The Inhibitory Role of Di-2-ethylhexyl Phthalate on Osteogenic Differentiation of Mesenchymal Stem Cells Via Down-regulation of RUNX2 and Membrane Function Impairment
International Journal of Medical Toxicology and Forensic Medicine,
Vol. 10 No. 2 (2020),
21 June 2020
,
Page 26673
https://doi.org/10.32598/ijmtfm.v10i2.26673
Abstract
Background: Blood contamination of di-2-ethyl hexyl phthalate (DEHP) has been reported due to its release following medical procedures such as blood transfusion and vital liquid injection. We investigated the effect of DEHP on osteogenic differentiation of mesenchymal stem cells and their viability.
Methods: The rat bone marrow mesenchymal cells (MSCs) were cultured three times, and the third passage kept in the differentiation medium with the presence of DEHP. The viability of differentiated cells, sodium and potassium level, calcium concentration, total protein concentration, and the activity of lactate dehydrogenase, alkaline phosphatase, alanine transaminase, and aspartate transaminase were determined. Also, the concentration of malondialdehyde, total antioxidant capacity, the activity of superoxide dismutase and catalase were estimated. Finally, the level of matrix deposition and expression of alkaline phosphatase (ALP) and runt-related transcription factor 2 (RUNX2) genes were evaluated.
Results: We observed a concentration-dependent and significant reduction of matrix mineralization based on alizarin red and calcium analysis. Besides, the expression of ALP and RUNX2 gene was down-regulated, and alkaline-phosphatase activity reduced significantly. Also, we observed cell viability reduction but the elevation of lactate dehydrogenase activity and malondialdehyde level. Sodium level was elevated too, whereas the activity of transaminases, oxidative stress enzymes, potassium level, and total antioxidants decreased.
Conclusion: DEHP contamination reduced matrix mineralization due to the down-regulation of the genes involved in osteogenic differentiation and viability reduction via electrolyte and metabolic imbalance as well as induction of oxidative stress.
- Differentiation
- Di-2-ethylhexyl phthalate (DEHP)
- Metabolic enzymes
- Mesenchymal stem cell
- Oxidative stress
How to Cite
References
1. Huber WW, Grasl-Kraupp B, Schulte-Hermann R. Hepatocarcinogenic potential of di (2-ethylhexyl) phthalate in rodents and its implications on human risk. Crit Rev Toxicol. 1996; 26(4):365-481. [DOI:10.3109/10408449609048302] [PMID]
2. Stanford Research Associates (SRI) International. 1996. Plasticizers. Chemical Economics Handbook. Palo Alto, CA.
3. Jaeger R, Rubin R. Plasticizers from PVC. Lancet. 1970; 296(7676):778. [DOI:10.1016/S0140-6736(70)90261-8]
4. Jaeger RJ, Rubin RJ. Migration of a phthalate ester plasticizer from polyvinyl chloride blood bags into stored human blood and its localization in human tissues. N Engl J Med. 1972; 287(22):1114-8. [DOI:10.1056/NEJM197211302872203] [PMID]
5. Autian J. Toxicity and health threats of phthalate esters: Review of the literature. Environ Health Perspect. 1973; 4:3-26. [DOI:10.1289/ehp.73043] [PMID] [PMCID]
6. Hanno C Erythropel, Maric M, Nicell JA, Leask RL, Yargeau V. Leaching of the plasticizer di(2-ethylhexyl)phthalate (DEHP) from plastic containers and the question of human exposure. Appl Microbiol Biotechnol . 2014; 98(24):9967-81. [DOI: 10.1007/s00253-014-6183-8] [PMID]
7. Jaeger RJ, Rubin RJ. Extraction, localization, and metabolism of di-2-ethylhexyl phthalate from PVC plastic medical devices. Environ Health Perspect. 1973; 3:95-102. [DOI:10.1289/ehp.730395] [PMID] [PMCID]
8. Meeker JD, Sathyanarayana S, Swan SH. Phthalates and other additives in plastics: Human exposure and associated health outcomes. Philos Trans R Soc Lond B Biol Sci. 2009; 364(1526):2097-113. [DOI:10.1098/rstb.2008.0268] [PMID] [PMCID]
9. Calafat AM, Needham LL, Silva MJ, Lambert G. Exposure to di-(2-ethylhexyl) phthalate among premature neonates in a neonatal intensive care unit. Pediatrics. 2004; 113:429-34. [DOI:10.1542/peds.113.5.e429] [PMID]
10. Gibson TP, Briggs WA, Boone BJ. Delivery of di-2-ethylhexyl phthalate to patients during hemodialysis. J Lab Clin Med. 1976; 87(3):519-24. https://www.ncbi.nlm.nih.gov/pubmed/1249479
11. Shneider B, Schena J, Truog R, Jacobson M, Kevy S. Exposure to di (2-ethylhexyl) phthalate in infants receiving extracorporeal membrane oxygenation. N Engl J Med. 1989; 320(23):1563(letter). [DOI:10.1056/NEJM198906083202322]
12. Chawla AS, Hinberg I. Leaching of plasticizers from and surface characterization of PVC blood platelet bags. Biomater. Artif Cells Immobilization Biotechnol. 1991; 19(4):761-83. [DOI:10.3109/10731199109117853] [PMID]
13. Pearson SD, Trissel LA. Leaching of diethylhexyl phthalate from polyvinyl chloride containers by selected drugs and formulation components. Am J Health Syst Pharm. 1993; 50(7):1405-9. [DOI:10.1093/ajhp/50.7.1405]
14. Mazzo DJ, Nguyen-Huu JJ, Pagniez S, Denis P. Compatibility of docetaxel and paclitaxel in intravenous solutions with polyvinyl chloride infusion materials. Am J Health Syst Pharm. 1997; 54(5):566-9. [DOI:10.1093/ajhp/54.5.566] [PMID]
15. Cole RS, Tocchi M, Wye E, Villeneuve DC, Rock G. Contamination of commercial blood products by di-2-ethylhexyl Phthalate and mono-2-ethylhexyl Phthalate. Vox Sang. 1981; 40(5):317-22. [DOI:10.1111/j.1423-0410.1981.tb00715.x] [PMID]
16. Kluwe WM. Overview of phthalate ester pharmacokinetics in mammalian species. Environ Health Perspect. 1982; 45:3-10. [DOI:10.1289/ehp.82453] [PMID] [PMCID]
17. Jarosova A, Harazim J, Suchy P, Kratka L, Stancova V. The distribution and accumulation of phthalates in the organs and tissues of chicks after the administration of feedstuffs with different phthalate concentrations. Vet Med. 2009; 54:427-34. [DOI:10.17221/2/2009-VETMED]
18. Tickner JA, Schettler T, Guidotti T, McCally M, Rossi M. Health risks posed by use of Di‐2‐ethylhexyl phthalate (DEHP) in PVC medical devices: A critical review. Am J Ind Med. 2001; 39(1):100-11. [DOI:10.1002/1097-0274(200101)39:13.0.CO;2-Q]
19. Peters JM, Taubeneck MW, Keen CL, Gonzalez FJ. Di (2-ethylhexyl) phthalate induces a functional zinc deficiency during pregnancy and teratogenesis that is independent of peroxisome proliferator-activated receptor-α. Teratology. 1997; 56(5):311-6. [DOI:10.1002/(SICI)1096-9926(199711)56:53.0.CO;2-#]
20. Bhat FA, Ramajayam G, Parameswari S, Vignesh RC, Karthikeyan S, Senthilkumar K, et al. Di 2-ethyl hexyl phthalate affects differentiation and matrix mineralization of rat calvarial osteoblasts-in vitro. Toxicol In Vitro. 2013; 27(1):250-6. [DOI:10.1016/j.tiv.2012.09.003] [PMID]
21. Sobiesiak M, Sivasubramaniyan K, Hermann C, Tan C, Orgel M, Treml S, et al. The mesenchymal stem cell antigen MSCA-1 is identical to tissue non-specific alkaline phosphatase. Stem Cells Dev. 2010; 19(5):669-77. [DOI:10.1089/scd.2009.0290] [PMID]
22. Hartmann C. A Wnt canon orchestrating osteoblastogenesis. Trends Cell Biol. 2006; 16:151-8. [DOI:10.1016/j.tcb.2006.01.001] [PMID]
23. Baghban Islami Nejad MR, Aghbibi NM, Piriaiee A. Study of the bone produced from canine mesenchymal stem cell differentiation. Sci J Kurdistan Uni Med Sci. 2007; 12(3):8-22. http://sjku.muk.ac.ir/article-1-35-en.html
24. Lina JB, Stein GS, Stein JL, Wijnen AJ. Osteoclacin gene promoter: Unlocking the secrets for regulation of osteoblast growth and differentiation. J Cell Biochem. 1998; 31:62-72. [DOI:10.1002/(SICI)1097-4644(1998)72:30/31+3.0.CO;2-S]
25. Melton LJ. The worldwide problem of osteoporosis: Insights afforded by epidemiology. Bone. 1995; 5: 505S-511S. [DOI:10.1016/8756-3282(95)00258-4]
26. Anderson D, Yu TW, Hincal F. Effect of some phthalate esters in human cells in the comet assay. Teratog Carcinog Mutagen. 1999; 19:275-80. [DOI:10.1002/(SICI)1520-6866(1999)19:43.0.CO;2-1]
27. Gregory CA, Gunn WG, Peister A, Prockop DJ. An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. Anal Biochem. 2004; 329(1):77-84. [DOI:10.1016/j.ab.2004.02.002] [PMID]
28. Abnosi MH, Golami S. Cadmium chloride treatment of rats significantly impairs membrane integrity of mesenchymal stem cells via electrolyte imbalance and lipid peroxidation, a possible explanation of Cd related osteoporosis. Iran J Basic Med Sci. 2017; 20(3):280-7. https://www.ncbi.nlm.nih.gov/pubmed/28392900
29. Stockert JC, Blázquez-Castro A, Cañete M, Horobin RW, Villanueva Á. MTT assay for cell viability: Intracellular localization of the formazan product is in lipid droplets. Acta Histochem. 2012; 114(8):785-96. [DOI:10.1016/j.acthis.2012.01.006] [PMID]
30. Kanno S, Hirano S, Kayama F. Effects of phytoestrogens and environmental estrogens on osteoblastic differentiation in MC3T3-E1 cells. Toxicology. 2004; 196(1):137-45. [DOI:10.1016/j.tox.2003.12.002] [PMID]
31. Lazdunski M, Frelin C, Vigne P. The sodium/hydrogen exchange system in cardiac cells: Its biochemical and pharmacological properties and its role in regulating internal concentrations of sodium and internal pH. J Mol Cell Cardiol. 1985; 17(11):1029-42. [DOI:10.1016/S0022-2828(85)80119-X]
32. Blaustein MP. The interrelationship between sodium and calcium fluxes across cell membranes. Rev Physiol Biochem Pharmacol. 1974; 70:32-82 [DOI:10.1007/BFb0034293] [PMID]
33. Post RL, Merritt CR, Kinsolving CR, Albright CD. Membrane adenosine triphosphatase as a participant in the active transport of sodium and potassium in the human erythrocyte. J biol chem. 1960; 235:1796-802. https://www.ncbi.nlm.nih.gov/pubmed/14434402
34. Vassalle M. Contribution of the Na+/K+-pump to the membrane potential. Experientia. 1987; 43(11-12):1135-40. [DOI:10.1007/BF01945511] [PMID]
35. Cox MM, Nelson DL. Chapter 14: Glycolysis, gluconeogenesis, and the pentose phosphate pathway. In: Lehninger principles of biochemistry. 5th ed. New York: W. H. Freeman & Co; 2008. https://www.slideshare.net/RIZWANABBAS3/glycolysis-gluconeogenesis-and-the-pentose-phosphate-pathway-80935444
36. Siddiqui A, Srivastava SP. Effect of di (2-ethylhexyl) phthalate administration on rat sperm count and on sperm metabolic enzymes. Bull Environ Contam Toxicol. 1992; 48(1):115-9. [DOI:10.1007/BF00197492] [PMID]
37. Palleschi S, Rossi B, Diana L, Silvestroni L. Di (2-ethylhexyl) phthalate stimulates Ca 2+entry, chemotaxis and ROS production in human granulocytes. Toxicol Lett. 2009; 187(1):52-7. [DOI:10.1016/j.toxlet.2009.01.031] [PMID]
38. Houten SM, Wanders RJ. A general introduction to the biochemistry of mitochondrial fatty acid β-oxidation. J Inherit Metab Dis. 2010; 33(5):469-77. [DOI:10.1007/s10545-010-9061-2] [PMID] [PMCID]
39. Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O. Oxidative stress and antioxidant defense. World Allergy Organ J. 2012; 5(1):9-19. [DOI:10.1097/WOX.0b013e3182439613] [PMID] [PMCID]
40. Wang Y, Wang T, Ban Y, Shen C, Shen Q, Chai X, et al. Di-(2-ethylhexyl) phthalate exposure modulates antioxidant enzyme activity and gene expression in juvenile and adult daphnia magna. Arch Environ Contam Toxicol. 2018; 75 (1):145-56. [DOI:10.1007/s00244-018-0535-9] [PMID]
41. Birmingham E, Niebur GL, McHugh PE. Osteogenic differentiation of mesenchymal stem cells is regulated by osteocyte and osteoblast cells in a simplified bone niche. Eur Cell Mater. 2013; 23:13-27. [DOI:10.22203/eCM.v023a02] [PMID]
42. Hwang JK, Min KH, Choi KH, Hwang YC, Jeong IK, Ahn KJ, et al. Bisphenol A reduces differentiation and stimulates apoptosis of osteoclasts and osteoblasts. Life Sci. 2013; 93(9):367-72. [DOI:10.1016/j.lfs.2013.07.020] [PMID]
43. Golub EE, Boesze-Battaglia K. The role of alkaline phosphatase in mineralization. Curr Opin Orthop. 2007; 18(5):444-8. [DOI:10.1097/BCO.0b013e3282630851]
44. Kanatani N, Fujita T, Fukuyama R, Liu W, Yoshida CA, Moriishi T, et al. Cbf beta regulates RUNX2 function isoform-dependently in postnatal bone development. Dev Biol. 2006; 296(1):48-61. [DOI:10.1016/j.ydbio.2006.03.039] [PMID]
- Abstract Viewed: 387 times
- PDF Downloaded: 424 times