Stimulation of Nitric Oxide Production Contributes to the Antithrombotic Effect of Stromal Cell–Derived Factor-1α in Preventing Microsurgical Anastomotic Thrombosis

Abstract

Background Intimal injury plays a critical role in initiating the pathogenesis of thrombosis formation after microsurgical anastomosis. Application of stromal cell–derived factor-1α (SDF-1α) is reported to promote early regeneration of injured intima through migration of endothelial cells and mobilization of endothelial progenitor cells. We therefore hypothesized that local transfer of SDF-1α gene would inhibit microsurgical anastomotic thrombosis.

Methods Sixty Sprague-Dawley rats were used and divided randomly into three groups (SDF-1α group, plasmid group, and saline group) in this study. Plasmid DNA encoding SDF-1α, empty plasmid, and saline were injected into the left femoral muscles of rats from each group, respectively. Seven days after injection, peripheral blood samples were obtained to measure the plasma levels of SDF-1α and nitric oxide (NO). The left femoral artery of each rat was crushed, transected, and repaired by end-to-end microsurgical anastomosis. Vascular patency was assessed at 15, 30, and 120 minutes after reperfusion using milk test. Thrombosis formation was assessed with hematoxylin and eosin staining and scanning electron microscopy at 120 minutes postoperatively.

Results The plasma levels of SDF-1α and NO in SDF-1α group were significantly higher than those in plasmid group and saline group (p < 0.01). The patency rate in SDF-1α group was significantly higher than that in control groups at 120 minutes after reperfusion (p < 0.05). Treatment of SDF-1α significantly reduced the size of thrombotic occlusion when compared with controls (p < 0.05). All SDF-1α recipients exhibited decreased thrombosis under scanning electron microscopy.

Conclusions The current study demonstrated that local transfer of SDF-1α gene increases arterial patency and inhibits microsurgical anastomotic thrombosis in a crush model of femoral artery in rat. The antithrombotic effect of SDF-1α may be mediated through increased production of endogenous NO. These findings provide a novel approach for inhibition of microsurgical anastomotic thrombosis.

Keywords

intimal injury - microsurgical anastomotic thrombosis - stromal cell–derived factor-1α

• References

• 1 Zumiotti A, Ferreira MC. Replantation of digits: factors influencing survival and functional results. Microsurgery 1994; 15 (1) 18-21
• 2 Khouri RK, Cooley BC, Kunselman AR , et al. A prospective study of microvascular free-flap surgery and outcome. Plast Reconstr Surg 1998; 102 (3) 711-721
• 3 Fufa D, Calfee R, Wall L, Zeng W, Goldfarb C. Digit replantation: experience of two U.S. academic level-I trauma centers. J Bone Joint Surg Am 2013; 95 (23) 2127-2134
• 4 Cho EH, Ligh C, Hodulik KL, Hollenbeck ST. Role of platelet inhibition in microvascular surgery. J Reconstr Microsurg 2014; 30 (9) 589-598
• 5 Isogai N, Tanaka H, Asamura S. Thrombosis and altered expression of intercellular adhesion molecule-1 (ICAM-1) after avulsion injury in rat vessels. J Hand Surg [Br] 2004; 29 (3) 230-234
• 6 Acland RD, Trachtenberg L. The histopathology of small arteries following experimental microvascular anastomosis. Plast Reconstr Surg 1977; 60 (6) 868-875
• 7 Itoh Y, Toriumi H, Yamada S, Hoshino H, Suzuki N. Resident endothelial cells surrounding damaged arterial endothelium reendothelialize the lesion. Arterioscler Thromb Vasc Biol 2010; 30 (9) 1725-1732
• 8 Lin HH, Chen YH, Yet SF, Chau LY. After vascular injury, heme oxygenase-1/carbon monoxide enhances re-endothelialization via promoting mobilization of circulating endothelial progenitor cells. J Thromb Haemost 2009; 7 (8) 1401-1408
• 9 Pi X, Wu Y, Ferguson III JE, Portbury AL, Patterson C. SDF-1alpha stimulates JNK3 activity via eNOS-dependent nitrosylation of MKP7 to enhance endothelial migration. Proc Natl Acad Sci U S A 2009; 106 (14) 5675-5680
• 10 Gao Y, Wang Q, Cui X , et al. Controlled release of stromal cell-derived factor-1α from silk fibroin-coated coils accelerates intra-aneurysmal organization and occlusion of neck remnant by recruiting endothelial progenitor cells. Int J Clin Exp Pathol 2014; 7 (12) 8366-8380
• 11 Yates YJ, Farias CL, Kazmier FR, Puckett CL, Concannon MJ. The effect of tirofiban on microvascular thrombosis: crush model. Plast Reconstr Surg 2005; 116 (1) 205-208
• 12 Liu Y, Luo W, Yang H , et al. Stimulation of nitric oxide production contributes to the antiplatelet and antithrombotic effect of new peptide pENW (pGlu-Asn-Trp). Thromb Res 2015; 136 (2) 319-327
• 13 Ching S, Thoma A, Monkman S, Kelton JG. Inhibition of microsurgical thrombosis by the platelet glycoprotein IIb/IIIa antagonist SR121566A. Plast Reconstr Surg 2003; 112 (1) 177-185
• 14 Khouri RK. Free flap surgery. The second decade. Clin Plast Surg 1992; 19 (4) 757-761
• 15 Chung TL, Pumplin DW, Holton III LH, Taylor JA, Rodriguez ED, Silverman RP. Prevention of microsurgical anastomotic thrombosis using aspirin, heparin, and the glycoprotein IIb/IIIa inhibitor tirofiban. Plast Reconstr Surg 2007; 120 (5) 1281-1288
• 16 Enajat M, Aziz Mohammadi M, Debeij J, van der Hulst RR, Mureau MA. Effect of acetylsalicylic acid on microvascular thrombosis in autologous breast reconstruction. J Reconstr Microsurg 2014; 30 (1) 65-70
• 17 Drijkoningen T, van Weel V, Eberlin KR, Cetrulo Jr CL. Eptifibatide salvage of arterial anastomotic thrombosis in lower extremity free flap reconstruction: a case report. J Reconstr Microsurg 2015; 31 (7) 544-546
• 18 Lykoudis EG, Contodimos GB, Tsoutsos DA , et al. Microsurgical repair after crush-avulsion injury of the femoral vein in rats: prevention of microvascular thrombosis with recombinant human tissue-type plasminogen activator (rt-PA). Microsurgery 2001; 21 (8) 357-361
• 19 Wester J, Sixma JJ, Geuze JJ, Heijnen HF. Morphology of the hemostatic plug in human skin wounds: transformation of the plug. Lab Invest 1979; 41 (2) 182-192
• 20 Yin Y, Zhao X, Fang Y , et al. SDF-1alpha involved in mobilization and recruitment of endothelial progenitor cells after arterial injury in mice. Cardiovasc Pathol 2010; 19 (4) 218-227
• 21 Momi S, Caracchini R, Falcinelli E, Evangelista S, Gresele P. Stimulation of platelet nitric oxide production by nebivolol prevents thrombosis. Arterioscler Thromb Vasc Biol 2014; 34 (4) 820-829
• 22 Freedman JE, Loscalzo J. Nitric oxide and its relationship to thrombotic disorders. J Thromb Haemost 2003; 1 (6) 1183-1188
• 23 Nakayama T, Sato W, Yoshimura A , et al. Endothelial von Willebrand factor release due to eNOS deficiency predisposes to thrombotic microangiopathy in mouse aging kidney. Am J Pathol 2010; 176 (5) 2198-2208
• 24 Tan XL, Xue YQ, Ma T , et al. Partial eNOS deficiency causes spontaneous thrombotic cerebral infarction, amyloid angiopathy and cognitive impairment. Mol Neurodegener 2015; 10 (1) 24
• 25 Rosenbaum TJ, Sundt Jr TM. Thrombus formation and endothelial alterations in microarterial anastomoses. J Neurosurg 1977; 47 (3) 430-441
• 26 Wieslander JB, Aberg M, Dougan P. Accumulation of isotope labelled platelets in small arteries after end-to-end and end-in-end anastomoses in the rabbit. Br J Plast Surg 1982; 35 (2) 158-162
• 27 Platelet Receptor Inhibition in Ischemic Syndrome Management (PRISM) Study Investigators. A comparison of aspirin plus tirofiban with aspirin plus heparin for unstable angina. N Engl J Med 1998; 338 (21) 1498-1505