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.