Cryopreserved porcine hepatocytes: expression and induction of cytochrome P450, isoform CYP2E1.

Cryopreservation of porcine hepatocytes for their use in bioartificial liver devices may result in reduced cytochrome P450 (CYP) enzyme activity. The aim of this study was to assess the effects of several CYP inducers on the isoform CYP2E1 protein expression in cryopreserved porcine hepatocytes. Isolated porcine hepatocytes were cryopreserved for 1 month, thawed, and cultured for 3 days. During medium culture, the hepatocytes were exposed to the following CYP inducers: dimethyl sulfoxide, rifampicin, phenobarbital, 3-methylcholanthrene, and dexamethasone. CYP2E1 protein expression was determined by immunoblotting. CYP2E1 protein levels were constantly detected in cryopreserved porcine hepatocytes. CYP inducers did not modify CYP2E1 protein levels. Long-term cryopreserved porcine hepatocytes preserved their capacity for CYP2E1 protein expression, although exposure of these hepatocytes to CYP inducers did not modify the CYP2E1 protein expression.

Peritoneal implantation of cryopreserved encapsulated porcine hepatocytes in rats without immunosuppression: viability and function.

The bioartificial liver (BAL) represents a promising approach to cell transplantation without immunosuppression as a method to support patients with hepatic insufficiency. The aim of this study was to assess viability and function of cryopreserved encapsulated porcine hepatocytes implanted intraperitoneally in rats without immunosuppression. Isolated porcine hepatocytes were cryopreserved at -196 degrees C for 1 month. Four groups were created: group 1 (n=10), freshly encapsulated porcine hepatocytes cultured in albumin-free medium for 10 days; group 2 (n=10), freshly encapsulated porcine hepatocytes implanted in the rat peritoneum without immunosuppression for 1 month and cultured for 10 days after explantation; group 3 (n=10), cryopreserved encapsulated porcine hepatocytes cultured for 10 days; group 4 (n=10), cryopreserved encapsulated porcine hepatocytes implanted in the rat peritoneum without immunosuppression for 1 month and cultured for 10 days after explantation. We assessed urea and albumin production and hepatocyte viability. The hepatocytes of all groups retained the capacity to produce urea and albumin, although the albumin synthesis was significantly decreased among hepatocytes of group 4 (P< .01). Encapsulated cryopreserved porcine hepatocytes explanted from rat peritoneum after 1 month appeared morphologically viable; their ultrastructure was preserved. In conclusion, long-term cryopreservation of porcine hepatocytes resulted in retention of their biological activity and in significant viability when transplanted into the rat peritoneum without immunosuppression.

[Experimental model for the evaluation of topical application of a solution to help myocutaneous flap transposition]. / Modèle expérimental pour l'évaluation d'une solution topique améliorant la survie des lambeaux myocutanés.

The aim of this article is the evaluation of the topical application of a solution of hydrogen peroxide (H2O2) 8% and dimethyl sulphoxide (DMSO) 50% in order to reduce ischaemic failure in random skin flaps. This study was performed using a rabbit model. Two parallel, cephalad-based para-midline random cutaneous flap (10 cm x 2.5 cm) were elevated and resutured in place on the dorsum of 40 New Zealand rabbits. The 80 flaps thus obtained were then randomly divided into one control group and three experimental groups of 20 flaps each. Flaps from the control group (group A) were topically treated with saline, while flaps from experimental group B were treated with H2O2 8%, flaps from experimental group C with DMSO 50%, and flaps from experimental group D with a solution of 50% DMSO + 8% H2O2. Each solution was topically applied, 20 cc per three times a day, on the flaps for seven days, starting on the immediate postoperative period. Transcutaneous oxygen tension (Ptc O2) measurements were carried out in all flaps, 72 h after flap elevation. The percentage of surviving skin area of each flap was determined by planimetry 7 days after flap elevation. The mean surviving area of the group A (control) flaps was 71%. The mean surviving area of the group B (H2O2-treated) flaps was 72%. The mean surviving area of the group C (DMSO-treated) flaps was 76%, and that of the group D (DMSO + H2O2-treated) flaps was 92%. While no statistically significant differences were found between the survival rates of both the flaps treated with H2O2 or DMSO alone and that of the control group, the mean surviving rate of the DMSO + H2O2 treated flaps (+20%) was statistically higher than that of the control flaps. Similarly, a statistically significant difference has been found between the mean Ptc O2 values of the DMSO + H2O2 flaps and those of the other three groups of flaps.