Artery-on-a-Chip: A Microfluidic Platform to Investigate Small Artery Function
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1 Artery-on-a-Chip: A Microfluidic Platform to Investigate Small Artery Function 1 C Lochovksy, 1 S Yasotharan, 1 A Vagaon, 1 D Lidington, 1,2 J Voigtlaender-Bolz, 1 S-S. Bolz, 1 A Günther 1 University of Toronto, 2 St. Michael s Hospital, Toronto Motivation, basic biology, current methods A microfluidic platform for investigating blood vessels Viability of vessels on chip New tools for vascular biology and beyond
2 Why Small Resistance Arteries Small resistance arteries: vessels prior to capillary beds Tissue Components: approx. 10% endothelium, 60% smooth muscle, 30% tissue Highly elastic compared to capillaries, which are fragile (95% endothelium) Inner Outer Diameters (for mice) R, resistance Resistance Arteries regulate flow and pressure drop in circulatory system Control of blood pressure key element in hypertension
3 Conventional Methods: Cannulation & Pressure Myographs pipettes 3cm Expensive, not scalable Platform is not versatile (spatially, temporally) Method is time consuming Requires high degree of skill and training Duling et al. Am J Physiol. 241(1), 1981
4 Present Microfluidic Strategies for Cardiovascular Applications Choi et al. Nat. Mater. 6 (11), 2007 Chiu et al, PNAS, 97(6) 2000 Poster PS-9A-147, Nano to Macro Fluidic Technology Advances Session I, Exhibit Hall C tomorrow, 9-1pm Vickerman et al. Lab Chip 8(9) 2008 Tanaka et al. Lab Chip 7(2) 2007 We present one of the first organ-based microfluidic approaches for cardiovascular research
5 Device Fabricated using Soft-Lithography Master fabrication Spincoat Su8 for ~150µm depth Verify depth using profilometer 1. Glass Slide 2. Su-8 3. Transparency 4. Expose Replica Molding, Integration Mold microchannel network in elastomer (poly-dimethylsiloxane or PDMS); the master acts as a stamp Degas (~2hr) and cure (~2hr) Plasma bond to glass slide with tubing inserts, pin device Xia et al, Annu. Rev. Mater. Sci., 37 (5) 1998
6 Microfluidic Platform Superfusion: flow around the vessel inflow mixing outflow (green) Perfusion: flow through lumen of the vessel inflow outflow (blue) Suction: vessel fixation (-45mmHg) At 8 points (orange) All lines are liquid filled Superfusion Inlets and Mixing Region of Interest (ROI) Loading Bath
7 Artery Vessel Loading Procedure on Chip Loading Channel Fixation Channel Perfusion Superfusion
8 Microfluidic Chip achieves Flow Separation between Superfusion and Perfusion Concentration Velocity Poster PS-9B-164, Nano to Macro Fluidic Technology Advances session II, Exhibit Hall C, tomorrow, 1-5pm
9 Small Artery Function Dose drug of interest to test smooth muscle cells (SMCs, outside vessel) and endothelial cells (ECs, lumen) We see a resulting change in vessel diameter
10 Phenylephrine Dose Response Consistent with Conventional Data Phenylephrine (PE) acts SMCs as a constrictor, delivered in 3-(N-morpholino)propanesulfonic acid (MOPS) solution
11 Chip is suitable for Endothelial cells and Longterm Culture Acetylcholine (AcH) is a vasodilator acting on ECs Vessels are kept at physiologically relevant pressure and temperature
12 Software system developed in MatLab containing: Automated/scheduled pumps and dosing Automated image processing/dose response measurement Automated Processes Flow Control Diameter Measurement Inverted Microscope Dose varies: Automated dose response of vessel with PE on chip, repeated 10 times Vessel response is diminished as more dose sequences are completed
13 Embryo Artery in Spatially Inhomogeneous Microenvironment 1 2 Lucchetta et al. Nature 434 (7037) 2005
14 Results show Uni-sided Constriction
15 Summary Scalable microfluidic platform for loading blood vessels developed Consistency of artery functional responses, and feasibility of longterm culture Demonstrated automated approach to dose response measurements Showed dose response in a inhomogeneous microenvironment
16 Acknowledgements Guenther Lab group (Prof. Axel Guenther) Vessel members: Sanjesh, Sascha, Calvin, Alfred, Xiao Physiology Lab (Prof. Steffen-Sebastian Bolz, Andrei, Julia, Meghan) OGS, U of T Open, Barbara & Frank Milligan Fellowship OCE (Champions of Innovation), NSERC (Idea to Innovation) Industrial Collaborators: OAI Technologies, Quorum Technologies (Guelph), Nanopoint Imaging (Honolulu)
A Microfluidic Platform for the Automated Multimodal Assessment of Small Artery Structure and Function.
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