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

IF 3 Q3 MATERIALS SCIENCE, BIOMATERIALS International Journal of Biomaterials Pub Date : 2024-09-18 eCollection Date: 2024-01-01 DOI:10.1155/2024/6148496
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
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Abstract

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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各种脱细胞方法对开发罗非鱼(Oreochromis niloticus)内脏脱细胞细胞外基质的影响。
罗非鱼是一种广泛养殖的水产鱼类,会产生大量废弃物,包括含有细胞外基质(ECM)的内脏,可用作组织再生应用的生物材料。从内脏中提取 ECM 需要特定的脱细胞方法,因为目前还没有标准化的方案。本研究使用浓度为 0.1% 和 0.3% 的 SDS 和 TX100 进行了三种脱细胞方法:超声处理、室温下的轨道振荡和 4°C 下的搅拌。通过 H&E 染色、dsDNA 定量和 SEM 成像评估每种方法的效果,以验证细胞成分的去除和 ECM 结构的保留。此外,还进行了其他分析,包括 ATR-傅立叶变换红外光谱、SDS-PAGE、蛋白质定量、HPLC 和洗涤剂残留测试,以检查脱细胞样本中的功能群、胶原蛋白组成、蛋白质含量、氨基酸谱和洗涤剂残留。H&E 染色结果表明,所有样品中的细胞成分都明显减少,DNA 定量也证实了这一点。使用 0.3% SDS 进行超声处理的 DNA 去除率最高(96.5 ± 1.1%),而 SEM 图像显示,在 4°C 下使用 0.3% TX100 进行搅拌能更好地保存 ECM 结构。ATR-FTIR 分析证实,所有样品中都含有胶原蛋白,该分析在酰胺 I、II、III、A 和 B 区域都发现了明显的光谱峰。使用 0.1% SDS 在 4°C 下搅拌处理的样品蛋白质含量最高(875 ± 15 µg/mg),而使用 TX100 处理的样品洗涤剂残留量较低。总之,脱细胞方法有效地降低了 DNA 含量,同时保留了 ECM 结构和成分,凸显了罗非鱼内脏作为生物支架的潜力,并为利用鱼类废弃物生产高价值产品提供了启示。
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来源期刊
International Journal of Biomaterials
International Journal of Biomaterials MATERIALS SCIENCE, BIOMATERIALS-
CiteScore
4.30
自引率
3.20%
发文量
50
审稿时长
21 weeks
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