Fig 1.
Wound closure of full-thickness wounds is delayed in HSA-/- mice.
The left picture in each panel represents the HSA+/+ mouse while the right is for the HSA-/-. A. t = 0, time of injury. B. t = 0, the wounds were stitched upon injury. C. t = 24 h. D. t = 24 h, with stitches. E. t = 7 days. F. t = 7 days, with stitches. G. A representative plot of the statistical differences between the groups. Each bar shows the fold change of the wound area compared to t = 0.
Fig 2.
Wound closure of bigger full-thickness wounds is faster in HSA+/+ mice.
The left picture in each panel represents the HSA+/+ mouse while the right is for the HSA-/-. A. t = 0, time of injury. B. t = 24 h. C. t = 72 h. D. t = 7 days. E. A representative plot of the statistical differences between the groups. Each bar shows the fold change of the wound area compared to t = 0.
Fig 3.
Tissue sections were stained with Hematoxylin and Eosin (H&E) and visualized on an Olympus AH light microscope at 400× magnification. The left picture in each panel represents the HSA+/+ mice while the right is for the HSA-/-. A- T = 0, time of injury; B- T = 72h; C- T = 2 weeks. D. Table of the scoring results.
Fig 4.
Tissue sections were stained with NovaUltraTM Picro-Sirius Red stain and visualized on an Olympus AH light microscope at 400× magnification. The left picture in each panel represents the HSA+/+ mouse while the right is for the HSA-/-. A. t = 0, time of injury, B. t = 72h, C. t = 14 days. The scoring is detailed in Table 2.
Table 1.
Table 2.
Fig 5.
Production of HIV-based viruses for gene delivery.
HEK 293T cells were co-transfected with the mentioned plasmids and after 48 h the expression of the transgene was evaluated by the mCherry marker (A). The infectivity of the virions, which was produced by the HEK293T helper cells, was tested on NIH-3T3 in vitro by fluorescence microscope (mCherry expression) (B). 72 h after the infection, cell lysates were prepared and 20 μg of total proteins was loaded. HSA was detected with the anti-HSA mAb, M1.69, and then the membrane was reprobed with anti-human tubulin (C). 1x106 infected cells were incubated with 10 μg/ml of M1.69 for 30 minutes at RT. FITC-labeled goat anti-rat antibody was used for the detection of bound antibody. The red curve represents the negative control (secondary antibody alone) and the green curve represents the binding of M1.69 (D).
Fig 6.
Imaging of the living organism.
The viruses were injected or dropped intothe wounded cells immediately after the injury, on t = 0.96 h after injection into the cells on the wound border (A) or dripping(B-C) of the mCherry-encoded viruses (n = 3) onto the wound area, mice were taken to in vivo imaging to detect the mCherry expression in the wounded cells using the Cri Maestro device. This imaging tool enables multiplexed in vivo fluorescence imaging of small animals with high sensitivity.
Fig 7.
A-The viruses were injected or dropped into the wounded cells on T = 0; B-HSA-/- mice 24 h after mCherry-encoded viruses injection (left panel) or dropping (right panel); C-HSA-/- mice 24 h after HSA encoded viruses injection (left panel) or dropping (right panel); D-HSA+/+ mice 24 h after mCherry-encoded viruses injection (left panel) or dropping (right panel); E-HSA+/+ mice 24 h after mCherry-encoded viruses injection (left panel) or dropping (right panel); F- 7 days after viruses injection; G- A representative plot of the statistical differences, in size of wound area, between the groups.
Table 3.
Fig 8.
Acceleration of wound healing upon over expression of HSA.
A-The wounds were infected with the viruses (right panel) or left un-treated (left panel) on T = 0; B- HSA+/+ mice 24 h after the injury (left panel), 24 h after infection, by dropping, with the mCherry-encoded viruses (middle panel), 24 h after mCherry-encoded viruses injection (right panel); C- HSA+/+ mice 24 h after infection, by dropping (left panel) or injection (right panel), with the HSA-encoded viruses; D- A representative plot of the statistical differences, in size of wound area, between the groups.
Table 4.
Fig 9.
Delayed wound healing upon down regulation of HSA expression.
A-The wounds were treated with anti-HSA antibody (right panel) or left un-treated (left and middle panel) on T = 0; B- HSA+/+ (left panel) and HSA-/- (middle panel) mice 48 h after the injury and HSA+/+ mice 48 h after antibody injection (right panel); C- B- HSA+/+ (left panel) and HSA-/- (middle panel) mice 72 h after the injury and HSA+/+ mice 72 h after antibody injection (right panel); D- A representative plot of the statistical differences, in size of wound area, between the groups. Student t-test was calculated in each time point between the tested group and the wt without treatment group.
Fig 10.
The rapid healing does not depend on mice strain.
Four groups of four mice were used in this study. Two of them were of C57/Bl while the other two were of Balb/c. The mice in one group from each strain were injected with the HSA-encoded viruses while the other group with the mCherry-encoded viruses. A- HSA+/+ mice 24 h after mCherry-encoded viruses injection; B- HSA+/+ mice 24 h after HSA-encoded viruses injection; C- HSA+/+ mice 48 h after mCherry-encoded viruses (the two pictures on the left) and HSA-encoded viruses injection (the two pictures on the right); D- HSA+/+ mice 6 days after mCherry-encoded viruses (the two pictures on the left) and HSA-encoded viruses injection (the two pictures on the right); E-A representative plot of the statistical differences, in size of wound area, between the groups. Student t-test was calculated in each time point between the HSA treated group and the mCherry treated group.