CPC based implants.

<p>Podoplanin immunopositive lymphatic vessels (arrow) were localized in the granulation tissue (gt) at the interface of the CPC implant (A-E) as well as at the strontium functionalized CPC (CPC-S in E-F) where an improved fragmentation was found (F). Lympatics were often associated with blood vessels that were identified by Microfil^® perfusion (star in C). Podoplanin also stained osteocytes (arrowhead in B-F) and mature osteoblasts (O). Nuclei were counterstained with hematoxylin. m = bone substitution material. Bars: 1 mm in A, F, 200 µm in B, G, and 20 µm in C-E.</p

Number of lymphatic vessels localized in the interconnected pores of iron-foams.

<p>A significant increase was calculated for the correlation of the counted number of lymphatic vessels localized in the Fe-S implant in comparison to the Fe (p = 0.004) as well as the Fe-BP implants (p = 0.001). Data are presented as box plots with the median indicated by solid line within the box. Small circles illustrate data beyond 3 x standard deviation (SD). ** = p ≤ 0.01, *** = p ≤ 0.001.</p

Number of lymphatic vessels in empty defects.

<p>The 4 mm sized empty defect contained a significant lower number of lymphatic vessels in comparison to the 3 mm and 5 mm defects. The results are present as box plot where the median is indicated by a solid line within the box, the 25^th and 75^th percentile as bottom and top of the box, the 0^th and 100^th percentile as lower and upper whiskers, respectively. Small circles illustrate data beyond 3 x standard deviation (SD). ** = p ≤ 0.01.</p

Podoplanin immunohistochemistry at the implant of iron foam with a strontium coating (Fe-S).

<p>Podoplanin labeled lymphatic vessels (arrow) were found in the granulation tissue (gt) at the implant (m) interface and in the interconnected pores (A-B). Higher magnification showed the adjacency of the bone substitution material and the lymphatics (C). A high amount of cells with yellow granular cytoplasm were localized in the granulations tissue (D). These cells are supposed to resorb the degraded implant. They are often found close to the lymphatic vessels that sometimes seem to contain lymphocytes (D). Nuclei were counterstained with hematoxylin. Bar represents 1 mm in A, 200 µm in B, 20 µm in C-D.</p

Podoplanin staining at the plain iron foam (Fe).

<p>The interface of the wedge-shaped implant (m) was covered with granulation tissue (gt) that infiltrated the interconnected pores (p) near the interface (A-B). Podoplanin immunopositive lymphatic vessels (arrow) were found in the granulation tissue often close to the yellow cells with the resorbed iron (C). Granula of resorbed iron (star) were found in lymphatic capillaries (D). Lymphatics were also situated next to iron fragments (arrowhead, E). Islets with podoplanin stained osteocytes were detected in the granulation tissue (F). Nuclei were counterstained with hematoxylin. Bar represents 1 mm in A, 200 µm in B, 20 µm in C-D.</p

Podoplanin immunoreactivity at composites assembled by silica, collagen (pB30, B30), and additional strontium (pB30S20).

<p>The implanted xerogel (B30, A-B) was surrounded by podoplanin immunopositive structures that were related to developing lymphatic capillaries (arrow in B) and additional myofibroblasts (f in B). Some immunoreactivity was also found in the implant (arrowhead and in higher magnification in the inset in A). No residues of the porous composite scaffolds (pB30, pB30S20) were detected (C-F). Podoplanin labeled lymphatic vessels (arrow) were found in the granulation tissue (gt) of the fracture gap (C, E). The lymphatics (arrow) were often localized close to blood vessels (stars in B, D) that were identified by their filling with Microfil® (star). Nuclei were counterstained with hematoxylin. m = bone substitution material. Bar represents 200 µm in A, C, E and 20 µm in inset of A, B, D, and F. </p

Expression of podoplanin in empty defects.

<p>No regulation of the podoplanin expression was detected for the empty defects by means of real-time RT-PCR. The results are presented as box plot with the median, the 0^th, 25^th, 75^th, and 100^th percentile. Small circles illustrate data beyond 3 x standard deviation (SD).</p

Number of lymphatic vessels at the interface of the implant.

<p>A significant up-regulation was found for the silica-collagen xerogel (B30), the strontium (Fe-S) and bisphosphonate (Fe-BP) functionalized iron foam in comparison to the negative control without implant. Data were presented as box plots where a solid line within the box indicated the median. Small circles show data beyond 3 x standard deviation (SD). CPC = calcium phosphate cement, CPC-S = strontium modified CPC, pB30 = composite scaffold, pB30S20 = strontium-modified composite scaffold, B30 = xerogel consisting of 70 wt% silica, 30 wt% collagen, Fe = iron foam. * = p ≤ 0.05.</p

Podoplanin immunohistochemical staining in the rat osteoporotic fracture model.

<p>The 3 mm (A) defect was narrowed by new built bone (b) but the remaining granulation tissue (gt) contains a high number of lymphatic vessels (arrow in B). Podoplanin positive osteocytes are localized in the new built bone (C). The 4 mm defect (D) is a typical critical size defect showing no bridging of the fracture gap that was filled with granulation tissue (gt) with only a few lymphatics (arrow in E-F) but a lot of blood vessels (stars). The vasculature was identified by Microfil^® perfusion (stars in D-F), immunohistochemistry for CD31 (inset in E), anatomical characteristics (F), and their absence of podoplanin immunreaction (F). The fracture gap of the 5 mm defect (C) was filled with granulation tissue (gt) that contained a high amount of lymphatic vessels (arrow) but only limited blood vessels. Higher magnification of lymphatics showed the irregular shape, the discontinuous endothelium, and thin walls in comparison to the thick muscle layer in the walls of arteries (Ar) and continuous endothelium and wider lumen of veins (V in F). The nuclei were counterstained with hematoxylin. Bar: 1 mm in A, D, G, 200 µm in B, E, H, 20 µm in C, F and inset in E.</p