Study of Recellularized Human Acellular Arterial Matrix Repairs Porcine Biliary Segmental Defects

BACKGROUND: With the popularity of laparoscopic cholecystectomy, common bile duct injury has been reported more frequently. There is no perfect method for repairing porcine biliary segmental defects.METHODS: After the decellularization of human arterial blood vessels, the cells were cultured with GFP⁺ (carry green fluorescent protein) porcine bile duct epithelial cells. The growth and proliferation of porcine bile duct epithelial cells on the human acellular arterial matrix (HAAM) were observed by hematoxylin-eosin (HE) staining, electron microscopy, and immunofluorescence. Then, the recellularized human acellular arterial matrix (RHAAM) was used to repair biliary segmental defects in the pig. The feasibility of it was detected by magnetic resonance cholangiopancreatography, liver function and blood routine changes, HE staining, immunofluorescence, real-time quantitative PCR (RT-qPCR), and western blot.RESULTS: After 4 weeks (w) of co-culture of HAAM and GFP? porcine bile duct epithelial cells, GFP⁺ porcine bile duct epithelial cells grew stably, proliferated, and fused on HAAM. Bile was successfully drained into the duodenum without bile leakage or biliary obstruction. Immunofluorescence detection showed that GFP-positive bile duct cells could still be detected after GFP-containing bile duct cells were implanted into the acellular arterial matrix for 8 w. The implanted bile duct cells can successfully resist bile invasion and protect the acellular arterial matrix until the newborn bile duct is formed.CONCLUSION: The RHAAM can be used to repair biliary segmental defects in pigs, which provides a new idea for the clinical treatment of common bile duct injury.

Mechanism of immune hyporesponsiveness induced by recipient- derived immature dendritic cells in liver transplantation rat / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the mechanism of immune hyporesponsiveness induced by donor-antigen- unloaded recipient-derived immature dendritic cell (imDC) of liver grafts in rats.</p><p><b>METHODS</b>Forty Sprague-Dawley rats (donor) and forty male Wistar rats (recipient) were randomly divided into 4 groups: control, cyclosporine A (CsA), mature DC (mDC), and imDC groups respectively, with 10 donor rats and 10 recipient rats in each group. Recipient rats in CsA group were treated with 10 mg•kg⁻¹•d⁻¹ CsA starting day 2 after the transplantation. Recipients in the mDC or imDC groups were given Wistar rat derived mDCs (1 × 10⁶/rat) or imDCs (1 × 10⁶/rat) via dorsal vein of the penis respectively 1 day before the transplantation. In each group, 5 recipients were kept for determination of survival time and the other 5 rats were executed at day 10 after transplantation. Blood samples were collected for the measurement of serum alanine aminotransferase (ALT), total bilirubin (TBIL), interleukin 2 (IL-2), interferon gamma (IFN-γ), IL-4, and IL-10 levels. Liver tissue was harvested for HE staining and acute rejection evaluation. Expression levels of Fas-L/Fas in the grafts were detected by immunohistochemical staining; and Western blot was used to detect the expression level of Scurfin.</p><p><b>RESULTS</b>The survival time of CsA and imDC groups was significantly longer than that of control and mDC groups (all P < 0.05). The levels of serum ALT and TBIL in the control group (2072.20 ± 217.93 IU/L and 147.42 ± 22.02 µmol/L) and mDC group (2117.00 ± 285.13 IU/L and 141.58 ± 20.82 µmol/L) were significantly higher than those in the CsA group (59.68 ± 13.48 IU/L and 15.40 ± 2.13 µmol/L) or imDC group (50.80 ± 9.63 IU/L and 14.44 ± 3.49 µmol/L) (all P < 0.05). In the CsA and imDC groups, the levels of IL-2 (22.52 ± 3.75 pg/mL and 22.12 ± 3.90 pg/mL) and IFN-γ (309.20 ± 25.19 pg/mL and 321.00 ± 21.64 pg/mL) were significantly lower, but the levels of IL-4 (297.60 ± 25.07 pg/mL and 277.00 ± 22.47 pg/mL) and IL-10 (1226.00 ± 140.49 pg/mL and 1423.00 ± 106.39 pg/mL) were higher than those of the control (IL-2: 147.78 ± 12.80 pg/mL, IFN-γ: 1758.60 ± 106.22 pg/mL, IL-4: 17.40 ± 4.77 pg/mL, IL-10: 81.00 ± 9.47 pg/mL) and mDC groups (IL-2: 142.34 ± 9.29 pg/mL, IFN-γ: 1835.00 ± 82.63 pg/mL, IL-4: 15.60 ± 3.96 pg/mL, IL-10: 68.80 ± 11.23 pg/mL) (all P < 0.01). The expression level of Scurfin protein on CD4+ CD25+ T cells of the imDC group (1.34 ± 0.29) was significantly higher than that in the control (0.72 ± 0.13), CsA (0.37 ± 0.11), and mDC groups (0.78 ± 0.17) (all P < 0.05).</p><p><b>CONCLUSION</b>Donor-antigen-unloaded recipient-derived imDC is an effective treatment in inducing immune hyporesponsiveness through induction of T cell apoptosis, shift in Thl/Th2 balance, and proliferation of regulatory T cell.</p>

A causal analysis of intra-abdominal hemorrhage after reduced-size liver transplantation in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the cause of abdominal hemorrhage after reduce-size liver transplantation in rats.</p><p><b>METHODS</b>Healthy female SD rats were used as the donors and male SD rats as the recipients (weighing 260-280 g), with the recipients weighing 10 g more than that of the donors. The donor operation was performed by the same surgeon under direct vision, and the liver graft size reduction procedure was completed in the donor operation. The recipient operation was performed by two surgeons under direct vision.</p><p><b>RESULTS</b>A total of 270 SD rats received reduce-size liver transplantation successfully, and 44 of the rats died from intra-abdominal hemorrhage. The abdominal hemorrhages, listed in the order of incidences, included anastomotic hemorrhage of the inferior vena cava of the superior liver (28 cases), subcapsular hemorrhage of the liver (9 cases), ligation hemorrhage of the left outboard lobe, the nipple lobe and the triangle lobe of the liver (9 , 7 and 7 cases, respectively), hemorrhage of the right suprarenal vein and lumbar veins (5 cases), hemorrhage of the mechanical injury (4 cases), cuff hemorrhage of the portal vein and inferior vein cava of the inferior liver (both 4 cases). Eight rats had anastomotic hemorrhage of the inferior vena cava of the superior liver and ligation hemorrhage of the left outboard lobe, 5 had hemorrhage of the two ligation points of the reduce-size liver; for management of the hemorrhage, 10 rats received suture or/and ligature, and 6 had washing and hot water bath.</p><p><b>CONCLUSION</b>The most common cause of hemorrhage after reduce-size liver transplantation in rats is the anastomotic hemorrhage of the inferior vena cava of the superior liver, and this finding may provide clues for improving the success rate of reduced size liver transplantation in rats.</p>