The effects of biphasic calcium phosphate and simvastatin on human dental pulp cells in vitro on dentin regeneration

Abdul Qader Al-Kukasih, Sarah Talib (2015) The effects of biphasic calcium phosphate and simvastatin on human dental pulp cells in vitro on dentin regeneration. Masters thesis, Universiti Sains Malaysia.

PDF - Submitted Version
Download (589kB) | Preview


This study was conducted to prepare an appropriate biphasic calcium phosphate (BCP) scaffold in combination with optimal concentration of simvastatin to induce human dental pulp cells differentiation and dentin tissue regeneration. BCP scaffold of 20/80 hydroxyapatite (HA) to β-tricalcium phosphate (β-TCP) ratio with micropores < 10 μm, macropores of 300 μm, and porosity of 65 % was successfully synthesized using wet precipitation method and polyethylene microspheres as pore-creating agents. The scaffold was characterized using x-ray diffraction (XRD), fourier transform infra-red spectroscopy (FTIR), x-ray fluorescence (XRF), field emission scanning electron microscope (FESEM), linear shrinkage, and total porosity measurements. BCP sample extract was prepared and combined with four different concentrations of simvastatin (2.0, 1.5, 1.0 and 0.5 μM) for the assessment of cell viability using MTT (3-(4,5- dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide) assay. The alkaline phosphatase activity was assessed using alkaline phosphatase assay. The groups which showed the best cell viability and alkaline phosphatase activity were selected. They were assessed for both odontogenic differentiation potential of collagen type I alpha 1 (COL1A1), bone sialoprotein (BSP), dentin matrix protein-1 (DMP-1), dentin sialophosphoprotein (DSPP), and Runt-related transcription factor-2 (RUNX-2) genes expression analysis using reverse transcription-polymerase chain reaction (RT-PCR) and extracellular matrix mineralization detection using Alizarin Red S staining. The results showed that combination groups of BCP +1.5 μM, BCP + 1.0 μM, and BCP + 0.5 μM had higher mean cell viability index, except BCP + 2.0 μM that showed cytotoxicity. For alkaline phosphatase activity, the combination groups of BCP + 1.5 μM, BCP +1.0 μM, and BCP + 0.5 μM showed higher mean alkaline phosphatase activity index with BCP + 1.5 μM the highest. For the odontogenic differentiation potential, BCP + 1.5 μM combination group showed up-regulation of COL1A1, DMP-1, BSP, and DSPP genes and down-regulation of RUNX-2 gene. For the extracellular matrix mineralization, BCP + 1.5 μM combination group showed the highest ability to induce the mineral deposition. In conclusion, the combination of BCP and 1.5 μM simvastatin achieved together a preferable induction of human dental pulp cells differentiation toward dentin tissue regeneration.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Dental pulp
Subjects: R Medicine > RK Dentistry
Divisions: Kampus Kesihatan (Health Campus) > Pusat Pengajian Sains Perubatan (School of Medical Sciences) > Thesis
Depositing User: Mr Abdul Hadi Mohammad
Date Deposited: 10 Feb 2019 07:50
Last Modified: 10 Feb 2019 07:50

Actions (login required)

View Item View Item