Background Bisphosphonates are a class of pharmacologic compounds that are commonly used to treat postmenopausal osteoporosis and malignant osteolytic processes. Studies have shown that bone marrow-derived endothelial progenitor cells (EPCs) play a significant role in postnatal neovascularization. Whether the nitrogen-containing bisphosphonate zoledronate inhibits ischemia-induced neovascularization by modulating EPC functions remains unclear. Methodology/Principal Findings Unilateral hindlimb ischemia was surgically induced in wild-type mice after 2 weeks of treatment with vehicle or zoledronate (low-dose: 30 μg/kg; high-dose: 100 μg/kg). Doppler perfusion imaging demonstrated that the ischemic limb/normal side blood perfusion ratio was significantly lower in wild-type mice treated with low-dose zoledronate and in mice treated with high-dose zoledronate than in controls 4 weeks after ischemic surgery (control vs. low-dose vs. high-dose: 87±7% vs. *61±18% vs. **49±17%, *p<0.01, **p<0.005 compared to control). Capillary densities were also significantly lower in mice treated with low-dose zoledronate and in mice treated with high-dose zoledronate than in control mice. Flow cytometry analysis showed impaired mobilization of EPC-like cells (Sca-1+/Flk-1+) after surgical induction of ischemia in mice treated with zoledronate but normal levels of mobilization in mice treated with vehicle. In addition, ischemic tissue from mice that received zoledronate treatment exhibited significantly lower levels of the active form of MMP-9, lower levels of VEGF, and lower levels of phosphorylated eNOS and phosphorylated Akt than ischemic tissue from mice that received vehicle. Results of the in vitro studies showed that incubation with zoledronate inhibited the viability, migration, and tube-forming capacities of EPC. Conclusions/Significance Zoledronate inhibited ischemia-induced neovascularization by impairing EPC mobilization and angiogenic functions. These findings suggest that administration of zoledronate should be withheld in patients with ischemic events such as acute limb ischemia.
References
[1]
Folkman J (1995) Seminars in Medicine of the Beth Israel Hospital, Boston. Clinical applications of research on angiogenesis. N Engl J Med 333: 1757–1763.
[2]
Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, et al. (1997) Isolation of putative progenitor endothelial cells for angiogenesis. Science 275: 964–967.
[3]
Takahashi T, Kalka C, Masuda H, Chen D, Silver M, et al. (1999) Ischemia- and cytokine-induced mobilization of bone marrow-derived endothelial progenitor cells for neovascularization. Nat Med 5: 434–438.
[4]
Asahara T, Masuda H, Takahashi T, Kalka C, Pastore C, et al. (1999) Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circ Res 85: 221–228.
[5]
Kalka C, Masuda H, Takahashi T, Kalka-Moll WM, Silver M, et al. (2000) Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci U S A 97: 3422–3427.
[6]
Iwaguro H, Yamaguchi J, Kalka C, Murasawa S, Masuda H, et al. (2002) Endothelial progenitor cell vascular endothelial growth factor gene transfer for vascular regeneration. Circulation 105: 732–738.
[7]
Selander KS, Monkkonen J, Karhukorpi EK, Harkonen P, Hannuniemi R, et al. (1996) Characteristics of clodronate-induced apoptosis in osteoclasts and macrophages. Mol Pharmacol 50: 1127–1138.
[8]
Kobayashi Y, Hiraga T, Ueda A, Wang L, Matsumoto-Nakano M, et al. (2010) Zoledronic acid delays wound healing of the tooth extraction socket, inhibits oral epithelial cell migration, and promotes proliferation and adhesion to hydroxyapatite of oral bacteria, without causing osteonecrosis of the jaw, in mice. Journal of Bone and Mineral Metabolism 28: 165–175.
[9]
Hikita H, Miyazawa K, Tabuchi M, Kimura M, Goto S (2009) Bisphosphonate administration prior to tooth extraction delays initial healing of the extraction socket in rats. Journal of Bone and Mineral Metabolism 27: 663–672.
[10]
Yamashita J, Koi K, Yang DY, McCauley LK (2011) Effect of zoledronate on oral wound healing in rats. Clin Cancer Res 17: 1405–1414.
[11]
Russell RG (2006) Bisphosphonates: from bench to bedside. Ann N Y Acad Sci 1068: 367–401.
[12]
Zhao X, Xu X, Guo L, Ragaz J, Guo H, et al. (2010) Biomarker alterations with metronomic use of low-dose zoledronic acid for breast cancer patients with bone metastases and potential clinical significance. Breast Cancer Res Treat 124: 733–743.
[13]
Yamada J, Tsuno NH, Kitayama J, Tsuchiya T, Yoneyama S, et al. (2009) Anti-angiogenic property of zoledronic acid by inhibition of endothelial progenitor cell differentiation. J Surg Res 151: 115–120.
[14]
Woodward EJ, Coleman RE (2010) Prevention and treatment of bone metastases. Curr Pharm Des 16: 2998–3006.
[15]
Heissig B, Hattori K, Dias S, Friedrich M, Ferris B, et al. (2002) Recruitment of Stem and Progenitor Cells from the Bone Marrow Niche Requires MMP-9 Mediated Release of Kit-Ligand. Cell 109: 625–637.
[16]
Bergers G, Brekken R, McMahon G, Vu TH, Itoh T, et al. (2000) Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nat Cell Biol 2: 737–744.
[17]
Johnson C, Sung HJ, Lessner SM, Fini ME, Galis ZS (2004) Matrix metalloproteinase-9 is required for adequate angiogenic revascularization of ischemic tissues: potential role in capillary branching. Circ Res 94: 262–268.
[18]
Huang PH, Chen YH, Wang CH, Chen JS, Tsai HY, et al. (2009) Matrix metalloproteinase-9 is essential for ischemia-induced neovascularization by modulating bone marrow-derived endothelial progenitor cells. Arterioscler Thromb Vasc Biol 29: 1179–1184.
[19]
Giraudo E, Inoue M, Hanahan D (2004) An amino-bisphosphonate targets MMP-9-expressing macrophages and angiogenesis to impair cervical carcinogenesis. J Clin Invest 114: 623–633.
[20]
Rogers MJ, Crockett JC, Coxon FP, Monkkonen J (2011) Biochemical and molecular mechanisms of action of bisphosphonates. Bone 49: 34–41.
[21]
Fujita H, Kurokawa K, Ogino T, Ono M, Yamamoto M, et al. (2011) Effect of risedronate on osteoblast differentiation, expression of receptor activator of NF-kappaB ligand and apoptosis in mesenchymal stem cells. Basic Clin Pharmacol Toxicol 109: 78–84.
[22]
Matsumoto T, Nagase Y, Iwasawa M, Yasui T, Masuda H, et al. (2011) Distinguishing the proapoptotic and antiresorptive functions of risedronate in murine osteoclasts: Role of the Akt pathway and the ERK/Bim axis. Arthritis & Rheumatism 63: 3908–3917.
[23]
Itzstein C, Coxon FP, Rogers MJ (2011) The regulation of osteoclast function and bone resorption by small GTPases. Small Gtpases 2: 117–130.
[24]
Hirbe AC, Roelofs AJ, Floyd DH, Deng H, Becker SN, et al. (2009) The bisphosphonate zoledronic acid decreases tumor growth in bone in mice with defective osteoclasts. Bone 44: 908–916.
[25]
Cornish J, Bava U, Callon KE, Bai J, Naot D, et al. (2011) Bone-bound bisphosphonate inhibits growth of adjacent non-bone cells. Bone 49(4): 710–716.
[26]
Kuiper JW, Forster C, Sun C, Peel S, Glogauer M (2011) Zoledronate and Pamidronate negatively affect neutrophil effector functions and survival in mice. Br J Pharmacol 165(2): 532–539.
[27]
Sminia T, Dijkstra CD (1986) The origin of osteoclasts: an immunohistochemical study on macrophages and osteoclasts in embryonic rat bone. Calcif Tissue Int 39: 263–266.
[28]
Russell HV, Groshen SG, Ara T, Declerck YA, Hawkins R, et al. (2010) A phase I study of zoledronic acid and low-dose cyclophosphamide in recurrent/refractory neuroblastoma: A new approaches to neuroblastoma therapy (NANT) study. Pediatr Blood Cancer 57(2): 275–282.
[29]
Gutteridge DH, Retallack RW, Ward LC, Stuckey BG, Stewart GO, et al. (1996) Clinical, biochemical, hematologic, and radiographic responses in Paget's disease following intravenous pamidronate disodium: a 2-year study. Bone 19: 387–394.
[30]
Coukell AJ, Markham A (1998) Pamidronate. A review of its use in the management of osteolytic bone metastases, tumour-induced hypercalcaemia and Paget's disease of bone. Drugs Aging 12: 149–168.
[31]
Basi DL, Hughes PJ, Thumbigere-Math V, Sabino M, Mariash A, et al. (2011) Matrix metalloproteinase-9 expression in alveolar extraction sockets of Zoledronic acid-treated rats. J Oral Maxillofac Surg 69: 2698–2707.
[32]
Ferrara N (2004) Vascular endothelial growth factor: basic science and clinical progress. Endocr Rev 25: 581–611.
[33]
Stresing V, Fournier PG, Bellahcene A, Benzaid I, Monkkonen H, et al. (2011) Nitrogen-containing bisphosphonates can inhibit angiogenesis in vivo without the involvement of farnesyl pyrophosphate synthase. Bone 48: 259–266.
[34]
Li YY, Chang JW, Liu YC, Wang CH, Chang HJ, et al. (2011) Zoledronic acid induces cell-cycle prolongation in murine lung cancer cells by perturbing cyclin and Ras expression. Anticancer Drugs 22: 89–98.
[35]
Fournier P, Boissier S, Filleur S, Guglielmi J, Cabon F, et al. (2002) Bisphosphonates inhibit angiogenesis in vitro and testosterone-stimulated vascular regrowth in the ventral prostate in castrated rats. Cancer Res 62: 6538–6544.
[36]
Santini D, Vincenzi B, Dicuonzo G, Avvisati G, Massacesi C, et al. (2003) Zoledronic acid induces significant and long-lasting modifications of circulating angiogenic factors in cancer patients. Clin Cancer Res 9: 2893–2897.
[37]
Yamagishi S, Abe R, Inagaki Y, Nakamura K, Sugawara H, et al. (2004) Minodronate, a newly developed nitrogen-containing bisphosphonate, suppresses melanoma growth and improves survival in nude mice by blocking vascular endothelial growth factor signaling. Am J Pathol 165: 1865–1874.
[38]
Lacy MQ, Dispenzieri A, Gertz MA, Greipp PR, Gollbach KL, et al. (2006) Mayo clinic consensus statement for the use of bisphosphonates in multiple myeloma. Mayo Clin Proc 81: 1047–1053.
[39]
Aicher A, Heeschen C, Mildner-Rihm C, Urbich C, Ihling C, et al. (2003) Essential role of endothelial nitric oxide synthase for mobilization of stem and progenitor cells. Nat Med 9: 1370–1376.
[40]
Huang PH, Tsai HY, Wang CH, Chen YH, Chen JS, et al. (2010) Moderate intake of red wine improves ischemia-induced neovascularization in diabetic mice–roles of endothelial progenitor cells and nitric oxide. Atherosclerosis 212: 426–435.
[41]
Huang PH, Huang SS, Chen YH, Lin CP, Chiang KH, et al. (2010) Increased circulating CD31+/annexin V+ apoptotic microparticles and decreased circulating endothelial progenitor cell levels in hypertensive patients with microalbuminuria. J Hypertens 28: 1655–1665.
