Methods and means of optical non-invasive diagnostics of ... non-invasive... · Methods and means...

Methods and means of optical non-invasive

diagnostics of hemodynamics and metabolic activity of human tissues

Head of Biomedical Photonics Laboratory of University Clinic,

Orel State University named after I.S. Turgenev, Orel, Russia

Summer School on Optics & Photonics

June 1-3, 2017, Oulu, Finland

Dr Andrey Dunaev

http://www.bmecenter.ru/en

2 Summer School on Optics & Photonics


Orel State University named after I.S. Turgenev (founded in 1931, from 01.04.2016 is united with the

State University – Education-Science-Production Complex) is a unique and dynamically developing

education-science-production complex in the Central part of Russia (Central Black Earth Region) which

integrates qualitative education, perspective science and effective production. More 250 directions of

specialist training, more than 20 000 students.

Orel city is situated in the south of the central European

region of Russia, 382 km from Moscow. It was founded in

1566. The population of Orel is about 400 000.

The University began training of specialists in biomedical engineering in 1999, scientific-educational center

of “Biomedical Engineering” was founded in 2010.



Our Team: Biomedical Photonics Laboratory of University Clinic

The main topics of research:

• Technical and metrological support of devices for LDF;

• Methods and devices for diagnostics of the functional state of peripheral vessels (using

functional tests - occlusion, heating, cold pressor and other);

• Methodological and instrumentation provision of FS and DRS for biomedical applications

(bladder cancer, diabetes mellitus etc.);

• Non-invasive monitoring of drug delivery by fluorescence spectroscopy.

Research interests include the development of biomedical optical non-invasive methods and devices for

diagnostics, such as Laser Doppler Flowmetry (LDF), Tissue Reflectance Oximetry (TRO), Fluorescence

Spectroscopy (FS) and Diffuse Reflectance Spectroscopy (DRS).



Outline

1. Introduction. Diseases associated with the condition of hemodynamics

and metabolic activity of human tissues.

2. Application basics of optical non-invasive diagnostics (OND) for study of hemodynamics and

metabolic activity of human tissues.

3. Medical and technical features of OND devices:

a) Variability of registered signal in OND.

b) Substantiation of medical and technical requirements for OND devices.

c) Technical aspects and application possibilities of LDF for blood and lymph monitoring.

d) The study influence of local pressure on evaluation parameters of skin blood perfusion and

fluorescence.

e) The study of blood microcirculation parameters by combined use of LDF and video

capillaroscopy methods.

5

4. New application approaches of the OND for medical practice:

a) Combined use of LDF and skin thermometry for functional diagnostics in patients with

rheumatic diseases.

b) The study by DRS method of microcirculatory disorders and inflammatory activity in

patients with rheumatic diseases.

c) Complex analysis of metabolic and hemodynamic processes in patients with diabetes

mellitus.

5. Conclusions.

6. Acknowledgements.



6

• diabetes mellitus (~7% of the population*);

• Raynaud's syndrome (~3-5% of the population**);

• vibration disease (one of the common forms of professional

pathology – up to 25%);

• atherosclerosis;

• scleroderma.

Diabetes mellitus Scleroderma

Raynaud's syndrome Atherosclerosis

http://dfridapi.org/images/diabetic-foot.jpg

The prognosis of vascular complications and

monitoring of treatment effectiveness of diabetes is

one of the most significant healthcare problem.

* IDF Diabetes Atlas (7th edition);

** Alekperov RT. Raynaud‟s phenomenon in the rheumatologist‟s practice. Modern Rheumatology Journal. 2014;(2):48–57.

1. Diseases associated with the condition of hemodynamics

and metabolic activity of human tissues:







7

Average magnitude

Аctive and passive regulation processes

(frequency rhythms) in a microcirculation

system

Typical view of LDF-gram Laser Doppler Flowmetry (LDF)

2. Application basics of optical non-invasive diagnostics (OND) for

study of hemodynamics and metabolic activity of human tissues



Classification of the blood flow oscillations:

Currently, for diagnostic purposes, 5 rhythmic components (oscillations) were isolated from

LDF recordings with help of the wavelet analysis in accordance with the modern interpretation

of their sources (Stefanovska A., Bracic M. et al.,1999):

# Name of rhythm Frequency interval, Hz

1 Endothelial (E) 0.0095-0.02

2 Neurogenic (N) 0.02-0.06

3 Myogenic (M) 0.06-0.16

4 Breathing (B) 0.16-0.4

5 Cardiac (C) 0.4-1.6

Active and passive regulation processes

(frequency rhythms) in a microcirculation system



Fluorescence Spectroscopy (FS)

The method allows to determine the content

NADH, flavins (FAD), porphyrin in tissues;

evaluate the cell‟s metabolic activity, and others.

Laser

Tissue

Backscattering and fluorescent spectra

Tissue Reflectance Oximetry (TRO)

HbHbO

HbO

tCC

COS

2

2

2

otherblood

bloodb

CC

CV



Diffuse Reflectance Spectroscopy (DRS)

The method allows to

determine the content of

hemoglobin, oxygenation of

tissues, blood volume

fraction and others.

11

MLNDS “LAKK-M” (SPE “LAZMA” Ltd, Russia)

4 channels in one system:

• fluorescence spectroscopy (FS);

• absorption spectroscopy

(tissue reflectance oximetry – TRO);

• laser Doppler flowmetry (LDF);

• pulse oximetry.

“LAKK-M” system allows the following blood microcirculation

parameters to be obtained:

• index of blood microcirculation (Im),

• tissue oxygen saturation (StO2),

• relative blood volume (Vb),

• arterial blood saturation (SaO2),

• fluorescence spectra of tissue endogenous biomarkers.

This system is used for research and diagnostics

in various fields of biomedicine (cardiovascular

diseases, diabetes, cancer, cosmetic surgery,

etc.).

Principles of construction of multifunctional laser-based non-invasive

diagnostic systems (MLNDS)


June 1-3, 2017, Oulu, Finland 11

12

Variability of the blood flow oscillations:

2010 Makarov D.S., Rogatkin D.A.:

LDF and TRO measurements have high variability, at least in the range of ± 30% in terms of SD of the

average measured values for each parameter (Im, StO2, Vb).

Deviation of the oscillation amplitude (only for LDF) at a test area (forearm) for a homogeneous

group of subjects can be up to 15%.

2000 Krasnikov G. V., Matrusov S. G., Piskunova G. M., Sidorov V. V., Chemeris N. K.:

3. Medical and technical features of non-invasive optic diagnostic devices

a) Variability of registered signal in optical non-invasive diagnostics.

0.02 0.04 0.09 0.26 1.250.0

0.5

1.0

1.5

2.0

0.0

0.5

1.0

1.5

2.0

StO

2, V

b, %

PulseBreathingMyogenicNeurogenicEndothelial

I m

, P

U

Rhythm frequency (Hz)

Im

StO

2

Vb

Distribution histogram of the amplitudes of the oscillations (Im, StO2, Vb):

* Dunaev A.V. et al. (2013). Laser reflectance oximetry and Doppler flowmetry in assessment of complex physiological parameters of cutaneous blood

microcirculation. Proc. SPIE 8572, 857205.

Long-term individual variability of oscillation

amplitudes for tissue oxygen saturation amounted to

around 30-55%, almost as for the microvascular

blood flow – 25-45%.

13

Graphs of parameter changes Ibs() and Vb for

the two zones of the skin for one volunteer

* Dunaev A.V. et al. (2015). "Individual variability analysis of fluorescence parameters measured in skin with different levels of nutritive blood flow." Medical

Engineering and Physics 37(6): 574-583.

r1 = –0.67 for the finger;

r2 = –0.48 for the forearm.

The fluorescence spectra of skin with high levels of melanin, a – fingertip; b – forearm:

the UV (1), green (2) and red (3) light excitation wavelengths

Histogram plot of the distribution of average

intensities of fluorescence for the area with the AVA

Individual

variability

mostly up

to 30 %!

Variability of the fluorescence intensity:




b) Substantiation of medical and technical requirements for non-invasive optic

diagnostic devices.

One of the approaches for the substantiation of medical and technical requirements for optic non-

invasive diagnostic (OND) devices is based on study of the influence of the tissue blood volume on

the levels of registered signals as well as the sensitivity of such devices.

Dimensionless coding function B(λ):

2/120

2

2/320

220

20 )(exp

)(

1

2)( zr

zrzr

AzB d

d

where represents the effective path length of the light, A is the detector

area, r is the separation distance between the source-detector fibers,

, are the reduced scattering coefficient,

g = g(λ) is the anisotropy factor, s=s(λ) is the scattering coefficient, a=a(λ)

is the absorption coefficient.

'/10 sz

2/1)'(3 saad ss g )1('

DATA

d



The dependence of B (λ) on the level

of the tissue blood volume Vb

* Dunaev A.V. et al. (2013). "Substantiation of medical and technical requirements for non-invasive spectrophotomeric diagnostic devices." Journal of Biomedical

Optics 18(10): 107009.

The dependence of the SNR of the signal on

the level of the tissue blood volume

• Considering different levels of the tissue blood volume, the proposed approach allows the calculation of important

technical and metrological restrictions of the instruments, such as the sensitivity ranges and power related signal-

to-noise ratios for different spectral channels and biomedical parameters.

• It is clear that the nonlinearity of the measurements carried out with OND systems directly depends on the

characteristics of the examined object.

16


c) Technical aspects and application possibilities of LDF for blood and lymph

monitoring.

Perfusion (index of microcirculation):

K1 ,K2 - proportional coefficients; NRBC - number of red blood cells (erythrocytes); VRBC - velocity average of red blood cells (erythrocytes) in measured part of the tissue; ω – frequency of the Doppler shift; S(ω) – power spectrum of the photocurrent.

1 2

(

RBC RBC

dc

S dPerfusion K K N V

i



• Laser source: λ=1064 nm; • Frequency bands: - 60 to 400 Hz; - 400 to 800 Hz; - 800 to 1600 Hz; - 1600 to 3200 Hz; - 3200 to 6400 Hz.

Experimental LDF device:



* Dremin V.V., et al. (2017) Laser Doppler flowmetry in blood and lymph monitoring, technical aspects and analysis // Proc. SPIE 10063, 1006303.

Breath Holding Test #1

Example total LDF-gram of a breath test (a) and LDF-grams recorded using the

experimental LDF device on the 60-400 Hz and 400-800 Hz sub-band spectra (b)

18

Processing for Breath Holding Test #1

Distribution of index of microcirculation by frequency of Doppler shift for Breath Holding Test



3. Medical and technical features of non-invasive optic diagnostic devices:

d) The study influence of local pressure on evaluation parameters of skin blood

perfusion and fluorescence

* Zherebtsov E.A. et al. (2017) The influence of local pressure on evaluation parameters of skin blood perfusion and fluorescence // Proc. SPIE 10336, 2017,

1033608

* Zherebtsov E.A. et al. (2017) The influence of local pressure on evaluation parameters of skin blood perfusion and fluorescence // Proc. SPIE 10336, 2017,

1033608

Typical fluorescence spectra at pressures 0 kPa, 5 kPa and 40 kPa

for wavelengths 365 nm and 450 nm

Average peaks of fluorescence intensity for all volunteers for wavelengths 365 nm and 450 nm



21

* Volkov M.V., et al. (2017) Evaluation of blood microcirculation parameters by combined use of the laser Doppler flowmetry and the video capillaroscopy methods

// Proc. SPIE 10336, 1033607.

3. Medical and technical features of non-invasive optic diagnostic devices:

e) The study of blood microcirculation parameters by combined use of LDF and

video capillaroscopy methods

Laser Doppler flowmetry setup based on “LAKK-OP”

device and custom electronic board

Frames with

capillaries

Graphs with RBC velocities

into capillary and LDF-grams

4. New application approaches of the OND for medical practice: a) Possibilities of the combined methods to assess microcirculatory disorders and

inflammatory activity in patients with rheumatic diseases.

Phase duration, min 2 4 10 3 10 5-11

Environment Air Water

Temperature, °C 25 42 25 42

Occlusion test Before occlusion Occlusion Postocclusion

* Zherebtsov E.A., et al. (2017) Combined use of laser Doppler flowmetry and skin thermometry for functional diagnostics of intradermal finger vessels // J.

Biomed. Opt., 22(4):040502.



* Zherebtsov E.A., et al. (2017) Combined use of laser Doppler flowmetry and skin thermometry for functional diagnostics of intradermal finger vessels // J.

Biomed. Opt., 22(4):040502.

Boxplot representing linear discriminant

function scores for the groups of healthy

volunteers and rheumatological patients

Experimental ROC-curve for the

proposed diagnostic approach

• The „blood flow reserve‟ and „index of the temperature response‟ were measured and used as the primarily

parameters of the functional diagnostics of the peripheral vessels of skin.

• Utilizing these parameters, a simple phenomenological model has been suggested to identify patients with the

angiospastic violations in vascular system, and can be used concurrently to the existing diagnostic methods.



In this research 36 patients with rheumatic diseases and 31 conditionally healthy volunteers were

examined in rheumatologic department of Orel Regional Clinical Hospital.

4. New application approaches of the OND for medical practice: b) The study by DRS method of microcirculatory disorders and inflammatory activity in

patients with rheumatic diseases.

* Potapova E.V., et al. (2017) Evaluation of Microcirculatory Disturbances in Patients with Rheumatic Diseases by the Method of Diffuse Reflectance

Spectroscopy // Human Physiology, 43(2):222-228.

25

Averaged spectra of the diffuse reflection and curves of the optical density of the skin for

rheumatologic patients and a control group of conditionally healthy volunteers

The calculated index of erythema and tissue saturation for rheumatologic patients and a control group

of conditionally healthy volunteers



26

With the combined use of LDF, FS methods and heating test

* Dremin V.V. et al. (2016) “The blood perfusion and NADH/FAD content combined analysis in patients with diabetes foot.” Proc. SPIE 9698, Advanced

Biomedical and Clinical Diagnostic and Surgical Guidance Systems XIV, 969810.

4. New application approaches of the OND for medical practice: d) Complex analysis of metabolic and hemodynamic processes in patients with diabetes

mellitus.

28

* Under reviewing

Comparison of parameters between control, diabetic and diabetic with ulcers groups: the normalised

fluorescence amplitude (A) and the average perfusion in the stages of heating to 35 and 42 oC (B)

29

* Under reviewing

Comparison of parameters between control and diabetic groups:

myogenic rhythms (A) and the nutritive blood flow (B) in the stages of heating to 35 and 42 oC

The scatter plot with applied discriminant lines, obtained by linear discriminant analysis method (A) and

ROC-curves for assessing the effectiveness of the classifiers (B)



5. Conclusions

• The presented results demonstrate the relevance of instrumentation, methodological and metrological

provision for these technologies in general and especially for the medical application of the LDF-,

TRO-, FS-, DRS-devices and combined systems.

• Further problem solving of the outlined issues will bring OND closer to standardized diagnostic

technologies and to wider application in real medical practice.

• The obtained results demonstrate the ability of the combined OND techniques to provide useful

information for diseases classification (for functional diagnostics, rheumatologic diseases and

diabetes) in clinical practice, but the actual remains the search clearly expressed of new diagnostic

criteria.

• The combined using of OND technologies with functional (provocative) tests allows increasing the

repeatability of results and accuracy of diagnostics (for example, LDF and FS methods allows to predict

the development of trophic disorders and the diabetic foot syndrome on the more early stages).



6. Acknowledgements

www.medilase.eu

I would like to thank all of our volunteers and patients for their contribution to these research projects.

Part of presented results was obtained with the support by:

• the European Community‟s Seventh Framework Program (FP7-People-2011-IAPP) under Grant

Agreement no. 251531 “MEDILASE”;

• the European Community‟s Seventh Framework Program (FP7-People-2013-IAPP) under Grant

Agreement no. 324370 “ABLADE”;

• the state task of the Ministry of Education and Science of the Russian Federation for the State

University – Education-Science-Production Complex (basic part, №310);

• Foundation for Assistance to Small Innovative Enterprises in the framework of the «U.M.N.I.K.»

(7 projects, 2011 - present time);

• grant for startup support program еt the early stages of innovative activity "START“.

www.ablade.eu



Teams / Collaborators

The research were performed in collaboration with: Aston University (Birmingham, UK), University of

Oulu (Oulu, Finland), Orel Regional Clinical Hospital (Orel, Russia), ITMO University (Saint-Petersburg,

Russia), SPE “LAZMA” Ltd (Moscow, Russia) and Priorov Central Research Institute of Traumatology

and Orthopaedics (Moscow, Russia).

Special thanks to:

Dr V.V. Sidorov

Prof A.I. Krupatkin

Prof E.U. Rafailov,

Dr S.G. Sokolovski,

Dr K.S. Litvinova,

Mr I.E. Rafailov

A huge thanks to my team: Dr Evgeny Zherebtsov, Dr Angelina Zherebtsova, Dr Elena Potapova, Mr Victor Dremin, Mrs Irina Makovik,

Mrs Olga Stelmashchuk and all of our students – Igor Kozlov, Elena Zharkikh, Maria Filina, Ksenia

Kandurova, Evgeniya Seryogina and others.

Prof I. Meglinski,

Dr A. Popov,

Dr A. Bykov

Prof I.P. Gurov,

Dr N.B. Margaryants,

Dr M.V. Volkov

E.A. Alimicheva, MD

G.I. Masalygina, MD

L.S. Khakhicheva, MD

V.F. Muradyan, MD

Thank you for your attention!

Contact information:

Dr Andrey V. Dunaev

Biomedical Photonics Laboratory of University Clinic,

Scientific-Educational Center of “Biomedical Engineering”,

Orel State University named after I.S. Turgenev

29 Naugorskoe Shosse, Orel, Russia, 302020

E-mail: [email protected], [email protected]

Mob.phone: +79192619906

URL: www.bmecenter.ru/en



mailto:[email protected]



http://www.bmecenter.ru/en

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