Biological reaction control using topography regulation of nanostructured titanium

Mayuko Shiozawa,Nami Akiba,Haruka Takeuchi,Jun Mizuno,Katsumi Uoshima,Kenji Izumi,Masako Nagasawa,Hiroyuki Kuwae,Yosuke Akiba,Kaori Eguchi,Yujin Aoyagi

doi:10.1038/s41598-020-59395-4

Mayuko Shiozawa, Nami Akiba + Show 9 more

Open Access

https://doi.org/10.1038/s41598-020-59395-4

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Journal: Scientific Reports	Publication Date: Feb 12, 2020
Citations: 8	License type: open-access

Affiliation: Waseda University, Niigata University

Abstract

The micro- and nanosize surface topography of dental implants has been shown to affect the growth of surrounding cells. In this study, standardized and controlled periodic nanopatterns were fabricated with nanosized surface roughness on titanium substrates, and their influence on bone marrow stromal cells investigated. Cell proliferation assays revealed that the bare substrate with a 1.7 nm surface roughness has lower hydrophilicity but higher proliferation ability than that with a 0.6 nm surface roughness. Further, with the latter substrate, directional cell growth was observed for line and groove patterns with a width of 100 nm and a height of 50 or 100 nm, but not for those with a height of 10 or 25 nm. With the smooth substrate, time-lapse microscopic analyses showed that more than 80% of the bone marrow cells on the line and groove pattern with a height of 100 nm grew and divided along the lines. As the nanosized grain structure controls the cell proliferation rate and the nanosized line and groove structure (50–100 nm) controls cell migration, division, and growth orientation, these standardized nanosized titanium structures can be used to elucidate the mechanisms by which surface topography regulates tissue responses to biomaterials.

Highlights

Both ion beam sputtering (IBS) and electron beam (EB) evaporation result in titanium substrates with very smooth surfaces, as revealed by low-magnification scanning electron microscopy (SEM) images
The bone marrow stromal cell (BMSC) were seeded on the 100 nm high line and groove pattern or a flat pattern on the 0.6 nm surface roughness substrate
Titanium is commonly used for dental implants and the nanoscale surface topography of titanium dental implants may affect the activity of the attached cells

Summary

Introduction

Both ion beam sputtering (IBS) and electron beam (EB) evaporation result in titanium substrates with very smooth surfaces, as revealed by low-magnification scanning electron microscopy (SEM) images. Twenty-four or 48 hours after seeding the BMSCs on the line and groove pattern with a height of 100 nm or a flat pattern on the smooth (0.6 nm surface roughness) substrate, the cells were fixed and stained with anti-focal adhesion kinase (FAK) and/or anti-actin filament (F-Actin) antibodies. 24 hours after seeding, the BMSCs on the 100 nm high line and groove pattern on the rough substrate showed spindle and extended shapes (Fig. 5B) with F-actin filaments aligned along the line and groove pattern (Fig. 5B) Under both low and high concentration conditions, FAK at the edge of the cells, which has a fibrous structure, was observed to align with the line and groove pattern Spindle-shaped cells always migrated following the patterns, whereas some flat-shaped cells could move across the patterns

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