. Metformin Carbon Dots for promoting periodontal bone regeneration via activation of ERK/AMPK pathway. Advanced Healthcare Materials. 2021;10(12):2100196. https://doi.org/10.1002/adhm.202100196 Go to original source... Go to PubMed...
    14. Huang W, Castelino RL, Peterson GM. Metformin usage in type 2 diabetes mellitus: are safety guidelines adhered to? Internal medicine journal. 2014;44(3):266-272. https://doi.org/10.1111/imj.12369 Go to original source... Go to PubMed...
    15. de Lima Santos A, da Silva CG, de Sá Barreto LS, et al. A new decellularized tendon scaffold for rotator cuff tears-evaluation in rabbits. BMC Musculoskeletal Disorders. 2020;21(1):1-2. https://doi.org/10.1186/s12891-020-03680-w Go to original source... Go to PubMed...
    16. Merkhan MM. Effect of metformin, glibenclamide, and insulin on lipid profile in type 2 diabetic patients. Tikret Journal of Pharmaceutical Sciences. 2013;9(2). Go to original source...
    17. Ikewuchi CC, Ikewuchi JC, Ifeanacho MO. Restoration of plasma markers of liver and kidney functions/integrity in alloxan-induced diabetic rabbits by aqueous extract of Pleurotus tuberregium sclerotia. Biomedicine & Pharmacotherapy. 2017;95:1809-1814. https://doi.org/10.1016/j.biopha.2017.09.075 Go to original source... Go to PubMed...
    18. Ahirwar LK, Kalra P, Sharma S, et al. Linezolid shows high safety and efficacy in the treatment of Pythium insidiosum keratitis in a rabbit model. Experimental Eye Research. 2021;202:108345. https://doi.org/10.1016/j.exer.2020.108345. Go to original source... Go to PubMed...
    19. Jiron JM, Mendieta Calle JL, Castillo EJ, et al. Comparison of isoflurane, ketamine-dexmedetomidine, and ketamine-xylazine for general anesthesia during oral procedures in rice rats (Oryzomys palustris). Journal of the American Association for Laboratory Animal Science. 2019;58(1):40-49. https://doi.org/10.30802/AALAS-JAALAS-18-000032 Go to original source... Go to PubMed...
    20. Garcia-Donas JG, Dalton A, Chaplin I, et al. A revised method for the preparation of dry bone samples used in the histological examination: Five simple steps. Homo. 2017;68(4):283-288. https://doi.org/10.1016/j.jchb.2017.07.001. Go to original source... Go to PubMed...
    21. Ahmed HN, Ghada AT, Wael T. The Effect of Xylitol on Osteoclastogenesis in Experimentally Induced Bone Defect in Rabbits. Journal of Applied Veterinary Sciences. 2022;7(1):58-63. https://doi.org/10.21608/JAVS.2021.104726.1114 Go to original source...
    22. Abdo FS. Effects of Combination of Platelet Rich Plasma and OSTEON Material in Rabbits Bone Healing (A comparative study). Al-Rafidain Dental Journal. 2014;14(1):90-100. https://doi.org/10.33899/rden.2014.89258 Go to original source...
    23. Mohammed ZS, Taqa GA, Sulaiman MS. Evaluating the local and systemic effects of silicon dioxide on healing of mandibular bone in rabbits. InAIP Conference Proceedings 2022 (Vol. 2547, No. 1, p. 020006). AIP Publishing LLC. https://doi.org/10.1063/5.0116015 Go to original source...
    24. Shen M, Yu H, Jin Y, et al. Metformin facilitates osteoblastic differentiation and M2 macrophage polarization by PI3K/AKT/mTOR pathway in human umbilical cord mesenchymal stem cells. Stem Cells International. 2022;2022. https://doi.org/10.1155/2022/9498876 Go to original source... Go to PubMed...
    25. Zhou R, Ma Y, Qiu S, et al. Metformin promotes cell proliferation and osteogenesis under high glucose conditions by regulating the ROS AKT mTOR axis. Molecular Medicine Reports. 2020;22(4):3387-3395. https://doi.org/10.3892/mmr.2020.11391 Go to original source... Go to PubMed...
    26. Zhao R, Yang R, Cooper PR, et al. Bone grafts and substitutes in dentistry: A review of current trends and developments. Molecules. 2021;26(10):3007. https://doi.org/10.3390/molecules26103007. Go to original source... Go to PubMed...
    27. Son HJ, Lee J, Lee SY, et al. Metformin attenuates experimental autoimmune arthritis through reciprocal regulation of Th17/Treg balance and osteoclastogenesis. Mediators of inflammation. 2014. https://doi.org/10.1155/2014/973986 Go to original source... Go to PubMed...
    28. Crockett JC, Rogers MJ, Coxon FP, et al. Bone remodeling at a glance. Journal of cell science. 2011;124(7):991-998. https://doi.org/10.1242/jcs.063032 Go to original source... Go to PubMed...
    29. Chen X, Wang Z, Duan N, et al. Osteoblast-osteoclast interactions. Connective tissue research. 2018;59(2):99-107. https://doi.org/10.1080/03008207.2017.1290085 Go to original source... Go to PubMed...
    30. Owen R, Reilly GC. In vitro models of bone remodeling and associated disorders. Frontiers in bioengineering and biotechnology. 2018;6:134. https://doi.org/10.3389/fbioe.2018.00134 Go to original source... Go to PubMed...
    31. Qing L, Fu J, Wu P, et al. Metformin induces the M2 macrophage polarization to accelerate wound healing via regulating AMPK/mTOR/NLRP3 inflammasome singling pathway. American journal of translational research. 2019;11(2):655. Go to PubMed...
    32. Bahrambeigi S, Yousefi B, Rahimi M, et al. Metformin; an old antidiabetic drug with new potentials in bone disorders. Biomedicine & Pharmacotherapy. 2019;109:1593-1601. https://doi.org/10.1016/j.biopha.2018.11.032. Go to original source... Go to PubMed...
    33. ¦mieszek A, Tomaszewski KA, Kornicka K, et al. Metformin promotes osteogenic differentiation of adipose-derived stromal cells and exerts pro-osteogenic effect stimulating bone regeneration. Journal of clinical medicine. 2018;7(12):482. https://doi.org/10.3390/jcm7120482. Go to original source... Go to PubMed...
    34. Zhen D, Chen Y, Tang X. Metformin reverses the deleterious effects of high glucose on osteoblast function. Journal of Diabetes and its Complications. 2010;24(5):334-344. https://doi.org/10.1016/j.jdiacomp.2009.05.002. Go to original source... Go to PubMed...
    35. Seetharaman R, Mahmood A, Kshatriya P, et al. An overview of stem cells in tissue regeneration. Current pharmaceutical design. 2019;25(18):2086-2098. https://doi.org/10.2174/1381612825666190705211705. Go to original source... Go to PubMed...
    36. Chai X, Zhang W, Chang B, et al. GPR39 agonist TC-G 1008 promotes osteoblast differentiation and mineralization in MC3T3-E1 cells. Artificial Cells, Nanomedicine, and Biotechnology. 2019;47(1):3569-3576. https://doi.org/10.1080/21691401.2019.1649270. Go to original source... Go to PubMed...
    37. Chava S, Chennakesavulu S, Gayatri BM, et al. Novel phosphorylation by AMP-activated kinase regulates RUNX2 from ubiquitination in osteogenesis over adipogenesis. Cell death & disease. 2018;9(7):1-6. https://doi.org/10.1038/s41419-018-0791-7. Go to original source... Go to PubMed...
    38. Sun R, Liang C, Sun Y, et al. Effects of metformin on the osteogenesis of alveolar BMSCs from diabetic patients and implant osseointegration in rats. Oral Diseases. 2022;28(4):1170-1180. https://doi.org/10.1111/odi.13808. Go to original source... Go to PubMed...
    39. Gonnelli S, Caffarelli C, Giordano N, et al. The prevention of fragility fractures in diabetic patients. Aging Clinical and Experimental Research. 2015;27(2):115-124. https://doi.org/10.1007/s40520-014-0258-3 Go to original source... Go to PubMed...
    40. Jiating L, Buyun J, Yinchang Z. Role of metformin on osteoblast differentiation in type 2 diabetes. BioMed Research International. 2019;2019. https://doi.org/10.1155/2019/9203934. Go to original source... Go to PubMed...
    41. Almulathanon AA, Mohammad JA, Fathi FH. Comparative effects of metformin and glibenclamide on the redox balance in type 2 diabetic patients. Pharmacia. 2021;68(2):327-332. https://doi.org/10.3897/pharmacia.68.e63365 Go to original source...
    42. Pernicova I, Kelly S, Ajodha S, et al. Metformin to reduce metabolic complications and inflammation in patients on systemic glucocorticoid therapy: a randomized, double-blind, placebo-controlled, proof-of-concept, phase 2 trial. The lancet Diabetes & endocrinology. 2020;8(4):278-291. https://doi.org/10.1016/S2213-8587(20)30021-8 Go to original source... Go to PubMed...
    43. Zhu X, Yan H, Chang X, et al. Association between non-alcoholic fatty liver disease-associated hepatic fibrosis and bone mineral density in postmenopausal women with type 2 diabetes or impaired glucose regulation. BMJ Open Diabetes Research and Care. 2020;8(1):e000999. http://dx.doi.org/10.1136/bmjdrc-2019-000999 Go to original source... Go to PubMed...
    44. Bahrambeigi S, Yousefi B, Rahimi M, et al. Metformin; an old antidiabetic drug with new potentials in bone disorders. Biomedicine & Pharmacotherapy. 2019;109:1593-1601. https://doi.org/10.1016/j.biopha.2018.11.032. Go to original source... Go to PubMed...
    45. Merkhan MM, Shephard MT, Forsyth NR. Physoxia alters the human mesenchymal stem cell secretome. Journal of Tissue Engineering. 2021;12:20417314211056132. https://doi.org/10.1177/20417314211056132. Go to original source... Go to PubMed...
    46. Forsyth NR, Steeg R, Ahmad M, et al. Mimicking Physiological Oxygen in Cell Cultures. InCell Culture Technology 2018 (pp. 129-137). Springer, Cham. https://doi.org/10.1007/978-3-319-74854-2_8. Go to original source...
    47. Chen L, Merkhan MM, Forsyth NR, et al. Chorionic and amniotic membrane-derived stem cells have distinct and gestational diabetes mellitus independent, proliferative, differentiation, and immunomodulatory capacities. Stem Cell Research. 2019;40:101537. https://doi.org/10.1016/j.scr.2019.101537 Go to original source... Go to PubMed...
    48. Shephard MT, Merkhan MM, Forsyth NR. Human Mesenchymal Stem Cell Secretome Driven T Cell Immunomodulation Is IL-10 Dependent. International Journal of Molecular Sciences. 2022;23(21):13596. https://doi.org/10.3390/ijms232113596. Go to original source... Go to PubMed...
    49. Yildirim TT, Dündar S, Bozoğlan A, et al. The effects of metformin on the bone filling ration around of TiAl6Va4 implants in non diabetic rats. Journal of Oral Biology and Craniofacial Research. 2020;10(4):474-477. https://doi.org/10.1016/j.jobcr.2020.07.012. Go to original source... Go to PubMed...
    50. Bonnet F, Scheen A. Understanding and overcoming metformin gastrointestinal intolerance. Diabetes, Obesity and Metabolism. 2017;19(4):473-481. https://doi.org/10.1111/dom.12854 Go to original source... Go to PubMed...

    Return to the content