Cilostazol Mediates Immune Responses and Affects Angiogenesis During the Acute Phase of Hind Limb Ischemia in a Mouse Model

Abstract

Background: Cilostazol is a drug of choice for the treatment of intermittent claudication that also affects innate and adaptive immune cells. The purpose of our study was the evaluation of cilostazol’s impact on the immune and angiogenic response in murine models of hind limb ischemia. Methods: We used 108 immunodeficient NOD.CB17-Prkdcscid/J mice and 108 wild-type CB17 mice. At day 0 (D0), all animals underwent hind limb ischemia. Half of them in both groups received daily cilostazol starting at D0 and for the next 7 postoperative days, while the rest of them served as controls, receiving vehicle. Interleukin (IL) 2, IL-4, IL-6, IL-10, IL-17A, tumor necrosis factor α (TNF-α), and interferon γ (IFN-γ) serum concentrations were measured by flow cytometry on postsurgery days D1, D3, D5, and D7. On D7, both groups underwent positron emission tomography scan with 68Ga-RGD. Mice were euthanatized and gastrocnemius muscles were obtained for histological evaluation. Results: There was a statistically significant augmentation ( P < .05) in IL-4, IL-10, IL-6, and IFN-γ concentrations in treated CB17 animals, while IL-2 was significantly suppressed. Significant difference was detected between the CiBisch and Bisch groups on D1 and D7 ( P < .05) in CD31 staining. In treated NOD.CB17 animals, TNF-α, IL-6, and IFN-γ presented significant augmentation, while 68Ga-NODAGA-RGDfK uptake and CD31 expression were found significantly lower for both legs in comparison to the control. Conclusion: Cilostazol seems to significantly increase angiogenesis in wild-type animals during the first postoperational week. It also influences immune cells, altering the type of immune response by promoting anti-inflammatory cytokine production in wild-type animals, while it helps toward inflammation regression in immunodeficient animals.

Evolving applications of the egg: chorioallantoic membrane assay and ex vivo organotypic culture of materials for bone tissue engineering

“…The various experimental methods for stimulating and quantifying angiogenesis have been reviewed by Tahergorabi and Khazaei, 59 Hasan et al 60 and Norrby; 61 however, the CAM has not superseded these methods in some areas of research, as the ex vivo aortic ring assay 62 and hind limb ischaemia model 63 are still used, depending on the question being asked and the ultimate research goal. The hindlimb ischaemia model is focused on finding therapies for peripheral vascular disease using methods such as growth factors and stem cells, and would not be a suitable method for scaffold assessment as the model does not imitate the clinical scenario.…”

Transcriptomic analysis reveals Cilostazol's role in ameliorating cardiovascular disease: Inhibition of monocyte-to-macrophage differentiation and reduction of endothelial cell reactive oxygen species production

“…It potentiates adipocyte differentiation in 3T3-L1 cells [ 44 ], and its effect on plasma lipoproteins in patients with intermittent claudication has also been noted [ 45 ].Conversely, the influence of cilostazol on specific immune cells like dendritic cells and macrophages indicates reduced inflammation, a critical factor in CVD [ [46] , [47] , [48] , [49] ]. Its anti-inflammatory attributes are further affirmed in various scenarios, including hindlimb ischemia in a mouse model [ 50 ], potential COVID-19 treatment [ 51 ], and the inactivation of NF-κB in BV2 microglial cells [ 52 ]. This aligned evidence consolidates cilostazol's potential therapeutic value.…”

Cilostazol induces angiogenesis and regulates oxidative stress in a dose-dependent manner: A chorioallantoic membrane study

“…Similarly, increased angiogenesis and induced cell proliferation through cAMP activation were detected in the rat myocardial ischemia-reperfusion model. [ 33 ] In another experimental hindlimb ischemia study, Paronis et al[ 34 ] investigated immune responses, such as interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17A, tumor necrosis factor-alpha, and interferon-γ concentrations, in cilostazol-treated rats and found that cilostazol promoted anti-inflammatory cytokine production and induced postoperative angiogenesis. The present study is the first CAM model study to investigate physiological development of embryos treated with cilostazol.…”