Ozone Nanobubble Water Inhibits the Biological Aging and Maintains the Affinity of Titanium Disks: An in Vitro Study

Abstract

This study evaluated the aging phenomenon and cell affinity of titanium after storage in ozone nanobubble water (NBW3). JIS Class 2 pure titanium disks were mirror polished, treated with glow discharge under an argon atmosphere, and stored under four different storage conditions: atmosphere, distilled water, acetone, and NBW3. X-ray photoelectron spectroscopy (XPS) was used to measure the changes in the adhesion and contact angles of the hydrocarbon compounds. Human mesenchymal stem cells were seeded on titanium disks and stored for 24 h under the four different conditions. The number of adherent cells was measured, and their morphology was observed using scanning electron microscopy and confocal laser scanning microscopy. Alkaline phosphatase (ALP) staining was performed after 7 days of cell culture. XPS analysis revealed that samples stored in atmosphere exhibited a high C1s peak, indicative of the attachment of hydrocarbon compounds; however, the peak in NBW3 remained low. Titanium disks stored in NBW3 exhibited higher wettability, higher cell attachment counts and cell adhesion, and increased ALP activity compared to those stored in atmosphere. Storage of titanium disks in NBW3 may prove effective in inhibiting the aging process of the titanium surfaces and maintaining high wettability and cell affinity.