Effects of tacrolimus on morphology, proliferation and differentiation of mesenchymal stem cells derived from gingiva tissue

Ha,Dong‑Ho; Yong,Chul  Soon; Kim,Jong  Oh; Jeong,Jee‑Heon; Park,Jun‑Beom

doi:10.3892/mmr.2016.5217

Effects of tacrolimus on morphology, proliferation and differentiation of mesenchymal stem cells derived from gingiva tissue

Authors:
- Dong‑Ho Ha
- Chul Soon Yong
- Jong Oh Kim
- Jee‑Heon Jeong
- Jun‑Beom Park
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Affiliations: College of Pharmacy, Yeungnam University, Gyeongsan‑si, Gyeongsangbuk‑do 38541, Republic of Korea, Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea
Published online on: May 6, 2016 https://doi.org/10.3892/mmr.2016.5217
Pages: 69-76
Copyright: © Ha et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Tacrolimus is a 23-membered macrolide lactone with potent immunosuppressive activity that is effective in the prophylaxis of organ rejection following kidney, heart and liver transplantation. Tacrolimus also exerts a variety of actions on bone metabolism. The aim of the present study was to evaluate the effects of different concentrations of tacrolimus on the morphology and viability of human stem cells derived from the gingiva. Gingival‑derived stem cells were grown in the presence of tacrolimus at final concentrations ranging from 0.001 to 100 µg/ml. The morphology of the cells was viewed under an inverted microscope and the cell viability was analyzed using Cell Counting kit‑8 (CCK‑8) on days 1, 3, 5 and 7. Alizarin Red S staining was used to assess mineralization of treated cells. The control group showed spindle‑shaped, fibroblast‑like morphology and the shapes of the cells in 0.001, 0.01, 0.1, 1 and 10 µg/ml tacrolimus were similar to those of the control group. All groups except the 100 µg/ml group showed increased cell proliferation over time. Cultures grown in the presence of tacrolimus at 0.001, 0.01, 0.1, 1 and 10 µg/ml were not identified to be significantly different compared with the control at days 1, 3 and 5 using the CCK‑8 assays. Increased mineralized deposits were noted with increased incubation time. Treatment with tacrolimus from 0.001 to 1 µg/ml led to an increase in mineralization compared with the control group. Within the limits of this study, tacrolimus at the tested concentrations (ranging from 0.001 to 10 µg/ml) did not result in differences in the viability of stem cells derived from gingiva; however it did enhance osteogenic differentiation of the stem cells.

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1	Beyer Nardi N and da Silva Meirelles L: Mesenchymal stem cells: Isolation, in vitro expansion and characterization. Handb Exp Pharmacol:. 249–282. 2006. View Article : Google Scholar
2	Chen WC, Péault B and Huard J: Regenerative translation of human blood-vessel-derived MSC precursors. Stem Cells Int. 2015:3751872015. View Article : Google Scholar : PubMed/NCBI
3	Deng S, Huang R, Wang J, Zhang S, Chen Z, Wu S, Jiang Y, Peng Q, Cai X and Lin Y: Miscellaneous animal models accelerate the application of mesenchymal stem cells for cartilage regeneration. Curr Stem Cell Res Ther. 9:223–233. 2014. View Article : Google Scholar : PubMed/NCBI
4	Landa-Solís C, Granados-Montiel J, Olivos-Meza A, Ortega-Sánchez C, Cruz-Lemini M, Hernández-Flores C, Chang-González ME, García RG, Olivos-Díaz B, Velasquillo-Martínez MC, et al: Cryopreserved CD90+ cells obtained from mobilized peripheral blood in sheep: a new source of mesenchymal stem cells for preclinical applications. Cell Tissue Bank. July 29–2015.Epub ahead of print.
5	Caplan AI: Adult mesenchymal stem cells for tissue engineering versus regenerative medicine. J Cell Physiol. 213:341–347. 2007. View Article : Google Scholar : PubMed/NCBI
6	Lee SI, Yeo SI, Kim BB, Ko Y and Park JB: Formation of size-controllable spheroids using gingiva-derived stem cells and concave microwells: Morphology and viability tests. Biomed Rep. 4:97–101. 2016.PubMed/NCBI
7	Jeong SH, Lee JE, Kim BB, Ko Y and Park JB: Evaluation of the effects of Cimicifugae Rhizoma on the morphology and viability of mesenchymal stem cells. Exp Ther Med. 10:629–634. 2015.PubMed/NCBI
8	Tamura S, Ohike A, Ibuki R, Amidon GL and Yamashita S: Tacrolimus is a class II low-solubility high-permeability drug: The effect of P-glycoprotein efflux on regional permeability of tacrolimus in rats. J Pharm Sci. 91:719–729. 2002. View Article : Google Scholar : PubMed/NCBI
9	Scholten EM, Cremers SC, Schoemaker RC, Rowshani AT, van Kan EJ, den Hartigh J, Paul LC and de Fijter JW: AUC-guided dosing of tacrolimus prevents progressive systemic overexposure in renal transplant recipients. Kidney Int. 67:2440–2447. 2005. View Article : Google Scholar : PubMed/NCBI
10	Felipe CR, Garcia C, Moreira S, Olsen N, Silva HT and Pestana OM: Choosing the right dose of new immunossuppressive drugs for new populations: Importance of pharmacokinetic studies. Transplant Proc. 33:1095–1096. 2001. View Article : Google Scholar : PubMed/NCBI
11	van Hooff JP, Boots JM, van Duijnhoven EM and Christiaans MH: Dosing and management guidelines for tacrolimus in renal transplant patients. Transplant Proc. 31:54S–57S. 1999. View Article : Google Scholar : PubMed/NCBI
12	Taylor DO, Barr ML, Radovancevic B, Renlund DG, Mentzer RM Jr, Smart FW, Tolman DE, Frazier OH, Young JB and VanVeldhuisen P: A randomized, multicenter comparison of tacrolimus and cyclosporine immunosuppressive regimens in cardiac transplantation: Decreased hyperlipidemia and hypertension with tacrolimus. J Heart Lung Transplant. 18:336–345. 1999. View Article : Google Scholar : PubMed/NCBI
13	Busuttil RW, Klintmalm GB, Lake JR, Miller CM and Porayko M: General guidelines for the use of tacrolimus in adult liver transplant patients. Transplantation. 61:845–847. 1996. View Article : Google Scholar : PubMed/NCBI
14	Staatz CE and Tett SE: Clinical pharmacokinetics and pharmacodynamics of tacrolimus in solid organ transplantation. Clin Pharmacokinet. 43:623–653. 2004. View Article : Google Scholar : PubMed/NCBI
15	Laskow DA, Neylan JF III, Shapiro RS, Pirsch JD, Vergne-Marini PJ and Tomlanovich SJ: The role of tacrolimus in adult kidney transplantation: A review. Clin Transplant. 12:489–503. 1998.PubMed/NCBI
16	Kugimiya F, Yano F, Ohba S, Igawa K, Nakamura K, Kawaguchi H and Chung UI: Mechanism of osteogenic induction by FK506 via BMP/Smad pathways. Biochem Biophys Res Commun. 338:872–879. 2005. View Article : Google Scholar : PubMed/NCBI
17	Park KM, Hay JE, Lee SG, Lee YJ, Wiesner RH, Porayko MK and Krom RA: Bone loss after orthotopic liver transplantation: FK 506 versus cyclosporine. Transplant Proc. 28:1738–1740. 1996.PubMed/NCBI
18	Stempfle HU, Werner C, Echtler S, Assum T, Meiser B, Angermann CE, Theisen K and Gärtner R: Rapid trabecular bone loss after cardiac transplantation using FK506 (tacrolimus)-based immunosuppression. Transplant Proc. 30:1132–1133. 1998. View Article : Google Scholar : PubMed/NCBI
19	Tang L, Ebara S, Kawasaki S, Wakabayashi S, Nikaido T and Takaoka K: FK506 enhanced osteoblastic differentiation in mesenchymal cells. Cell Biol Int. 26:75–84. 2002. View Article : Google Scholar : PubMed/NCBI
20	Byun YK, Kim KH, Kim SH, Kim YS, Koo KT, Kim TI, Seol YJ, Ku Y, Rhyu IC and Lee YM: Effects of immunosuppressants, FK506 and cyclosporin A, on the osteogenic differentiation of rat mesenchymal stem cells. J Periodontal Implant Sci. 42:73–80. 2012. View Article : Google Scholar : PubMed/NCBI
21	Hoogduijn MJ, Crop MJ, Korevaar SS, Peeters AM, Eijken M, Maat LP, Balk AH, Weimar W and Baan CC: Susceptibility of human mesenchymal stem cells to tacrolimus, mycophenolic acid and rapamycin. Transplantation. 86:1283–1291. 2008. View Article : Google Scholar : PubMed/NCBI
22	Jin SH, Lee JE, Yun JH, Kim I, Ko Y and Park JB: Isolation and characterization of human mesenchymal stem cells from gingival connective tissue. J Periodontal Res. 50:461–467. 2015. View Article : Google Scholar
23	Lechler RI, Sykes M, Thomson AW and Turka LA: Organ transplantation - how much of the promise has been realized? Nat Med. 11:605–613. 2005. View Article : Google Scholar : PubMed/NCBI
24	Kaihara S, Bessho K, Okubo Y, Sonobe J, Kusumoto K, Ogawa Y and Iizuka T: Effect of FK506 on osteoinduction by recombinant human bone morphogenetic protein-2. Life Sci. 72:247–256. 2002. View Article : Google Scholar : PubMed/NCBI
25	Utheim TP, Raeder S, Utheim ØA, Cai Y, Roald B, Drolsum L, Lyberg T and Nicolaissen B: A novel method for preserving cultured limbal epithelial cells. Br J Ophthalmol. 91:797–800. 2007. View Article : Google Scholar
26	Bang SM, Moon HJ, Kwon YD, Yoo JY, Pae A and Kwon IK: Osteoblastic and osteoclastic differentiation on SLA and hydrophilic modified SLA titanium surfaces. Clin Oral Implants Res. 25:831–837. 2014. View Article : Google Scholar
27	Wang B, Bi M, Zhu Z, Wu L and Wang J: Effects of the antihypertensive drug benidipine on osteoblast function in vitro. Exp Ther Med. 7:649–653. 2014.PubMed/NCBI
28	Nakamura T, Shinohara Y, Momozaki S, Yoshimoto T and Noguchi K: Co-stimulation with bone morphogenetic protein-9 and FK506 induces remarkable osteoblastic differentiation in rat dedifferentiated fat cells. Biochem Biophys Res Commun. 440:289–294. 2013. View Article : Google Scholar : PubMed/NCBI
29	Quarles LD, Yohay DA, Lever LW, Caton R and Wenstrup RJ: Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: An in vitro model of osteoblast development. J Bone Miner Res. 7:683–692. 1992. View Article : Google Scholar : PubMed/NCBI
30	Park JB, Zhang H, Lin CY, Chung CP, Byun Y, Park YS and Yang VC: Simvastatin maintains osteoblastic viability while promoting differentiation by partially regulating the expressions of estrogen receptors α. J Surg Res. 174:278–283. 2012. View Article : Google Scholar