Islet Transplantation and Insulin Administration Relieve Long-Term Complications and Rescue the Residual Endogenous Pancreatic β Cells  Marina Figliuzzi,

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Islet Transplantation and Insulin Administration Relieve Long-Term Complications and Rescue the Residual Endogenous Pancreatic β Cells  Marina Figliuzzi, Roberto Bianchi, Cristina Cavagnini, Raffaella Lombardi, Carla Porretta-Serapiglia, Giuseppe Lauria, Federica Avezza, Annalisa Canta, Valentina Carozzi, Alessia Chiorazzi, Paola Marmiroli, Cristina Meregalli, Norberto Oggioni, Barbara Sala, Guido Cavaletti, Andrea Remuzzi  The American Journal of Pathology  Volume 183, Issue 5, Pages 1527-1538 (November 2013) DOI: 10.1016/j.ajpath.2013.07.032 Copyright © 2013 American Society for Investigative Pathology Terms and Conditions

Figure 1 Blood glucose (A) and body weight (B) in male Sprague–Dawley rats. Data are expressed as means ± SEM. P < 0.001 for control versus all other groups at all time points. P < 0.001 for hyperglycemia versus all other groups at all time point. ∗P < 0.05, ∗∗P < 0.001 for islet transplantation versus mild hyperglycemia. †P < 0.05 for control versus islet transplantation. ‡P < 0.05 for hyperglycemia versus mild hyperglycemia. §P < 0.05, §§P < 0.001 for moderate hyperglycemia versus islet transplantation. ¶P < 0.05, ¶¶P < 0.001 for moderate hyperglycemia versus mild hyperglycemia. The American Journal of Pathology 2013 183, 1527-1538DOI: (10.1016/j.ajpath.2013.07.032) Copyright © 2013 American Society for Investigative Pathology Terms and Conditions

Figure 2 OGTT blood glucose values in male Sprague–Dawley rats at 4 months (A) and 8 months (B). Data are expressed as means ± SEM. The American Journal of Pathology 2013 183, 1527-1538DOI: (10.1016/j.ajpath.2013.07.032) Copyright © 2013 American Society for Investigative Pathology Terms and Conditions

Figure 3 A and B: Thermal (A) and mechanical (B) nociceptive threshold over time. C and D: Nerve conduction velocity in the tail vein at 8 months (C) and Na+-K+-ATPase activity at 8 months (D). Data are expressed as means ± SEM. ∗P < 0.05 for control versus moderate hyperglycemia, mild hyperglycemia, and islet transplantation. ∗∗P < 0.001 for control versus hyperglycemia, moderate hyperglycemia, and mild hyperglycemia. †P < 0.05 for hyperglycemia versus moderate hyperglycemia, mild hyperglycemia, and islet transplantation. ††P < 0.001 for hyperglycemia versus mild hyperglycemia and islet transplantation. The American Journal of Pathology 2013 183, 1527-1538DOI: (10.1016/j.ajpath.2013.07.032) Copyright © 2013 American Society for Investigative Pathology Terms and Conditions

Figure 4 Representative images of sciatic nerve in control, hyperglycemic, and islet transplantation groups of rats. Degenerating fibers in hyperglycemic animals are indicated by arrows. Scale bar = 25 μm. Original magnification, ×40. The American Journal of Pathology 2013 183, 1527-1538DOI: (10.1016/j.ajpath.2013.07.032) Copyright © 2013 American Society for Investigative Pathology Terms and Conditions

Figure 5 Representative images of stained glucagon cells in the five groups of rats at the end of the study. Two images for islet transplantation are shown Scale bar = 25 μm. Original magnification, ×20. The American Journal of Pathology 2013 183, 1527-1538DOI: (10.1016/j.ajpath.2013.07.032) Copyright © 2013 American Society for Investigative Pathology Terms and Conditions

Figure 6 β-Cell analysis at 4 and 8 months in Sprague–Dawley rats for volume density of β cells (A), mean area of positive insulin cells (B), and β-cell density per unit of volume (C). Data are expressed as means ± SEM. ∗P < 0.05 for control versus hyperglycemia and islet transplantation. ∗∗P < 0.001 for control versus all other groups. ‡P < 0.05 for hyperglycemia versus moderate hyperglycemia, mild hyperglycemia, and islet transplantation. ‡‡P < 0.001 for hyperglycemia versus moderate hyperglycemia and islet transplantation. ‡P < 0.05 for moderate hyperglycemia versus islet transplantation. §P < 0.05 for islet transplantation versus mild hyperglycemia. The American Journal of Pathology 2013 183, 1527-1538DOI: (10.1016/j.ajpath.2013.07.032) Copyright © 2013 American Society for Investigative Pathology Terms and Conditions

Figure 7 Representative images of stained insulin cells in the groups of rats at 8 months (end of study). Scale bars: 200 μm (left); 50 μm (right). The American Journal of Pathology 2013 183, 1527-1538DOI: (10.1016/j.ajpath.2013.07.032) Copyright © 2013 American Society for Investigative Pathology Terms and Conditions

Figure 8 Correlation between volume density of β cells and log value of blood glucose at 4 months (A) and 8 months (B). The American Journal of Pathology 2013 183, 1527-1538DOI: (10.1016/j.ajpath.2013.07.032) Copyright © 2013 American Society for Investigative Pathology Terms and Conditions