Tissue regeneration observed in a basic fibroblast growth factor–loaded porous acellular bovine pericardium populated with mesenchymal stem cells

Ultrastructures of a porous acellular bovine pericardium (a) before and (b) after slicing off the dense layers on its surfaces using a cryostat microtome; (c) SEM micrograph of the bFGF/MSC patch, 18 hours after cell seeding; (d) photomicrograph of a porous acellular bovine pericardium (the control patch) before implantation stained with H&E; (e) photomicrograph of the bFGF/MSC patch before implantation stained with H&E; (f) photomicrograph of the bFGF/MSC patch stained with a monoclonal antibody against BrdU. Scale bars, 100 μm (d) and 50 μm (e and f).

Photomicrographs of the inner (endocardial) and outer (epicardial) surfaces of the control, bFGF, and bFGF/MSC patches retrieved at 4 weeks postoperatively stained for factor VIII and with van Gieson, respectively. Scale bars, 30 μm.

Photomicrographs of the middle layers of the control, bFGF, and bFGF/MSC patches retrieved at 4 weeks postoperatively stained with Masson’s trichrome and for α-SMA, respectively. The positive stained cells for α-SMA is indicated by. Scale bars, 50 μm.

Photomicrographs of the middle layer → of the bFGF/MSC patch retrieved at 12 weeks postoperatively (a) stained with H&E; (b and c) the insets 1 and 2 of (a) showed the additional staining with Safranin-O or for factor VIII; (d) stained with Masson’s trichrome; (e, f, and g) the insets 1, 2, and 3 of (d) showed the additional staining with EVG for α-SMA and double-immunostaining for α-sacromeric actin (blue) and anti-BrdU (brown); (h) the negative data for the bFGF patch retrieved at 12 weeks postoperatively. Scale bars, 50 μm (a and b), 30 μm (b, c, e, and f), and 10 μm (g and h).

Representative results from 5 independently performed experiments of flow cytometric analysis of (a) 5-aza-induced MSCs and (b) BrdU-labeled MSCs.

Photographs of the inner (endocardial) and outer (epicardial) surfaces of the control, bFGF, and bFGF/MSC patches retrieved at 12 weeks postoperatively. The implanted patch is indicated by arrow. →

Photomicrographs of the inner (endocardial) and outer (epicardial) layers of the control, bFGF, and bFGF/MSC patches retrieved at 4 weeks postoperatively stained with H&E; insets of the bFGF/MSC patch magnified an additional fourfold photomicrographs that stained for factor VIII. Scale bars, 100 μm.

Photomicrographs of the bFGF/MSC patch retrieved at 4 weeks postoperatively double-immunostained for: (a) α-sacromeric actin (blue) and anti-BrdU (brown); (b) α-SMA (blue) and anti-BrdU (brown); (c) factor VIII (blue) and anti-BrdU (brown). Scale bars, 10 μm.