Appraisal of state-of-the-art
Isolated heart perfusion according to Langendorff—Still viable in the new millennium

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Abstract

The isolated perfused mammalian heart preparation was established in 1897 by Oscar Langendorff. The method was developed on the basis of the isolated perfused frog heart established by Elias Cyon at the Carl Ludwig Institute of Physiology in Leipzig, Germany in 1866. Observations made using both methods at the end of the 19th and at the beginning of the 20th century led to important discoveries, forming the basis for our understanding of heart physiology. This included the role of temperature, oxygen and calcium ions for heart contractile function, the origin of cardiac electrical activity in the atrium, the negative chronotropic effect of vagus stimulation and the chemical transmission of impulses in the vagus nerve by acetylcholine. Langendorff himself demonstrated that the heart receives its nutrients and oxygen from blood via the coronary arteries and that cardiac mechanical function is reflected by changes in the coronary circulation.

The method underwent many modifications but its general principle remains the same today. Blood, or more commonly crystalloid perfusates, are delivered into the heart through a cannula inserted in the ascending aorta, either at constant pressure or constant flow. Retrograde flow in the aorta closes the leaflets of the aortic valve and as a consequence, the entire perfusate enters the coronary arteries via the ostia at the aortic root. After passing through the coronary circulation the perfusate drains into the right atrium via the coronary sinus.

The simplicity of the isolated mammalian heart preparation, the broad spectrum of measurements which can be done using this method, its high reproducibility and relatively low cost make it a very useful tool in modern cardiovascular and pharmacological research, in spite of a few shortcomings. In the last decade the method has brought many important advances in many areas including ischemia–reperfusion injury, cell-based therapy and donor heart preservation for transplant.

Introduction

Only few isolated organ experimental models have been used as extensively as the isolated heart preparation according to Langendorff. It was first applied by physiologists, biochemists and morphologists for the study of heart biology. It was further applied by pharmacologists to test the effect of different cardiovascular drugs on the coronary vasculature, muscle contraction and heart rate. Today a variety of cardiovascular researchers still use this vital technique in myriad ways to investigate the heart, from the study of the effect of a single gene alteration on heart physiology, to novel therapeutic means to protect the heart from ischemia and other insults.

Although there are some limitations of the method including the absence of normal humoral influences and neuronal regulation, as well as high coronary flow and oedema when using cell free perfusate, the impact of these shortcomings is limited by the very advantage that much important first hand information can be gained by virtue of the elegant simplicity of this technique. Moreover, methods of recording some biophysical and biochemical parameters have been improved over the decades and the number of parameters we can now obtain using this method, the accuracy of these measurements, their high reproducibility and the relatively low costs predominate over the shortcomings, making this method a very useful tool in modern cardiovascular and pharmacological research.

This review will first provide a brief historical background of the method as well as details regarding organ isolation, perfusion solutions, set-ups for different modalities of perfusion, methods of recording various cardiovascular parameters and finally illustrate the utility of the isolated heart preparation according to Langendorff with a few select examples of recent research.

Section snippets

The isolated perfused frog heart

The isolated perfused mammalian heart preparation was developed on the basis of the isolated perfused frog heart. This method, widely used in the last part of the 19th century, was a starting point for the study of heart physiology. The scientist who deserves credit for first devising the isolated perfused frog heart preparation is Elias Cyon who established the method at the Carl Ludwig Institute of Physiology in Leipzig, Germany in 1866. For excellent reviews and original references in

General principle

Isolated perfused heart preparations nowadays are largely based on adaptations of the method originally described by Langendorff (Langendorff, 1898). The Langendorff heart preparation involves the cannulation of the aorta. The heart, once removed from a terminally anaesthetized animal, is held with forceps by the opened aortic root and is slipped over a fixed cannula on the perfusion apparatus. This cannula is attached to the outflow of a reservoir containing an oxygenated perfusion solution

Anaesthesia

Surgery on the animal requires general anaesthesia to be performed prior to the excision of the heart. Anaesthesia can be induced either by inhalation of volatile agents such as halothane or injection with barbiturates (Doring and Dehnert, 1987, Sutherland and Hearse, 2000). Ether should be avoided due to its ability to stimulate the sympathetic nervous system as well as its flammability. None of these methods are ideal for rendering an animal unconscious. Although both methods have their

Measurements

The isolated perfused heart preparation is extremely valuable for assessing the direct effects of drugs on contractile function, heart rate, coronary vascular tone, cardiac metabolism and electrical activity of the heart. In addition it may be utilized for morphological studies—hearts can be processed for examination by various microscopical techniques following microbiopsies or whole heart preparation and sectioning at the completion of the perfusion protocol. By switching the perfusate to one

Limitations of isolated heart perfusion

Limitations of the method include those mentioned above: lack of the normal humoral background and neuronal regulation of the heart, high coronary flow, vulnerability of the isolated heart to injury during instrumentation, increased possibility of preconditioning during instrumentation of the excised heart, higher oxidative stress, and deterioration of contractile function. Another criticism of isolated perfused heart studies is that, in addition to inducing a very simplistic form of ischemia

Specific applications of the method

The original Langendorff preparation underwent many modifications which permitted studies under numerous pathological conditions. Below, some of the possible applications of the method are described.

A few selected examples to illustrate that the method is thriving in the new millennium

The isolated perfused heart technique has evolved for over 100 years. Hardly any other procedure has resulted in such a great contribution to cardiovascular physiology and pharmacology and is still actively being used today as a valuable tool in cardiovascular research.

Conclusions

The isolated perfused heart according to Langendorff has served as a robust model for many fundamental discoveries in cardiac physiology, pathology and pharmacology for more than 100 years. Only few experimental models have enjoyed such an undiminished popularity over such a long time. It is still one of the most popular experimental designs in cardiovascular research and cardiovascular pharmacology. The disadvantages the model may suffer are outweighed by its benefits and the technique

Acknowledgements

Studies described here from R. Schulz were supported by grants from the Canadian Institutes for Health Research, the Heart and Stroke Foundation of Alberta, NWT and Nunavut and the Alberta Heritage Foundation for Medical Research (AHFMR). R. Schulz is an AHFMR Scientist.

M. Skrzypiec-Spring wishes to thank the University of Alberta for visiting fellowship support, and the Medical University of Wroclaw and the Foundation of the Medical University of Wroclaw for travel support. The study sponsors

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