Characterization of Indonesia Decellularized Liver Cubes Scaffold using Scanning Electron Microscopy

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Abstract:

Liver biological scaffold was developed in order to resemble native liver tissue environment. It can be achieved by decellularizing native liver tissue that will remove cells and preserve extracellular matrix (ECM). Furthermore, ECM fibers are arranged in a special pattern, which affect liver cell polarity and topography that are important for cells’ implantation, proliferation and differentiation. Therefore, the aim of this study was to evaluate liver cube scaffold topography that was decellularized with fixed multiple sites syringe injection (Indonesia patent number: S00201907930).Rat liver cubes (n=3) underwent decellularization with Ethylene Glycol Tetraacetic Acid (EGTA) immersion and increased Sodium Dodecyl Sulfate (SDS) concentrations using previous multiple sites syringe injection protocol study. Deoxyribonucleic Acid (DNA) concentrations were measured to confirm less DNA materials remaining in scaffolds. Scanning Electron Microscope (SEM) analysis of scaffolds were conducted for topographic characterization compared to undecellularized liver control. Molecular analysis of DNA concentration showed complete removal of DNA material. SEM analysis gave appearance of intact liver cube scaffold microarchitecture. Liver cubes decellularization using multiple sites syringe injection showed good topographic liver scaffold characterization.

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[1] Z. Heydari, M. Najimi, H. Mirzaei, A. Shpichka, M. Ruoss, Z. Farzaneh, L. Montazeri, A. Piryaei, P. Timashev, R. Gramignoli, Tissue engineering in liver regenerative medicine: insights into novel translational technologies, Cells. 9 (2020) 304.

DOI: 10.3390/cells9020304

Google Scholar

[2] G. Mazza, W. Al‐Akkad, K. Rombouts, M. Pinzani, Liver tissue engineering: From implantable tissue to whole organ engineering, Hepatol. Commun. 2 (2018) 131–141.

DOI: 10.1002/hep4.1136

Google Scholar

[3] P. Maghsoudlou, F. Georgiades, H. Smith, A. Milan, P. Shangaris, L. Urbani, S.P. Loukogeorgakis, B. Lombardi, G. Mazza, C. Hagen, Optimization of liver decellularization maintains extracellular matrix micro-architecture and composition predisposing to effective cell seeding, PLoS One. 11 (2016) e0155324.

DOI: 10.1371/journal.pone.0155324

Google Scholar

[4] D. Habka, Mann D. Landes R, Soto-Gutierrez A. Future economics of liver transplantation: a 20-year cost modeling forecast and the prospect of bioengineering autologous liver grafts, PLoS One. 10 (2015) e0131764.

DOI: 10.1371/journal.pone.0131764

Google Scholar

[5] S. Caddeo, M. Boffito, S. Sartori, Tissue engineering approaches in the design of healthy and pathological in vitro tissue models, Front. Bioeng. Biotechnol. 5 (2017) 40.

DOI: 10.3389/fbioe.2017.00040

Google Scholar

[6] R.V. Langer, JP (1993).", Tissue Eng. Sci. 260 (n.d.) 920–926.

Google Scholar

[7] A. Gilpin, Y. Yang, Decellularization strategies for regenerative medicine: from processing techniques to applications, Biomed Res. Int. 2017 (2017).

DOI: 10.1155/2017/9831534

Google Scholar

[8] G. Mazza, W. Al-Akkad, A. Telese, L. Longato, L. Urbani, B. Robinson, A. Hall, K. Kong, L. Frenguelli, G. Marrone, O. Willacy, M. Shaeri, A. Burns, M. Malago, J. Gilbertson, N. Rendell, K. Moore, D. Hughes, I. Notingher, G. Jell, A. Del Rio Hernandez, P. De Coppi, K. Rombouts, M. Pinzani, Rapid production of human liver scaffolds for functional tissue engineering by high shear stress oscillation-decellularization., Sci. Rep. 7 (2017) 5534. https://doi.org/10.1038/s41598-017-05134-1.

DOI: 10.1038/s41598-017-05134-1

Google Scholar

[9] R. Grant, D. Hay, A. Callanan, From scaffold to structure: the synthetic production of cell derived extracellular matrix for liver tissue engineering, Biomed. Phys. Eng. Express. 4 (2018) 65015.

DOI: 10.1088/2057-1976/aacbe1

Google Scholar

[10] G. Mazza, K. Rombouts, A.R. Hall, L. Urbani, T.V. Luong, W. Al-Akkad, L. Longato, D. Brown, P. Maghsoudlou, A.P. Dhillon, Decellularized human liver as a natural 3D-scaffold for liver bioengineering and transplantation, Sci. Rep. 5 (2015) 1–15.

DOI: 10.1038/srep13079

Google Scholar

[11] P.M. Crapo, T.W. Gilbert, S.F. Badylak, An overview of tissue and whole organ decellularization processes, Biomaterials. 32 (2011) 3233–3243.

DOI: 10.1016/j.biomaterials.2011.01.057

Google Scholar

[12] R.D. Antarianto, A.A.A.A.P. Dewi, A. Pragiwaksana, J.A. Pawitan, Decellularization of liver cubes using multiple site syringe injection for generating native liver scaffold: Preliminary report, in: AIP Conf. Proc., AIP Publishing LLC, 2019: p.40005.

DOI: 10.1063/1.5139367

Google Scholar

[13] F. Mußbach, U. Settmacher, O. Dirsch, C. Xie, U. Dahmen, Bioengineered livers: a new tool for drug testing and a promising solution to meet the growing demand for donor organs, Eur. Surg. Res. 57 (2016) 224–239.

DOI: 10.1159/000446211

Google Scholar

[14] E. Moc, J. Thai, Scanning Electron Microscopy and Histology Imaging and Analysis of Decellularized Porcine Vessel, (2012).

Google Scholar

[15] M. Lorvellec, F. Scottoni, C. Crowley, R. Fiadeiro, P. Maghsoudlou, A.F. Pellegata, F. Mazzacuva, A. Gjinovci, A.-M. Lyne, J. Zulini, Mouse decellularised liver scaffold improves human embryonic and induced pluripotent stem cells differentiation into hepatocyte-like cells, PLoS One. 12 (2017) e0189586.

DOI: 10.1371/journal.pone.0189586

Google Scholar

[16] M.M. Bobrova, L.A. Safonova, O.I. Agapova, M.E. Krasheninnikov, M.Y. Shagidulin, I.I. Agapov, Liver Tissue Decellularization as a Promising Porous Scaffold Processing Technology for Tissue Engineering and Regenerative Medicine., Med. Technol. Med. Tehnol. v Med. 7 (2015).

DOI: 10.17691/stm2015.7.4.01

Google Scholar

[17] E.S. Mirdamadi, D. Kalhori, N. Zakeri, N. Azarpira, M. Solati-Hashjin, Liver tissue engineering as an emerging alternative for liver disease treatment, Tissue Eng. Part B Rev. 26 (2020) 145–163.

DOI: 10.1089/ten.teb.2019.0233

Google Scholar