High resolution contrast ultrasound and NADH fluorescence imaging of myocardial perfusion in excised rat hearts

Kay, Matthew W.; Swift, Luther; Sangave, Amit; Zderic, Vesna

doi:10.1109/iembs.2008.4649316

2008 30th Annual International Conference of the IEEE Engineering in Medicine and Biology Society 2008

DOI: 10.1109/iembs.2008.4649316

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High resolution contrast ultrasound and NADH fluorescence imaging of myocardial perfusion in excised rat hearts

Matthew W. Kay

Luther Swift

Amit Sangave

et al.

Abstract: Simultaneous imaging of myocardial flow and hypoxia could be vital for identifying acute ischemic mechanisms that may trigger an arrhythmia. We have studied the distribution of flow and hypoxia in excised locally ischemic rat hearts using simultaneous contrast ultrasound imaging and beta-nicotinamide adenine dinucleotide (NADH) fluorescence imaging. Local ischemia was induced by controlling flow within a major coronary artery. Intra-myocardial flow was imaged using contrast high-resolution ultrasound (linear p… Show more

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Cited by 3 publications

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“…Oxygen can be a limiting factor in heart tissue that becomes ischemic and/or hypoxic, and insufficient oxygen availability can lead to an accumulation of NADH. Demand-induced ischemia may also occur during an episode of tacchycardia, when a fast heart rate is accompanied by a drop in aortic pressure that is inadequate for full coronary perfusion (23,30).Fluorescent imaging of epicardial NADH (fNADH) has been demonstrated as a useful tool to gain insight into the redox state of cardiac tissue (1,2,14,15,17,18,20). However, the effect of heart preparation on fNADH within the context of myocardial work load and oxygen supply is not well understood.…”

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NADH changes during hypoxia, ischemia, and increased work differ between isolated heart preparations

Wengrowski

Kuzmiak-Glancy

Jaimes

et al. 2014

American Journal of Physiology-Heart and Circulatory Physiology

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Wengrowski AM, Kuzmiak-Glancy S, Jaimes R 3rd, Kay MW. NADH changes during hypoxia, ischemia, and increased work differ between isolated heart preparations. Am J Physiol Heart Circ Physiol 306: H529 -H537, 2014. First published December 13, 2013; doi:10.1152/ajpheart.00696.2013.-Langendorff-perfused hearts and working hearts are established isolated heart preparation techniques that are advantageous for studying cardiac physiology and function, especially when fluorescence imaging is a key component. However, oxygen and energy requirements vary widely between isolated heart preparations. When energy supply and demand are not in harmony, such as when oxygen is not adequately available, the imbalance is reflected in NADH fluctuations. As such, NADH imaging can provide insight into the metabolic state of tissue. Hearts from New Zealand white rabbits were prepared as mechanically silenced Langendorffperfused hearts, Langendorff-perfused hearts, or biventricular working hearts and subjected to sudden changes in workload, instantaneous global ischemia, and gradual hypoxia while heart rate, aortic pressure, and epicardial NADH fluorescence were monitored. Fast pacing resulted in a dip in NADH upon initiation and a spike in NADH when pacing was terminated in biventricular working hearts only, with the magnitude of the changes greatest at the fastest pacing rate. Working hearts were also most susceptible to changes in oxygen supply; NADH was at half-maximum value when perfusate oxygen was at 67.8 Ϯ 13.7%. Langendorff-perfused and mechanically arrested hearts were the least affected by low oxygen supply, with halfmaximum NADH occurring at 42.5 Ϯ 5.0% and 23.7 Ϯ 4.6% perfusate oxygen, respectively. Although the biventricular working heart preparation can provide a useful representation of mechanical in vivo heart function, it is not without limitations. Understanding the limitations of isolated heart preparations is crucial when studying cardiac function in the context of energy supply and demand.Langendorff; biventricular working; blebbistatin; nadh imaging; workload THE HEART IS RESPONSIBLE FOR pumping blood with vital supplies to the entire body and therefore requires a constant high rate of energy supply. Additionally, the heart has a large dynamic range with the capacity to increase its workload 10-fold and maintain this increase for extended periods of time. This metabolic requirement necessitates acute regulation of energy supply to match demand, as well as a constant adequate oxygen supply. Any limitation in energy supply is rapidly detrimental to cardiac function.The demand for ATP and oxygen varies, depending on the amount of work being performed (12a, 17). The amount of available ATP to the heart at any given time is enough for only a few beats and must constantly be resynthesized to keep up with utilization (16). In the heart, 80 -90% of energy production occurs via oxidative phosphorylation (25). Fats and carbohydrates are broken down and electrons are harvested, reducing NAD ϩ to NADH, (and, to a lesser exte...

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mentioning

confidence: 99%

“…Fluorescent imaging of epicardial NADH (fNADH) has been demonstrated as a useful tool to gain insight into the redox state of cardiac tissue (1,2,14,15,17,18,20). However, the effect of heart preparation on fNADH within the context of myocardial work load and oxygen supply is not well understood.…”

mentioning

confidence: 99%

NADH changes during hypoxia, ischemia, and increased work differ between isolated heart preparations

Wengrowski

Kuzmiak-Glancy

Jaimes

et al. 2014

American Journal of Physiology-Heart and Circulatory Physiology

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show abstract

“…This can be done by imaging the fluorescence of NADH (the reduced form of nicotinamide adenine dinucleotide) [9][10][11] , a coenzyme found in large quantities in the mitochondria. NADH fluorescence (fNADH) displays a near linearly inverse relationship with local oxygen concentration 12 and provides a measure of mitochondrial redox state 13 .…”

mentioning

confidence: 99%

NADH Fluorescence Imaging of Isolated Biventricular Working Rabbit Hearts

Asfour

Wengrowski

Jaimes

et al. 2012

JoVE

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Since its inception by Langendorff 1 , the isolated perfused heart remains a prominent tool for studying cardiac physiology 2 . However, it is not well-suited for studies of cardiac metabolism, which require the heart to perform work within the context of physiologic preload and afterload pressures. Neely introduced modifications to the Langendorff technique to establish appropriate left ventricular (LV) preload and afterload pressures 3 . The model is known as the isolated LV working heart model and has been used extensively to study LV performance and metabolism [4][5][6] . This model, however, does not provide a properly loaded right ventricle (RV). Demmy et al. first reported a biventricular model as a modification of the LV working heart model 7,8 . They found that stroke volume, cardiac output, and pressure development improved in hearts converted from working LV mode to biventricular working mode 8 . A properly loaded RV also diminishes abnormal pressure gradients across the septum to improve septal function. Biventricular working hearts have been shown to maintain aortic output, pulmonary flow, mean aortic pressure, heart rate, and myocardial ATP levels for up to 3 hours 8 .When studying the metabolic effects of myocardial injury, such as ischemia, it is often necessary to identify the location of the affected tissue. This can be done by imaging the fluorescence of NADH (the reduced form of nicotinamide adenine dinucleotide) [9][10][11] , a coenzyme found in large quantities in the mitochondria. NADH fluorescence (fNADH) displays a near linearly inverse relationship with local oxygen concentration 12 and provides a measure of mitochondrial redox state 13. fNADH imaging during hypoxic and ischemic conditions has been used as a dye-free method to identify hypoxic regions 14,15 and to monitor the progression of hypoxic conditions over time 10.The objective of the method is to monitor the mitochondrial redox state of biventricular working hearts during protocols that alter the rate of myocyte metabolism or induce hypoxia or create a combination of the two. Hearts from New Zealand white rabbits were connected to a biventricular working heart system (Hugo Sachs Elektronik) and perfused with modified Krebs-Henseleit solution 16 at 37 °C. Aortic, LV, pulmonary artery, and left & right atrial pressures were recorded. Electrical activity was measured using a monophasic action potential electrode. To image fNADH, light from a mercury lamp was filtered (350±25 nm) and used to illuminate the epicardium. Emitted light was filtered (460±20 nm) and imaged using a CCD camera. Changes in the epicardial fNADH of biventricular working hearts during different pacing rates are presented. The combination of the heart model and fNADH imaging provides a new and valuable experimental tool for studying acute cardiac pathologies within the context of realistic physiological conditions. Video LinkThe video component of this article can be found at https://www. 10.0 glucose, 2.00 NaPyruvate, and 20.0 mg/L albumin). The soluti...

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Signal Decomposition of Transmembrane Voltage-Sensitive Dye Fluorescence Using a Multiresolution Wavelet Analysis

Asfour

Swift

Sarvazyan

et al. 2011

IEEE Trans. Biomed. Eng.

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Fluorescence imaging of transmembrane voltage-sensitive dyes is used to study electrical activation in cardiac tissue. However, the fluorescence signals, typically, have low SNRs and may be contaminated with motion artifact. In this report, we introduce a new processing approach for fluoresced transmembrane potentials (fTmps) that is based upon a discrete wavelet transform. We show how fTmp signals can be decomposed and reconstructed to form three subsignals that contain signal noise (noise signal), the early depolarization phase of the action potential (rTmp signal), and motion artifact (rMA signal). A coiflet4 wavelet is used for fTmp decomposition and reconstruction of these subsignals. Results using fTmp signals that are contaminated with motion artifact indicate that the approach is a useful processing step to remove baseline drift, reduce noise, and reveal wavefronts. It streamlines the preprocessing of fTmps for the subsequent measurement of activation times and conduction velocities. It is a promising approach for studying wavefronts without aggressive mechanical tissue constraint or electromechanical uncoupling agents and is, useful for single-camera systems that do not provide for ratiometric imaging.

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High resolution contrast ultrasound and NADH fluorescence imaging of myocardial perfusion in excised rat hearts

Cited by 3 publications

References 15 publications

NADH changes during hypoxia, ischemia, and increased work differ between isolated heart preparations

NADH changes during hypoxia, ischemia, and increased work differ between isolated heart preparations

NADH Fluorescence Imaging of Isolated Biventricular Working Rabbit Hearts

Signal Decomposition of Transmembrane Voltage-Sensitive Dye Fluorescence Using a Multiresolution Wavelet Analysis

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