Sterilization using Electrolyzed Water Highly Retains the Biological Properties in Tissue-Engineered Porcine Liver Scaffold
Abstract
Purpose: The aims of this study were to investigate the effects of sterilization with peracetic acid (PAA) and ethanol on the biological activity of porcine liver scaffolds and to develop a new technique for sterilization using slightly acidic electrolyzed water (SAEW). Methods: Decellularization of liver slices was performed using 0.1% sodium-dodecyl-sulfate, then evaluated by histological and polymerase chain reaction analyses. Decellularized slices were treated with either PAA or ethanol or SAEW, and then DNA content was quantified. We determined sterilization efficiency by culturing scaffolds in culture medium and on blood agar. We next analyzed the glycosaminoglycan and collagen contents of the scaffolds. Finally, we tested the cytotoxicity of the scaffolds as well as the effects of sterilization on host cell attachment and proliferation. Results: Complete cell and antigenic epitopes removal emphasized the decellularization efficiency. PAA and SAEW treatments achieved the highest efficiency of sterilization compared to that of the ethanol treated scaffolds, and were able to remove a considerable fraction of DNA from decellular-ized livers. The retained glycosaminoglycan content decreased in all treatments in the following order: SAEW, ethanol, and PAA. Ethanol caused a significant loss in collagen content compared to the other groups. A cytotoxicity evaluation revealed that all scaffolds were nontoxic. SAEW-treated scaffolds supported cell attachment and proliferation at a significantly higher rate than other groups. Conclusions: These data suggest that SAEW is highly efficient for sterilizing scaffolds and allowed the scaffolds to retain their bioactivity in addition to its high efficiency for cell remnant removal.
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Ethanol-based decellularization methods have drawbacks including tissue fixation, damage to ECM ultrastructure, and precipitation of cellular proteins.
“However, ethanol-based decellularization methods have drawbacks, including tissue fixation, damage to the ECM ultrastructure, and the precipitation of cellular proteins [104,112,113].”
From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [104]
Hussein et al. observed that PAA and SAEW treatments yielded the highest sterilization efficiency and effectively removed a considerable amount of DNA from decellularized livers.
“They observed that PAA and SAEW treatments yielded the highest sterilization efficiency and effectively removed a considerable amount of DNA from decellularized livers [104].”
From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [104]
SAEW retained more GAGs compared to other sterilization methods.
“SAEW also retained more GAGs compared to other sterilization methods.”
From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [104]
In our previous study, both SAEW and PAA effectively sterilized decellularized pig liver slices without affecting collagen, whereas ethanol decreased collagen amount and did not efficiently sterilize the scaffold.
“In one of our previous studies, we compared between 0.1 % PAA, 70 % ethanol, and slightly acidic electrolyzed water for sterilizing decellularized pig liver slices for 2 h [104]. Both SAEW and PAA effectively sterilized the scaffold without affecting the collagen, whereas ethanol decreased the collagen amount and did not efficiently sterilize the scaffold upon incubation in DMEM supplemented with”
From Liver tissue engineering using decellularized scaffolds: Current progress, challenges, and opportunities (2024) · cites this paper as [104]
Cites
The use of whole organ decellularization for the generation of a vascularized liver organoid
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Rana, D., Zreiqat, H., Benkirane-Jessel, N., Ramakrishna, S., & Ramalingam, M. (2017) · Journal of Tissue Engineering and Regenerative Medicine · doi:10.1002/term.2061 · Cited by 243 · see citing works
Biocompatibility evaluation of tissue-engineered decellularized scaffolds for biomedical application
Hussein, K. H., Park, K.-M., Kang, K.-S., & Woo, H.-M. (2016) · Materials Science and Engineering: C · doi:10.1016/j.msec.2016.05.068 · Cited by 218 · see citing works
Self-citationTao, M., Ao, T., Mao, X., Yan, X., Javed, R., Hou, W., Wang, Y., Sun, C., Lin, S., Yu, T., et al. (2021) · Bioactive Materials · doi:10.1016/j.bioactmat.2021.02.010 · Cited by 119 · see citing works
A sterilization method for decellularized xenogeneic cardiovascular scaffolds
Fidalgo, C., Iop, L., Sciro, M., Harder, M., Mavrilas, D., Korossis, S., Bagno, A., Palù, G., Aguiari, P., & Gerosa, G. (2018) · Acta Biomaterialia · doi:10.1016/j.actbio.2017.11.035 · Cited by 71 · see citing works
Hussein, K. H., Ahmadzada, B., Correa, J. C., Sultan, A., Wilken, S., Amiot, B., & Nyberg, S. L. (2024) · Bioactive Materials · doi:10.1016/j.bioactmat.2024.06.001 · Cited by 28 · see citing works
“In one of our previous studies, we compared between 0.1 % PAA, 70 % ethanol, and slightly acidic electrolyzed water for sterilizing decellularized pig liver slices for 2 h [104].”
BackgroundMethodInfluentialSelf-citation
- Authors
- Hussein, K. H., Park, K. M., Teotia, P. K., Hong, S. H., Yang, S. R., Park, S. M., Ahn, C., & Woo, H. M.
- Affiliations
- Kangwon National University (KR)
- Brigham and Women's Hospital (US)
- Corresponding author
- Woo, Heung M.
- Venue
- The International Journal of Artificial Organs
- Publisher
- SAGE Publications
- Published
- 2013-02-10
- Volume
- 36
- Issue
- 11
- Pages
- 781-792
- ISSN
- 0391-3988 (ISSN-L)
- 1724-6040
- OpenAlex
- W2091374305
- Cited by
- 34
- References
- 1
- Citations by year
- 2025: 4 · 2024: 5 · 2023: 3 · 2022: 1 · 2021: 3 · 2020: 1 · 2019: 2 · 2018: 4 · 2017: 3 · 2016: 2 · 2015: 5 · 2014: 1
- FWCI
- 2.62
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- The International Journal of Artificial Organs (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Life Sciences › Immunology and Microbiology › Microbiology › Medical Device Sterilization and Disinfection
- Physical Sciences › Engineering › Biomedical Engineering › Bone Tissue Engineering Materials
- Keywords
- sterilization, electrolyzed water, peracetic acid, ethanol, porcine liver, decellularization, glycosaminoglycans, collagen, cytotoxicity, cell attachment, cell proliferation, DNA removal
- MeSH terms
- Animals, Humans, Liver, Sterilization, Swine, Water, Tissue Engineering, Tissue Scaffolds
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