Kaempferol-immobilized titanium dioxide promotes formation of new bone: effects of loading methods on bone marrow stromal cell differentiation in vivo and in vitro

Abstract

Background

Surface modification of titanium dioxide (TiO₂) implants promotes bone formation and shortens the osseointegration period. Kaempferol is a flavonoid that has the capacity to promote osteogenic differentiation in bone marrow stromal cells. The aim of this study was to promote bone formation around kaempferol immobilized on TiO₂ implants.

Methods

There were four experimental groups. Alkali-treated TiO₂ samples (implants and discs) were used as a control and immersed in Dulbecco’s phosphate-buffered saline (DPBS) (Al-Ti). For the coprecipitation sample (Al-cK), the control samples were immersed in DPBS containing 50 µg kaempferol/100% ethanol. For the adsorption sample (Al-aK), 50 µg kaempferol/100% ethanol was dropped onto control samples. The surface topography of the TiO₂ implants was observed by scanning electron microscopy with energy-dispersive X-ray spectroscopy, and a release assay was performed. For in vitro experiments, rat bone marrow stromal cells (rBMSCs) were cultured on each of the TiO₂ samples to analyze cell proliferation, alkaline phosphatase activity, calcium deposition, and osteogenic differentiation. For in vivo experiments, TiO₂ implants placed on rat femur bones were analyzed for bone–implant contact by histological methods.

Results

Kaempferol was detected on the surface of Al-cK and Al-aK. The results of the in vitro study showed that rBMSCs cultured on Al-cK and Al-aK promoted alkaline phosphatase activity, calcium deposition, and osteogenic differentiation. The in vivo histological analysis revealed that Al-cK and Al-aK stimulated new bone formation around implants.

Conclusion

TiO₂ implant-immobilized kaempferol may be an effective tool for bone regeneration around dental implants.

Keywords:

• kaempferol
• titanium implant
• surface treatment
• biomaterial

Acknowledgments

This work was supported in part by grants from the Japanese Ministry of Education, Culture, Sports, Science, and Technology (Kakenhi Kiban C, 17K11802 to Shuhei Tsuchiya; Kakenhi Kiban B, 16H05540 to Hideharu Hibi; and Kakenhi Kiban B, 00283408 to Kensuke Kuroda).

Disclosure

The authors report no conflicts of interest in this work.