Table 1.
List of Antibodies.
Figure 1.
SECPBMC leads to enhanced wound closure and re-epithelialization.
(A) Wound areas were measured during the first 3 days after wounding. Treatment with SECPBMC significantly enhanced wound closure. Error bars represent one standard deviation calculated from 15 animals for each set of values (*: p<0.05). (B) Representative photographs from mouse wounds (n = 15 from each group) immediately after wounding and at day 7 after wounding are shown. (C) H&E staining of wounds treated with medium or SECPBMC 7 days after wounding is shown. While medium treated wounds still show a thick crust and little re-epithelialization, SECPBMC treated wounds are fully re-epithelialized. C = crust, E = newly formed epidermis, G = granulation tissue. Scale bars: 100 µm. One representative animal of 15 is shown.
Figure 2.
SECPBMC induces migration of human primary fibroblasts and keratinocytes.
Scratch wounds of FB (A) and KC (D) are shown. One representative experiment of three each done in triplicates is shown. The mean-width of the gaps of nine scratch wounds after 18 h was measured and the percentage of closure for FB (B) and KC (E) was calculated. (*: p<0.01). Cells cycle analyses revealed no significant differences in FB (C) and in KC (F). One representative experiment of three each done in triplicates is shown. (G) Ki67 staining of medium and SECPBMC treated wounds showed no significant alterations in proliferating cells. Photographs were taken at the wound-edge of wounds 7 days after wounding. Ki67 staining is shown in red and nuclear staining is shown in blue. C = crust, E = newly formed epidermis, G = granulation tissue HF = hair follicle. One representative animal of 15 is shown. Scale bars: 50 µm. (H) The mean from 15 animals per group of Ki67 positive cells at the wound edge of one high power image was calculated.
Figure 3.
SECPBMC induces formation of new blood vessels.
(A) Representative CD31 stainings of wounds underneath the original wound edge are shown. Scale bars: 50 µm (B) The numbers of CD31 positive cells underneath the newly formed epidermis were evaluated. The graph represents the mean of 15 animals in each group (*: p<0.01). (C) The area of the granulation tissue taken up by CD31+ cells was evaluated. The graph represents the mean of 15 animals in each group. (D) Cell cycle analysis of SECPBMC treated microvascular EC shows a strong increase in proliferating cells. VEGF treatment served as positive control. One representative experiment of three each done in triplicates is shown (*: p<0.01). (E) A tube formation assay is shown. Compared to medium alone SECPBMC strongly induced tube formation in a matrigel assay. One representative experiment of three is shown.
Figure 4.
SECPBMC leads to activation of several signaling cascades.
KC, FB and EC were treated for 1 h with medium or SECPBMC. Western blot analyses of several signaling factors are shown. One representative experiment of three is shown. The graphs in the right panel represent the mean band intensity of all three experiments. The increase in expression of the phosphorylated proteins was calculated in relation to the respective non-phosphorylated proteins (*: p<0.01).