In-Vitro Biocompatibility And Osteogenic Potential Of Synthetic Calcium Phosphate Apatite

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dc.contributor.authorIbrahim, Shirin
dc.date.accessioned2017-11-09T01:13:47Z
dc.date.available2017-11-09T01:13:47Z
dc.date.issued2014-07
dc.description.abstractSynthetic calcium phosphate apatite (CaP) has unique properties such as biocompatible, bioresorbable and the ability to promote bone formation. Among the bioceramics, CaP has excellent osteoconductivity, bioactivity and has ability to form a strong bone-cell interface by excellent cell bonding onto the material. It has been used widely in non- or low-load bearing applications due to its low mechanical properties. The purpose of this study was to evaluate the biocompatibility and osteogenesis between two different CaP, the triphasic calcium phosphate apatite (TCaP) which is locally synthesised using commercial chemical and calcium phosphate bone replacement product, MIMIX™ (CMM) which is commercially available. Two types of cells were used as an in vitro model. A model for biocompatibility evaluation is L929 mouse fibroblast cells, while human osteoblast cells for osteogenic potential studies. According to the ISO 10993-5:2009(E) Biological evaluation of medical devices – Part 5: Tests for in vitro cytotoxicity, one of the recommended cell lines is CCL-1 (NCTC clone 929 - American Type Culture Collection (ATCC)). The human osteoblast cell line is the one of the appropriate cell to evaluate the functioning properties on osteogenic potential of the material as the material will be a bone graft and implanted into the bone. Three different methods were applied for biocompatibility evaluation. Cytotoxicity study was performed using MTT assay. Both TCaP and CMM extract were non toxic on L929 cells. The levels in MTT assay of both materials were more than 70% cell viability at the highest concentration (200 mg/ml). Cell proliferation conducted using Trypan Blue Exclusion Assay (TBEA) showed CMM extract proceeded more significantly with unexposed cell, than on TCaP extract. Meanwhile cellular morphology observation on L929 cells for both materials were viable, excellently differentiated, proliferated and grew well at day 7 of incubation. Cell density of TCaP material was observed to be higher compared to CMM on day 5 and day 7 with a direct contact method using VPSEM. These results showed TCaP and CMM were non toxic and biocompatible on L929 cells. Osteogenenic potential evaluation was performed using Immunoblot analysis with ALP, BSP and Col I as osteoblastic differentiation markers. The analysis of bone formation at protein level showed there were expressions of ALP and BSP on day 3 and day 7 of incubation with TCaP, CMM and biological apatite-dried human bone (DHB) extract on osteoblast cell. There was no expression from Col I as it was undetectable in all materials and unexposed cell. In conclusion, TCaP and CMM are biocompatible and have the ability to promote osteoblastic differentiation of human osteoblast. These results suggested TCaP may be used as bone filler and bone replacement for bone regeneration or drug delivery agent.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/5254
dc.language.isoenen_US
dc.publisherUniversiti Sains Malaysiaen_US
dc.subjectSynthetic calcium phosphate apatiteen_US
dc.subjectpromote bone formationen_US
dc.titleIn-Vitro Biocompatibility And Osteogenic Potential Of Synthetic Calcium Phosphate Apatiteen_US
dc.typeThesisen_US
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