Milkfish (Chanos chanos) is an abundant fish commodity in the Philippines that generates a large number of wastes such as skin, scales, viscera, and bones, which, upon disposal, cause environmental pollution. The abundance of these wastes, such as fish skin, rich in bioactive natural products such as collagen, elicits interest in their conversion into high-market-value products. The decellularization of milkfish skin waste can extract its extracellular matrix (ECM), a potential raw material for biomedical applications such as the repair of damaged skin tissues. In particular, this study characterized the developed decellularized ECM with different concentrations (0.1%, 1.0%) of the decellularizing agents (Triton X-100, SDS) and temperature (4 °C, room temperature) using milkfish skin. The decellularized ECM structure was better preserved using Triton X-100, while SDS was more effective in cell component removal, especially at 1% concentration and 4 °C temperature. There were significant effects of varying the temperatures and concentrations on the physical and mechanical properties of the decellularized ECM. Future studies could explore more variables to further establish protocols and more analyses to better characterize the decellularized milkfish skin.

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.

Tilapia (Oreochromis niloticus) is a widely cultivated fish in tropical and subtropical regions such as the Philippines, generating substantial waste during processing, including bones that are a valuable source of extracellular matrix (ECM). However, the extraction of ECM from fish bones requires an essential step of demineralization. This study aimed to assess the efficiency of tilapia bone demineralization using 0.5 N HCl at different time durations. By evaluating the residual calcium concentration, reaction kinetics, protein content, and extracellular matrix (ECM) integrity through histological analysis, composition assessment, and thermal analysis, the effectiveness of the process was determined. Results revealed that after 1 h of demineralization, the calcium and protein contents were 1.10 ± 0.12% and 88.7 ± 0.58 μg/mL, respectively. The study found that after 6 h, the calcium content was almost completely removed, but the protein content was only 51.7 ± 1.52 μg/mL compared to 109.0 ± 1.0 μg/mL in native bone tissue. Additionally, the demineralization reaction followed second-order kinetics with an R² value of 0.9964. Histological analysis using H&E staining revealed a gradual disappearance of the basophilic components and the emergence of lacunae, which can be attributed to decellularization and mineral content removal, respectively. As a result, organic components such as collagen remained in the bone samples. ATR-FTIR analysis showed that all demineralized bone samples retained collagen type I markers, including amide I, II, and III, amides A and B, and symmetric and antisymmetric CH₂ bands. These findings provide a route for developing an effective demineralization protocol to extract high-quality ECM from fish bones, which could have important nutraceutical and biomedical applications.

The exploration of alternative sources of extracellular matrix (ECM) is driven by increasing demand and the need for sustainable biomaterials. Tuna (Thunnus albacares) bones, a by-product of the fishing industry, represent a potential ECM source due to their abundance, collagen-rich composition, and biocompatibility. This study investigated the effects of demineralization using hydrochloric acid (HCl) at concentrations of 0.5 M and 1.0 M for varying durations (0.5, 1, 6, 12, and 24 h), and decellularization using sodium dodecyl sulfate (SDS) and Triton X-100 (TX100) at concentrations of 0.1%, 0.5%, and 1.0%. Demineralization effectively reduced inorganic content, with residual calcium levels dropping below 5% after 6 h. Kinetic analysis indicated a second-order reaction, and ATR–FTIR spectra confirmed the disappearance of phosphate and carbonate peaks alongside the preservation of amide bands. Demineralized bone matrix (dBM) retained good thermal stability and mechanical properties. Decellularization efficiency was assessed through H&E staining, dsDNA quantification, and SDS-PAGE analysis. Decellularized extracellular bone matrix (dEBM) treated with 1.0% SDS showed the lowest dsDNA levels (14.00 ± 7.94 ng/mg) and absence of cellular material. SDS-PAGE confirmed type I collagen preservation, particularly in samples treated with ≤0.5% SDS or TX100. This study establishes that tuna fish bones can be effectively processed into ECM, supporting their potential as a sustainable biomaterial for advanced biomedical applications.