pkh-26 and Necrosis

Improvement of flap survival represents an ongoing challenge in reconstructive surgery. The angiogenic potential of adipose-derived stem cells (ASCs) offers a promising approach to improve the viability of random pattern flaps. Recently, to maximize the therapeutic effects of ASCs, increasing focus is being placed on how to deliver the stem cells to target lesions. The purpose of the present study was to compare the effectiveness of different administration routes of ASCs to improve the viability of the random pattern skin flap. ASCs labelled with PKH26 were applied via four methods to the cranially-based random pattern skin flaps of rats: (a) intravenous injection; (b) subcutaneous injection; (c) application with collagen sponge seeding; and (d) application with fibrin glue seeding. ASCs led to a significant increase in flap viability in the subcutaneous injection group and the collagen sponge group. Cutaneous blood flow was increased in the intravenous injection, subcutaneous injection and collagen sponge groups. Capillary density in the intravenous injection group and collagen sponge group was significantly greater than in the control group (no treatment). PKH26-positive cells via the collagen sponge were distributed more densely within the flap than in other groups. This study demonstrated that the collagen sponge method delivered ASCs most effectively within the flap and increased flap vascularity. The clinical therapeutic effects of ASCs can therefore be maximized when the optimal delivery route is chosen.

Topics: Adipose Tissue; Animals; Capillaries; Drug Administration Routes; Female; Flow Cytometry; Humans; Immunohistochemistry; Ischemia; Laser-Doppler Flowmetry; Necrosis; Neovascularization, Physiologic; Organic Chemicals; Platelet Endothelial Cell Adhesion Molecule-1; Rats, Sprague-Dawley; Skin; Stem Cell Transplantation; Surgical Flaps

Stem cell tracking is essential for evaluation of its migration, transplantation and therapeutic response. The aim of this study was to evaluate early distribution of intravenously transplanted rat bone marrow mesenchymal stem cells (BMSCs) in rats with acute cerebral trauma by labeling with (99m)Tc-hexamethylpropyleneamine oxime ((99m)Tc-HMPAO).. (99m)Tc-HMPAO-labeled BMSCs were injected intravenously to trauma rats (n=14) and sham-operated controls (n=13). Gamma camera images were acquired at 4 h after injection, and then organs were removed for gamma counting. Confocal microscope was used to confirm the migration of (99m)Tc-BMSCs by co-labeling with PKH26. Cytometric analysis was performed to evaluate apoptotic or necrotic change until the seventh day after labeling.. (99m)Tc-BMSCs were distributed mostly to lungs, liver and spleen at 4 h, and uptake of these organs was not significantly different between traumatic rats and controls. Meanwhile, the cerebral uptake of (99m)Tc-BMSCs was significantly higher in the traumatic rats than in controls (0.40% vs. 0.20%; P=.0002). Additionally, (99m)Tc-BMSCs' uptake of traumatic hemisphere was significantly higher than that of contralateral ones (0.27% vs. 0.13%; P=.0001) in traumatic rats. Regardless of radiolabeling, BMSCs migrated to traumatic regions, but not to nontraumatic hemispheres. However, gamma camera failed to demonstrate (99m)Tc-BMSCs in traumatic hemispheres. No significant apoptotic or necrotic change was observed until 7 days after radiolabeling.. Early distribution of BMSCs in traumatic brain disease could be monitored by (99m)Tc-labeling, which does not induce cellular death. However, our data showed that the amount of migrated (99m)Tc-BMSCs was not enough to be demonstrated by clinical gamma camera.

Topics: Animals; Apoptosis; Brain; Brain Injuries; Case-Control Studies; Female; Fluorescent Dyes; Gamma Cameras; Liver; Lung; Mesenchymal Stem Cells; Microscopy, Polarization; Necrosis; Organic Chemicals; Radionuclide Imaging; Radiopharmaceuticals; Rats; Rats, Sprague-Dawley; Spleen; Technetium Tc 99m Exametazime; Tissue Distribution

Up until now, research on fat cells has been unable to prove their survival rate objectively in vivo. In this article, the first application of the cell surface marker PKH26 in the fat cells of rats is reported. In a study of 48 Lewis rats, this method enabled the objective stereometry of viable and necrotic grafts after variable follow-up times in groups of eight animals each. The best survival rate was 30.41 percent, and the best implantation site was the interscapular subcutis. During follow-up, a characteristic change in size of the viable fat cells matched the in vitro findings of various investigators. Because of the surface marking, it could be proved that the viable cells found after 6 months were transplanted cells that had undergone a cycle of fat deprivation and regaining. This is proof of the cell survival theory postulated by Peer in 1950.

Topics: Adipocytes; Animals; Cell Survival; Cell Transplantation; Fluorescent Dyes; Male; Microscopy, Fluorescence; Microscopy, Interference; Necrosis; Organic Chemicals; Rats; Rats, Inbred Lew