# Enhanced Osseointegration, Osteogenic Differentiation and Adherence Behaviour of Healthy Human Osteoblasts on a Roughened Titanium Surface by Vitamin K2 and Vitamin D3

## TL;DR

This study shows that rough titanium surfaces and vitamin K2 improve bone cell adhesion and growth, enhancing implant integration.

## Contribution

The first in vitro study using primary human osteoblasts to show vitamin K2's role in enhancing osseointegration on rough titanium surfaces.

## Key findings

- Rough titanium surfaces promote better cell adhesion and network formation compared to smooth surfaces.
- Vitamin K2 enhances long-term osteogenic differentiation, while combined vitamins boost initial differentiation.
- Vitamin K2's role in osseointegration is more significant than previously recognized.

## Abstract

The number of endoprosthetic implants is constantly increasing. Successful osseointegration of the inserted material into the bone is essential for a prosthesis to remain in the bone as long as possible. In the clinical setting, a roughened titanium surface of implants is used as standard to enable the best possible osseointegration. Vitamin K2 and vitamin D3 play a decisive role in dynamic bone metabolism and therefore also influence osseointegration. For the first time, we carried out in vitro investigations with clinically relevant cells, primary healthy human osteoblasts (hOBs). We qualitatively compared the adhesion behaviour of hOBs on a plastic surface, a smooth, regular titanium surface structure and a roughened, irregular titanium surface structure by scanning electron microscopy and fluorescence microscopy. The osteogenic behaviour and the osteogenic differentiation capacity were quantitatively investigated by analysing the activity of alkaline phosphatase and the alizarin red S assay under the influence of vitamin K2, vitamin D3 and the combination of both vitamins. It was shown that more adhesion points formed between the cells and the titanium on the rough surface structure. In addition, a solid cell network developed more quickly on this side, with cell runners forming in three-dimensional space, which means the interactions between the cells across different cell layers. On the other hand, a structured cell network also appeared on the regular smooth surface structure, which means that the network seems to be formed and built up along a defined structure. The addition of vitamins further increased the osteogenic differentiation capacity on the rough titanium surface structure. In particular, the isolated addition of vitamin K2 showed an improved osteogenic differentiation in the long-term observation, whereas the combined addition of both vitamins promoted the initial osteogenic differentiation. Vitamin K2, therefore, plays a greater role in osseointegration than previously assumed. This opens up new possibilities for the use of vitamin K2 during and after the surgical insertion of an implant. The use of vitamin K2 should be reconsidered for clinical applications in implant care and further investigated clinically.