Study on 3D Bioprinted Liver Structure Implanted in the Fibrosis Liver of Rabbits
Tang Liang1, Luo Tao2, Yang Yadong2, Yang Geng2, Xu Yimeng2, Zhang Wenyuan2*
1(Institute of Health Food,Zhejiang Academy of Medical Sciences,Hangzhou 310013,China) 2(Institute of Bioengineering,Zhejiang Academy of Medical Sciences,Hangzhou 310013,China)
Abstract Three dimensional (3D) bioprinted liver-like hydrogel structures were implanted into the fibrosis liver of rabbit models by the liver surface cladding method to study its feasibility of improving the liver fibrosis and of generating liver-like tissue. Primary hepatocytes were extracted from the liver tissue of rabbit by 0.1% collagenase type IV digestion, and the hepatic fibrosis model was established by subcutaneous injection of carbon tetrachloride (CC14) into rabbits. Thirty rabbits with liver fibrosis were randomly divided into experimental group, control group, and sham operation group, with 10 rabbits in each group. The structure lamella containing rabbit primary hepatocytes/alginate/gelatin blend was constructed using 3D bioprinting technique. Then the 3D structure lamellas were implanted into the liver surface of hepatic fibrosis rabbits, as an experimental group. Sixteen days after implantation, liver functional biochemical index and liver histopathological changes of the liver were examined as well as the developmental degrees of hepatic fibrosis and the formation of liver-like tissue structure. The 3D structures with the grid hydrogel composing of multilayer staggered cylindrical microfilament were obtained and live/dead cell fluorescent staining results showed that the hepatocytes’ survival rate was about 82% ± 3% after printing. Most of the implants were not degraded 16 days after the implantation. The undegraded implants were tightly integrated into the liver. The biochemical indexes of liver function in the experimental group were ALT (90.26±13.05)U/L, AST (75.37±13.45)U/L, γGT (16.62±6.72)U/L, ALB (32.48±4.43)g/L. The significance between any two groups was P>0.05. The pathological examination scores of liver fibrosis degree in the experimental group, control group and sham operation group were 2.95±0.50, 3.11±0.58 and 3.02±0.62, respectively, and the significance between any two groups was P>0.05. The results showed that the fibrosis degree and the function of the rabbits’ liver had improvements to some extents in experimental group though there was no statistical significance. Histological observations from the experimental group showed that hepatocytes were evenly distributed, and that cell degeneration and death were not discovered, succeeding in generating liver-like tissue structure. In conclusion, the bioprinted hepatic structure has potentials of regenerating liver-like tissue structure and rebuilding liver functions.
Tang Liang,Luo Tao,Yang Yadong, et al. Study on 3D Bioprinted Liver Structure Implanted in the Fibrosis Liver of Rabbits[J]. Chinese Journal of Biomedical Engineering, 2019, 38(6): 726-732.
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