ATP is an important second messenger in cells involved in wound healing. In particular, ATP levels are known to regulate osteoblast function after surgical trauma. Oxidative stress is a natural inflammatory consequence of surgically induced bone trauma and ATP levels are negatively regulated by oxidative stress. The purpose of this study was to evaluate the potential for negative regulation of osteoblast ATP levels by oxidative stress after the placement of implants. The hypothesis to be tested is that titanium oxide interacts with hydrogen peroxide (oxidative molecules) to inhibit ATP production in osteoblasts, which would potentially interfere with osseointegration.

Objective: to evaluate ATP production in an in vitro model of titanium implant healing.

Methods: MG-63 human osteoblastic cells were cultured in DMEM-10. Cells were cultured (3000/well) with various concentrations of H2O2 (hydrogen peroxide) (0, 32.1, 62.5, 125.0, 250.0μM) and TiO2 (Wako Pure Chemical Industries) (0%, 0.01%, 0.1%). UV (using Supercure-203S) radiation exposure for 2 seconds was used to induce reactive oxygen species (ROS) by the photocatalytic reaction of TiO2. The ATP measurement was carried out by utilizing Cell Titer-Glo Luminescent Cell Viability Assay Kit (Promega, WI, USA).

Results: Exposure of cells to UV radiation in the presence of 0.1% TiO2 and higher reduced ATP production in osteoblasts an order of magnitude. Differences were significant at P-value<0.05.

Conclusion: TiO2 in the presence of reactive oxygen species reduces ATP production in a human osteoblastic cell line suggesting that surgically induced inflammation may have a negative impact on osseointegration. Control of inflammation after surgery may provide a therapeutic a therapeutic opportunity for improvement of osseointegration of TiO2 implants.

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