Abstract
Vasoactive liabilities are typically assayed using wire myography, which is limited by its high cost and low throughput. To meet the demand for higher throughput in vitro alternatives, this study introduces a magnetic 3D bioprinting-based vasoactivity assay. The principle behind this assay is the magnetic printing of vascular smooth muscle cells into 3D rings that functionally represent blood vessel segments, whose contraction can be altered by vasodilators and vasoconstrictors. A cost-effective imaging modality employing a mobile device is used to capture contraction with high throughput. The goal of this study was to validate ring contraction as a measure of vasoactivity, using a small panel of known vasoactive drugs. In vitro responses of the rings matched outcomes predicted by in vivo pharmacology, and were supported by immunohistochemistry. Altogether, this ring assay robustly models vasoactivity, which could meet the need for higher throughput in vitro alternatives.
Highlights
Throughput and offering kinetic analysis compared to traditional well-by-well imaging under a microscope[24,27]
The rings were exposed to a small panel of compounds with known vasoactive responses
The contractile responses of rings were measured over 24 h of exposure, as well as their viability. Their expression of α-smooth muscle actin, a cytoskeletal component and smooth muscle marker[28], and phosphorylated myosin light chain, a precursor for vascular smooth muscle contraction were analyzed with immunohistochemistry[29]
Summary
Throughput and offering kinetic analysis compared to traditional well-by-well imaging under a microscope[24,27]. Taken together, this assay has the potential to meet the needs for an in vitro assay for high-throughput vasoactivity screening. The goal of this study was to validate this ring assay as an in vitro measure of vasoactivity. The contractile responses of rings were measured over 24 h of exposure, as well as their viability Their expression of α-smooth muscle actin (αSMA), a cytoskeletal component and smooth muscle marker[28], and phosphorylated myosin light chain (pMLC), a precursor for vascular smooth muscle contraction were analyzed with immunohistochemistry[29]. The results of this study validate contraction in magnetically 3D bioprinted rings as an endpoint for vasoactivity, and form the foundation for high-throughput screening of vasoactivity
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