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DOI: 10.1055/s-2005-836533
© Georg Thieme Verlag Stuttgart · New York
Stimulation of Human Endothelial Cells by Amino Acids: Expression of Nitric Oxide Synthases and Endothelial Growth Factors
Publication History
Publication Date:
23 August 2005 (online)
![](https://www.thieme-connect.de/media/osteotrauma/200503/lookinside/thumbnails/10.1055-s-2005-836533-1.jpg)
Abstract
Background/Aims: The complex process of fracture healing relies on the reciprocal action of microvascular endothelial cells and osteoblasts. We hypothesized that, in addition to revascularization, stimulated microendothelial cells secrete growth factors known to influence osteoblasts in vitro and in animal trials. Methods: The study describes the effect of L-glycine, L-arginine, and L-lysine on human microvascular endothelial cells (HDMEC) with respect to the in-vitro expression of basic fibroblast growth factor (b-FGF), vascular endothelial growth factor (VEGF), and endothelial and inducible nitric oxide synthases (e-NOS, i-NOS). The amino acids were added on day 0, and b-FGF, VEGF, e-NOS, and i-NOS were measured by ELISA on days 0, 1, 3, and 6. Results: The number of HDMEC was not significantly influenced by the addition of amino acids, suggesting that the observed effects were not related to proliferation. HDMEC immediately released b-FGF into the cellular supernatant when L-glycine was added. In addition, HDMEC were stimulated to produce i-NOS and e-NOS after a single addition of L-arginine. Conclusion: These findings suggest that L-arginine and L-glycine stimulate HDMEC to express NOS and b-FGF, compounds well-known for their osteoblast modulating activities. If confirmed in animal studies, supplementation with L-arginine or L-glycine may be an attractive therapeutic approach to enhance fracture repair.
Key words
L-arginine - L-glycine - human microvascular endothelial cells - fracture healing
References
- 1 Barnes G L, Kostenuik P J, Gerstenfeld L C, Einhorn T A. Growth factor regulation of fracture repair. J Bone Miner Res. 1999; 15 1805-1815
- 2 Binion D G, Fu S, Ramanujam K S, Chai Y C, Dweik R A, Drazba J A, Wade J G, Ziats N P, Erzurum S C, Wilson K T. iNOS expression in human intestinal microvascular endothelial cells inhibits leukocyte adhesion. Am J Physiol. 1998; 275 G 592-G 603
- 3 Collin-Osdoby P, Nickols G A, Osdoby P. Bone cell function, regulation, and communication: a role for nitric oxide. J Cell Biochem. 1995; 57 399-408
- 4 Corbett S A, Hukkanen M, Batten J, McCarthy I D, Polak J M, Hughes S P. Nitric oxide in fracture repair. Different localisation, expression and activity of nitric oxide synthases. J Bone Joint Surg [Br]. 1999; 81 531-537
- 5 Deckers M M, Bezooijen R L van, Horst G van der, Hoodgendam J, Bent C van der, Papapoulos S E, Löwik C W. Bone morphogenetic proteins stimulate angiogenesis through osteoblast-derived vascular endothelial growth factor A. Endocrinology. 2002; 143 1545-1553
- 6 Fini M, Torricelli P, Giavaresi G, Carpi A, Nicolini A, Giardino R. Effect of L-lysine and L-arginine on primary osteoblast cultures from normal and osteopenic rats. Biomed Pharmacother. 2001; 55 213-220
- 7 Flores C, Rojas S, Aguayo C, Parodi J, Mann G, Pearson J D, Casanello P, Sobrevia L. Rapid stimulation of L-arginine transport by D-glucose involves p42/44mapk and nitric oxide in human umbilical vein endothelium. Circ Res. 2003; 92 64-72
- 8 Furchgott R F, Zawadzki J V. The obligatory role of the endothelium in the relaxation of artifical smooth muscle by acetylcholine. Nature. 1980; 286 373-376
- 9 Harada S, Nagy J A, Sullivan K A, Thomas K A, Endo N, Rodan G A, Rodan S B. Induction of vascular endothelial growth factor expression by prostaglandin E2 and E1 in osteoblasts. J Clin Invest. 1994; 93 2490-2496
- 10 Irie K, Tsukahara F, Fujii E, Uchida Y, Yoshioka T, He W R, Shintashige M, Murota S, Muraki T. Cationic amino acid transporter-2 mRNA induction by tumor necrosis factor-α in vascular endothelial cells. Eur J Pharmakol. 1997; 339 289-293
- 11 Kdolsky R, Mohr W, Beer R, Donath K. Oral administered L-arginine improves fracture healing. European J Trauma. 2002; 28 (Suppl 1) 139
- 12 Keller C, Fullerton J, Mobley C. Supplemental and complementary alternatives to hormone replacement therapy. J Am Acad Nurse Pract. 1999; 11 187-198
- 13 MacPherson H, Noble B S, Ralston S H. Expression and functional role of nitric oxide synthase isoforms in human osteoblast-like cells. Bone. 1999; 24 179-185
- 14 Matziolis G, Rau H M, Klever P, Erli H J, Paar O. Beeinflussung humaner Osteoblasten durch verschiedene Analgetika. [Modification of human osteoblasts by various analgesics]. Unfallchirurg. 2002; 105 527-531
- 15 Mayr-Wohlfahrt U, Waltenberger J, Hausser H, Kessler S, Günther K P, Dehio C, Puhl W, Brenner R E. Vascular endothelial growth factor stimulates chemotactic migration of primary human osteoblasts. Bone. 2002; 30 472-477
- 16 Nakajima F, Ogasawara A, Goto K, Moriya H, Ninomiya Y, Einhorn T A, Yamazaki M. Spatial and temporal gene expression in chondrogenesis during fracture healing and the effects of basic fibroblast growth factor. J Orthop Res. 2001; 19 935-944
- 17 Reher P, Doan N, Bradnock B, Meghji S, Harris M. Effect of ultrasound on the production of IL-8, basic FGF and VEGF. Cytokine. 1999; 11 416-423
- 18 Rodan S B, Wesolowski G, Thomas K A, Yoon K, Rodan G A. Effects of acidic and basic fibroblast growth factors on osteoblastic cells. Connect Tissue Res. 1989; 20 283-288
- 19 Saadeh P B, Mehrara B J, Steinbrech D S, Spector J A, Greenwald J A, Chin G S, Ueno H, Gittes G K, Longaker M T. Mechanisms of fibroblast growth factor-2 modulation of vascular endothelial growth factor expression by osteoblastic cells. Endocrinology. 2000; 141 2075-2083
- 20 Sandberg M M, Aro H, Vuorio E I. Gene expression during bone repair. Clin Orthop. 1993; 289 292-312
- 21 Schäffer M R, Tantry U, Wesep R A van, Barbul A. Nitric oxide metabolism in wounds. J Surg Res. 1997; 71 25-31
- 22 Seghezzi G, Patel S, Ren C I, Gualandris A, Pintucci G, Robbins E S, Shapiro R L, Galloway A C, Rifkin D B, Mignatti P. Fibroblast growth factor-2 (FGF-2) induces vascular endothelial growth factor (VEGF) expression in the endothelial cells of forming capillaries: an autocrine mechanism contributing to angiogenesis. J Cell Biol. 1998; 141 1659-1673
- 23 Tokuda H, Hirade K, Wang X, Oiso Y, Kozawa O. Involvement of SAPK/JNK in basic fibroblast growth factor-induced vascular endothelial growth factor release in osteoblasts. J Endocrinol. 2003; 177 101-107
- 24 Torricelli P, Fini M, Giavaresi G, Giardino R, Gnudi S, Nicolini A, Carpi A. L-Arginine and L-Lysine stimulation on cultured human osteoblasts. Biomed Pharmacother. 2002; 56 492-497
- 25 Tsukahara H, Miura M, Tsuchida S, Hata I, Hata K, Yamamoto K, Ishii Y, Muramatsu I, Sudo M. Effect of nitric oxide synthase inhibitors on bone metabolism in growing rats. Am J Physiol. 1996; 270 E 840-E 845
- 26 Turner C H, Takano Y, Owan I, Murrell G A. Nitric oxide inhibitor L-NAME suppresses mechanically induced bone formation in rats. Am J Physiol. 1996; 270 E 634-E 639
- 27 Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitsui Y, Yazaki Y, Goto K, Masaki T. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature. 1988; 332 411-415
- 28 Yeung H Y, Lee S KM, Fung K P, Leung K S. Expression of basic fibroblast growth factor during distraction osteogenesis. Clin Orthop. 2001; 385 219-229
Ass. Prof. Dr. R. Kdolsky
Department of Traumatology · University of Vienna Medical School · AKH
Währinger Gürtel 18-20
1090 Vienna
Austria
Phone: +43/1/4 04 00 56 19
Fax: +43/1/4 04 00 59 49
Email: richard.kdolsky@meduniwien.ac.at