Anti-bacterial properties of WED.

<p><b>(A)</b> Energy Dispersive X-Ray Spectroscopy (EDS) elemental analysis of Ag/Zn WED. <b>a.</b> Scanning Electron Microscope (SEM) image; <b>b.</b> Light Microscope Image; <b>c</b>. and <b>d.</b> Close-up view of gold and silver dots under light microscope. <b>e-h.</b> EDS element maps of Zn (blue), Silver (red), Oxygen (green) and Carbon (Magenta). <b>(B,C)</b> Absorbance and CFU measurement from planktonic PAO1 culture in presence of placebo or Ag/Zn WED, n = 4 <b>(D) (i)</b> schematic diagram for experimental design showing dressing embedded in the agar plate. <b>(ii, iii)</b> Zone of inhibition above WED is marked with red dotted line, while no such zone was observed over the placebo dressing, n = 4 <b>(E)</b> Biofilm formation measured by absorbance at 540 nm, n = 4 <b>(F)</b> Biofilm co-aggregation observed in the placebo treated overnight PAO1 culture was not observed in WED treated overnight cultures, n = 3.</p

WED inhibits glycerol-3-phosphate dehydrogenase activity.

<p>Measurement of glycerol-3-Phosphate dehydrogenase enzyme activity from mature PAO1 biofilms treated with placebo or WED <b>A.</b> OD measured in the kinetic mode. <b>B.</b> GPDH activity calculated using the formula: Glycerol-3-Phosphate dehydrogenase activity = B/(ΔT X V) x Dilution Factor = nmol/min/ml, where: B = NADH amount from Standard Curve (nmol). ΔT = reaction time (min). V = sample volume added into the reaction well (ml), n = 3.</p

WED generates superoxide (A) EPR spectra using 20mM DIPPMPO demonstrates spin adduct generation upon exposure to dressings for 40 mins in PBS.

<p>Addition of 500U Catalase does not attenuate EPR signal but is attenuated using both 500U SOD and 500U Catalase, n = 3</p

WED impairs cell viability.

<p>CLSM micrographs of mature PAO1 biofilm stained with live-dead stain after treatment with placebo, WED or Ag control. The green fluorescence indicates live bacteria while the red indicates dead bacteria, n = 3.</p

WED represses redox-sensitive multidrug efflux genes in <i>P</i>. <i>aeruginosa</i> Real-time PCR was performed to assess mex gene expression post-treatment with placebo, WED or 3mM DTT, n = 3.

<p>WED represses redox-sensitive multidrug efflux genes in <i>P</i>. <i>aeruginosa</i> Real-time PCR was performed to assess mex gene expression post-treatment with placebo, WED or 3mM DTT, n = 3.</p

Confocal laser scanning microscopy images showing three dimensional presence of staphylococci in infected debrided wound tissue.

<p>Z-stack image created by merging serial scans of thick tissue section (20 µm), viewed under 600x magnification confocal laser scanning microscopy. Showing in the <i>x/y</i> plane clumps of staphylococci colonizing the debrided tissues (red), while the <i>x/z</i> and <i>y/z</i> planes display the depth of the colonization throughout the tissue section. Three-dimensional orthogonal projections of z-stack image in panel (oriented in two different planes) showing of staphylococci aggregates biomass within the debrided tissues.</p

Demographic characteristics of patients (n = 9) and SWI status.

<p>M, male, F, female; AKI, acute kidney disease; BMI, body mass index, CAD, coronary artery disease; CGH, coronary heart disease; DM, diabetes mellitus; END, endocarditis; GERD, gastro esophageal reflux disease; HTN, hypertension; HTN- P, Pulmonary hypertension; HLD, hyperlipidemia; RD, renal dysfunction; COPD, chronic obstructive pulmonary disease; PVD, peripheral vascular disease; OSA, obstructive sleep apnea; RHD, rheumatic heart disease; CABG, coronary artery bypass graft; MVR, mitral valve replacement; LVAD, left ventricular assisted device; PM, pace maker; RV, right ventricle; N, negative; MSSA, Methicillin-sensitive <i>Staphylococcus aureus</i>; MRSA, Methicillin-resistant <i>Staphylococcus aureus;</i> SVT, supraventricular tachycardia.</p

Presence of staphylococci within the infected debrided wound tissues.

<p>Representative confocal microscopy images of debrided tissue using immunofluorescence staining (debrided tissues was counterstained red with Phalloidin). Note large aggregates of staphylococci (intense green granular stain) colonizing the debrided tissues of infected sternal wound (lower panels), while tissues taken from a non-infected sternal wound during resternotomy (upper panels) show no colonization with staphylococci. Scale bar = 50 µm, 400x magnification. (SWI: sternal wound infection). Right panel is the zoom of the dashed boxed area in the left panel (scale bar = 20 µm).</p

MRSA Strain USA300 biofilm exhibits enhanced tolerance to tobramycin when grown as a biofilm on surgical wires.

<p>USA300 was used to inoculate in vitro wells containing sections of wire. Planktonic bacteria and wire-associated biofilms were challenged with 10 ug/ml of tobramycin for 2 hours. Bacteria tolerant to antibiotic challenge were enumerated using viability plating and compared to untreated parallel controls. Percent survivability of triplicate cultures is represented. nd, not detected, ns, not significant. Data are mean±SD (n = 3), *p<0.05 compared to untreated planktonic (Mann Whitney test).</p

Digital photos and gram staining of deep sternal wound infection in two patients scheduled for a debridement procedure.

<p>(A) Digital photos of the infected sternal wounds. Note the signs of active infection with localized erythema, exudates, friable wound edges and sternal instability. Sternal wires were removed before the debridement procedure. (B) Gram-Twort staining of debrided tissues taken of infected sternal wound showing clumps of Gram-positive cocci (arrows in right panel). Left panel, scale bar = 50 µm, 400x magnification. Right panel is the zoom of the dashed boxed area in the left panel (scale bar = 50 µm).</p