Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Nov 17:10:1021827.
doi: 10.3389/fbioe.2022.1021827. eCollection 2022.

Novel dalbavancin-PLLA implant coating prevents hematogenous Staphylococcus aureus infection in a minimally invasive mouse tail vein model

Marlen Kloss  1 , Caroline Moerke  2 , Franziska Woitschach  1 , Katharina Wulf  3 , Sabine Illner  3 , Steffen Schulz  4 , Viktoria I Pauker  5 , Katharina Riedel  6 , Niels Grabow  3 , Hüseyin Ince  7 , Emil C Reisinger  1 , Martina Sombetzki  1
Affiliations

Novel dalbavancin-PLLA implant coating prevents hematogenous Staphylococcus aureus infection in a minimally invasive mouse tail vein model

Marlen Kloss et al. Front Bioeng Biotechnol. .

Abstract

Infective/bacterial endocarditis is a rare but life-threatening disease with a hospital mortality rate of 22.7% and a 1-year mortality rate of 40%. Therefore, continued research efforts to develop efficient anti-infective implant materials are of the utmost importance. Equally important is the development of test systems that allow the performance of new materials to be comprehensively evaluated. In this study, a novel antibacterial coating based on dalbavancin was tested in comparison to rifampicin/minocycline, and the suitability of a recently developed mouse tail vein model for testing the implant coatings was validated. Small polymeric stent grafts coated with a poly-L-lactic acid (PLLA) layer and incorporated antibiotics were colonized with Staphylococcus (S.) aureus before implantation into the tail vein of mice. The main assessment criteria were the hematogenous spread of the bacteria and the local tissue reaction to the contaminated implant. For this purpose, colony-forming units (CFU) in the blood, spleen and kidneys were determined. Tail cross sections were prepared for histological analysis, and plasma cytokine levels and expression values of inflammation-associated genes were examined. Both antibiotic coatings performed excellently, preventing the onset of infection. The present study expands the range of available methods for testing the anti-infectivity of cardiovascular implants, and the spectrum of agents for effective surface coating.

Keywords: antibiotic coating; cardiovascular implants; dalbavancin; hematogenous implant-related infections; in vivo biofilm model; rifampicin/minocycline.

PubMed Disclaimer

Conflict of interest statement

VP was employed by the company Ceva Animal Health GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Schematic representation of (A) the experimental set-up including (1) contamination of the implant using dipping protocol (2) application of the implants into the tail vein of mice and (3) analysis of the mice local and systemic reaction; (a) part of the tail for qPCR analysis and (b) histology and (B) the insertion system containing (I) the implant (II) 21 1/2 G needle (III) guide wire, 0.4 mm diameter (IV) 5 ml syringe body (V) 1 ml syringe plunger.
FIGURE 2
FIGURE 2
Characterization of implants samples (A) Schematic representation of implant design, (B) exemplary measurement of the implant average diameter by biaxial laser scanner, (C) Scanning electron micoscropy (SEM) image of implant cross section with PLLA coating containing rifampicin/minocycline; magnification ×200, (D) SEM detailed image of the PLLA coating containing rifampicin/minocycline; magnification ×800.
FIGURE 3
FIGURE 3
Rifampicin/minocycline and dalbavancin inhibit the biofilm formation of Staphylococcus aureus FR20. The effectiveness of rifampicin/minocycline and dalbavancin against Staphylococcus aureus FR20 biofilm formation was analyzed in a flow system. Inhibition of biofilm formation was monitored by (A) measuring green fluorescent protein (GFP) fluorescence intensity and (B) confocal laser scanning micoscropy (CLSM) of randomly selected areas (spanning 100 μm × 100 µm) after 16 h p values <0.05 were considered significant. *p < 0.05. Rif/Mino: rifampicin/minocycline; Dalba: dalbavancin; au: arbitrary unit.
FIGURE 4
FIGURE 4
Antibiotic coatings reduce the bacteria-induced systemic immune response in the host. The concentration of cytokines (CRP, TNF-α, IL-6 and G-CSF) in pooled blood plasma was quantified using ELISA after 6 and 48 h. For CRP detection, samples were diluted 1:106 in recommended reaction buffer. Data are represented as mean + SD. Rif/Mino: rifampicine/minocycline; Dalba: dalbavancin.
FIGURE 5
FIGURE 5
Antibiotic activity decreases the local inflammatory tissue response towards the contaminated implants. Relative gene expression levels of vegfa, icam1, tnf-α, vwf and tlr2 were determined by qPCR. Data are represented in scattered dot plots with mean + SD. p values <0.05 were considered significant. *p < 0.05; **p < 0.02.
FIGURE 6
FIGURE 6
Histological trichrome staining of tail cross sections was performed after 6 and 48 h. Representative images of mice tails that received implants without antibiotic coating (excerpted from Moerke et al. (Moerke et al., 2021)), coated with rifampicin/minocycline, and dalbavancin are shown; bars 100 µm or 50 μm; Magnification, × 200 or x 400. Different color expression is due to performing the staining on different days. Rif/Mino: rifampicine/minocycline; Dalba: dalbavancin.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Abram M., Vu ković D., Wraber B., Dorić M. (2000). Plasma cytokine response in mice with bacterial infection. Mediat. Inflamm. 9, 229–234. 10.1080/09629350020025746 - DOI - PMC - PubMed
    1. Alfonso F., Byrne R. A., Rivero F., Kastrati A. (2014). Current treatment of in-stent restenosis. J. Am. Coll. Cardiol. 63, 2659–2673. 10.1016/j.jacc.2014.02.545 - DOI - PubMed
    1. Almangour T. A., Perry G. K., Terriff C. M., Alhifany A. A., Kaye K. S. (2019). Dalbavancin for the management of gram-positive osteomyelitis: Effectiveness and potential utility. Diagn. Microbiol. Infect. Dis. 93, 213–218. 10.1016/j.diagmicrobio.2018.10.007 - DOI - PubMed
    1. Ammerlaan H. S. M., Harbarth S., Buiting A. G. M., Crook D. W., Fitzpatrick F., Hanberger H., et al. (2013). Secular trends in nosocomial bloodstream infections: Antibiotic-resistant bacteria increase the total burden of infection. Clin. Infect. Dis. 56, 798–805. 10.1093/cid/cis1006 - DOI - PubMed
    1. Anderl J. N., Franklin M. J., Stewart P. S. (2000). Role of antibiotic penetration limitation in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob. Agents Chemother. 44, 1818–1824. 10.1128/AAC.44.7.1818-1824.2000 - DOI - PMC - PubMed

LinkOut - more resources