Implantation of a Tissue-Engineered Neo-Bile Duct in Domestic Pigs

Abstract

Background: Extrahepatic bile duct injuries remain severe complications during cholecystectomies and often require reconstruction by bilioenteric anastomosis (i.e., hepaticojejunostomy), which comes with further long-term complications (e.g., recurring ascending cholangitis, secondary biliary cirrhosis). In the case of inherent extrahepatic biliary atresia or during liver transplant, artificial or engineered bile ducts could allow novel surgical strategies without the need for hepaticojejunostomy. Methods: We present data on the implantation of in vitro-generated neo-bile ducts in 5 domestic pigs. The neo-bile ducts were engineered through decellularization of allogeneic blood vessels and recellularization with autologous cholangiocytes. On postoperative days 0, 1, 7, and 14, blood samples were taken and analyzed (aspartate aminotransferase, alanine aminotransferase, bilirubin, alkaline phosphatase, creatinine, and leukocytes). Magnetic resonance cholangiopancreatography was performed on postoperative day 14 on 1 pig. Fourteen days after implantation, the pigs were sacrificed and the bile ducts were explanted. Results: All pigs survived the complete study period without severe complications. None of the pigs showed signs of biliary leakage or peritonitis. The neo-bile ducts were infiltrated by neutrophils, and neoangiogenesis was observed around and into the implanted tissue. Conclusion: We present a novel strategy for extrahepatic bile duct replacement by implantation of an autologous neo-bile duct generated ex vivo. Whether the presented technique allows the long-term replacement of native bile ducts must be further evaluated.

Scaffolds obtained from decellularized human extrahepatic bile ducts support organoids to establish functional biliary tissue in a dish

“…Cheng et al (2016) successfully transplanted a decellularized ureter, splinted by a silicone stent or a T‐tube, in guinea pigs. In parallel, Struecker et al (2016) transplanted porcine abdominal aorta, which was recellularized with autologous cholangiocytes, in a porcine model. However, ureter or aorta ECM does not resemble the architecture of human EBD as these structures are lacking the complex PBG architecture.…”

“…Furthermore, maintaining viability of cholangiocytes after transplantation would require the development of a blood vessel network. Struecker et al (2016) showed feasibility of transplanting a bile duct construct solely with cholangiocytes in a large animal model without forming blood vessels (before transplantation). This could suggest that a preformed blood vessel network is not required, as formation of blood vessel after implantation appeared to be adequate.…”

Biliary Atresia – emerging diagnostic and therapy opportunities

“…On the tissue repair side, the technology for engineering endodermal tissues in vitro on decellularized matrices has improved during the last few years [103] , and may be applied to engineering bile ducts. Engineered neo-bile ducts were successfully transplanted into pigs and well tolerated [104] . Recently, Sampaziodis et al., (2021) showed that extrahepatic organoids could successfully be transplanted to repair intrahepatic biliary ducts in human livers ex vivo and organoid-derived cells regenerated 40-85% of the injected ducts [28] .…”

3D Printed Model of Extrahepatic Biliary Ducts for Biliary Stent Testing

“…Many tissue engineering protocols rely on multi-day culture timelines and numerous reagents to evaluate products [34][35][36][37]. Reactors housing organs or vessels require calibrated perfusion [38][39][40]. Scale-up and product transport remain challenges for tissue engineered products [41][42][43].…”