Establishment of human induced pluripotent stem cell-derived hepatobiliary organoid with bile duct for pharmaceutical research use.

In vitro models of the human liver are promising alternatives to animal tests for drug development. Currently, primary human hepatocytes (PHHs) are preferred for pharmacokinetic and cytotoxicity tests. However, they are unable to recapitulate the flow of bile in hepatobiliary clearance owing to the lack of bile ducts, leading to the limitation of bile analysis. To address the issue, a liver organoid culture system that has a functional bile duct network is desired. In this study, we aimed to generate human iPSC-derived hepatobiliary organoids (hHBOs) consisting of hepatocytes and bile ducts. The two-step differentiation process under 2D and semi-3D culture conditions promoted the maturation of hHBOs on culture plates, in which hepatocyte clusters were covered with monolayered biliary tubes. We demonstrated that the hHBOs reproduced the flow of bile containing a fluorescent bile acid analog or medicinal drugs from hepatocytes into bile ducts via bile canaliculi. Furthermore, the hHBOs exhibited pathophysiological responses to troglitazone, such as cholestasis and cytotoxicity. Because the hHBOs can recapitulate the function of bile ducts in hepatobiliary clearance, they are suitable as a liver disease model and would be a novel in vitro platform system for pharmaceutical research use.

Simultaneous gradual lengthening of proximal and distal nerve stumps for repair of chronic peripheral nerve defect in rats.

We investigated nerve regeneration of rat sciatic nerves after chronic injury of 15 mm-defect by the gradual lengthening of proximal and distal nerve stumps. Thirty days after the primal injury, both stumps were grasped and lengthened at a rate of 1 mm per day using external nerve-lengthening devices for 15 days. Then end-to-end neurorrhaphy was performed. After the lengthening, both stumps were evaluated by immunohistochemical analysis. Nerve regeneration was evaluated by electrophysiological and histological studies at 12 weeks after the repair. In the lengthened proximal stump, Schwann cells and axons existed along the whole nerve stump. In the lengthened distal stump, Schwann cells exist along the overall length. The whole nerve trunk was lengthened. The nerve regeneration was comparable with the delayed end-to-end suture without nerve defect. We showed the feasibility of direct gradual lengthening of proximal and distal nerve stumps for the treatment of chronic segmental nerve defect.

New treatment for peripheral nerve defects: reconstruction of a 2 cm, monkey median nerve gap by direct lengthening of both nerve stumps.

We have developed a new treatment for peripheral nerve defects: nerve-lengthening method, and confirmed the efficacy and safety of our method using cynomolgus monkeys. A 20-mm defect in the median nerve of monkey's forearms was repaired through the simultaneous lengthening of both nerve stumps with original nerve-lengthening device. To evaluate nerve regeneration after neurorrhaphy, electrophysiological, histological, and functional recovery were examined and compared to the standard autografting. Nerve conduction velocity, axon maturation, and the result of functional test were superior in the nerve-lengthening method than in the autografting. And there were no adverse events associated with our method. We concluded that this method is practical for clinical application.

The salvage of graft occlusion in a maintenace hemodialysis patient with tuberous sclerosis by percutaneous transluminal angioplasty using intravascular ultrasound: case report.

End-stage renal failure due to tuberous sclerosis is rare and there is no previous report of a patient with tuberous sclerosis undergoing long-term hemodialysis for over 18 years. The patency rate for a dialysis prosthetic graft is low, however, our patient's graft survived over 16 years. For thrombotic occlusion of a graft, we performed percutaneous thrombectomy and balloon angioplasty and salvaged graft occlusion. Moreover, this case is the first report in which the interventional procedure (mechanical thrombectomy and balloon angioplasty) could be observed by intravascular ultrasound in addition to angiography.