Surface Characterization of Titanium Plates with Different Surface Treatments and Cellular Proliferation and Expression of Osteoblast-like Cells in vitro on their Surfaces

DOI 20 Citations 62 References
  • FUJIMORI Shinya
    Department of Oral Biomterials and Technology, Showa University School of Dentistry

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  • 種々の微小表面形状を付与したチタン板の表面分析並びにその表面上での骨芽細胞様細胞の増殖と分化に関する研究

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Abstract

Titanium plates(TPs)were prepared with either an irregular and rough or a regular and rather smooth surface morphology.TPs with an irregular surface were produced by either wire-type electric discharge machining(Wire-EDM)or plasma coating(Plasma).TPs with a regular surface were produced by polishing with 1, 500-grit emery papers(Emery).The surfaces of TPs were characterized by surface profilometry, scanning electron microscopy(SEM), X-ray photoelectron spectroscopy(XPS)and X-ray diffraction(XRD).Mouse osteoblast-like cells(MC3T3-E1)were exposed to the surfaces of the prepared TPs and cultured for 15 days.Glass plates for tissue culture(TCP)were used as a control.Total DNA of the cultured cells was evaluated as a marker of cell proliferation.Alkaline phosphatase(ALP)enzyme activity was evaluated as a marker of cell expression.The amounts of Ca and P at day 12 and 15 were also measured.The surface roughness(Rmax)of TPs prepared with Wire-EDM, Plasma and Emery was 20.5, 31.4 and 0.80μm respectively.The surfaces with Wire-EDM were irregularly oriented in morphology with many small depressions and indentations, but the height of depression was rather controlled.On the other hand, the surfaces with Plasma were highly irregular and porous.XPS and XRD appeared that the surface oxide layer of the surface of the TPs with Wire-EDM were composed of TiO2, TiO and Ti2O and were far thicker than those with Plasma and Emery.The amount of DNA of cultured cells gradually increased on all surfaces, and became significantly higher with Wire-EDM and Plasma than with Emery and TCP after day 9.The phenotypic characteristic of bone cells in vitro are often characterized by relatively high ALP enzyme activities.ALP activities/μg DNA of cultured cells on the titanium surfaces with Wire-EDM and Plasma were significantly higher than those with Emery and TPC after day 9.The Ca/P ratios of the cultured cells on the titanium surfaces with Wire-EDM and Plasma at day 15 were 1.26 and 1.28 respectively.Therefore, the cells prolifierated more abundantly and expressed their phenotypic characteristics earlier on TPs with irregular and rough surfaces than with regular and smooth surfaces.SEM observation of the cultured cells appeared that the body of the cells changed the shape extending the flopodial-like processes connecting irregular edges, which suggested the alterations in cell shape inducing cellular expression.These findings suggest that the micromorphology of the bone contact surface of implants influenced the biological responses and further investigations are reguired to optimize the surface micromorphology of the implants.

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