Experimental Study Of The Feasibility Of MSCs Transplantation Via The Portal Vein By Interventional Technique For Management Of The Liver Cirrhosis

Objective: Liver Cirrhosis is the common end stage of most liver diseases, for which, unfortunately, there are few effective treatments available currently. Recent reports have shown the capacity of mesenchymal stem cells (MSCs) to differentiate into hepatocytes in vitro and in vivo. MSCs administration could repair injured liver through reducing inflammation, collagen deposition, and remodeling. These results provide a new clue to treatment of liver fibrosis. The aim of this study was to observe the MSCs' homing in the fibrotic liver in rabbit after MSCs transplanted via the portal vein slectively using interventional techniques, to evaluate the technical feasibility and safety, to observe the therapeutic effect to the fibrotic liver, and to provide the evidence of the clinical application. Materials and Methods:1. There were 25 New Zealand rabbits, whose weight ranged from 2.5kg to 3.5kg; An animal model of liver fibrosis was established using the New Zealand rabbit, induced by subcutaneous injection of hypodermic inject the CCl₄-olive oil.2. MSCs were isolated from the rabbit bone marrow and purified by combining gradient density centrifugation with plastic adherence, and then the MSCs were labelled with BrdU.3. Transcatheter intra-portal vein selectively transplantation of the labelled MSCs was performed using a 3-French microcatheter under digital subtractionangiograpy.4. The liver function testing, including the TP, ALB, AST, ALT, TB and DB, were performed before and after the transplantation. The rabbits liver specimens were obtained 3 weeks after intra-portal transplantation and fluorescence immunohistochemistry was used for tracing MSCs resident in the liver. Results:1, The rabbits' liver specimens were randomly taken out after hypodermic inject the CCl₄-olive oil. The liver looked brown, and it was smaller and tougher than normal liver. The HE stained slice showed that the hepatocytes were varicose, denaturalized, and flake necrosis, and the hyperplastic fibrous tissue arround the fake lobules was presented. The liver function testing showed that ALT value were significantly higher than before(69.14± 14.78U/L vs 144.44 ± 53.20U/L, t=6.539, P<0.05), but AST(43.48±15.08U/L vs 44.61±8.05 U/L), TB(21.27± 6.48 umol/L vs 20.28 ± 4.75umol/L), DB(11.84± 2.80 umol/L vs 11.52 ± 2.95 umol/L),TP(68.47±3.53g/Lvs 68.22±2.87 g/L),ALB(43.57± 5.83 g/L vs 43.10 ± 6.61 g/L)were not significantly changed(t=0.317, 0.583, 0.385, 0.262, 0.255, p>0.05).2, MSCs were isolated and purified by combining gradient density centrifugation with plastic adherence, At passage 3, MSCs' appearance and distribution looked well in the inverted microscope. The immunohistochemistry showed that the cell labeling efficiency of BrdU was more than 90%, which could satisfied the request of the cell tracing.3, The fluorescence immunohistochemistry showed that the engrafted MSCs' positive ratio was about 81.2%(13/16) 3w after transplantation, which proved the engrafted MSC could alive in the pathological liver.4, The liver function testing showed that ALT value were significantly decreased after transplantation(146.96 ±63.12U/L vs 101.26±39.23U/L, t=2.459, P<0.05), but AST (44.89 ± 7.95U/L vs 42.35 ± 16.35U/L), TB (19.66± 4.32umol/L vs 20.87 ± 5.32umol/L), DB (11.37 ± 2.70umol/L vs 11.79 ±2.72umol/L,) TP (68.57±3.07g/L vs 69.13 ±3.38g/L,) ALB (41.93± 5.95g/L vs 42.04±5.37g/L)were not significantly changed(t=0.558, 0.691 , 0.437, 0.488, 0.059, p>0. 05).Conclusion: The transplanted MSCs could be resident and alive in the fibrotic recipient liver after transplantation via the portal vein by interventional technique, the positive ratio of the engrafted MSCs was about 81.2%(13/16) 3w after the transplantation, The damaged liver function could be improved partially after transplantation of the MSCs. The presented evidence indicated that transplantation of the MSCs via the portal vein by interventional technique could be used as a new way to treat the liver cirrhosis, but some problems such as improving the alive ratio of the engrafted MSCs and objective analyse the therapeutic effect still needed more investigation.