Primary Human Lung Alveolus‐on‐a‐chip Model of Intravascular Thrombosis for Assessment of Therapeutics

Abstract

Pulmonary thrombosis is a significant cause of patient mortality; however, there are no effective in vitro models of thrombi formation in human lung microvessels that could also assess therapeutics and toxicology of antithrombotic drugs. Here, we show that a microfluidic lung alveolus-on-a-chip lined by human primary alveolar epithelium interfaced with endothelium and cultured under flowing whole blood can be used to perform quantitative analysis of organ-level contributions to inflammation-induced thrombosis. This microfluidic chip recapitulates in vivo responses, including platelet-endothelial dynamics and revealed that lipopolysaccharide (LPS) endotoxin indirectly stimulates intravascular thrombosis by activating the alveolar epithelium, rather than acting directly on endothelium. This model is also used to analyze inhibition of endothelial activation and thrombosis due to a protease activated receptor-1 (PAR-1) antagonist, demonstrating its ability to dissect complex responses and identify antithrombotic therapeutics. Thus, this methodology offers a new approach to study human pathophysiology of pulmonary thrombosis and advance drug development.

Biomimetics of the pulmonary environment in vitro: A microfluidics perspective

“…Such efforts have principally focused on airway disease models such as a micro-engineered ACB platform for pulmonary edema, 16 pulmonary thrombosis, 20 and a small airway-on-a-chip model for COPD studies 24 . The former study utilizes the same device and alveolar epithelium (i.e., type II A549 cell line) as in Huh et al 23 to investigate barrier integrity as a result of cytotoxicity arising from therapeutics (e.g., chemotherapy), showing that stretching of the epithelium compromises the pulmonary barrier.…”

“…This is exemplified in the broad range of pulmonary-related research currently explored in vitro with the support of microfluidic platforms (Fig. 5): notable efforts in the above-mentioned areas include amongst other (i) exploring ALI and ACB barrier characteristics, 23 (ii) conducting therapeutic drug screens, 20 (iii) mapping aerosol deposition patterns in airways, 41 (iv) quantifying the dynamics of liquid plug transport, 86 and (v) assessing airway epithelial injury 84 . In further advancing the field to deliver respiratory platforms that meet the complex integration of anatomical, physiological, and biological constraints, microfluidic designs must strive to recreate more faithfully whole-organ functions; a condition that would encourage expanding the breadth (i.e., scaling-up) of microfluidic platforms [e.g., Figs.…”

“…Copyright 2010 American Association for the Advancement of Science. (b) Jain et al 20 demonstrated that a microfluidic alveolus-on-a-chip lined with both human primary alveolar epithelial and endothelial cells cultured under whole blood perfusion can be leveraged to identify antithrombotic therapeutics. Reproduced with permission from Clin.…”

“…Concurrently, the accessibility to a vast tissue surface offers a potent non-invasive gateway for inhalation therapeutics in the context of both topical 10,11 (e.g., antibiotics and chemotherapy) and systemic delivery 12–14 (e.g., gene therapy and vaccination). Alongside, in vitro microfluidic platforms of the respiratory airways are undergoing rapid development motivated amongst others by lung disease modeling, 15–20 cytotoxicity studies, 16,21 cancer diagnosis, 22 and drug screening assays 20,23–25 . Among the arguments advocated for perseverance lies, for example, the prospect of alternatives to animal testing 21,26,27 both as a testing platform for the above-mentioned applications as well as for individualized disease models, allowing an optimized match between patient and treatment 18 .…”

A Synthetic Circuit for Mercury Bioremediation Using Self-Assembling Functional Amyloids

“…3 In utero exposure results in severe developmental abnormalities, resulting in EPA-FDA advisories against eating fish during pregnancy. 4 Recent studies have found that mercury contamination in the environment is much more prevalent than previously thought, 5 hence the need for innovative -binding curli biofilm circuit. The reporter gene in the MerR-based mercury biosensing circuit is replaced with a curli operon encoding the synthesis and export of self-assembling functional amyloids that are able to bind mercury ions.…”