Projects

KP06-N28/11/14.12.18 „Novel bio-optical methods for determination of chemical and conformational reorganization of collagen-based tissue structures in vitro and in vivo“

Funded by NSF-MES-Bulgaria

Partners:
Institute of Electronics, Bulgarian Academy of Sciences

Institute of Organic Chemistry with Centre for Phytochemistry, Bulgarian Academy of Sciences

Scientific team:
Assoc. Prof. Ekaterina Georgieva Borisova, PhD-Coordinator of the project, Institute of Electronics, BAS
Latchezar Avramov, Prof., D.Sci., PhD, IE-BAS
Petranka Troyanova, Prof. PhD, MD – Dermatology, University Hospital “Tsaritsa Yoanna-ISUL”
Alexander Gisbrecht, Assoc. Prof., PhD, IE-BAS
Petya Pavlova, Prof., PhD, TU-Sofia, branch Plovdiv 
Ivan Angelov, Assoc. Prof., PhD, IOChCP-BAS
Vanya Mantareva, Assoc. Prof., PhD, IOChCP-BAS
Meliha Aliosman, assistant, IOChCP-BAS
Plamen Hristov, biologist, IOChCP-BAS
Tsanislava Genova, PhD, IE-BAS 
Deyan Ivanov, PhD student, IE-BAS
Viktoria Mircheva, BS student in Medical Physics
Stoyan Ilyov, BS student in Medical Physics
Ivan Bratchenko, Assoc. Prof., PhD, Samara State University, Russian Federation, external expert
Evgeny Shirshin, Assoc. Prof. PhD, Moscow State Unviersity, Russian Federation, external expert

PROJECT OBJECTIVES

The main aim of the project is to develop novel bio-optical methods for determination of the chemical and conformational reorganization of collagen-based tissue structures in vitro and in vivo using the methods of fluorescent spectroscopy with frequency and time resolution. Also planned is the increase in the possibilities of the existing experimental spectroscopic practices for fluorescence spectroscopy of bio-molecules and model systems in a frequency and time-resolved modes of detection, as well as the enhancement of the collected databases for the fluorescence properties of the different types of collagen and collagen-based tissue structures. Investigation and characterization of the basic fluorescent characteristics of different types of collagen and modified (glycated) collagen – spectra of excitation and emission, fluorescence lifetime (time decay signal), which will allow creating of a generalized spectral database for the possible forms of collagen in the human body for normal and for abnormal states during the development of pathology, to be used as indicators of the tissue condition.
Development of collagen-based optical phantoms, mimicking the spectral characteristics of the biological tissues for the purposes of fluorescence diagnosis, as well as obtaining new knowledge and optimization of diagnostic algorithms with applicability in clinical practice. We envisage training of young researchers and exchange of good practices for the development and application of spectroscopic techniques for analysis of skin pathologies in vivo for the needs of dermatology and oncology.

METHODS APPLIED

The specific fluorescent characteristics of the different types of collagen typical for the human tissues will be evaluated, using the methods of fluorescent analysis in a frequency- and time-resolved regime of detection.
The specific spectral features will be determined both for the pure forms of the different types of collagen, and for the mixtures thereof, in proportions typical for the biological tissues, in order to mimic in a best way the quantitative ratios observed under the fluorescent measurements in vivo. Collagen-based structures with conformational and chemically modified collagen will be developed and tested for the purpose of tissue optical biopsy using fluorescence diagnostics in order to model the regenerative (wound healing, therapeutic changes) and degenerative (collagenosis, sclerodermia, diabetes, pre-cancer, etc.) conditions in the tissues in vivo, as well as to develop and create optical phantoms for fluorescence diagnosis of tissues.
The characteristic changes in the optical spectral properties of different types of collagen will be investigated, related to the modification of their biochemical composition and changes in the conformation of collagen molecules and collagen matrices (supramolecular structures, such as collagen x-links). The methods of spectral analysis will include fluorescence spectroscopy in steady-state regime with frequency resolution, namely obtaining the spectra of excitation and emission, the development of the excitation-emission matrices (EEM), synchronous fluorescence spectroscopy (SFS), and with a temporal resolution – TCSPC (time-correlated single photon counting) – fluorescence spectroscopy in a mode of counting the first photon detected from sample emission, and FLIM (fluorescence lifetime imaging) – amplitude-modulated fluorescence decay time technique, where the amplitude demodulation and phase shift of the scanning optical signal is detected and evaluated. The FLIM technique allows to obtain two-dimensional images, which will be used in the analysis of the collagen-based structures, gels, optical phantoms and etc.

графика

Excitation–emission spectral data of normal (a) and cancerous (b) GIT mucosa for excitation range from 280 to 440 nm and synchronous fluorescence spectral data of normal (c) and cancerous (d) GIT mucosa for spectral offset varying from 10 to 200 nm between excitation and emission spectra respectively.

EXPECTED RESULTS

An upgrade of the technical and analytic skills of the project teams members will be achieved in the field of fluorescent analysis in steady-state and time-resolved regime of detection. New knowledge for the spectral peculiarities of the different forms of degenerative and regenerative cutaneous alterations will be obtained, related to the re-structural changes of collagen extracellular matrix in the frequency and time-domain.
New and original experimental results for the spectral properties of different types of collagen and chemically and conformationally modified forms of collagen will be obtained, for the needs of their diagnostic and prognostic applications in clinical dermatology for regenerative and degenerative skin alterations. The expected results are related to enhancement of the diagnostic accuracy of a noninvasive and real time determination of skin changes.
The research results will be published in specialized international journals and conference proceedings, necessary for their promotion and will contribute to the overall research growth of the participants, members of the project team. The developed methodologies and algorithms will be used in the dissertation thesis of one PhD student in the field of Physics, as well in the Master degree theses of two students from IE-BAS, specialty Medical Physics. The research carried out will be a prerequisite for the start of other PhD studies of these young scientists, which will allow achieving a sustainable development of the IE- BAS human resources in the field of biophotonics and its applications.

  1. Borisova E., Troyanova P.., Optical Spectroscopy for Skin Cancer Detection , 4th International Mediterranean Science and Engineering Congress , 25.04.2019 – 27.04.2019 , Alanya, Turkey (Oral presentation);
  2. Borisova E., Genova Ts., Pavlova E., Terziev I., Pavlova P., Troyanova P.., Autofluorescence and diffuse-reflectance spectroscopy for skin cancer diagnosis 4th International Mediterranean Science and Engineering Congress , 25.04.2019 – 27.04.2019 , Alanya, Turkey (Oral presentations);
  3. Deyan Ivanov, Razvigor Ossikovski, Tatyana Novikova, Pengcheng Li, Ekaterina Borisova, Tsanislava Genova, Lian Nedelchev, Dimana Nazarova, Tissue polarimetric study I: In search of reference parameters and depolarizing Mueller matrix model of ex vivo colon samples, European Conference on Biomedical Optics, 23.06.2019 – 27.06.2019, Munich, Germany (Poster Presentation);
  4. Borisova E., Genova Ts., Khorovodov A., Agranovich I., Angelov I., Mantareva V., Kanevsky M., Navolokin N., Semyachkina-Glushkovskaya O., Avramov L.., Photodetection of Stress-induced Gastrointestinal Neoplasia in Laboratorial Animals, European Conference on Biomedical Optics, 23.06.2019 – 27.06.2019, Мюнхен, Германия (Oral presentation);
  5. Borisova E., Khorovodov A., Agranovich I., Kanevskiy M., Konnova S., Genova Ts., Angelov I., Mantareva V., Navolokin N., Semyachkina-Glushkovskaya O., Exogenous fluorescence diagnostics of stress-induced gastric cancer – current state and future perspectives, Topical Problems of Biophotonics , 27.07.2019 – 31.07.2019 , Nijni Novgorod, Russian Federation (Oral presentation);
  6. Deyan Ivanov, Ekaterina Borisova, Razvigor Ossikovski, Alexander Bykov, Victor Dremin, A full Mueller matrix measurement of ex vivo colon samples with Stokes polarimeter, V Summer School “Photonics meet Biology”16.09.2019 – 20.09.2019, Heraklion, Greece (Poster presentation);
  7. Borisova E., Genova Ts., Bratashov D., Lomova M., Semyachkina-Glushkovskaya O., Vladimirov B., Valkov Ch. Fluorescence spectroscopy and microscopy of colon benign and malignant lesions – comparative study Saratov Fall Meeting 2019 , 23.09.2019 – 27.09.2019 , Saratov, Russian Federation (Oral presentation);
  8. Borisova E., Ivanov D., Kolev B., Genova Ts., Mircheva V., Ilyov S., Zaharieva L., Lihachova I., Lihachovs A., Spigulis J., Troyanova P., Autofluorescence spectroscopy of cutaneous neoplasia under ultraviolet, visible and near infrared excitation, Photonics Europe 2020 – Conference “Tissue Optics and Photonics”, 29.03.2020 – 03.04.2020, Strassbourg, France (Oral presentation).
  9. Borisova E., Genova Ts., Troyanova P., Bratchenko I., Bratchenko L., Zakharov V., Lihacova I., Lihacovs A., Spigulis J.”Multispectral fluorescence detection and imaging of skin tumours”, 3rd International Confenrece “Biophotonics-Riga”, 24.08.2020 – 26.08.2020, Riga, Latvia (Oral presentations);
  10. Mircheva V., Zaharieva L., Ilyov S., Troyanova P., Lihacova I., Lihacovs A., Bratchenko I., Bratchenko L., Khristoforova Yu., Zakharov V., Avramov L., Borisova E.. Near Infrared Autofluorescence Spectroscopy and Photobleaching Detection of Melanin-Pigmented Cutaneous Neoplasia”, 21st International Conference and School on Quantum Electronics “Laser Physics and Applications”, 21.09.2020 – 24.09.2020, Sofia, Bulgaria (virtual forum) (Poster presentations);
  11. Ivanov D., Borisova E., Novikova T., Ossikovski R.. Experimental validation of depolarizing Mueller matrix model via ex vivo colon samples. 21st International Conference and School on Quantum Electronics “Laser Physics and Applications”, 21.09.2020 – 24.09.2020, Sofia, Bulgaria (virtual forum) (Poster presentation);
  12. Genova, Ts., Valkov, H., Vladimirov, B., Kolev, B., Angelov, I., Avramov, L., Borisova, E.. Endogenous and Exogenous Fluorescence Diagnosis of Lower Part Gastrointestinal Tract Tumours. 21st International Conference and School on Quantum Electronics “Laser Physics and Applications”, 21.09.2020 – 24.09.2020, Sofia, Bulgaria (virtual forum) (Poster presentation);
  13. Borisova E., Genova Ts., Troyanova P., Pavlova E., Vladimirov B.., “Fluorescence diagnostics of soft tissues neoplasia”, Saratov Fall Meeting – SFM’2020, 28.09.2020 – 02.10.2020, Saratov, Russian Federation (Oral presentation);
  14. Borisova E., Genova Ts., Mircheva V., Ilyov S., Zaharieva L., Ivanov D., Gisbrecht A., Avramov L., Lihachova I., Lihachovs A., Spigulis J., Bratchenko I., Myakinin O., Zakharov V., Terziev I., Troyanova P., “Skin optical spectroscopy – diagnostics of cancerous and degenerative diseases”, 19th International Conference Laser Optics, 02.11.2020 – 06.11.2020, Sankt Petersburg, Russian Federation (virtual forum) (Poster presentation);
    1. Borisova E., Genova T., Bratashov D., Lomova M., Terziev I., Vladimirov B., Avramov L., Semyachkina-Glushkovskaya O.. Macroscopic and microscopic fluorescence spectroscopy of colorectal benign and malignant lesions – diagnostically important features. Biomedical Optics Express, 10, 6, Optical Society of America, 2019, DOI:https://doi.org/10.1364/BOE.10.003009, 3009-3017. JCR-IF (Web of Science) :3.91, Q1 link;
    2. Agranovich I., Borisova E., Navolokin N., Bucharskaya A., Maslyakova G., Shirokov A., Abdurashidov A., Angelov I., Khorovodov A., Terskov A., Mamedova A., Klimova M., Semyachkina-Glushkovskaya O.. Phenomenon of atypical vascular effects of epinephrine and an increase of photodynamic response by nitroglycerin in rats with colon adenocarcinoma: adrenergic and nitrergic mechanisms and novel applied aspects. Biomedical Optics Express, 10, 8, OSA, 2019, ISSN:2156-7085, DOI:10.1364/BOE.10.004115, 4115-4125. JCR-IF (Web of Science) :3.91, Q1 link;
    3. Borisova E., Genova Ts., Bratashov D., Lomova M., Terziev I., Vladimirov B., Valkov H., Semyachkina-Glushkovskaya O.. Fluorescence spectroscopy and confocal fluorescence microscopy of colon benign and malignant lesions – comparative study. Proceedings SPIE, 11457, SPIE, 2020, DOI:https://doi.org/10.1117/12.2559559, 1145702. SJR (Scopus):0.24 link;
    4. Borisova E., Ivanov D., Kolev B., Genova Ts., Mircheva V., Ilyov S., Zaharieva L., Lihachova I., Lihachovs A., Spigulis J., Troyanova P.. Autofluorescence spectroscopy of cutaneous neoplasia under ultraviolet, visible and near infrared excitation. Proceedings SPIE, 11363, SPIE, 2020, DOI:https://doi.org/10.1117/12.2555982, 113630Z-1-113630Z-5. SJR (Scopus):0.24 link;
    5. Borisova E., Kanevskiy M., Konnova S., Khorovodov A., Agranovich I., Genova Ts., Avramov L., Navolokin N., Smyachkina-Glushkovskaya O.. 5-ALA/PpIX photodiagnosis of stress-induced gastrointestinal metastatic tumours in laboratorial animals. Proceedings SPIE, 11363, SPIE, 2020, ISSN:0277-786X, DOI:https://doi.org/10.1117/12.2555992, 113630U-1-113630U-7. SJR (Scopus):0.24 link;
    6. Zaharieva L., Mircheva V., Uzunov Ts., Borisova E., Autofluorescent Characteristics of Different Stages of Dental Carious Lesions. Journal of Physics and Technology, 3, 2, Plovdiv University Press “Paisii Hilendarski”, 2019, ISSN:2535-0536, 20-25 link;
    7. Ivanov D., Dremin V., Bykov A., Borisova E., Genova Ts., Popov A., Ossikovski R., Novikova T., Meglinski I.. Colon cancer detection by using Poincar? sphere and 2D polarimetric mapping of ex vivo colon samples. Journal of Biophotonics, 13, WILEY?VCH Verlag GmbH, 2020, ISSN:1864-0648, DOI:https://doi.org/10.1002/jbio.202000082, e202000082. JCR-IF (Web of Science):3.763, Q1 link;
    8. Kanevskiy M., Borisova E., Mironova I., Konnova S., Galitskaya A., Khorovodov A., Agranovich I., Pavlova P., Avramov L., Semyachkina-Glushkovskaya O.. Stress-induced gastric adenocarcinoma: fluorescent and electrical measurements. JOAM, 22, 5-6, INOE Publishing House, 2020, 316-322. JCR-IF (Web of Science):0.588, Q4 link;
  1. Borisova E., Troyanova P.. Chapter 7: Diagnostics of Pigmented Skin Tumors Based on Light-Induced Autofluorescence and Diffuse Reflectance Spectroscopy. Multimodal Optical Diagnostics of Cancer, Springer Nature Switzerland AG, 2020, ISBN:978-3-030-44594-2, DOI:10.1007/978-3-030-44594-2_7, 25, 245-270 link
  2. Genova, Ts., Valkov, H., Vladimirov, B., Kolev, B., Angelov, I., Avramov, L., Borisova, E.. Endogenous and Exogenous Fluorescence Diagnosis of Lower Part Gastrointestinal Tract Tumours. Journal of Physics: Conference Series, IOP Publishing Ltd., приета за печат: 2020, SJR (Scopus):0.23 SJR, непопадащ в Q категория (Web of Science)link
  3. Ivanov D., Borisova E., Novikova T., Ossikovski R.. Experimental validation of depolarizing Mueller matrix model via ex vivo colon samples. Journal of Physics: Conference Series, IOP Publishing Ltd., приета за печат: 2020, SJR (Scopus):0.23 SJR, непопадащ в Q категория (Web of Science)link
  4. Mircheva V., Zaharieva L., Ilyov S., Troyanova P., Lihacova I., Lihacovs A., Bratchenko I., Bratchenko L., Khristoforova Yu., Zakharov V., Avramov L., Borisova E.. Near Infrared Autofluorescence Spectroscopy and Photobleaching Detection of Melanin-Pigmented Cutaneous Neoplasia. Journal of Physics: Conference Series, IOP Publishing Ltd., приета за печат: 2020, SJR (Scopus):0.23 SJR, непопадащ в Q категория (Web of Science) link