Here - EWMA 2013

POSTER PRESENTATIONS
P 214
Poster: Basic Science
NMR-based metabolomics as a novel approach to diagnostics of
biofilm presence in chronic wounds – the pilot study
Adam Feliks Junka 1 , Stanislaw Deja 2 , Marzenna Bartoszewicz 1 , Piotr Mlynarz 3 ,
Danuta Smutnicka 1 , Beata Maczynska 1 , Patrycja Szymczyk 4
1 Departament of Microbiology (Wroclaw, Poland);
2 Faculty of Chemistry (Opole, Poland);
3 Departament of Bioorganic Chemistry (Wroclaw, Poland);
4 Mechanics Departament (Wroclaw, Poland).
Aim: To develop reliable NMR-based method useful for rapid detection of biofilm formed
on biomaterials and wound tissues.
Methods: A strain* was allowed to form a biofilm on the surface of surgical polystyrene
mesh. Stage of biofilm development was evaluated with a use of quantitative cultures
and Scanning Electron Microscopy. Samples of forming, maturing and mature biofilm
were frozen at -80°C till the time of further analysis. Assessment of bacterial foot and
fingerprint was performed by means of Nuclear Magnetic Resonance Spectroscopy
(NMR) with Bruker 600 MHz Spectrometer. Obtained set of 1H NMR spectra was
subjected to chemometric analysis involving multivariate statistical approach.
Results: Investigated staphylococcal strain was able to form biofilm on surface of the
surgical mesh. Trends of biofilm formation were found in Principal Component Analysis
(PCA) score plot. Essential metabolites related to bacterial cell functions were detected.
Among them, acetone, acetate, 2,3-butanediol and two unassigned resonances
(chemical shift: 1.38 and 4.42 ppm) were positively correlated to biofilm formation,
whereas glycine betaine was positively correlated with planktonic forms of bacteria.
When established, entire assay, including sample preparation and analysis of NMR
spectra, took less than 1 hour.
Conclusions: Identified metabolites related to biofilm formation may be used as
biomarkers of staphylococcal biofilm presence in wound. However experiments
concerning biofilms formed by other wound pathogens are required, presented pilot
results reveal promising potency of NMR spectroscopy for diagnostics of biofilm
presence.
Key words: NMR, staphylococcus, biofilm, metabolomics
* ATCC6538 S. aureus
POSTER: BASIC SCIENCE
P 215
Poster: Basic Science
EXPERIMENTAL JUSTIFICATION FOR CLINICAL USE OF PLATELET RICH
PLASMA
Vladimir Obolenskiy 1 , Darya Ermolova 2 , Leonid Laberko 2 , Maxim Makarov 3 ,
Natalia Borovkova 3
1 City Hospital # 13 (Moscow, Russia);
2 RNRMU (Moscow, Russia);
3 N.V. Sklifosovskiy RIEM (Moscow, Russia).
Aim: To asses in vitro the efficiency of platelet rich plasma (PRP) for clinical use.
Methods: Blood collected from a peripheral vein of the patients with chronic wounds of
various etiologies was usedas a biological material. PRP was obtained by separating
blood with the use of BTI equipment. Whole blood platelets were stained with vital
fluorochrome dye for further morpho-functional analysison a fluorescence microscope.
Concentration of platelet-derived growth factor (PDGF) in blood serum was determined
by enzyme-linked immunosorbent assay (ELISA) using reagents «Qantikine, Human
PDGF-BB Immunoassay» and system «Multiskan ascent». Proliferative activity of
human fibroblast culture M-22 was assessed in the wells containing different amount of
serum PDGF and also using fluorochrome vital dyes.
Results: A direct correlation between concentration and morpho-functional parameters
of whole blood platelets and concentration of PDGF in the serum has been observed.
Degranulation of platelets is associated with the massive release of PDGF. The study
finding was that the content of PDGF in one well equal to 150 pg is associated with the
maximum gain of proliferative activity of fibroblastswith their viability being preserved; at
the higher PDGF concentrations fibroblasts viability decreased and cells death was
observed.
Conclusions: These data allow recommending the clinical use of PRP for stimulation of
the regenerative-reparative processes.
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