Poster U3 - Atherosclerotic three-layer nanomatrix vascular sheets for high-throughput therapeutic evaluation

Introduction: Atherosclerosis is a major cause of cardiovascular disease, and effective therapeutics require reliable in vivo and in vitro models. However, current models have limitations, such as high cost, long induction time, and lack of accurate replication of human atherosclerosis pathogenesis. To overcome these challenges, we developed an innovative in vitro platform called tissue-engineered vascular sheets with atherosclerosis (VSA). The VSA platform features a 3-layered artery structure and replicates disease characteristics such as endothelial dysfunction, macrophages, and foam cells. Additionally, the VSA platform can be easily scaled up for drug evaluation using multi-well plates, allowing for high-throughput screening of potential therapeutics in a cost-effective way.

Materials and

Methods: VSA are fabricated as follows:1) hAAF and hAoSMC sheets were obtained by encapsulating human aortic adventitial fibroblasts (hAAFs) and aortic smooth muscle cells (hAoSMCs) by nanomatrix thin sheet technology using peptide amphiphile (PA-RGDS) gel; 2) human aortic endothelial cells (hAECs) were seeded on hAoSMC sheet; 3) The hAAF sheet was assembled with hAEC-hAoSMC sheet to form a 3-layered vascular sheet; 4) Atherosclerosis was induced by adding pro-atherogenic cytokines, Ox-LDL, and monocytes.

Results, Conclusions, and Discussions: Fig.1A-C show the cross-sectional fluorescent image of hAEC, hAoSMC and the 3-layer of the vascular sheet indicates an excellent three-layered structure which mimics the native vascular structure. The hAEC, hAoSMC and fibroblasts of the vascular sheet show proper phenotype in sheet (Fig.E-F). Moreover, the positive immunostaining of ICAM-1 on the VSA indicate successful induction of EC dysfunction (Fig.1G). Monocyte recruitment and macrophage formation were also achieved on the VSA (Fig.1H-I). Other atherosclerotic features, such as aggregated foam cells and lipid pool (Fig.J-K) and various types of human atherosclerosis related inflammatory cytokines were also observed (Fig.1M). Notably, the production of VSA can be upscaled in a 48-well plate for drug evaluation. Statin and sirolimus were used as postive controls for demonstrating the feasibility of using VSA for drug evaluation (Fig.1N-O). Promisingly, the statin or sirolimus-treated VSA showed significantly less foam cells and various inflammatory cytokine production than the untreated VSA, which agrees with the therapeutic effect of statin or sirolimus reported in vivo studies.

Conclusions: We have developed VSA and VSA plates. The VSA plate shows excellent potential for drug evaluation.

Acknowledgements (Optional):