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Research Papers

Pulsatile Perfusion Bioreactor for Biomimetic Vascular Impedances

[+] Author and Article Information
David A. Prim

College of Engineering and Computing,
Biomedical Engineering Program,
University of South Carolina,
Columbia, SC 29208

Jay D. Potts

School of Medicine,
Department of Cell Biology and Anatomy,
College of Engineering and Computing,
Biomedical Engineering Program,
University of South Carolina,
Columbia, SC 29208

John F. Eberth

School of Medicine,
Department of Cell Biology and Anatomy,
College of Engineering and Computing,
Biomedical Engineering Program,
University of South Carolina,
Columbia, SC 29208
e-mail: john.eberth@uscmed.sc.edu

1Corresponding author.

Manuscript received January 7, 2018; final manuscript received June 14, 2018; published online September 21, 2018. Assoc. Editor: Yaling Liu.

J. Med. Devices 12(4), 041002 (Sep 21, 2018) (10 pages) Paper No: MED-18-1003; doi: 10.1115/1.4040648 History: Received January 07, 2018; Revised June 14, 2018

Pulsatile waves of blood pressure and flow are continuously augmented by the resistance, compliance, and inertance properties of the vasculature, resulting in unique wave characteristics at distinct anatomical locations. Hemodynamically generated loads, transduced as physical signals into resident vascular cells, are crucial to the maintenance and preservation of a healthy vascular physiology; thus, failure to recreate biomimetic loading in vitro can lead to pathological gene expression and aberrant remodeling. As a generalized approach to improve native and engineered blood vessels, we have designed, built, and tested a pulsatile perfusion bioreactor based on biomimetic impedances and a novel five-element electrohydraulic analog. Here, the elements of an incubator-based culture system were formulaically designed to match the vascular impedance of a brachial artery by incorporating both the inherent (systemic) and added elements of the physical system into the theoretical approach. Freshly harvested porcine saphenous veins were perfused within a physiological culture chamber for 6 h and the relative expression of seven known mechanically sensitive remodeling genes analyzed using the quantitative polymerase chain reaction (qPCR) method. Of these, we found plasminogen activator inhibitor-1 (SERPINE1) and fibronectin-1 (FN1) to be highly sensitive to differences between arterial- and venous-like culture conditions. The analytical approach and biological confirmation provide a framework toward the general design of long-term hemodynamic-mimetic vascular culture systems.

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Figures

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Fig. 1

Visualization of Fourier analysis illustrating (a) the steady and composite waveforms of volumetric blood flow (solid) and blood pressure (dash-dot) of the brachial artery (BrA). The fundamental and first two harmonics are shown for the (b) volumetric blood flow rate and (c) blood pressure. The impedance (d) magnitude and (e) phase for the BrA.

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Fig. 2

(a) Electro-hydraulic analog of the cardiovascular system and bench top schematic with (b) equivalent impedances

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Fig. 3

Parameter sensitivity of the five-element electrohydraulic analog demonstrating impedance magnitude (left) and phase (right). (a) and (b) R1∈[10,110] (mmHg⋅s/mL), (c) and (d) R2∈[0,24], (e) and (f) C∈[0.005,0.025] (mL/mmHg), and (g) and (h) La∈[0,0.8] (mmHg⋅s2/mL). Default values of R1 = 60 (mmHg⋅s/mL), R2 = 5 (mmHg⋅s/mL), C = 0.01 (mL/mmHg), La = 0.33 (mmHg⋅s2/mL), and Ls = 0.33 (mmHg⋅s2/mL). Arrows indicate the direction of increasing values of the given parameter.

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Fig. 4

Hardware schematic for the pulsatile perfusion bioreactor

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Fig. 5

Values of isolated design elements in the pulsatile perfusion bioreactor: (a) measured systemic resistance and the resistances at discrete positions of a custom designed plate (linearized) valve and (b) resistances at discrete positions of the tubing pinch valve, (c) measured systemic compliance and measured versus added compliance using a chamber with different preloaded volumes of air. (d) Measured systemic inductance and added inductances from a coil of tubing using different tubing diameters.

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Fig. 6

(a) Customized three channel peristaltic pump with (b) synchronous gearing system and timing belts that generate the fundamental (k = 1) and first two harmonics (k = 2, 3) of the frequency. (c) Illustration of a single roller pump head containing three rollers where the volumetric flow phase angle was set using a cam mechanism and the displaced volume adjusted by using tubing with different lumen diameters.

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Fig. 7

Simultaneous brachial artery (BrA) (a) volumetric blood flow rate and (b) blood pressure for one cardiac cycle. Solid lines indicate the desired waveform and (×) the values recorded in our bioreactor system. (c) The impedance modulus and (d) phase for the desired (+), theoretical (○), and measured response (×).

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Fig. 8

Simultaneous (a) and (b) GSV and left anterior descending coronary artery (LCA) (c) and (d) volumetric blood flow rate and pressure for one cardiac cycle. Solid lines indicate the desired waveform and (×) the values recorded in our bioreactor system. The arrow indicates the shifted coronary artery blood pressure from the desired value.

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Fig. 9

Gene expression profiles for GSVs exposed to GSV, BrA, or LCA-like pulsatile media pressure and flow waveforms for 6 h. Data are reported as fold change relative to the GSV conditions. Error bars indicate±standard error of the mean. (*) and (**) denote differences between GSVs exposed to LCA or BrA conditions at p < 0.1 and p < 0.05, respectively.

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Fig. 10

Relative expression profiles of select vascular remodeling genes for the GSV after 6 h of in vitro culture at in situ pressure, flow and axial loading conditions represented as fold change from freshly harvested vessels. (*) and (**) denote significant differences between the measured gene after 6 h of culture and freshly harvested at p < 0.1 and p < 0.05, respectively.

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