Bio-functionalized titanium surfaces with modified silk fibroin carrying
titanium binding motif to enhance the ossific differentiation of
MC3T3-E1.
Mai Watanabe†‡, Ujjal K. Bhawala¶,
Shinji Takemoto&, Norihiro
Nishiyama%, Yuichi Nakahara∞,
Ken-ichiro Tatematsu∞, Hideki
Sezuts∞, Nobuo Kuwabaraę, Tamiko
Minamisawa†, Kiyotaka Shiba†* and
Tetsuo Asakura‡*
†Division of Protein Engineering, Cancer Institute,
Japanese Foundation for Cancer Research, 3-8-31 Ariake, Koto, Tokyo
125-8550, Japan, ‡Department of Biotechnology, Tokyo
University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei,
Tokyo 184-8588, Japan, ¶Department of Biochemistry and
Molecular Biology, and Research Institute of Oral Science, Nihon
University School of Dentistry at Matsudo, 2-870-1 Sakae-cho Nishi,
Matsudo, Chiba 271-8587, Japan, &Department of
Biomedical Engineering, Iwate Medical University, Iwate 028-3694, Japan,%Department of Dental Biomaterials, Nihon University
School of Dentistry at Matsudo, 2-870-1 Sakae-cho Nishi, Matsudo, Chiba
271-8587, Japan, ∞Transgenic Silkworm Research Unit,
National Agriculture and Food Research Organization,1-2 Owashi, Tsukuba,
Ibaraki 305-8634, Japan, ęGunma Sericultural
Technology Center, 2326osteo-2 Soja-machi, Maebashi, Gunma 371-0852,
Japan
ABSTRACT. min-TBP-1 modified silk fibroin
Silk fibroin (SF) from Bombyx mori has superior properties as
both a textile and a biomaterial, and has been used to functionalize the
surfaces of various medical inorganic materials including titanium (Ti).
In this paper, we endowed SF with reversible binding ability to Ti by
embedding a titanium binding motif (minTBP-1, RKLPDA). Artificial SF
proteins were first created by conjugating gene cassettes for SF motif
(AGSGAG) and minTBP-1 motif with different ratios, which have been shown
to bind reversibly to Ti surfaces in quartz crystal microbalance
analyses. Based on these results, the functionalized SF (TiBP-SF)
containing the designed peptide
[TS[(AGSGAG)3AS]2RKLPDAS]8was prepared from the cocoon of transgenic B. mori , which
accelerates the ossific differentiation of MC3T3-E1 cells when coated on
titanium substrates. Thus, TiBP-SF presents an alternative for endowing
the surfaces of titanium materials with osseointegration functionality,
which would allow the exploration of potential applications in the
medical field.
KEYWORDS. Peptide aptamer, transgenic silk fibroin, artificial
protein, biomaterial, 13C solid-state NMR
Running title. minTBP-1 functionalized silk fibroin.