Observation on the process of vascular reconstruction in acellular hepatic scaffolds of rat after transplantation into the greater omentum
HUANG Junjie1, 3, WANG Zhibin1, CHEN Na2, ZHANG Jianse1, WANG Zhiyi1, LI Ting1, Dai Kaiyu1, MEI Jin1, LOU Xinfa1.
1.Institute of Bioscaffold Transplantation and Immunology, Wenzhou Medical University, Wenzhou, 325035; 2.School of the Second Clinical Medical Sciences, Wenzhou Medical University, Wenzhou, 325035; 3.Department of General Surgery, the First Affiliated Hospital of Zhejiang University, Hangzhou, 310058
HUANG Junjie,WANG Zhibin,CHEN Na, et al. Observation on the process of vascular reconstruction in acellular hepatic scaffolds of rat after transplantation into the greater omentum[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2017, 47(10): 703-707.
Abstract：Objective: To observe the process of vascular reconstruction in acellular hepatic scaffolds of rat after transplantation into the greater omentum. Methods: Acellular hepatic scaffolds were harvested from adult SD rats by perfusing with TritonX-100. The scaffolds and native livers were observed through genomic DNA content analysis, scanning electron microscopy, HE stain, immunofluorescence and vascular cast. Then the acellular hepatic scaffolds were transplanted into the greater omentum in rats to investigate the vascular reconstruction. Rats were sacrificed respectively at 1, 3, 7, 14 or 28 days post-implantation and sections from the explanted grafts were analyzed by HE stain for morphology and microstructures. Results: Scanning electron microscope, HE stain and immunofluorescence showed most collagen fibers but no visible cell nuclei remained after decellularization in the scaffolds compared with the native livers. Vascular cast showed the pipeline structures of acellular hepatic scaffolds were intact and clear. Quantitative analysis of DNA content within the scaffolds showed only (211.6 ± 25.3) ng/µL. Then the blood sinus were observed on border of grafts at 3 days after transplantation into greater omentum. Afterwards the neovessels appeared, got into the internal of scaffolds, and formed microvascular network gradually in hepatic scaffolds after 14 days. The de novo microvascular network were still existed in hepatic scaffolds on 28 days. Conclusion: The implantation of greater omentum can promote the recellularization and eventually formation of vascularized reticular structures in the decellularized hepatic scaffolds.
[1] REN H, SHI X, TAO L, et al. Evaluation of two decellularization methods in the development of a whole-organ decellularized rat liver scaffold[J]. Liver Int, 2013, 33(3): 448-458.
[2] BAHARVAND H, HASHEMI S M, SHAHSAVANI M, et al. Differentiation of human embryonic stem cells into functional hepatocyte-like cells in a serum-free adherent culture condition[J]. Differentiation, 2008, 76(5): 465-477.
[3] JI R, ZHANG N, YOU N, et al. The differentiation of MSCs into functional hepatocyte-like cells in a liver biomatrix scaffold and their transplantation into liver-fibrotic mice[J].Biomaterials, 2012, 33(35): 8995-9008.
[4] OTT H C, MATTHIESEN T S, GOH S K, et al. Perfusion-decellularized matrix: using nature’s platform to engineer a bioartificial heart[J]. Nat Med, 2008, 14(2): 213-221.
[5] PRICE A P, ENGLAND K A, MATSON A M, et al. Development of a decellularized lung bioreactor system for bioengineering the lung: the matrix reloaded[J]. Tissue Eng Part A, 2010, 16(8): 2581-2591.
[6] 陈婵, 王志斌, 翁杰, 等. 全肺AHS的制备与鉴定[J]. 温州医科大学学报, 2016, 46(2): 88-91.
[7] YU Y L, SHAO Y K, DING Y Q, et al. Decellularized kidney scaffold-mediated renal regeneration[J]. Biomaterials, 2014, 35(25): 6822-6828.
[8] BADYLAK SF, TAYLOR D, UYGUN K. Whole-organ tissue engineering: decellularization and recellularization of three-dimensional matrix scaffolds[J]. Annu Rev Biomed Eng, 2011, 13: 27-53.
[9] CHOORAMANI G S, SINGH G K, SRIVASTAVA R N, et al. Chooramani technique: A novel method of omental transposition in traumatic spinal cord injury[J]. Asian J Eurosurg, 2013, 8(4): 179-182.
[10] BIGHAM-SADEGH A, ORYAN A, MIRSHOKRAEI P, et al. Bone tissue engineering with periosteal-free graft and pedicle omentum[J]. ANZ J Surg, 2013, 83(4): 255-261.
[11] SAKO A, KITAYAMA J, YAMAGUCHI H, et al. Vascular endothelial growth factor synthesis by human omental mesothelial cells is augmented by fibroblast growth factor-2: possible role of mesothelial cell on the development of peritoneal metastasis[J]. J Surg Res, 2003, 115(1): 113-120.
[12] CARALT M, VELASCO E, LANAS A, et al. Liver bioengineering: from the stage of liver decellularized matrix to the multiple cellular actors and bioreactor special effects[J]. Organogenesis, 2014, 10(2): 250-259.
