Modeling of human liver development, especially cellular organization and the mechanisms underlying it, is fundamental for studying liver organogenesis and congenital diseases, yet there are no reliable models that mimic these processes ex vivo.
Design:
Using an organ engineering approach and relevant cell lines, we designed a perfusion system that delivers discrete mechanical forces inside an acellular liver extracellular matrix scaffold to study the effects of mechanical stimulation in hepatic tissue organization.
Results:
We observed a fluid flow rate-dependent response in cell distribution within the liver scaffold. Next, we determined the role of nitric oxide (NO) as a mediator of fluid flow effects on endothelial cells. We observed impairment of both neovascularization and liver tissue organization in the presence of selective inhibition of endothelial NO synthase. Similar results were observed in bioengineered livers grown under static conditions.
Conclusion:
Overall, we were able to unveil the potential central role of discrete mechanical stimulation through the NO pathway in the revascularization and cellular organization of a bioengineered liver. Last, we propose that this organ bioengineering platform can contribute significantly to the identification of physiological mechanisms of liver organogenesis and regeneration and improve our ability to bioengineer livers for transplantation.
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References
1.
MatsumotoK., YoshitomiH., RossantJ., and ZaretK.S.Liver organogenesis promoted by endothelial cells prior to vascular function. Science, 294, 559, 2001.
2.
LammertE., CleaverO., and MeltonD.Induction of pancreatic differentiation by signals from blood vessels. Science, 294, 564, 2001.
3.
LammertE., CleaverO., and MeltonD.Role of endothelial cells in early pancreas and liver development. Mech Dev, 120, 59, 2003.
4.
AbshagenK., EipelC., and VollmarB.A critical appraisal of the hemodynamic signal driving liver regeneration. Langenbecks Arch Surg, 397, 579, 2012.
5.
SatoY., KoyamaS., TsukadaK., and HatakeyamaK.Acute portal hypertension reflecting shear stress as a trigger of liver regeneration following partial hepatectomy. Surg Today, 27, 518, 1997.
6.
SatoY., TsukadaK., and HatakeyamaK.Role of shear stress and immune responses in liver regeneration after a partial hepatectomy. Surg Today, 29, 1, 1999.
BaptistaP.M., SiddiquiM.M., LozierG., RodriguezS.R., AtalaA., and SokerS.The use of whole organ decellularization for the generation of a vascularized liver organoid. Hepatology, 53, 604, 2011.
9.
Soto-GutierrezA., ZhangL., MedberryC., FukumitsuK., FaulkD., JiangH., ReingJ., et al.A whole-organ regenerative medicine approach for liver replacement. Tissue Eng Part C Methods, 17, 677, 2011.
10.
BaoJ., ShiY., SunH., YinX., YangR., LiL., ChenX., et al.Construction of a portal implantable functional tissue-engineered liver using perfusion-decellularized matrix and hepatocytes in rats. Cell Transplant, 20, 753, 2011.
11.
UygunB., Soto-GutierrezA., YagiH., IzamisM., GuzzardiM., ShulmanC., MilwidJ., et al.Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med, 16, 814, 2010.
12.
BarakatO., AbbasiS., RodriguezG., RiosJ., WoodR.P., OzakiC., HolleyL.S., et al.Use of decellularized porcine liver for engineering humanized liver organ. J Surg Res, 173, E11, 2012.
13.
YagiH., FukumitsuK., FukudaK., KitagoM., ShinodaM., ObaraH., ItanoO., et al.Human-scale whole-organ bioengineering for liver transplantation: a regenerative medicine approach. Cell Transplantat, 22, 231, 2013.
14.
PollackG.M., BrouwerK.L., DembyK.B., and JonesJ.A.Determination of hepatic blood flow in the rat using sequential infusions of indocyanine green or galactose. Drug Metab Dispos, 18, 197, 1990.
15.
DaviesB., and MorrisT.Physiological parameters in laboratory animals and humans. Pharm Res, 10, 1093, 1993.
16.
MoranE.C., BaptistaP.M., EvansD.W., SokerS., and SparksJ.L.Evaluation of parenchymal fluid pressure in native and decellularized liver tissue. Biomed Sci Instrum, 48, 303, 2012.
17.
FukuchiT., HiroseH., OnitsukaA., HayashiM., SengaS., ImaiN., ShibataM., et al.Effects of portal-systemic shunt following 90% partial hepatectomy in rats. J Surg Res, 89, 126, 2000.
18.
BaptistaP.M., VyasD., MoranE., WangZ., and SokerS.Human liver bioengineering using a whole liver decellularized bioscaffold. Methods Mol Biol, 1001, 289, 2013.
19.
YangY.Z., YangY.S., XuY., LickS.D., AwasthiY.C., and BoorP.J.Endothelial glutathione-S-transferase A4-4 protects against oxidative stress and modulates NOS expression through NF-kappa B translocation. Toxicol Appl Pharmacol, 230, 187, 2008.
20.
WangL., YangY., DwivediS., XuY., ChuE.T., LiJ., FitchettK., et al.Manipulating glutathione-S-transferases may prevent the development of tolerance to nitroglycerin. Cardiovasc Toxicol, 6, 131, 2006.
21.
HuchM., GehartH., van BoxtelR., HamerK., BlokzijlF., VerstegenM.M.A., EllisE., et al.Long-term culture of genome-stable bipotent stem cells from adult human liver. Cell, 160, 299, 2015.
22.
KnowlesB.B., HoweC.C., and AdenD.P.Human hepatocellular-carcinoma cell-lines secrete the major plasma-proteins and hepatitis-B surface-antigen. Science, 209, 497, 1980.
23.
SchoenJ.M., WangH.H., MinukG.Y., and LauttW.W.Shear stress-induced nitric oxide release triggers the liver regeneration cascade. Nitric Oxide, 5, 453, 2001.
24.
Schoen SmithJ.M., and LauttW.W.The role of prostaglandins in triggering the liver regeneration cascade. Nitric Oxide, 13, 111, 2005.
25.
ColganS.P., TaylorC.T., NarravulaS., SynnestvedtK., and BlumeE.D.Endothelial COX-2 induction by hypoxia liberates 6-keto-PGF1 alpha, a potent epithelial secretagogue. Adv Exp Med Biol, 507, 107, 2002.
26.
InoueH., TabaY., MiwaY., YokotaC., MiyagiM., and SasaguriT.Induction of cyclooxygenase-2 expression by fluid smear stress in vascular endothelial cells. Adv Exp Med Biol, 525, 141, 2003.
27.
WuG., LuoJ., RanaJ.S., LahamR., SellkeF.W., and LiJ.Involvement of COX-2 in VEGF-induced angiogenesis via P38 and JNK pathways in vascular endothelial cells. Cardiovasc Res, 69, 512, 2006.
28.
ZhuZ., and HuangfuD.Human pluripotent stem cells: an emerging model in developmental biology. Development, 140, 705, 2013.
29.
TakebeT., SekineK., EnomuraM., KoikeH., KimuraM., OgaeriT., ZhangR.-R., et al.Vascularized and functional human liver from an iPSC-derived organ bud transplant. Nature, 499, 481, 2013.
30.
Collardeau-FrachonS., and ScoazecJ.-Y.Vascular development and differentiation during human liver organogenesis. Anat Rec, 291, 614, 2008.
31.
UcuzianA.A., and GreislerH.P.In vitro models of angiogenesis. World J Surg, 31, 654, 2007.
32.
MorinK.T., and TranquilloR.T.In vitro models of angiogenesis and vasculogenesis in fibrin gel. Exp Cell Res, 319, 2409, 2013.
33.
CantreD., SchuettH., HildebrandtA., DoldS., MengerM.D., VollmarB., and EipelC.Nitric oxide reduces organ injury and enhances regeneration of reduced-size livers by increasing hepatic arterial flow. Br J Surg, 95, 785, 2008.
34.
MeiY., and ThevanantherS.Endothelial nitric oxide synthase is a key mediator of hepatocyte proliferation in response to partial hepatectomy in mice. Hepatology, 54, 1777, 2011.
35.
Diaz-JuarezJ., and Hernandez-MunozR.The role of calcium and nitric oxide during liver enzyme release induced by increased physical forces as evidenced in partially hepatectomized rats. Liver Transplant, 17, 334, 2011.
36.
YoshidaD., AkahoshiT., KawanakaH., YamaguchiS., KinjoN., TaketomiA., TomikawaM., et al.Roles of vascular endothelial growth factor and endothelial nitric oxide synthase during revascularization and regeneration after partial hepatectomy in a rat model. Surg Today, 41, 1622, 2011.
37.
Garcia-TrevijanoE.R., Martinez-ChantarM.L., LatasaM.U., MatoJ.M., and AvilaM.A.NO sensitizes rat hepatocytes to proliferation by modifying S-adenosylmethionine levels. Gastroenterology, 122, 1355, 2002.
38.
TanakaY., YamatoM., OkanoT., KitamoriT., and SatoK.Evaluation of effects of shear stress on hepatocytes by a microchip-based system. Meas Sci Technol, 17, 3167, 2006.
39.
AnadaT., FukudaJ., SaiY., and SuzukiO.An oxygen-permeable spheroid culture system for the prevention of central hypoxia and necrosis of spheroids. Biomaterials, 33, 8430, 2012.
40.
HaC.H., KimS., ChungJ., AnS.H., and KwonK.Extracorporeal shock wave stimulates expression of the angiogenic genes via mechanosensory complex in endothelial cells: mimetic effect of fluid shear stress in endothelial cells. Int J Cardiol, 168, 4168, 2013.
41.
SearlesC.D.Transcriptional and posttranscriptional regulation of endothelial nitric oxide synthase expression. Am J Physiol Cell Physiol, 291, C803, 2006.
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