title: Side view thrombosis microfluidic device with controllable wall shear rate and transthrombus pressure gradient.
abstract: Hemodynamic conditions vary throughout the vasculature, creating diverse environments in which platelets must respond. To stop bleeding, a growing platelet deposit must be assembled in the presence of fluid wall shear stress (τw) and a transthrombus pressure gradient (ΔP) that drives bleeding. We designed a microfluidic device capable of pulsing a fluorescent solute through a developing thrombus forming on collagen ± tissue factor (TF), while independently controlling ΔP and τw. Computer control allowed step changes in ΔP with a rapid response time of 0.26 mm Hg s(-1) at either venous (5.2 dynes cm(-2)) or arterial (33.9 dynes cm(-2)) wall shear stresses. Side view visualization of thrombosis with transthrombus permeation allowed for quantification of clot structure, height, and composition at various ΔP. Clot height was reduced 20% on collagen/TF and 28% on collagen alone when ΔP was increased from 20.8 to 23.4 mm Hg at constant arterial shear stress. When visualized with a platelet-targeting thrombin sensor, intrathrombus thrombin levels decreased by 62% as ΔP was increased from 0 to 23.4 mm Hg across the thrombus-collagen/TF barrier, consistent with convective removal of thrombogenic solutes due to pressure-driven permeation. Independent of ΔP, the platelet deposit on collagen had a permeability of 5.45 × 10(-14) cm(2), while the platelet/fibrin thrombus on collagen/TF had a permeability of 2.71 × 10(-14) cm(2) (comparable to that of an intact endothelium). This microfluidic design allows investigation of the coupled processes of platelet deposition and thrombin/fibrin generation in the presence of controlled transthrombus permeation and wall shear stress.
authors.affiliation: Institute for Medicine and Engineering, Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Ryan W
authors.initials: RW
authors.lastname: Muthard
authors.forename: Scott L
authors.initials: SL
authors.lastname: Diamond
mesh_terms: D001792:Blood Platelets / Q000378:metabolism; D003094:Collagen / Q000737:chemistry / Q000378:metabolism; D005337:Fibrin / Q000378:metabolism; D020342:Finite Element Analysis; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D010539:Permeability; D033081:Shear Strength; D013917:Thrombin / Q000737:chemistry / Q000378:metabolism; D013925:Thromboplastin / Q000737:chemistry / Q000378:metabolism; D013927:Thrombosis / Q000378:metabolism / Q000503:physiopathology*
chemical_list: D005456:Fluorescent Dyes; D005337:Fibrin; D003094:Collagen; D013925:Thromboplastin; D013917:Thrombin
references.citation: J Thromb Haemost. 2012 Nov;10(11):2344-53
references.pmid: 22978514
references.citation: Arterioscler Thromb Vasc Biol. 2001 Jan;21(1):142-8
references.pmid: 11145946
references.citation: Blood. 2013 Mar 7;121(10):1875-85
references.pmid: 23303817
references.citation: J Clin Invest. 1962 Aug;41:1591-8
references.pmid: 14040228
references.citation: Anal Chem. 2010 Feb 1;82(3):808-16
references.pmid: 20050596
references.citation: FASEB J. 2009 Jul;23(7):2155-64
references.pmid: 19246488
references.citation: Bioconjug Chem. 2011 Oct 19;22(10):2104-9
references.pmid: 21902184
references.citation: Mol Immunol. 1984 Jan;21(1):89-94
references.pmid: 6200769
references.citation: Am J Physiol. 1984 Nov;247(5 Pt 2):H784-91
references.pmid: 6496759
references.citation: J Appl Physiol (1985). 1991 Dec;71(6):2469-75
references.pmid: 1778948
references.citation: Blood. 2007 Jan 1;109(1):130-8
references.pmid: 16940416
references.citation: Nature. 1979 Jun 14;279(5714):636-8
references.pmid: 313016
references.citation: Biophys J. 2002 Sep;83(3):1650-60
references.pmid: 12202388
references.citation: J Thromb Haemost. 2008 Dec;6(12):2193-201
references.pmid: 18983510
references.citation: Arterioscler Thromb Vasc Biol. 2012 Jun;32(6):1466-76
references.pmid: 22516070
references.citation: N Engl J Med. 2007 Aug 9;357(6):535-44
references.pmid: 17687129
references.citation: J Thromb Haemost. 2004 Jul;2(7):1155-62
references.pmid: 15219199
references.citation: Crit Rev Biomed Eng. 1990;18(1):27-54
references.pmid: 2204514
references.citation: Nat Rev Cancer. 2004 Oct;4(10):806-13
references.pmid: 15510161
references.citation: Arterioscler Thromb Vasc Biol. 2012 Dec;32(12):2938-45
references.pmid: 23087356
references.citation: Nat Med. 2002 Oct;8(10):1175-81
references.pmid: 12244306
references.citation: Am J Pathol. 1989 May;134(5):1087-97
references.pmid: 2719077
references.citation: Microvasc Res. 1979 Jan;17(1):38-54
references.pmid: 459932
references.citation: Biophys J. 1986 Nov;50(5):937-45
references.pmid: 3790695
references.citation: Microvasc Res. 2007 Sep-Nov;74(2-3):72-84
references.pmid: 17560615
references.citation: Lab Chip. 2008 May;8(5):701-9
references.pmid: 18432339
references.citation: Lab Chip. 2008 Sep;8(9):1468-77
references.pmid: 18818801
references.citation: Blood. 2012 Jul 5;120(1):190-8
references.pmid: 22517902
references.citation: Anal Chem. 1998 Dec 1;70(23):4974-84
references.pmid: 21644679