Nazir, Nurshuhada Mohd
(2016)
Biocompatibility and osteogenesis of locally produced B-tricalcium phosphate.
Masters thesis, Universiti Sains Malaysia.
Abstract
β-Tricalcium Phosphate (β-TCP) with molecular formula [β-Ca3(PO4)2] is a synthetic ceramic biomaterial that have been extensively used as a bone graft material in orthopaedic and dental field due to its biocompatibility and properties. The purpose of this study was to evaluate the biocompatibility and osteogenesis of locally produced β-TCP in-vitro using osteoblast cell line. β-TCP ceramic was locally prepared using same starting material of Ca(OH)2 and H3PO4 with Ca/P ratio of 1.5 but synthesis by two different methods namely, hydrothermal and precipitation.
The characteristics of β-TCP samples were observed using vernier caliper, Fourier Transform Infrared Spectroscope (FTIR) and Atomic Force Microscope (AFM) for its dimension, functional group and surface roughness. Osteoblast cells were cultured with extracts of β-TCP at various concentration to determine the cytotoxicity using alamar blue and MTS assay. Cell attachment and viability of osteoblast cell with β- TCP were observed using Confocal Laser Scanning Microscope (CLSM) after staining with calcein-AM and ethidium homodimer to produce fluoresecence colour. The presence of calcium deposition and extracellular matrix (ECM) protein such as Collagen 1 and fibronectin that responsible during osteogenesis were determine using alizarin red staining and western blot analysis. Chronos β-TCP as a comercial produced β-TCP was used as controls.
Hydrothermal and precipitation β-TCP showed the similar characteristic in dimension, functional group and surface roughness. The results of in-vitro studies indicated the non-cytotoxicity of hydrothermal and precipitation β-TCP at the tested concentrations (6.25 mg/ml to 200 mg/ml) and images from optical microscopy showed that the cells were able to proliferate and retain its morphology after incubation with the extracts. Images from CLSM showed the viable cells had grown and able to attach on the surface of hydrothermal and precipitation β-TCP. Calcium deposition observation and ECM protein expression showed the positive osteogenesis from osteoblast cell cultured with hydrothermal and precipitation β-TCP.
The results showed that the locally produced β-TCP using hydrothermal or precipitation method are non-cytotoxic and biocompatible. This biomaterials support cells attachment and cells proliferation, expressing protein related to osteogenesis and have the potential to be used as biomaterial in dental and orthopedic.
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