Cytokine profile of wound healing.

<p>Serum levels of VEGF-A, TGF-β1 and IL-6 from mice of different treatment groups in both acute thermal injury at day 7 and excision wound at day 15 were examined by ELISA method.</p><p>Results are expressed as the mean (pg/ml) ± standard deviation (SD) from three independent experiments of each group.</p>*<p>Statistically significant as compared to the Staph group (<i>P</i><0.01, Student’s <i>t</i> test).</p

mRNA levels of <i>VEGF-A, TGF-β1</i> and <i>IL-6</i> in different treatment groups during wound healing.

<p>AgNP/NSP treatment also modulates the expression of cytokine mRNAs in wounded skin. The levels of <i>VEGF-A</i>, <i>TGF-β1</i> and <i>IL-6</i> mRNA expression from mice of different treatment groups in both acute thermal injury at day 7 (a) and excision wound at day 15 (b) were examined by RT-PCR. Relative band intensities of different groups were calculated by a densitometer and are demonstrated by the values under the bands. The data shown are representative of three independent experiments.</p

IL-6 induced Mcl-1L expression in rat hepatocytes.

<p>(A) Serum levels of IL-6 in PH rats or flavopiridol (2.5 mg/kg) treated for 24 hours before PH group or control group were determined by EIA. Data are provided as mean +/− S.D. *P<0.05. (B) Rat hepatocytes were serum-starved for 24 hours prior treated with recombinant rat IL-6 (0.1, 1 and 10 ng/ml), under serum-starved conditions, and after 4 hours, Mcl-1L protein levels were determined by Western blot analysis. (C) Rat hepatocytes were serum-starved for 24 hours prior treated with recombinant rat IL-6 ( 1 ng/ml), under serum-starved conditions, at the indicated time periods, Mcl-1L protein levels were determined by Western blot analysis. (D) Rat hepatocytes were treated with rat recombinant IL-6 (10 ng/ml) at the indicated time periods. Mcl-1 mRNA expression in rat hepatocytes was determined by q-RT-PCR. *P<0.05.</p

Mcl-1L chemical inhibitor flavopiridol inhibits liver regeneration in rats.

<p>(A) Rats were pretreated with flavopiridol (2.5 mg/kg) or vehicle (control) 24 hours before partial hepatectomy (PH). (A) The remnant liver weight (RLW) of each group was measured at the indicated time periods after PH and expressed as a percentage of the original liver weight (OLW). Data are compared between the vehicle and flavopiridol-treated groups. *P<0.05. (B) The expression of Mcl-1L in remnant liver tissue was determined by IHC. (C) ki67 staining. (D) TUNEL staining. Magnification, 400×.</p

Cytotoxicity toward the human foreskin fibroblast (Hs68) cell line after 12 h of incubation with AgNP/NSP, polymer dispersed AgNP (Poly-Ag) and silver sulfadiazine (SS) using the MTT assay (a).

<p>The values are expressed as the mean ± SD from 4 independent experiments. DNA damage in Chinese hamster ovary (CHO) cells by the comet assay showed the average lengths of the cell tails after incubation with different concentrations of AgNP/NSP and Poly-Ag (b). Data are shown as the mean ± SD (at least 200 cell tails were counted in each sample). *<i>P</i><0.05 at each concentration, Student’s <i>t</i> test.</p

JAK/PI3K/Akt/CREB signaling is involved in IL-6-induced Mcl-1L expression in hepatocytes.

<p>(A) Rat hepatocytes were pretreated with chemical inhibitors, JAK Inhibitor InSolution™ 420097 (20 nM), for 1 h prior to IL-6 (10 ng/ml) treatment. After 4 h, the Mcl-1L protein levels were determined by Western blotting. Between-group comparisons are as indicated with *p<0.05. (B) Rat hepatocytes were pretreated with chemical inhibitors, LY294002 (50 µg/mL), PD98059 (50 µg/mL), and staurosporine (20 nM) for 1 h prior to IL-6 (10 ng/ml) treatment. After 4 h, Mcl-1L expression was analyzed by Western blotting. Between-group comparisons are as indicated. *p<0.05. (C) Rat hepatocytes were pretreated with the NF-κB and CREB decoy ODN (10 µM) for 24 h prior to IL-6 treatment. After 4 h, mcl-1 mRNA expression was analyzed by q-RT-PCR. Data are fold-induction relative to the control (Lane 1). Between-group comparisons are as indicated. *p<0.05. (D) Rat hepatocytes were pretreated with JAK Inhibitor InSolution™ 420097 (20 nM), or LY294002 (50 µg/mL) for 1 h prior to IL-6 treatment. After 30 minutes, p-Akt, Akt, p-CREB and CREB were determined by Western blotting. Between-group comparisons are as indicated. *p<0.05.</p

Changes in the ratio of remnant liver weight to original liver weight (RLW/OLW) after 70% partial hepatectomy (PH).

<p>(A) OLW was estimated retrospectively from the excised liver weight after 70% PH. Data are presented as mean ± S.D., and comparisons were made between groups as indicated. *P<0.05. (B) ki67 staining of remnant liver tissue. Magnification, 400×.</p

Schematic of the proposed signal transduction pathway of Mcl-1L induction in hepatocytes during liver regeneration.

<p>IL-6 regulates Mcl-1L expression through the IL-6 dependent JAK/PI3K/Akt/CREB activation pathway in rat hepatocytes.</p

IL-6 induced Mcl-1L expression plays an anti-apoptotic role in hepatocytes.

<p>(A) Rat hepatocytes were pretreated with mcl-1L siRNA (25 nM) or control siRNA (25 nM) for 24 h prior to IL-6 (10 ng/ml) treatment; four hours later, Mcl-1L levels were determined by Western blot analysis. Between-group comparisons are as indicated. *p<0.05. (B) Rat hepatocytes were pretreated with mcl-1L siRNA or control siRNA for 24 h prior to IL-6 treatment or not in serum-free conditions for another 48 hours. The percentage of apoptotic hepatocytes were determined by annexin-V staining. The illustration shown represents one of three independent experiments. (C) Quantitative result of annexin-V staining. Between-group comparisons are as indicated. *p<0.05. (D) Similar treatment protocol as described in (C), another 72 hours. DNA ladder analysis was performed. The illustration shown is representative of three independent experiments.</p

Wound healing rates on (a) acute burn model (b) excision wound model and the photographs of wound appearance on day 5 after (c) acute burn injury and (d) excision injury.

<p>The rate of infected wound healing was compared in animals treated with staphylococcus only (Staph), staphylococcus and NSP (Staph+NSP), staphylococcus and Poly-Ag (Staph+Poly-Ag), staphylococcus and AgNP/NSP (Staph+AgNP/NSP), staphylococcus and Aquacel® (Staph+AQ), staphylococcus and silver sulfadiazine (Staph+SS), and control (untreated). The results are expressed as the mean ± SD from three independent experiments in each group. *<i>P</i><0.05, comparison between AgNP/NSP and the other 6 groups at each time point; † <i>P</i><0.05, comparison between SS treatment with the untreated, Staph, Staph+NSP, and Staph+Poly-Ag groups; ‡ <i>P</i><0.05, comparison between AQ treatment with the untreated, Staph, Staph+NSP, and Staph+Poly-Ag groups; # <i>P</i><0.05, comparison between NSP treatment with untreated, Staph and Staph+Poly-Ag groups; § <i>P</i><0.05, comparison between Poly-Ag treatment with untreated and Staph groups. All the comparisons were confirmed by Student’s <i>t</i> test.</p