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We extracted magnesium-rich calcium phosphate bioceramics from tilapia bone using a gradient thermal treatment approach and investigated their chemical and physicochemical properties. X-ray diffraction showed that tilapia fish bone-derived hydroxyapatite (FHA) was generated through the first stage of thermal processing at 600-800 °C. Using FHA as a precursor, fish bone biphasic calcium phosphate (FBCP) was produced after the second stage of thermal processing at 900-1200 °C. The beta-tricalcium phosphate content in the FBCP increased with an increasing calcination temperature. The fact that the lattice spacing of the FHA and FBCP was smaller than that of commercial hydroxyapatite (CHA) suggests that Mg-substituted calcium phosphate was produced via the gradient thermal treatment. Both the FHA and FBCP contained considerable quantities of magnesium, with the FHA having a higher concentration. In addition, the FHA and FBCP, particularly the FBCP, degraded faster than the CHA. After one day of degradation, both the FHA and FBCP released Mg2+, with cumulative amounts of 4.38 mg/L and 0.58 mg/L, respectively. Furthermore, the FHA and FBCP demonstrated superior bone-like apatite formation; they are non-toxic and exhibit better osteoconductive activity than the CHA. In light of our findings, bioceramics originating from tilapia bone appear to be promising in biomedical applications such as fabricating tissue engineering scaffolds.

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In this study, a calcium-binding peptide was obtained by hydrolyzing tilapia bone and its osteogenic activity was evaluated. Animal protease was selected from nine enzymes, and its hydrolysate was purified through preparative and semi-preparative reverse phase high-performance liquid chromatography. The purified peptide was identified as DGPSGPK (656.32 Da) and its calcium-binding capacity reached 111.98 µg/mg. The peptide calcium chelate (DGPSGPK-Ca) was obtained, and its structure was characterized through Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and mass spectrometry (MS). The results of XRD and SEM showed that DGPSGPK-Ca was formed as a new compound. The carboxyl and amino groups of Lys and Asp residues may be the chelating sites of DGPSGPK according to the FTIR and MS results. The molecular simulation showed the carbonyl groups of Asp, Pro, Ser, and Lys residues involved in the binding of calcium. The interaction of DGPSGPK and different integrins was evaluated by molecular docking simulation, and the main forces involved were electrostatic interaction forces, hydrogen bonding and hydrophobic interactions. Furthermore, DGPSGPK could inhibit the differentiation of osteoclast and promote the proliferation, differentiation and mineralization of osteoblasts.

RESUMEN

Background: Water pollution has become a major threat to the environment and the living so an eco-friendly bio-filter was chosen for its merits over conventional techniques. Aim: Investigating the purifying activities of the Tilapia bone powder against inorganics, heavy metals, and microbial water pollutants and its impacts on performance, biochemical and antioxidant levels, cortisol and immunoglobulin concentrations, and intestinal microbiota in challenged broiler chickens. Methods: The in-vitro activity of Tilapia bone powder was evaluated against magnesium chloride and lead nitrate using tube minimal inhibitory concentration (MIC), as well as against Escherichia coli O1527:H7, Salmonella Typhimurium, Mycoplasma gallisepticum, Aspergillus niger, Trichophyton mentagrophytes, and Candida albicans using a 96-micro-well MIC. A total of 250 1-day-old Hubbard chicks were divided into five groups on a deep litter system. Chicks were supplemented daily with Tilapia bone powder (1 g/l) for 4-6 hours from the 3rd day. Challenges were served on the 7th, 14th, 21st, 28th, and 35th days for four broiler groups using magnesium chloride (100 mg/l), lead nitrate (350 mg/l), E. coli (2.4 × 1012 CFU/ml), S. Typhimurium (1.8 × 108 CFU/ml), respectively, and the 5th group was assigned as a control. A total of 2,250 samples (90 Tilapia-pollutants mixes, 480 Tilapia-microbial mixes, 240 sera, 240 intestinal swabs, and 1,200 tissue samples) were collected. Results: Tilapia bone powder 1% reveals a 100% reduction in the lead after 1 hour, total and calcium hardness after 0.5 hours, as well as 100% killing efficacy against E. coli O1527:H7, S. Typhimurium, M. gallisepticum, A. niger, T. mentagrophytes, and C. albicans after 0.5, 1, 1, 1, 1, and 1 hour, respectively. Tilapia bone powder 1% treated water reveals highly significant (p < 0.01) increases in dissolved oxygen and declines in physicochemical and microbial parameters compared with tap water. Challenged treated broilers revealed highly significant (p < 0.01) increases in weight gains, performance index, body weights, carcasses, and organs weights, immunoglobulin concentrations, and antioxidant levels, as well as highly significant (p < 0.01) improvements in feed conversions, feed and water intakes, biochemical profile, cortisol hormone, and intestinal microbiota. Conclusion: Tilapia bone powder provided significant in-vitro adsorptive and antimicrobial actions, as well as supported the broiler chickens to mitigate the polluted water stress accompanied by enhanced performance, carcass quality, immunity, and intestinal microbiota.

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RESUMEN

In this study, we used reversed-phase high performance liquid chromatography (LC) to isolate three novel peptides with calcium-chelating capacity from tilapia bone collagen hydrolysate. Using LC-tandem mass spectrometry, we determined the amino acid sequences to be GPAGPHGPVG, FDHIVY, and YQEPVIAPKL. We then synthesized the three peptides and verified their calcium-chelating activity. Results showed that the calcium-chelating activity of GPAGPHGPVG, FDHIVY, and YQEPVIAPKL reached 18.80 ± 0.49, 35.73 ± 0.74, and 28.4 ± 0.94 mg/g, respectively. We next investigated how each peptide enhanced intestinal calcium absorption using Caco-2 cell monolayers. Compared with the control group, GPAGPHGPVG, FDHIVY, and YQEPVIAPKL potently enhanced calcium transport within 30 min by 89 ± 9, 202 ± 12, and 130 ± 7%, respectively. Results suggest that these peptides isolated from tilapia bone hydrolysate can be used as dietary supplements to increase calcium absorption.

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