Exploring smooth muscle phenotype and function in a bioreactor model of abdominal aortic aneurysm

Journal of Translational Medicine

Karen E Porter³  D Julian A Scott⁴  Peter G Walker¹  Sotirios Korossis⁵  Soroush Sohrabi⁴  Marc A Bailey⁴  Emily Clark¹  Jean Kaye²  Gurprit S Mudhar²  Timothy G Angelini²  Kirsten Riches³ 

[1] Institute of Medical and Biological Engineering, School of Mechanical Engineering, University of Leeds, Leeds, UK;Division of Cardiovascular and Diabetes Research, Leeds Institute of Genetics, Health and Therapeutics (LIGHT), University of Leeds, Leeds, UK;Multidisciplinary Cardiovascular Research Centre (MCRC), University of Leeds, Leeds, UK;Leeds Vascular Institute, Leeds General Infirmary, Leeds, UK;Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany

关键词: MMP-2;    Apoptosis;    Senescence;    Proliferation;    Morphology;    Bioreactor;    Porcine;    Human;    Smooth muscle cell;    Abdominal aortic aneurysm;   

Others  :  826680
DOI  :  10.1186/1479-5876-11-208

 received in 2013-03-13, accepted in 2013-09-04,  发布年份 2013

【 摘 要 】

Background

Vascular smooth muscle cells (SMC) are central to arterial structure and function yet their involvement in the progression of abdominal aortic aneurysm (AAA) disease is not well studied. The progressive and silent nature of AAA in man essentially restricts research to the use of “end-stage” tissue recovered during surgical repair. This study aimed to generate an ex vivo model of AAA using protease-treated porcine carotid arteries maintained in a novel bioreactor, and to compare the structural and functional changes in SMC cultured from the recovered vessels with those from human tissue acquired at elective surgical repair.

Methods

Freshly isolated porcine arteries were pretreated with collagenase and/or elastase before culturing under flow in a bioreactor for 12 days. Human end-stage aneurysmal tissue and saphenous veins from age-matched controls were collected from patients undergoing surgery. SMC were cultured and characterised (immunocytochemistry, measurement of spread cell area) and assessed functionally at the level of proliferation (cell-counting) and matrix-metalloproteinase (MMP) secretion (gelatin zymography). Cellular senescence was investigated using β-galactosidase staining and apoptosis was quantified using a fluorescence-based caspase 3 assay.

Results

Co-expression of alpha-smooth muscle actin and smooth muscle myosin heavy chain confirmed all cell populations as SMC. Porcine SMC harvested and cultivated after collagenase/elastase pretreatment displayed a prominent “rhomboid” morphology, increased spread area (32%, P < 0.01), impaired proliferation (47% reduction, P < 0.05), increased senescence (52%, P < 0.001), susceptibility to apoptosis and reduced MMP-2 secretion (60% decrease, P < 0.01) compared with SMC from vehicle, collagenase or elastase pre-treated vessels. Notably, these changes were comparable to those observed in human AAA SMC which were 2.4-fold larger than non-aneurysmal SMC (P < 0.001) and exhibited reduced proliferation (39% reduction, P < 0.001), greater apoptosis (4-fold increase, P < 0.001), and increased senescence (61%, P < 0.05).

Conclusions

Combined collagenase/elastase exposure of porcine artery maintained in a bioreactor under flow conditions induced a SMC phenotype characteristic of those cultured from end-stage AAA specimens. This model has potential and versatility to examine temporal changes in SMC biology and to identify the molecular mechanisms leading to early aberrancies in SMC function. In the longer term this may inform new targets to maintain aortic SMC content and drive cells to a “reparative” phenotype at early stages of the disease.

【 授权许可】

   
2013 Riches et al.; licensee BioMed Central Ltd.

【 图 表 】

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