Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (Oreochromis niloticus) Viscera

Tilapia, a widely farmed aquaculture fish, produces substantial waste, including viscera that contain extracellular matrix (ECM) utilized as a biomaterial for tissue regeneration applications. Extracting ECM from viscera requires a specific decellularization method, as no standardized protocol exist...

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Main Authors: Jemwel Aron, Ronald Bual, Johnel Alimasag, Fernan Arellano, Lean Baclayon, Zesreal Cain Bantilan, Gladine Lumancas, Michael John Nisperos, Marionilo Labares, Kit Dominick Don Valle, Hernando Bacosa
Format: Article
Language:English
Published: Wiley 2024-01-01
Series:International Journal of Biomaterials
Online Access:http://dx.doi.org/10.1155/2024/6148496
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author Jemwel Aron
Ronald Bual
Johnel Alimasag
Fernan Arellano
Lean Baclayon
Zesreal Cain Bantilan
Gladine Lumancas
Michael John Nisperos
Marionilo Labares
Kit Dominick Don Valle
Hernando Bacosa
author_facet Jemwel Aron
Ronald Bual
Johnel Alimasag
Fernan Arellano
Lean Baclayon
Zesreal Cain Bantilan
Gladine Lumancas
Michael John Nisperos
Marionilo Labares
Kit Dominick Don Valle
Hernando Bacosa
author_sort Jemwel Aron
collection DOAJ
description Tilapia, a widely farmed aquaculture fish, produces substantial waste, including viscera that contain extracellular matrix (ECM) utilized as a biomaterial for tissue regeneration applications. Extracting ECM from viscera requires a specific decellularization method, as no standardized protocol exists. This study performed three decellularization methods: sonication, orbital shaking at room temperature, and agitation at 4°C, using SDS and TX100 at concentrations of 0.1% and 0.3%. The effectiveness of each method was assessed through H&E staining, dsDNA quantification, and SEM imaging to verify cellular content removal and ECM structure preservation. Additional analyses, including ATR-FTIR, SDS-PAGE, protein quantification, HPLC, and detergent residue tests, were performed to examine functional groups, collagen composition, protein content, amino acid profiles, and detergent residues in the decellularized samples. The results of H&E staining showed a significant reduction in cellular components in all samples, which was confirmed through DNA quantification. Sonication with 0.3% SDS achieved the highest DNA removal rate (96.5 ± 1.1%), while SEM images revealed that agitation at 4°C with 0.3% TX100 better preserved ECM structure. Collagen was present in all samples, as confirmed by ATR-FTIR analysis, which revealed pronounced spectral peaks in the amide I, II, III, A, and B regions. Samples treated with agitation at 4°C using 0.1% SDS exhibited the highest protein content (875 ± 15 µg/mg), whereas those treated with TX100 had lower detergent residue. Overall, the decellularization methods effectively reduced DNA content while preserving ECM structure and components, highlighting the potential of tilapia viscera as bioscaffolds and offering insights into utilizing fish waste for high-value products.
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spelling doaj-art-ef859371f16243b7bd22de07e7e96f092025-08-20T03:21:13ZengWileyInternational Journal of Biomaterials1687-87952024-01-01202410.1155/2024/6148496Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (Oreochromis niloticus) VisceraJemwel Aron0Ronald Bual1Johnel Alimasag2Fernan Arellano3Lean Baclayon4Zesreal Cain Bantilan5Gladine Lumancas6Michael John Nisperos7Marionilo Labares8Kit Dominick Don Valle9Hernando Bacosa10Environmental Science Graduate Program-Department of Biological SciencesCenter for Sustainable PolymersCenter for Sustainable PolymersEnvironmental Science Graduate Program-Department of Biological SciencesEnvironmental Science Graduate Program-Department of Biological SciencesCenter for Sustainable PolymersEnvironmental Science Graduate Program-Department of Biological SciencesEnvironmental Science Graduate Program-Department of Biological SciencesCenter for Sustainable PolymersCenter for Sustainable PolymersEnvironmental Science Graduate Program-Department of Biological SciencesTilapia, a widely farmed aquaculture fish, produces substantial waste, including viscera that contain extracellular matrix (ECM) utilized as a biomaterial for tissue regeneration applications. Extracting ECM from viscera requires a specific decellularization method, as no standardized protocol exists. This study performed three decellularization methods: sonication, orbital shaking at room temperature, and agitation at 4°C, using SDS and TX100 at concentrations of 0.1% and 0.3%. The effectiveness of each method was assessed through H&E staining, dsDNA quantification, and SEM imaging to verify cellular content removal and ECM structure preservation. Additional analyses, including ATR-FTIR, SDS-PAGE, protein quantification, HPLC, and detergent residue tests, were performed to examine functional groups, collagen composition, protein content, amino acid profiles, and detergent residues in the decellularized samples. The results of H&E staining showed a significant reduction in cellular components in all samples, which was confirmed through DNA quantification. Sonication with 0.3% SDS achieved the highest DNA removal rate (96.5 ± 1.1%), while SEM images revealed that agitation at 4°C with 0.3% TX100 better preserved ECM structure. Collagen was present in all samples, as confirmed by ATR-FTIR analysis, which revealed pronounced spectral peaks in the amide I, II, III, A, and B regions. Samples treated with agitation at 4°C using 0.1% SDS exhibited the highest protein content (875 ± 15 µg/mg), whereas those treated with TX100 had lower detergent residue. Overall, the decellularization methods effectively reduced DNA content while preserving ECM structure and components, highlighting the potential of tilapia viscera as bioscaffolds and offering insights into utilizing fish waste for high-value products.http://dx.doi.org/10.1155/2024/6148496
spellingShingle Jemwel Aron
Ronald Bual
Johnel Alimasag
Fernan Arellano
Lean Baclayon
Zesreal Cain Bantilan
Gladine Lumancas
Michael John Nisperos
Marionilo Labares
Kit Dominick Don Valle
Hernando Bacosa
Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (Oreochromis niloticus) Viscera
International Journal of Biomaterials
title Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (Oreochromis niloticus) Viscera
title_full Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (Oreochromis niloticus) Viscera
title_fullStr Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (Oreochromis niloticus) Viscera
title_full_unstemmed Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (Oreochromis niloticus) Viscera
title_short Effects of Various Decellularization Methods for the Development of Decellularized Extracellular Matrix from Tilapia (Oreochromis niloticus) Viscera
title_sort effects of various decellularization methods for the development of decellularized extracellular matrix from tilapia oreochromis niloticus viscera
url http://dx.doi.org/10.1155/2024/6148496
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