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Laser Diagnostics in Cardiosurgery

#1858


Research into the Creation of the Laser-induced Fluorescence Diagnostics of the Condition of Heart Tissues, Transplants and Allografts in Cardiosurgery

Tech Area / Field

  • PHY-OPL/Optics and Lasers/Physics
  • MED-DID/Diagnostics & Devices/Medicine

Status
8 Project completed

Registration date
23.04.2000

Completion date
26.12.2006

Senior Project Manager
Novozhilov V V

Leading Institute
Siberian Branch of RAS / Institute of Theoretical and Applied Mechanics (ITPMech), Russia, Novosibirsk reg., Novosibirsk

Supporting institutes

  • Research Institute of Blood Circulation Pathology, Russia, Novosibirsk reg., Novosibirsk\nState Research Center of Virology and Biotechnology VECTOR, Russia, Novosibirsk reg., Koltsovo

Collaborators

  • University of Michigan/ College of Engineering/Spase Physics Laboratory, USA, MI, Ann Arbor\nLawrence Livermore National Laboratory, USA, CA, Livermore

Project summary

The goal of the proposed Project is to study the reaction of biological structures to laser irradiation and to develop methods of integral evaluation of changes in the state and viability of biological tissues based on laser-induced fluorescence (LIF).

Evaluation of viability of donor tissues and organs used for transplantation is an important factor influencing the transplantation result. Initial unsatisfactory state of the donor heart tissues causes 15-20 % of intraoperative deaths. Thus, the problem of preliminary control of viability of tissues and organs supplied for operation, either being preserved or restored from conservation, and used for transplantation has become especially urgent. Unfortunately, no universal system for evaluation of the quality of transplants. That’s why development of new methods for appropriate evaluation of the state of donor tissues and organs is urgent, necessary and practically important.
Control of the state of a transplant in the postoperative period is of great importance. In spite of on-going studies for development of methods of noninvasive evaluation of the transplant rejection crises, which is especially important during the first year post operation, there is no alternative to endomyocardial biopsy. Performing endomyocardial biopsy itself can cause a number of complications associated with surgical manipulations. Taking into account the need for multiple operations of the type, it becomes obvious that the risk of complications developing in a patient becomes extremely high. The above data once again point to necessity, urgency and practical importance of search for new methods to evaluate the state of the transplanted (donor) organ and development of low-invasive methods of diagnostics of rejection crises post operation.
Many cardiac pathologies of different geneses are combined with mineralization (calcinosis) of tissues, especially in patients with valvular disease of the heart. Carrying out any surgical manipulations on the valvular system of a calcinosis-affected heart can be accompanied by fragmentation of tissues, which in its turn, is a prerequisite for a dangerous postoperative complication – material embolism. The process of mineralization of valves is a limiting factor for plastic cardiosurgery. It is the mineralization degree of valvular endocard and paravalvular structure that plays the decisive role in predicting the results of therapy and the outcome for a patient. That’s why quick and timely intraoperative diagnostics of fragments of calcinated tissues in the heart cavity can become a method for prophylaxis of this complication.
New appropriate and quick methods for diagnostics of the state of donor heart tissues are available. At the same time, development of fundamental research in the field of creating laser sources and studying their effect on biological samples opens new broad prospects for the use of lasers in biology and experimental and practical medicine. Laser technologies are already widely used in clinical practice. The recent experience of research work shows that lasers can used for therapy of many complicated diseases, including oncologic ones. At the same time, the general possibility of use of lasers for diagnostics of vascular sclerosis and atheromatosis and intestinal neoplasms has been reported for the first time. However, the method of laser diagnostics of the state of tissues and organs in practical medicine, including cardiosurgery, has not been developed yet. Based on the above considerations, it seems more promising to develop a method for quick and exact diagnostics of the state of donor hearts using technologies involving the use of laser irradiation.
At the same time, it seems promising to use not only tissues and organ cultures having a complicated morphology but also inpidual structural components in a cell culture in vitro in order to develop the reaction of the biological sample to laser irradiation and to create a data bank.
Favorable conditions are now available at IT&AM, SB of RAS; SRC VB “Vector” and RI BCP, RF MH for carrying out such studies. Radiation sources and experience of research on the use of laser-induced fluorescence in medical studies in cooperation with RI BCP, RF MH, are available at IT&AM. A base for carrying out works on creation, certification and cultivation of cells of different animal organs and tissues has been created at SRC VB “Vector”. The Center has a group of highly-qualified specialists and necessary equipment and premises.
Taking into all the above, it appears to be necessary and expedient to carry out studies on development of methods based on laser-induced fluorescence, diagnostics of viability state of donor heart tissues before operation (including the state of allografts and transplants) and that of low-invasive methods of evaluation of rejection crises post operation as well as intraoperative diagnostics of fragments of mineralized tissues in operations on valves.

Basic activities:

1. To investigate the effect of laser irradiation on cell cultures in vitro, animal and human tissues or organs ex vivo experiments.
2. To determine the parameters of laser irradiation which does not injure cells and tissues but allow to evaluate their state and to develop methods to control the state of live tissues.
3. To create a data bank on LIF spectra of cell cultures cultivated in vitro and being at different viability states and different cardiac tissues, both healthy and pathology-affected ones. To decode molecular mechanisms of photobiological reactions. To identify the most active enzymes and macromolecules of in the cell and to study the key stages of formation of LIF spectrum accompanied by identification of the most active components, radicals, molecular bonds, etc.
4. To carry out comparative studies on identification of the state of cells and tissues by LIF spectra and the data of traditional clinical and laboratory analysis. To study the mechanisms of functionally active light absorption. To study the mechanisms of the effect of ultraviolet irradiation on protein (cells) - their inactivation, i.e. loss of enzymatic, regulatory, hormonal, transporting and immunological activities. To determine the quantum outputs of photoinactivation of proteins. To investigate the energy migration in proteins. To determine contact rates of reactions of the photochemical, luminescent and thermodynamical way of deactivation of electron-excited state of molecules absorbing ultraviolet quantum of light.
5. To develop methods for:
· appropriate diagnostics of viability state of donor heart tissues before and post operation (including the state of allografts at different technological stages);
· intraoperative diagnostics of fragments of mineralized tissues at operations on valves;
· low-invasive evaluation of rejection crises after operation of heart transplantation.
6. To obtain safety for the method being used for diagnostics in clinical practice and to develop methodological recommendations on the method application.

To provide the proposed Project implementation it is planned :
· to develop methods to control the cells exposed to laser irradiation;
· to obtain data on the effects of different doses and conditions of laser irradiation on model cell cultures in vitro and animal and human tissues and organs in vivo;
· to identify conditions of laser exposure which provide appropriate evaluation of viability of cells in vitro and tissues and organs in vivo;
· to develop a method for diagnostics of viability of tissues and organs in vitro and vivo;
· to investigate the method’s safety in laboratory animals;
· to obtain comparative data on the method’s efficiency in patients as compared to traditional methods of control of tissues and organs.

The studies to be performed will allow to obtain for the first time data on spectra of tissues and organs, both normal and with affection of different nature, and to develop new methods for diagnostics of the state of donor issues and organs. Besides, the Project implementation will provide new fundamental knowledge of reaction of biological objects to laser irradiation. This knowledge is required not only for development of methods for diagnostics of the tissues viability and vital activities but, possibly, for development of new therapeutic methods as well. As a result of the Project implementation, the question will be answered about the general possibility of LIF being used for integral evaluation of the state of donor tissues before operation, development of low-invasive evaluation of crises of transplants rejection and intraoperative diagnostics of mineralized fragments of cardiac tissues.
Thus, urgency and scientific novelty of the Project consists in obtaining fundamental knowledge of reaction of live tissues and cells to laser irradiation. This knowledge will allow to work out approaches to the development of a technology for diagnostics of affected tissues of the heart and other organs, which will be new not only for Russia. The Project implementation will provide new possibilities for practical medicine in Russia and abroad and will open new promising trends of research work. Laser diagnostic methods for evaluation of viability of donor tissues and organs (allografts and donor hearts for transplantation) will be commercially valuable. These methods will serve as a basis for achieving better results of surgical treatment of patients with cardiac pathologies, which is of scientific and practical interest for medicine both in Russia and abroad.
Similar studies are underway in Lawrence Livermore National Laboratory (LLNL), USA, collaborator in these activities.
The Project activities will allow to use experience and knowledge of specialists, who have dealt with the problems of biological safety (SRC VB “Vector) and have been involved in research for development of rocket and laser weapons (IT&AM, SB of RAS), for developing a new technology which will be very important both for fundamental science and the needs of public health service. Besides, involvement of specialists of SRC VB “Vector” and IT&AM, SB of RAS, will help to solve the social problem of changing the character of the personnel’s activities, which meets ISTC goals and objectives.


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