Transcript Presentation

Detailed spectral analysis
of decellularized skin
implants
E.V.Timchenko
Coauthors of work: P.E.Timchenko, L.T.Volova1,
D.A.Dolgushkin, P.Y. Shalkovsky, S.V. Pershutkina.
1
Samara State Medical University
supported by the Ministry of Education and Science of
the Russian Federation
Samara 2016
THE PROBLEM RESEARCHES
BREACH OF THE
STRUCTURE
OF THE SKIN
BURN SKIN
LONG
REHABILITATION
DISABILITY
MORTALITY
Bone Grafts BURN THIS
DECISION
PROBLEMS
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THE RESEARCH METHOD
OPTICAL METHODS OF CONTROL OF
HEART VALVES
IR- spectroscopy1
The differential
backscattering 2
Fluorescence
analysis3
Raman spectroscopy4
1 Jastrzebska, M. Atomic Force Microscopy and FT-IR Spectroscopy Investigations of Human Heart Valves [Текст] /M. Jastrzebska,
J. Zalewska-Rejdak, I. Mróz et al. // Gen. Physiol. Biophys. – 2006 – № 25 – С. 231-244.
2 Zakharov, VP Backscatter spectroscopy to assess skin lesions [Text] / VP Zakharov, EV Timchenko, PE Timchenko, LA Taskina,
SV Kozlov AA Starved // Bulletin of Samara Scientific Center of the Russian Academy of Sciences. - 2013 - 15 m, №6.. - S. 126130.
3 Jonak, C. Intradermal Indocyanine Green for In Vivo Fluorescence Laser Scanning Microscopy of Human Skin: A Pilot Study
[Текст]/ Constanze Jonak ,Hans Skvara, Rainer Kunstfeld, Franz Trautinger, J. A. Schmid// PLoS One. – 2011. - №6. С. 378-390.
4 Ali, S. M. Raman Spectroscopic Analysis of Human Skin Tissue Sections Ex-vivo: Evaluation of the Effects of Tissue Processing
and Dewaxing [Текст]/ S. M. Ali, F. Bonnier, A.Tfayli, H. Lambkin, K. Flynn, // Journal of Biomedical Optics. – 2013. - №8. – С.
1-39.
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GOAL AND TASKS
GOAL:
Purpose - to use Raman spectroscopy to measure
the quality of a person's skin implants, made in
different ways.
TASKS:
1. Set the characteristics of the Raman spectra
for dermal implants, made by different
protocols;
2. By analyzing the input optical coefficients to
estimate the optimal protocol for producing
cutaneous implants.
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EXPERIMENTAL SETUP
Stand Features:
The wavelength range from 190 nm to 1200 nm;
The exposure time of 0.01 to 10c;
Camera resolution of 1024 * 255 pixels
The laser power of 50-400 mW
Accuracy: δ = 4,25% (according to GOST 8.20776)
Figure 1. Experimental setup
1 - the object under study;
2 - Raman Probe RPB785;
3 - spectrometer Shamrock sr-303i;
4 - Digital camera ANDOR DV420A-OE;
5 - Laser Module LuxxMaster Raman Boxx;
6 - the source of the laser power supply module;
7 - PC;
8, 9, 10 - electric cables;
11 - coordinate table
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RESEARCHES FACILITIES
HUMAN SKIN IMPLANTS
The test implants made by two different protocols using the "Lioplast" technology1
Figure 2 - Microscopic picture of the samples: a) a sample with the epidermis treated with the
protocol number 1; b) the sample without the epidermis treated with the protocol number 2; c) A
control sample of untreated human skin. H & E stain. The increase of 400.
1-
Federal License for harvesting and conservation allotkaney number 99-01-002104 from 09.02.2006 Resolution on the mass
production and sale Registration certificate FS 01032004 / 1567-05 from 29.04.05.
TU was 9398-001-0196143-2005
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THE SPECTRAL FEATURES
Figure 3 - Averaged Raman
spectrum
of
bio-implants
manufactured by 1.2 protocols,
and control samples; where №12 -Numbers protocols, which are
made implants
Wave number, cm-1
863,975
1062
1202
1260,1268,1271
1410,1412,1415
1440,1446
1553, 1556-1557
1645
Fragment, oscillation
Vibration ribose (RNA)
Glycosaminoglycans (α-glycans)
Hydroxyproline, tyrosine
Amide III (collagen)
ν (C-O) from COO (amino acid glutamic acid)
СН2, СН3, С-Н vibrations (proteins and lipids)
NH deformation Amide II
Amide I
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DETAILED ANALYSIS OF SPECTRA
Figure 4 - Software division lines in the spectrum
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DETAILED ANALYSIS OF SPECTRA
Фирменный шрифт внедрен в файл презентации для отображения на компьютерах,
на которых данный шрифт отсутствует.
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QUALITY CONTROL OF THE USE OF OPTICAL COEFFICIENT
;
Optical coefficient:
k
I
i
I
1553
Where Ii - the intensity values at wavenumbers 1062 cm-1,
1645 cm -1, 1260 cm -1, 850cm -1, 863 cm -1, corresponding to a
significant
quality
implant
components:
glycosaminoglycans, amide I type amide III type asymmetricc
ommunication
C-O-S
vibration glycosaminoglycans, tyrosine and C-C stretch
of proline ring, ribose. When the denominator of 1553 cm 1, corresponding to collagen type II.
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FOR TWO-DIMENSIONAL ANALYSIS OPTICAL COEFFICIENT
Figure 5 - Two-dimensional diagram of the optical input coefficients
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RESULTS
Figure 6 - Diagrams substances as defined by the surface of the skin implants: a
sample with the epidermis treated with the protocol number 1; b) the sample without
the epidermis treated with the protocol number 2; c) A control sample of untreated
human skin.
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CONCLUSIONS
1) The analysis of the spectra from the surface of skin
implants, obtained by different methods. He showed
differences between them at wavenumbers 1062 cm-1,
1645 cm-1, 1260 cm-1, 850 cm-1, 863 cm -1, corresponding
to a significant quality for the implant components.
2) Determined that:
a) Raman spectroscopy allows for quick non-invasive control
of the structure of the skin bio-implants;
b) obtain an estimate of the composition of components
biomatriksov surfaces;
c) to carry out the choice of optimal bionositeley and monitor
the effectiveness of methods of processing;
d) optimize the creation of cellular and tissue products for
regenerative medicine.
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THANK YOU
FOR YOUR
ATTENTION
Elena Vladimirovna Timchenko,
Candidate of Physico-Mathematical Sciences
e-mail: [email protected]
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