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Confocal fluorescence microspectroscopy
in human cartilage autotransplantation
Wojciech Ciszek
1
, Zofia Drzazga
1
, and Jarosław Paluch
2
1
Department of Medical Physics, Chelkowski’s Institute of Physics, Chorzów, Poland,
e-mail: ciszek.wojciech@us.edu.pl
2
Department of Laryngology, Silesian Medical University, Katowice, Poland
Human cartilage plays important role in body, because it keeps organs’ shape and in case of
articular cartilage it facilitates motion of joints. Recently, the autotransplantation of human
cartilage, especially within the laryngological region has increasing application. Emitted light
analysis based on confocal autofluorescence microspectroscopy brings new insight into the
biocompatibility of cartilage which takes into account comparison of tissue biochemical signals
and could be used as a biochemical tool to assist transplantation surgery.
Nasal, laryngeal, ear, meniscus and articular cartilage tissue samples were taken from
healthy corpse and during standard laryngological surgical intervention. Prepared samples were
fixed by ethanol and then dried. Standard confocal imaging using 400x and 800x zoom was
performed. The excitation source was 404 nm diode laser. Autofluorescence measurements
(400–700 nm range) were made using circular ROI within extracellular matrix (ECM).
Differences in spectra of ECM were analyzed regarding to collagen II and metabolites.
Moreover, specificity ratio R was calculated as spectrum area ratio in green range of emission in
order to compare compatibility between cartilage types.
Confocal imaging revealed differentiation of cartilage seen between tissue types. Each of
studied samples showed autofluorescence with maximum in 460–480 nm range and some
differences in shape of spectra. Analysis of R factor suggests which cartilage type has the most
similar to the crucial laryngological cartilage tissues.
Our study shows that using spectroscopic characteristics of chemical state of bio-
fluorophores is useful in reconstructive surgery. Human cartilage tissue existing in different
places seems to have different metabolism. Further investigation could lead to develop advanced
procedures to measure the real biocompatibility of cartilage for patients individually.
Keywords: confocal microscopy; fluorescence microspectroscopy; cartilage; biocompatibility
Acknowledgment
The authors send thanks to Krzysztof Jasik, Grzegorz Bajor and Artur Pałasz from Silesian Medical
University for tissue preparation and consultation of results.
References
[1] M. Zellweger, P. Grosjean, D. Goujon, P. Monnier, H. van den Bergh, G. Wagnières, J. Biomed. Opt.
6(1): 41.
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Cancerous neck tissues in the FTIR and Raman mapping
W.M. Kwiatek
1
, G. Lisowska
2
, M. Misiołek
2
, C. Paluszkiewicz
1
,
E. Pięta
1
, N. Piergies
1
, W. Ścierski
2
1
Institute of Nuclear Physics Polish Academy of Sciences, PL-31342 Krakow, Poland,
e-mail: czeslawa.paluszkiewicz@ifj.edu.pl
2
Medical University of Silesia, Department of Otorhinolaryngology and Laryngological Oncology in
Zabrze, Poland
One of the head cancer type is inverted papilloma. The neoplastic lesion has a high risk of
regress after surgery with a recurrence rate of 20–47% [1, 2]. Due to the still not fully known
etiology of inverted papilloma treatment and prevention of recurrence is a big challenge for
doctors and scientists. The incidence of this type of cancer is 1.5 cases per 100 000 with a
predominance of males in the age group between 40–60 years of age [3, 4]. Most often it is
located in the side wall of the nasal cavity and the maxillary sinus.
The vibrational spectroscopic methods, i.e. RS, FTIR combined with the microscope can be
applied in various field of medicine, including biomedical materials characterizations. In this
work we present data for the tumor tissues (inverted papilloma) studied by these spectroscopic
methods (see Fig. 1 for an overview of the exemplary RS spectrum and Amide I intensity map).
The experimental materials were obtained from patients during routine surgical intervention
performed due to cancer disease.
Fig. 1. The RS spectrum of inverted papilloma together with the Amide I intensity map.
Keywords: inverted papilloma; Raman spectroscopy (RS);
Fourier transform infrared absorption spectroscopy (FTIR)
Acknowledgment
This project has been supported by the National Science Centre Poland under decision no. DEC-
2012/05/B/ST4/01150. This research was performed using equipment purchased in the frame of the project
co-funded by the Małopolska Regional Operational Program Measure 5.1 Krakow Metropolitan Area as an
important hub of the European Research Area for 2007-2013, project No. MRPO.05.01.00-12-013/15.
References
[1] N. Sadeghi, S. Al-Dhahri, J.J. Manoukian, The Laryngoscope 113 (2003) 749.
[2] W.S. Kim, D.W. Hyun, C.H. Kim, J.H. Yoon, Acta Otolaryngol 130 (2010) 493.
[3] C. Durucu, T. Baglam, E. Karatas, S. Mumbuc, M. Kanlikama, J. Craniofac. Surg. 20 (2009) 1985.
[4] C. Von Buchwald, P.J. Bradley, Curr. Opin. Otolaryngol. Head Neck Surg. 15 (2007) 95.
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