A new rabbit model of impaired wound healing in an X-ray-irradiated field - Fig 1

<p>A: Schematic diagram of shielding. Irradiation was restricted to a hindlimb. B: An anesthetized rabbit in a shielding apparatus was placed in an X-ray irradiation unit.</p

Tracking of implanted cells.

<p>DiI-labeled cells were fluorescently identified (red) by CD31/PECAM1 in sections stained with DAB and counterstained with hematoxylin. Objective lens: 40x; bar: 50 micro meters. pb: pb-PRP injected, bm: bm-PRP injected, f: fluorescent view, m: merged with bright-field view. A: 2 weeks after injection. In both pb and bm, fluorescence was identified in wound bed muscular layer. B: 4 weeks after injection. In pb, fluorescence was hardly observed, whereas in bm, robust fluorescence was confirmed. Correlation between fluorescence and DAB stain was unclear.</p

Histological view of the wounds 2 weeks after the surgery.

<p>Serial sections were prepared and stained; 2x objective lens; bar: 1 mm. R-: without irradiation, PBS injected. R+: irradiated, PBS injected. pb: irradiated, peripheral blood-derived PRP injected. bm: irradiated, bone marrow aspirate-derived PRP injected. HE: hematoxylin-eosin stain. CD31: DAB stained with anti-CD31/PECAM1 antibody, counterstained with hematoxylin. PR: Picrosirius red stain, bright-field view. PP: Picrosirius red stain, polarized view. Black triangles in first row (HE) indicate epidermal edges.</p

Percentage of colored areas in picrosirius red-stained, polarized images of normal and post-irradiated skin.

<p>Percentage of colored areas in picrosirius red-stained, polarized images of normal and post-irradiated skin.</p

Picrosirius red staining just below epidermal defect.

<p>Four weeks after wound creation; 20x objective lens; bar: 100 micro meters. R-: without irradiation, PBS injected. R+: irradiated, PBS injected. pb: irradiated, peripheral blood-derived PRP injected. bm: irradiated, bone marrow aspirate-derived PRP injected. PR: Picrosirius red stain, bright-field view. PP: Picrosirius red stain, polarized view. See also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184534#pone.0184534.t002" target="_blank">Table 2</a>.</p

Percentage of CD31/PECAM1-positive area in neoplastic granulation tissue.

<p>Percentage of CD31/PECAM1-positive area in neoplastic granulation tissue.</p

Picrosirius red staining just below epidermal defect.

<p>Two weeks after wound creation; 20x objective lens; bar: 100 micro meters. R-: without irradiation, PBS injected. R+: irradiated, PBS injected. pb: irradiated, peripheral blood-derived PRP injected. bm: irradiated, bone marrow aspirate-derived PRP injected. PR: Picrosirius red stain, bright-field view. PP: Picrosirius red stain, polarized view. Green areas indicate fine fibers; yellow areas indicate thick fibers. Marked collagen fiber decrease can be observed in irradiated wounds. See also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184534#pone.0184534.t002" target="_blank">Table 2</a>.</p

Percentage of colored areas in picrosirius red-stained polarized images.

<p>Percentage of colored areas in picrosirius red-stained polarized images.</p