428 BONE MARROW MONONUCLEAR CELLS THERAPY IMPROVES LIVER PERFUSION IN CIRRHOTIC PATIENTS

POSTERS Results: Sorafenib increased serum aminotransferases and the number of S-phase nuclei at baseline. Relative liver weights were decreased 20 and 24 hours after PH and protein levels of pERK slightly decreased 20–24 hours after PH. In the second experiment the number of S-phase nuclei and mitotic indices were significantly decreased 48 hours after surgery, but significantly increased 7 days after surgery in animals on continuous sorafenib treatment. Relative liver weights were restored 5 days after surgery in controls, 7 days after surgery in animals treated with sorafenib only prior to PH, 10 days in those treated before and three days after PH, and 14 days in rats on continuous treatment with sorafenib. Conclusion: Sorafenib exerted an inhibiting effect on liver cell proliferation 24–48 hours after PH and a compensatory increase in cell proliferation during prolonged treatment, indicating an adaptation to the drug. In spite of this, the time for restoration of liver weight increased from 5 to 14 days in rats continuously exposed to sorafenib compared with controls. The effects of sorafenib on liver regeneration in humans needs to be further evaluated. 427 PROCESS OF NEO-LIVER REGENERATION WHILE RECRUITING DONOR-DERIVED NON-PARENCHYMAL CELLS K. Ohashi, R. Utoh, T. Okano. Institute of Adv Biomed Eng and Sci, Tokyo Women’s Medical University, Tokyo, Japan E-mail: [email protected] Background: Liver tissue engineering is an emerging field in which a functional liver system is created in vivo using isolated hepatocytes. Our lab has succeeded in achieving functional persistency of the ectopically engineered liver tissues and had them grow in response to the regenerative stimulus derived by partial hepatectomy. The present study was conducted to create complex 3-D neo-livers under the kidney capsule by inducing progressive proliferative growth in response to continuous regeneration stimulus derived by chornic liver injury. We further discuss the process of neo-liver construction in terms of non-parenchymal recruitment. Methods: Hepatocytes were isolated and purified from FVB/N mice. For induction of chronic liver injury to recipient FVB/N mice, mice were inoculated twice with monocrotaline (MCT) at an interval of 2 weeks. Two weeks later, mice were subjected to partial hepatectomy (PH) and received liver tissue engineering procedure to left subrenal capsule space by transplanting 1×106 hepatocytes. Histological investigations and tissue volume of the engineered livers were assessed as a function of time. Results: The engineered liver tissue volume showed continuous increase in MCT/PH treated mice throughout 180 days experimental period. H&E staining at day 90 revealed that the engineered liver tissues were significantly thicker in MCT-PH group (~300 mm) (Fig) than that of no MCT-PH control group (~30 mm). The liver tissues were initially made by purified hepatocytes, but it is surprising that numerous liver-specific non-parenchymal cells (stellate, sinusoidal endothelial, and kupffer cells) emerged in the engineered livers at day 30 or later. At day 60, these non-parenchymal cells formed complete hepatic plate structure in coordination with hepatocytes. Detailed investigation revealed that these non-parenchymal cells were derived from recipients. Finally, the engineered livers were found to be amazingly enlarged to form complex and thicker neolivers (Fig).

Fig. 3-D structured neo-livers engineered at kidney capsule. (A) H&E staining of the neo-liver at day 90. Bar, 100 mm. (B) Gross appearance of the neo-liver at day 180. Bar, 1 mm. S170

Conclusions: The present study demonstrated that the ability of ectopically engineered liver tissues to perform progressive regenerative growth while recruiting non-parencymal cells from donor. This 3-D neo-liver regeneration process structured with complete hepatic plate could provide new insights into the field of liver regeneration. 428 BONE MARROW MONONUCLEAR CELLS THERAPY IMPROVES LIVER PERFUSION IN CIRRHOTIC PATIENTS R. Bica1,2 , C. Resende3 , A. Alves1,2 , A. Torres1,2,4 , B. Couto1,2,4 , F. Azevedo5 , D. Mercante6 , J. Dias1,2 , T. Kasai-Brunswick7 , H.S. Coelho1,8 , A.C. Campos-de-Carvalho7,9 , R. Goldenberg7 , G. Rezende1,4,8 . 1 Internal Medicine, Faculty of Medicine, Federal University of Rio de Janeiro, Rio de Janeiro, 2 Coordenac˜ ¸ ao de Aperfeicoamento ¸ de Pessoal de N´ıvel Superior (CAPES-MEC), Bras´ılia, 3 Radiology, Hospital Universit´ ario Clementino Fraga Filho, Federal University of Rio de Janeiro, 4 Fundac˜ ¸ ao Carlos Chagas Filho de Amparo a ` Pesquisa (FAPERJ), 5 Radiology, Faculty of Medicine, Federal University of Rio de Janeiro, 6 Hematology, Hospital Universit´ ario Clementino Fraga Filho, Federal University of Rio de Janeiro, 7 Instituto de Biof´ısica Carlos Chagas Filho, Federal University of Rio de Janeiro, 8 Hepatology, Hospital Universit´ ario Clementino Fraga Filho, Federal University of Rio de Janeiro, Rio de Janeiro, 9 Conselho Nacional de Desenvolvimento Cient´ıfico e Tecnol´ ogico (CNPq), Bras´ılia, Brazil E-mail: [email protected] Background and Aims: The effect of bone marrow mononuclear cells (BMMC) therapy in cirrhotic portal hypertension has not been clinically investigated. This study was conducted to analyze liver morphology and hemodynamic changes by abdominal ultrasound with color Doppler fluxometry (DUS) of portal vein, splenic and hepatic arteries of cirrhotic patients after BMMC infusion. Methods: Autologous BMMC were isolated from 8 patients presenting cirrhosis due to different etiologies and moderate liver dysfunction. Then 2.0–15.0×108 cells were infused via the common hepatic artery. DUS was performed on days 0, 2, 14, 90, 180 and 360, splenic and hepatic artery resistance and pulsatility indexes (SA-RI, SA-PI, HA-RI, HA-PI) were calculated and compared to MELD score. Results: Morphologic aspects of liver, spleen and related vessels did not change over follow-up. SA-RI, SA-PI, HA-RI, HA-PI showed a transient reduction, the lowest value was on day 14 and the average declines were, respectively, 8.4%, 14.0%, 11.5% and 22.0%. MELD score also decreased (p = 0.018) and it correlated to SA-RI and SA-PI trends during the first 90 days. Conclusions: Reduction of the splenic and hepatic artery resistance improves liver perfusion. This effect occurred concurrently to MELD score improvement, suggesting an association between them. If the causative effect between stem cell infusion and lowering of the arterial resistance is confirmed, the mechanisms involved must be clarified as well as its potential to contribute to liver function improvement. 429 MELATONIN ATTENUATES INFLAMMATION AND PROMOTES REGENERATION IN RABBITS WITH FULMINANT HEPATITIS OF VIRAL ORIGIN ´ B. San Miguel, I. Crespo, A. Laliena, M. Alvarez, J. Gonzalez-Gallego, ´ M.J. Tun˜ on. ´ University of Le´ on, Le´ on, Spain E-mail: [email protected] Impaired liver regeneration is one of the most critical issues in the prognosis of fulminant hepatic failure (FHF). The objective of the present study was to investigate the effect of melatonin on inflammatory and regenerative response in an animal model of FHF of viral origin. Rabbits were experimentally infected with 2×104 hemagglutination units of a rabbit hemorrhagic

Journal of Hepatology 2012 vol. 56 | S71–S224