Dermawan, Siti Khadijah (2023) Bioactivity (in vitro) of halal hydroxyapatite from black tilapia fish bone for biomedical applications. Masters thesis, Universiti Tun Hussein Onn Malaysia.

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Abstract

Hydroxyapatite (HAp), Ca10(PO4)6(OH)2, is extensively utilized in biomedical field because it mimics inorganic part of human bone and teeth. Synthetic HAp is difficult to produce and costly. Large volumes of by-product waste from fishery factories have a negative impact on the environment. Hence, this research has extracted the halal HAp from waste black tilapia fish bones, characterized the properties of that HAp powder and determined their bioactivity. The bones were calcined at 600 °C to 1000 °C with a heating rate of 10 °C/min. Characterized by using Thermogravimetric Analysis (TGA) for thermal stability, Scanning Electron Microscopy (SEM) for morphology, X-ray Diffraction (XRD) for mineralogy, Fourier-transform Infrared Spectroscopy (FTIR) for chemical bonding and Energy Dispersive Spectroscopy (EDX) for elemental analysis. The extracted HAp was immersed in a simulated body fluid (SBF) solution for 3, 7, and 14 days to evaluate their bioactivity. The Kirby- Bauer test assessed the antimicrobial behaviour of the samples against Staphylococcus aureus and Escherichia coli bacteria. The cytotoxicity effect of the sample was tested using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylte tetrazolium bromide (MTT) assay with human fetal osteoblast cells (hFOB 1.19). There are three stages of weight loss found by TGA which are dehydration, decomposition of organic compounds, and decarbonization. The derived HAp was compatible with standard HAp and the biphasic material was found at 900 and 1000 °C. Raw samples have denser and less porous microstructures than calcined samples, with grain size rising as temperature increases. Organic chemicals were gradually eliminated during the calcination process. The presence of magnesium and sodium showed as their trace elements and the Ca/P molar ratio is 1.63 (1000 °C). Then, apatite formed on the surface of pellet after the immersion in SBF. The extracted HAp does not possess any antimicrobial properties against both bacteria. The sample is non-toxic as the cell viability were 120.10 % and 162.62 % for Day 1 and Day 2 of incubation respectively. Thus, these findings have potential as biomaterial for biomedical applications

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