In vitro assessment of alendronate toxic and apoptotic effects on human dental pulp stem cells

Document Type : Original Article

Authors

1 Department of Oral and Maxillofacial Surgery, Dental Research Center, School of Dentistry, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Iran

3 Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s): Osteonecrosis of the jaw, as an exposed necrotic bone in the oral cavity, is one of the adverse effects of bisphosphonates, which have an affinity for bone minerals. This study investigates the cytotoxic effects of alendronate (ALN) as a nitrogen-containing bisphosphonate, on human dental pulp stem cells (hDPSCs).
Materials and Methods: The mesenchymal stem cells (MSCs), obtained from third molar tooth pulps were characterized by immunophenotyping assay in order to identify surface markers to evaluate their expression. To detect multipotency hDPSCs, they were differentiate into osteocytes and adipocytes. Cell proliferation was measured by MTT assay. PI staining of DNA fragmentation by flowcytometry (sub-G1 peak) was performed for determination of apoptotic cells and Bax, Bcl-2, and cleaved caspase 3 expressions. Protein expression was detected by Western blotting.
Results: As the results revealed, ALN decreased viable cells (in 0.8–100 µM) after 72 hr and 168 hr (P<0.001), significantly. ALN could lower cell proliferation in hDPSCs in a concentration and time-dependent manner. Sub-G1 peak as an indicator of flowcytometry histogram of treated cells by ALN, showed apoptosis was involved in ALN-induced cytotoxicity. Expressions of cleaved caspase 3 and Bax protein, as pro-apoptotic proteins, were increased and Bcl-2 protein as anti-apoptotic protein was decreased in response to increases in the concentration of ALN (0.8–25 µM).
Conclusion: Long-term effects of ALN on cell proliferation and apoptosis in hDPSCs can result in either initiation or potentiation of ALN-induced osteonecrosis.

Keywords

Main Subjects

References

1. Wellington K and Goa KL., Zoledronic Acid. Drugs, 2003; 63: 417-437.
2. Green JR. Zoledronic acid: pharmacologic profile of a potent bisphosphonate. J Organomet Chem, 2005; 690: 2439-2448.
3. Sato M, Grasser W, Endo N, Akins R, Simmons H, Thompson DD, et al. Bisphosphonate action. Alendronate localistaion in rat bone and effects on osteoclast ultrastructure. J Clin Invest 1991; 88: 2095.
4. Li EC and Davis LE. Zoledronic acid: a new parenteral bisphosphonate. Clin Ther 2003; 25: 2669-2708.
5. Sun J, Song F, Zhang W, Sexton BE, Windsor LJ. Effects of alendronate on human osteoblast-like MG63 cells and matrix metalloproteinases. Arch Oral Biol 2012; 57: 728-736.
6. Nussbaum SR, Warrell RP Jr, Rude R, Glusman J, Bilezikian JP, Stewart AF, et al. Dose-response study of alendronate sodium for the treatment of cancer-associated hypercalcemia. J Clin Oncol 1993; 11:1618-1623.
7. Walter C, Pabst A, Ziebart T, Klein M, Al-Nawas B. Bisphosphonates affect migration ability and cell viability of HUVEC, fibroblasts and osteoblasts in vitro. Oral Dis 2011; 17: 194–199.
8. Reid IR, Bolland MJ, Grey AB. Is bisphosphonate-associated osteonecrosis of the jaw caused by                 soft tissue toxicity? Bone 2007; 41: 318-320.
9. Scheper MA, Badros A, Chaisuparat R, Cullen KJ, Meiller TF. Effect of zoledronic acid on oral fibroblasts and epithelial cells: a potential mechanism of bisphosphonate‐associated osteonecrosis. Br J Haematol 2009; 144: 667-676.
10. Soydan SS, Araz K, Senel FV, Yurtcu E, Helvacioglu F, Dagdeviren A, et al. Effects of alendronate and pamidronate on apoptosis and cell proliferation in cultured primary human gingival fibroblasts. Hum Exp Toxicol 2015; 34: 1073–1082.
11. Sloan AJ, Smith AJ. Stem cells and the dental pulp: potential roles in dentine regeneration and repair. Oral Dis 2007; 13: 151-157.
12.    Yu J, Wang Y, Deng Z, Tang L, Li Y, Shi J, Jin Y, et al. Odontogenic capability: bone marrow stromal stem cells versus dental pulp stem cells. Biol Cell 2007; 99: 465-474.
13.    Graziano A, d’Aquino R, Laino G, Papaccio G. Dental pulp stem cells: a promising tool for bone regeneration. Stem Cell Rev 2008; 4: 21-26.
14.    Yu J, Deng Z, Shi J, Zhai H, Nie X, Zhuang H, et al. Differentiation of dental pulp stem cells into regular-shaped dentin-pulp complex induced by tooth germ cell conditioned medium. Tissue Eng 2006; 12: 3097-3105.
15. Pourgonabadi S, Mousavi SH, Tayarani-Najaran Z, Ghorbani A, Effect of zoledronate, a third-generation bisphosphonate, on proliferation and apoptosis of human dental pulp stem cells. Can J Physiol Pharmacol, 2017. 3: 1-8.
16.    Ghorbani A, Jalali SA, Varedi M. Isolation of adipose tissue mesenchymal stem cells without tissue destruction: a non-enzymatic method. Tissue Cell 2014; 46: 54-58.
17.    Feizpour A, Boskabady MH, Ghorbani A. Adipose-derived stromal cell therapy affects lung inflammation and tracheal responsiveness in guinea pig model of COPD. PloS one 2014; 9.
18.    Alinejad B, Shafiee-Nick R, Sadeghian H, Ghorbani A. Metabolic effects of newly synthesized phosphodiesterase-3 inhibitor 6-[4-(4-methylpiperidin-1-yl)-4-oxobutoxy]-4-methylquinolin-2 (1H)-one on rat adipocytes. Daru 2015; 23: 19.
19.    Ghorbani A, Feizpour A, Hashemzahi M, Gholami L, Hosseini M, Soukhtanloo M, et al. The effect of adipose derived stromal cells on oxidative stress level, lung emphysema and white blood cells of guinea pigs model of chronic obstructive pulmonary disease. Daru 2014; 22: 26.
20. Zaker A, Asili J, Abrishamchi P, Tayarani-Najaran Z, Mousavi SH, Cytotoxic and apoptotic effects of   root extract and tanshinones isolated from Perovskia abrotanoides Kar. Iran J Basic Med Sci. 2017; 20: 1377-84.
21.    Mousavi SH, Naghi zadeh B, Pourgonabadi S, Ghorbani A, Protective effect of Viola tricolor and Viola odorata extracts on serum/glucose deprivation-induced neurotoxicity: role of reactive oxygen species. Avicenna J Phytomed. 6: 434-41.
22. Mortazavian SM, Parsaee H, Mousavi SH, Tayarani-Najaran Z, Ghorbani A, Sadeghnia HR. Acetylcholinesterase inhibitors promote angiogenesis in chick chorioallantoic membrane and inhibit apoptosis of endothelial cells. Int J Alzheimers Dis 2013; 2013 :121068.
23.    Pourgonabadi S, Amiri MS, Mousavi SH, Cytotoxic and apoptogenic effects of Bryonia aspera root extract against Hela and HN-5 cancer cell lines. Avicenna J Phytomed, 2017. 7: 66-72.
24.    Correia Vde F, Caldeira CL, Marques MM. Cytotoxicity evaluation of sodium alendronate of cultured human periodontal ligament fibroblast. Dent Traumatol 2006; 22: 312-317.
25.    Moreira MS, Katayama E, Bombana AC, Marques MM. Cytotoxicity analysis of alendronate on cultured endothelial cells and subcutaneous tissue. A pilot study. Dent Traumatol 2005; 21: 329-335.
26.    Sato M, Grasser W, Endo N, Akins R, Simmons H, Thompson DD, et al. Bisphosphonate action. Alendronate localistaion in rat bone and effects on osteoclast ultrastructure. J Clin Invest 1991; 88: 2095.
27.    Porras AG, Holland SD, Gertz BJ. Pharmacokinetics of alendronate. Clin Pharmacokinet 1999; 36: 315-328.
27.    Farese JP, Ashton J, Milner R, Ambrose LL, Van Gilder J. The effect of the bisphosphonate alendronate on viability of canine osteosarcoma cells in vitro. In vitro Cell Dev Biol Anim 2004; 40: 113-117.
28.    McLeod NM, Moutasim KA, Brennan PA, Thomas G, Jenei V, In vitro Effect of Bisphosphonates on Oral Keratinocytes and Fibroblasts. J Oral Maxillofac Surg 2014; 72: 503-509.
29.    Pabst AM, Koch FP, Taylor KY, Al-Nawas B, WalterC. The influence of bisphosphonates on viability,migration, and apoptosis of human oral keratinocytes-in vitro study. Clin Oral Investig 2012; 16: 87–93.
30.    Sloan AJ, Smith AJ. Stem cells and the dental pulp: potential roles in dentine regeneration and repair. Oral Dis 2007; 13: 151-157.
31.    Aguirre JI, Altman MK, Vanegas SM, Franz SE, Bassit AC, Wronski TJ. Effects of alendronate on bone healing after tooth extraction in rats. Oral Dis 2010; 16: 674-685.
Iranian Journal of Basic Medical Sciences
Volume 21, Issue 9
September 2018
Pages 905-910

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