Effect of a wash-step on aPDI with tetracyclines.

<p>Bacterial cells were incubated for 30 min in PBS with stated concentration of tetracycline and either exposed (or not “dark”) to 10 J/cm2 of the appropriate light (“no wash”), or centrifuged and resuspended in fresh PBS and exposed to 10 J/cm2 (“wash”). (A) <i>E</i>. <i>coli</i> UTI89, DMCT and BL; (B) <i>E</i>. <i>coli</i> UTI89, DOCT and UVA; (C) MRSA, DMCT and BL; (D) MRSA, DOTC and UVA.</p

Chemical structures of tetracyclines together with methylene blue.

<p>Chemical structures of tetracyclines together with methylene blue.</p

Histological evaluation of wound healing effects of the peptides in CY-treated Balb/c mice.

<p>Animals were either wounded and treated with CMC (A) or injected with CY 1 and 4 days prior to injury, wounded and treated with CMC alone (B), with Regranex (C), with UN3 (D), comb1 (E) or a combination of UN3 and comb1 (F). Wounds were excised at day 10 post-injury, formalin-fixed, sectioned and stained with haematoxylin and eosin. Dotted line delineates the wound bed. Scale bar 500 µm.</p

Characterization of Lot 1-derived fractions.

<p>A – silver stained SDS-PAGE demonstrating reduction of composition complexity following gel filtration and ion exchange chromatography. 1 - MW markers, 2 – unfractionated lot 1, 3- fraction 22, 4 – proteins eluted with 0.5 M NaCl. B – fractionated platelet extracts retain their biological activity toward epithelial cells. For proliferation assay NeoNHEK were plated in keratinocyte growth medium the presence or absence of unfractionated proteins from Lot 1 or fractions of Lot 1 (0.1–200 ng/mL) as indicated. HB-EGH (10 ng/mL) was used as positive control. Relative proliferation compared to control is shown. Data are presented as mean +/− standard error.</p

ECM-and platelet extract-derived peptides stimulate in vitro angiogenesis and epithelial proliferation.

<p>A – In vitro angiogenesis assay. Bovine capillary endothelial cells were plated on the surface of growth factor reduced Matrigel in the presence or absence of 100 nM comb1 or 250 nM UN3, or the two peptides combined. DMEM supplemented with 1% BCS was used as control, cells that have received DMEM/1% supplemented with 10 ng/ml bFGF or VEGF served as positive control. Total tube length was measured at 7 h post-plating. Relative tube length compared to control is shown. B – In vitro epithelial proliferation assay. Hacat cells were plated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032146#s2" target="_blank">Materials and Methods</a>. The peptides were added at 100 or 250 nM (comb1 and UN3 respectively). Cell counting was performed at 5 days post-plating. Relative cells numbers as compared to control are shown. Data are presented as mean +/− standard error; * - indicates p<0.05.</p

Characterization of protein composition of human platelet extracts.

<p>A – Coomassie Blue stained SDS-PAGE demonstrating the complexity of protein composition of platelet lysate and extracts. B – Western blot analysis of platelet lysate and extracts revealing the presence of myosin in lots 2 and 3, but not lot 1 extract or platelet lysate. 1- platelet lysate; 2 – platelet extract lot 1, 15 µg/mL; 3 - platelet extract lot 1, 50 µg/mL; 4 - platelet extract lot 2, 15 µg/mL; 5 - platelet extract lot 2, 50 µg/mL; 6 - platelet extract lot 3, 15 µg/mL; 7 - platelet extract lot 3, 50 µg/mL.</p

Platelet extracts stimulate epithelial proliferation in vitro.

<p>Hacat cells were plated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032146#s2" target="_blank">Materials and Methods</a> in the presence or absence of at 1 µg/mL of proteins from platelet extracts (Lots 1–3). Relative proliferation compared to control is shown. Data presented as mean +/− standard error; * - indicates p<0.05.</p