Study of Recellularized Human Acellular Arterial Matrix Repairs Porcine Biliary Segmental Defects
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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.
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.
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.
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.
KeywordsTissue engineering Human acellular arterial matrix Common bile duct defect
The authors thank Doc. Li Li (Department of hepatobiliary surgery, Ganmei Hospital affiliated to Kunming Medical University, Kunming, Yunnan, 650500, China) for the critical comment and discussion on this study.
Wei Liu drafting the article, a substantial contribution to the conception and design of the study. Sheng-Ning Zhang, Zong-Qiang Hu, Shi-Ming Feng, Zhen-Hui Li, Shu-Feng Xiao, and Hong-Shu Wang performed the research and contribution to acquisition, interpretation, and analysis of data. Li Li revising the draft critically and final approval of the version to be published.
The study was supported by National Natural Science Foundation of China (No. 81560089) and Yunnan Province Health Science and Technology Project (No.2014NS198).
Compliance with ethical standards
Conflicts of interest
The authors have no financial conflicts of interest.
The study protocol was approved by the institutional review board of Ganmei Hospital affiliated to Kunming Medical University. Informed consent was confirmed by the Ganmei Hospital affiliated to Kunming Medical University. The animal studies were performed after receiving approval of the Ethics Committee of Ganmei Hospital affiliated to Kunming Medical University (2015-01).