Interaction between titanium and phosphoproteins revealed by chromatography column packed with titanium beads

Article type: Research Article

Authors: Kuboki, Yoshinori^; | Furusawa, Toshitake | Sato, Masaaki | Sun, Yongkun | Unuma, Hidero | Fujisawa, Ryuichi | Abe, Shigeaki | Akasaka, Tsukasa | Watari, Fumio | Takita, Hiroko | Sammons, Rachel

Affiliations: Hokkaido University, Sapporo, Japan | Graduate School of Bio-Medical Engineering, Tohoku University, Sendai, Japan | Graduate School of Environmental Science, Hokkaido University, Sapporo, Japan | Graduate School of Science and Engineering, Yamagata University, Yamagata, Japan | Graduate School of Dental Medicine, Hokkaido University, Sapporo, Japan | School of Dentistry, Birmingham University, Birmingham, AL, USA

Note: [] Address for correspondence: Yoshinori Kuboki, Professor Emeritus, Hokkaido University, N-13, W-7, Kita-ku, Sapporo, 060-8586, Japan. Tel.: +81 11 706 4251; Fax: +81 11 706 4251; E-mail: [email protected].

Abstract: The biochemical mechanism behind the strong binding between titanium and living bone has not been fully elucidated, in spite of worldwide clinical application of this phenomenon. We hypothesized that one of the core mechanisms may reside in the interaction between certain proteins in the host tissues and the implanted titanium. To verify the interaction between titanium and proteins, we chose the technique of chromatography in that titanium spherical beads (45 μm) were packed into a column to obtain a bed volume of 16×50 mm, which was eluted with phosphate buffered saline (PBS) and a straight gradient system made by using PBS and 25 mM NaOH. Fetal calf serum, albumin, lysozyme, casein, phosvitin and dentin phosphoprotein (phosphophoryn) were applied to the column. Most part of albumin and lysozyme eluted with the breakthrough peak, indicating practically no affinity to titanium. Fetal bovine serum also eluted mostly as the breakthrough peak, but distinct retained peak was observed. On the other hand, α-casein, phosvitin and phosphophoryn exhibited a distinct retained peak separated from the breakthrough peak. We proposed that phosphate groups (phosphoserines) in the major phosphoproteins, α-casein, phosvitin and phosphophoryn may be involved in the binding of these proteins with titanium.

Keywords: Titanium binding proteins, chromatography, phosphoprotein, caseins, phosvitin, dentin phosphoprotein

DOI: 10.3233/BME-2012-0718

Journal: Bio-Medical Materials and Engineering, vol. 22, no. 5, pp. 283-288, 2012