Effect of L-lysine in culture medium on nodule formation by bone marrow cells

Masataka Yoshikawa, Yasunori Shimomura,, Hideyuki Kakigi, Norimasa Tsuji, Takayoshi Yabuuchi, Hiroyuki Hayashi

Department of Endodontics, Osaka Dental University, Osaka, Japan.

DOI: 10.4236/jbise.2012.510072                

Abstract

The aim of this study was to estimate the effect of L-lysine on nodule formation by rat bone marrow cells in vitro. In this study, L-lysine was added to medium for mesenchymal stem cell culture to promote proliferation and differentiation of the cells, and then nodule formation was estimated in an in vitro rat bone marrow cell culture. Bone marrow cells from the bone shafts of the femora of Fischer 344 rats were cultured in minimum essential medium with 20 μl of L-lysine solution at 10^﹣4, 10^﹣5, 10^﹣6, 10^﹣7 or 10^﹣8 M. Dexamethasone was also added to the medium at 10 nM for differentiation of stem cells from bone marrow into osteoblast progenitor cells. The subculture was performed for 2 weeks. The quantity of osteocalcin in rat bone marrow cell culture with dexamethasone was 392 ng/ml. In the medium with dexamethasone and 10^﹣8 M L-lysine, the quantity of osteocalcin was 437 ng/ml. Nodules only formed upon addition of 20 μl of L-lysine at 10^﹣8 M. It was indicated that 10^﹣8 M L-lysine should be the optimal concentration for calcification. For nodule formation by rat bone marrow cells in vitro, the optimum concentration of L-lysine in culture medium might be 20 μl of 10^﹣8 M. L-lysine could play an important role in matrix production for bone formation in vitro.

Keywords

L-Lysine; Bone marrow Cells; Calcium Deposition; Dexamethasone; In Vitro

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Glucose uptake mediated by glucose transporter 1 is essential for early tooth morphogenesis and size determination of murine molars. Developmental biology, 363, 52-61. doi: 10.1016/ j.ydbio.2011.12.020
[2]	Traphagen, S. and Yelick, P.C. (2009) Reclaiming a natural beauty: whole-organ engineering with natural extracellular materials. Regenerative Medicine, 4, 747-758. doi: 10.2217/ rme.09.38
[3]	Thesleff, I. and Tummers, M. (2003) Stem Cells and Tissue Engineering: Prospects for Regenerating Tissues in Dental Practice. Medical Principles and Practice, 12, 43-50. doi: 10.1159/ 000069840
[4]	Caplan, A.I. and Bruder, S.P. (2001) Mesenchymal stem cells: building blocks for molecular medicine in the 21st century. Trends in Molecular Medicine, 7, 259-264. doi: 10.1016/S1471-4914(01)02016-0
[5]	Derubeis, A.R. and Cancedda, R. (2004) Bone marrow stromal cells (BMSCs) in bone engineering: limitations and recent advances. Annals of Biomedical Engineering, 32, 160-165. doi: 10. 1023/B:ABME.0000007800.89194.95
[6]	Canalis, E., McCarthy, T.L. and Centrella, M. (1991) Growth factors and cytokines in bone cell metabolism. Annual Review of Medicine, 42, 17-24. doi:0.1146/annurev.me.42.020191.000313
[7]	D’Ippolito, G., Schiller, P.C., Ricordi, C., Roos, B.A. and Howard, G.A. (1999) Age-related osteogenic potential of mesenchymal stromal stem cells from human vertebral bone marrow. Journal of Bone and Mineral Research, 14, 1115-1122. doi: 10.1359/jbmr. 1999.14.7.1115
[8]	Chevalley, T., Rizzoli, R., Manen, D., Caverzasio, J. and Bonjour, J.P. (1998) Arginine increases insulin-like growth factor-I production and collagen synthesis in osteoblast-like cells. Bone, 23, 103-109. doi: 10.1016/S8756-3282 (98)00081-7
[9]	Baylink, D.J., Finkelma, R.D. and Mohan, S. (1993) Growth factors to stimulate bone formation. Journal of Bone and Mineral Reseach, 8, 565-572. doi: 10.1002/jbmr.5650081326
[10]	Yoshikawa M, Tsuji N, Shimomura Y, Hayashi H, Oh-gushi H (2008) Osteogenesis depending on geometry of porous hydroxyapatite scaffolds. Clcified Tissue International, 83, 139-145. doi: 10.1007/s00223-008-9157-y
[11]	Pittenger MF, Martin BJ (2004) Mesenchymal stem cells and their potential as cardiac therapeutics. Circulation Research, 95, 9-20. doi: 10.1161/01.RES.0000135902.99383.6f
[12]	Froesch ER, Schmid C, Schwander J, Zapf J (1985) Actions of insulin-like growth factors. Annual Review of Physiology, 47, 443-467. doi: 10.1146/annurev.ph.47.030185.002303
[13]	Fiore CE, Pennisi P, Cutuli VM, Prato A, Messina R, Clementi G (2000) L-arginine prevents bone loss and bone collagen breakdown in cyclosporin A-treated rats. European Journal of Pharmacology, 408, 323-326. doi: 10.1016/S0014-2999(00)00800-1
[14]	Torricelli P, Fini M, Giavaresi G, Giardino R (2003) Osteoblasts cultured from osteoporotic bone: a comparative investigation on human and animal-derived cells. Artificial Cells, Blood Substitutes and Biotechnology, 31, 263-277. doi: 10.1081/BIO-120023157
[15]	Torricelli P, Fini M, Giavaresi G, Giardino R (2001) Bone tissue cultures: an in vitro model for the evaluation of bone defect healing after L-arginine and L-lysine administration. Artificial Cells, Blood Substitutes and Bio-technology, 29, 325-334. doi: 10.1081/BIO-100104234
[16]	Visser JJ, Hoekman K (1994) Arginine supplementation in the prevention and treatment of osteoporosis. Medical Hypotheses, 43, 339-342. doi: 10.1016/0306-9877(94)90113-9
[17]	Barry FP, Murphy JM (2004) Mesenchymal stem cells: clinical applications and biological characterization. The International Journal of Biochemistry & Cell Biology, 36, 568-584. doi: 10.1016/j.biocel.2003.11.001
[18]	Petite H, Viateau V, Bensa?d W, Meunier A, de Pollak C, Bourguignon M, Oudina K, Sedel L, Guillemin G (2000) Tissue-engineered bone regeneration. Nature Biotechnology, 18, 959- 963. doi: 10.1038/79449
[19]	Ikeda E, Yagi K, Kojima M, Yagyuu T, Ohshima A, Sobajima S, Tadokoro M, Katsube Y, Isoda K, Kondoh M, Kawase M, Go MJ, Adachi H, Yokota Y, Kirita T, Ohgushi H (2008) Multipotent cells from the human third molar: feasibility of cell-based therapy for liver disease. Differentiation, 76, 495-505. doi: 10.1111/j.1432-0436.2007.00245.x
[20]	Conconi MT, Tommasini M, Muratori E, Parnigotto PP. (2001) Essential amino acids increase the growth and al-kaline phosphatase activity in osteoblasts cultured in vitro. ll.Farmaco, 56, 755-761. doi: 10.1016/S0014-827X(01)01126-0
[21]	Mostaqul Huq MD, Tsai NP, Khan SA, Wei LN (2007) Lysine trimethylation of retinoic acid receptor-alpha: a novel means to regulate receptor function. Molecular & Cellular Proteomics, 6, 677-688. doi: 10.1074/mcp.M600223-MCP200
[22]	Marie P (1997) Growth factors and bone formation in osteoporosis: roles for IGF-I and TGF-beta. Revue du rhumatisme English edition, 64, 44-53. ISSN: 1169-8446
[23]	Tumber A, Meikle MC, Hill PA (2000) Autocrine signals promote osteoblast survival in culture. Journal of Endocrinology, 167, 383-390. doi: 10.1677/joe.0.1670383