Skeletal Effects of a Prolonged Oral Metformin Treatment in Adult Wistar Rats

Abstract

Introduction We previously showed that a 3-week oral metformin (MET) treatment enhances the osteogenic potential of bone marrow stromal cells (BMSCs) and improves several bone histomorphometric parameters in Wistar rats with metabolic syndrome (MetS). However, the skeletal effects of extended periods of MET need to be completely elucidated. Hence, in this study, the impact of a prolonged (3-month) MET treatment was investigated on bone architecture, histomorphometric and biomechanics variables, and osteogenic potential of BMSCs in Wistar rats with or without MetS.

Materials and Methods Young male Wistar rats (n=36) were randomized into four groups (n=9) that received either 20% fructose (F), MET (MET), F plus MET treatments (FMET), or drinking water alone (Veh). Rats were euthanized, blood was collected, and bones were dissected and processed for peripheral quantitative computed tomography (pQCT) analysis, static and dynamic histomorphometry, and bone biomechanics. In addition, BMSCs were isolated to determine their osteogenic potential.

Results MET affected trabecular and cortical bone, altering bone architecture and biomechanics. Furthermore, MET increased the pro-resorptive profile of BMSCs. In addition, fructose-induced MetS practically did not affect the the structural or mechanical variables of the skeleton.

Conclusion A 3-month treatment with MET (with or without MetS) affects bone architecture and biomechanical variables in Wistar rats.

Keywords

bone architecture - biomechanics - bone marrow stromal cells - metformin - metabolic syndrome

• References

• 1 Grundy SM, Cleeman JI, Daniels SR. et al. Diagnosis and management of the metabolic syndrome: An American Heart Association/National Heart, Lung, and Blood Institute Scientific Statement. Circulation 2005; 112: 2735-2752
• 2 Alberti KG, Eckel RH, Grundy SM. et al. Harmonizing the metabolic syndrome: A joint interim statement of the International Diabetes Federation Task Force on Epidemiology and Prevention; National Heart, Lung, and Blood Institute; American Heart Association; World Heart Federation; International Atherosclerosis Society; and International Association for the Study of Obesity. Circulation 2009; 120: 1640-1645
• 3 Chin KY, Wong SK, Ekeuku SO. et al. Relationship between metabolic syndrome and bone health - an evaluation of epidemiological studies and mechanisms involved. Diabetes Metab Syndr Obes 2020; 13: 3667-3690
• 4 Chen DZ, Xu QM, Wu XX. et al. The combined effect of nonalcoholic fatty liver disease and metabolic syndrome on osteoporosis in postmenopausal females in Eastern China. Int J Endocrinol 2018; 2018: 2314769
• 5 Hwang DK, Choi HJ. The relationship between low bone mass and metabolic syndrome in Korean women. Osteoporos Int 2020; 21: 425-431
• 6 Kim H, Oh HJ, Choi H. et al. The association between bone mineral density and metabolic syndrome: A Korean population-based study. J Bone Miner Metab 2013; 31: 571-578
• 7 Muka T, Trajanoska K, Kiefte-de Jong JC. et al. The Association between metabolic syndrome, bone mineral density, hip bone geometry and fracture risk: The Rotterdam Study. PLoS One 2015; 10: e0129116
• 8 Von Muhlen D, Safii S, Jassal SK. et al. Associations between the metabolic syndrome and bone health in older men and women: The Rancho Bernardo Study. Osteoporos Int 2007; 18: 1337-1344
• 9 Qin L, Yang Z, Zhang W. et al. Metabolic syndrome and osteoporotic fracture: A population-based study in China. BMC Endocr Disord 2016; 16: 27
• 10 McCarthy AD, Molinuevo MS, Cortizo AM. AGEs and bone aging in diabetes mellitus. J Diabetes Metab 2013; 4: 6
• 11 Panchal SK, Brown L. Rodent models for metabolic syndrome research. J Biomed Biotechnol 2011; 2011: 351982
• 12 Felice JI, Gangoiti MV, Molinuevo MS. et al. Effects of a metabolic syndrome induced by a fructose-rich diet on bone metabolism in rats. Metabolism 2014; 63: 296-305
• 13 Felice JI, Schurman L, McCarthy AD. et al. Effects of fructose-induced metabolic syndrome on rat skeletal cells and tissue, and their responses to metformin treatment. Diabetes Research and Clinical Practice 2017; 126: 202-213
• 14 Bahrambeigi S, Yousefi B, Rahimi M. et al. Metformin; an old antidiabetic drug with new potentials in bone disorders. Biomed Pharmacother 2019; 109: 1593-1601
• 15 Mu W, Liang G, Feng Y. et al. The potential therapeutic role of metformin in diabetic and non-diabetic bone impairment. Pharmaceuticals 2022; 15: 1274
• 16 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 Dis 2022; 28: 1170-1180
• 17 Loh DKW, Kadirvelu A, Pamidi N. Effects of metformin on bone mineral density and adiposity-associated pathways in animal models with type 2 diabetes mellitus: A systematic review. J Clin Med 2022; 11: 4193
• 18 McCarthy AD, Cortizo AM, Sedlinsky C. Metformin revisited: Does this regulator of AMP-activated protein kinase secondarily affect bone metabolism and prevent diabetic osteopathy?. World J Diabetes 2016; 7: 122-133
• 19 Tolosa MJ, Chuguransky SR, Sedlinsky C. et al. Insulin-deficient diabetes-induced bone microarchitecture alterations are associated with a decrease in the osteogenic potential of bone marrow progenitor cells: Preventive effects of metformin. Diabetes Res Clin Pract 2013; 101: 177-186
• 20 Cortizo AM, Sedlinsky C, McCarthy AD. et al. Osteogenic actions of the antidiabetic drug metformin on osteoblasts in culture. Eur J Pharmacol 2006; 536: 38-46
• 21 Molinuevo MS, Schurman L, McCarthy AD. et al. Effect of metformin on bone marrow progenitor cell differentiation: In vivo and in vitro studies. J Bone Miner Res 2010; 25: 211-221
• 22 Kaya YE, Karaarslan N, Yilmaz I. et al. A study of the effects of metformin, a biguanide derivative, on annulus fibrosus and nucleus pulposus cells. Turk Neurosurg 2020; 30: 434-441
• 23 Marycz K, Tomaszewski KA, Kornicka K. et al. Metformin decreases reactive oxygen species, enhances osteogenic properties of adipose-derived multipotent mesenchymal stem cells in vitro, and increases bone density in vivo. Oxid Med Cell Longev 2016; 2016: 9785890
• 24 Wang C, Li H, Chen SG. et al. The skeletal effects of thiazolidinedione and metformin on insulin-resistant mice. J Bone Miner Metab 2012; 30: 630-637
• 25 La Fontaine J, Chen C, Hunt N. et al. Type 2 diabetes and metformin influence on fracture healing in an experimental rat model. J Foot Ankle Surg 2016; 55: 955-960
• 26 Jeyabalan J, Viollet B, Smitham P. et al. The antidiabetic drug metformin does not affect bone mass. in vivo or fracture healing. Osteoporos Int 2013; 24: 2659-2670
• 27 UFAW. The UFAW handbook on the care and management of laboratory and other research animals. 8th ed. United Kingdom: Wiley-Blackwell; 2011
• 28 Mamikutty N, Thent ZC, Sapri SR. et al. The establishment of metabolic syndrome model by induction of fructose drinking water in male Wistar rats. Biomed Res Int 2014; 2014: 263897
• 29 Dupas J, Feray A, Goanvec C. et al. Metabolic syndrome and hypertension resulting from fructose enriched diet in Wistar rats. Biomed Res Int 2017; 2017: 2494067
• 30 Maiztegui B, Borelli MI, Raschia MA. et al. Islet adaptive changes to fructose-induced insulin resistance: beta-cell mass, glucokinase, glucose metabolism and insulin secretion. J Endocrinol 2009; 200: 139-149
• 31 Choi YH, Lee MG, Lee I. Effects of diabetes mellitus induced by alloxan on the pharmacokinetics of metformin in rats: Restoration of pharmacokinetic parameters to the control state by insulin treatment. J Pharm Pharm Sci 2008; 11: 88-103
• 32 Cacho J, Sevillano J, de Castro J. et al. Validation of simple indexes to assess insulin sensitivity during pregnancy in Wistar and Sprague-Dawley rats. Am J Physiol Endocrinol Metab 2008; 295: E1269-E1276
• 33 da Luz PL, Favarato D, Faria-Neto JR. et al. High ratio of triglycerides to HDL cholesterol predicts extensive coronary disease. Clinics 2008; 63: 427-432
• 34 Aguirre JI, Franz SE, Altman MK. et al. Skeletal effects of fibroblast growth factor mimetic (F2A) in ovariectomized rats. J Musculoskelet Neuronal Interact 2009; 9: 38-43
• 35 Yarrow JF, Conover CF, Beggs LA. et al. Testosterone dose-dependently prevents bone and muscle loss in rodents after spinal cord injury. J Neurotrauma 2014; 31: 834-845
• 36 Dempster DW, Compston JE, Drezner MK. et al. Standardized nomenclature, symbols, and units for bone histomorphometry: A 2012 update of the report of the ASBMR Histomorphometry Nomenclature Committee. J Bone Miner Res 2013; 28: 2-17
• 37 UK Prospective Diabetes Study (UKPDS) Group. Effect of intensive blood-glucose control with metformin on complications in overweight patients with type 2 diabetes (UKPDS 34). Lancet 1998; 352: 854-865
• 38 Rask Larsen J, Dima L, Correll CU. et al. The pharmacological management of metabolic syndrome. Exp Rev Clin Pharmacol 2018; 11: 397-410
• 39 Meier C, Schwartz AV, Egger A. et al. Effects of diabetes drugs on the skeleton. Bone 2016; 82: 93-100
• 40 Colhoun HM, Livingstone SJ, Looker HC. et al. Hospitalised hip fracture risk with rosiglitazone and pioglitazone use compared with other glucose-lowering drugs. Diabetologia 2012; 55: 2929-2937
• 41 Kolovou GD, Anagnostopoulou KK, Salpea KD. et al. The prevalence of metabolic syndrome in various populations. Am J Med Sci 2007; 333: 362-371
• 42 Bray GA, Popkin BM. Dietary sugar and body weight: Have we reached a crisis in the epidemic of obesity and diabetes?: Health be damned! Pour on the sugar. Diabetes Care 2014; 37: 950-956
• 43 Wong SK, Chin KY, Suhaimi FH. et al. Effects of metabolic syndrome on bone mineral density, histomorphometry and remodelling markers in male rats. PLoS One 2018; 13: e0192416
• 44 Tsanzi E, Light HR, Tou JC. The Effect of feeding different sugar-sweetened beverages to growing female Sprague-Dawley rats on bone mass and strength. Bone 2008; 42: 960-968
• 45 Monami M, Cresci B, Colombini A. et al. Bone fractures and hypoglycemic treatment in type 2 diabetic patients: A case-control study. Diabetes Care 2008; 31: 199-203
• 46 Oh TK, Song IA. Metformin therapy and hip fracture risk among patients with type II diabetes mellitus: A population-based cohort study. Bone 2020; 135: 115325
• 47 Schurman L, McCarthy AD, Sedlinsky C. et al. Metformin reverts deleterious effects of advanced glycation end-products (AGEs) onosteoblastic cells. Exp Clin Endocrinol Diabetes 2008; 116: 333-340
• 48 Kanazawa I, Yamaguchi T, Yano S. et al. Metformin enhances the differentiation and mineralization of osteoblastic MC3T3-E1 cells via AMP kinase activation as well as eNOS and BMP-2 expression. Biochem Biophys Res Commun 2008; 375: 414-419
• 49 Jang WG, Kim EJ, Bae IH. et al. Metformin induces osteoblast differentiation via orphan nuclear receptor SHP mediated transactivation of Runx2. Bone 2011; 48: 885-893
• 50 Mai QG, Zhang ZM, Xu S. et al. Metformin stimulates osteoprotegerin and reduces RANKL expression in osteoblasts and ovariectomized rats. J Cell Biochem 2011; 112: 2902-2909
• 51 Sedlinsky C, Molinuevo MS, Cortizo AM. et al. Metformin prevents anti-osteogenic in vivo and ex vivo effects of rosiglitazone in rats. Eur J Pharmacol 2011; 668: 477-485
• 52 Jang WG, Kim EJ, Lee KN. et al. AMP-activated protein kinase (AMPK) positively regulates osteoblast differentiation via induction of Dlx5-dependent Runx2 expression in MC3T3E1 cells. Biochem Biophys Res Commun 2011; 404: 1004-1009