Induction of human aortic myofibroblast-mediated extracellular matrix dysregulation: A potential mechanism of fluoroquinolone-associated aortopathy

Abstract

Objectives: Fluoroquinolone (FQ) antibiotics are associated with adverse aortic clinical events. We assessed human aortic myofibroblast-mediated extracellular matrix (ECM) dysregulation as a possible cellular mechanism underlying FQassociated aortopathy. Methods: Human aortic myofibroblasts were isolated from patients with aortopathy undergoing elective ascending aortic resection (N ¼ 9). The capacity for extracellular matrix degradation in cells exposed to FQ was assessed by multiplex analysis of secreted matrix metalloproteinases relative to tissue inhibitors of matrix metalloproteinases (TIMPs). Direct evaluation of extracellular matrix degradation was investigated in human aortic cells using a 3-dimensional gelatin-fluorescein isothiocyanate fluorescence microgel assay. Aortic cellular collagen-1 expression following FQ exposure was determined by immunoblotting and immunofluorescent staining. Cell apoptosis, necrosis, and metabolic viability was determined by annexin-V, propidium iodide staining, and water-soluble tetrazolium salt (WST1) assay. Results: FQ exposure significantly decreased aortic cell TIMP-1 (P ¼ .004) and TIMP-2 (P ¼ .0004) protein expression compared with vehicle control. The ratio of matrix metalloproteinase-9/TIMP-2 was increased suggesting an increased capacity for extracellular matrix degradation (P ¼ .01). In collagen gels, we show a trend toward increased aortic myofibroblast-mediated collagen fiber degradation with FQ exposure (P ¼ .09). Similarly, FQ exposure attenuated collagen-1 expression as assessed by immunoblotting (P ¼ .002) and immunofluorescence (P ¼ .02). Cell apoptosis, necrosis, and metabolic viability was not significantly influenced by FQ exposure. Conclusions: For the first time, we document a putative mechanism underlying FQ-associated aortopathy whereby decreased TIMP expression with impaired compensatory collagen-1 expression results in human aortic myofibroblastmediated extracellular matrix dysregulation. These novel data may provide a cellular and molecular mechanism to explain the established clinical association between FQ exposure and acute aortic events.

Fluoroquinolones and the Risk of Aortic Aneurysm or Aortic Dissection: A Systematic Review and Meta-Analysis

“…Cell apoptosis, necrosis, and metabolic viability were not significantly affected by ciprofloxacin exposure. These findings suggest rapid extracellular matrix degradation as the main underlying mechanism behind fluoroquinolone-associated aortic event, which is consistent with the observed early-onset characteristic of the association with respect to the exposure to fluoroquinolones [29]. Further studies are needed to strengthen this observation.…”

Insights on the Pathogenesis of Aneurysm through the Study of Hereditary Aortopathies

“…Consistent with these latter observations, genetic or pharmacological inactivation of lysyl oxidases, enzymes necessary for collagen and elastin cross-linking, cause or exacerbate aneurysm in patients and animal models [197,198,246,255,269,273,[395][396][397]. The detrimental effects of fluoroquinolones on TAA pathogenesis have also been attributed to excess ECM degradation and reduced levels of collagen [398][399][400][401].…”

The dark side of fluoroquinolones

“…6 The samples were procured from only 9 patients with aortopathy, and bicuspid aortic valves were present in most of the patients; aortopathy may genetically differ in some patients according to aortic valve morphology. 7 As Guzzardi and colleagues 4 state, this is clearly a preliminary work.…”

“…The study presented in this issue of the Journal by Guzzardi and colleagues 4 sheds fascinating light on the mystery association of fluoroquinolones and aortopathy. Human aortic myofibroblasts were procured from patients undergoing surgery for the ascending aorta.…”