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.

Tilapia (Oreochromis niloticus), which is extensively farmed globally and ranks as the second most cultivated fish in the Philippines, generates significant amounts of waste that are often underutilized. One specific type of waste material consists of fish heads, which contain a valuable source of extracellular matrix (ECM). This study aims to evaluate the effects of sonication as a viable decellularization method for the extraction of ECM from tilapia fish heads. Particularly, two treatments were tested on the head samples: sonication-assisted decellularization (dWS) using a water bath sonicator, and decellularization without sonication (dNS), each with different contact times (5 min and 10 min). Histological analysis with H and E staining and DNA quantification revealed that sonication-assisted samples (dWS) showed a greater reduction in basophilic components and DNA content, achieving a 93.7% removal rate. These dWS samples also had the highest protein loss, retaining only 33.86% of the original protein. SDS–PAGE analysis indicated that both dWS and dNS samples maintained similar collagen structures, as evidenced by identical subunit bands. ATR–FTIR spectra confirmed the presence of collagen type I in all samples, detecting characteristic amides A, B, I, II, and III. The results revealed that varying treatments and contact times had significant effects on the physical and mechanical properties of the decellularized extracellular matrix (ECM). These findings highlight the effectiveness of sonication in the decellularization process, particularly for utilizing waste tilapia heads.

Tilapia, the second most extensively farmed fish in the Philippines, generates significant amounts of waste that are often underutilized, leading to environmental concerns. One specific waste material is tilapia heads, which contain a valuable extracellular matrix (ECM) and an abundant source of bioactive materials that have a potential application in various fields, including biomaterials for wound healing and nutraceuticals. This study aims to evaluate the effects of sonication as a viable decellularization method for extracting ECM from tilapia fish heads. Histological analyses, employing H&E staining, revealed a higher reduction in basophilic components in the sonication-assisted (dWS) samples compared to those without sonication (dNS), as confirmed by DNA quantification. Notably, dWS samples subjected to 1% SDS sonication for 10 minutes exhibited the lowest DNA content with a removal rate of 93.7%. However, the same dWS sample also manifested the highest protein removal with a retained protein of 33.86%. The SDS-PAGE analysis of both dWS and dNS samples exhibited a uniform pattern, featuring identical four subunit bands—namely, the β-dimer (225-227 kDa), γ bands (228-230 kDa), and two alpha chains (α1 and α2 chains) (116-120 kDa)—indicating that sonication did not introduce discernible alterations in the helical structure of collagen. ATR-FTIR spectra further confirmed the presence of collagen as indicated by the detection of collagen type I indicators, including amides A, B, I, II, and III, in all decellularized samples. The above findings highlight the potential of sonication to aid in the decellularization of waste tilapia heads with future research targeting practical applications in wound healing, nutraceuticals, and sustainable environmental preservation through waste utilization.