الفهرس 
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Abstract
This study was carried out to provide further evidence for the effect of recombinant human bone morphogenetic protein-7 (rhBMP-7) on the osteoblast-like cells cultured on implant by isolation of osteoclast cells and culturing them on titanium surfaces under specific conditions.
The present study was performed on four groups. The investigations were done in the laboratory of the medical biochemistry and molecular biology Department, faculty of Medicine, Cairo University.
Groups were divided as group 1 were osteoclast like cells only, group 2 implanted osteoclast like cells on titanium stated as modified, group 3 osteoclast like cells supplemented with BMP7 known as machined +BMP7 while group 4 were implanted osteoclast like cells on titanium and supplemented with BMP7 known as modified +BMP7, all groups were cultured for one and 3 weeks.
Cells were then proliferated; measurement of cell viability and proliferation were done and comprises the underlying basis for numerous in vitro assays directed towards the quantitation of a cell population’s response to external factors. RNA extraction was then done, the required amount of lysis buffer was supplemented with β- mercaptoethanol. 20 μL of 14.3 M β-mercapto ethanol was added to each 1 mL volume of Lysis Buffer used. Lysis Buffer was checked for salt precipitation before each use. Any precipitate was re-dissolved by warming the solution at 37°C, and then cooled back down to 25°C before use.
After the RT-PCR run the data were expressed in Cycle threshold (Ct). The PCR data sheet included Ct values of assessed gene (collagen type I) and the reference gene (GAPDH). The relative quantitation (RQ) of each target gene is quantified according to the calculation of delta-delta Ct (ΔΔCt).
The comet assay was conducted under alkaline conditions, two slides were prepared for each sample, and randomly chosen 50 cells were measured by Comet Assay automatic image analysis system fitted with Leica fluorescence microscope. All results were evaluated in terms of nine image-analysis parameters.
Our results showed that significance difference between Mod+BMP7 and all groups in terms of cell proliferation, while in collagen gene expression despite the significant increase in mod+BMP7 group in 3 weeks compared to 1week duration but there was no significant difference among all groups in 1 and 3 weeks duration.
On the other hand, same results and outcome was denoted for the alkaline phosphatase enzyme activity among the 6 weeks duration investigation with increase in enzyme activity especially in mod+BMP7 group with no significant difference between the studied groups.
We concluded that implanting on titanium with rhBMP7 supplementation not only had a great effect on osteoblast proliferation and differentiation but also, it reduces DNA damage and increase cellular viability during bone remodeling. Allkaline phosphatase reach its maximum activity after 6 weeks of rhBMP7 supplementation. For all these reasons rhBMP7 supplemented on titanium considered a crucial and important to bone remodeling process during implantation.