Effect of Bioactivation on Traditional Surfaces and Zirconium Nitride: Adhesion and Proliferation of Preosteoblastic Cells and Bacteria

Purpose: The aim of this vitro study was to reproduce and evaluate the response of bone and bacteria to traditional and innovative implant surfaces with difference wettability. Materials and Methods: Two hundred fifty-two samples made of grade 4 titanium with different coating (machined [MAC]; double-etched, Ti-AE; zirconium nitride [Ti-ZrN]) were used for this in vitro study. Disks were divided into test (bioactivated using plasma of argon) and control group (untreated). To assess the surface morphology of the specimens, representative images were acquired via scanning electron microscopy (SEM). Murine preosteoblasts (MC3T3-E1) were used to study the biologic response in vitro, while the quantification of protein adsorption was achieved through the incubation of the titanium samples in a 2% solution of fetal bovine serum (FBS) in phosphate-buffered saline (PBS). The sterilized titanium disks were then colonized by bacterial species from a single sputum sample obtained from a healthy volunteer. For every analysis, 24 disks were used (12 for each group). Results: SEM and topographic analyses demonstrated a Sa value of 0.33 (Ti-ZrN), 0.34 (MAC), and 0.62 (Ti-AE). Compared with the control groups, plasma treatment significantly increased the protein adsorption level on all the different titanium surfaces (5.88 ± 0.21 vs 7.85 ± 0.21, 7.13 ± 0.14 vs 9.74 ± 0.65, 4.41 ± 0.62 vs 6.13 ± 0.52, respectively, for MAC, Ti-treated, and Ti-ZrN). Similar behavior was described for cell adhesion (27.67 ± 2.03 vs 58.00 ± 20.13, 116.67 ± 12.02 vs 159.33 ± 8.09, 52.00 ± 4.73 vs 78.33 ± 4.67, respectively, for MAC, Ti-treated, and Ti-ZrN). Plasma treatment significantly augmented the number of CFU only in MAC and ZrN samples. Conclusion: With the limitations of this in vitro study, the following conclusions could be drawn: (1) rough implant surfaces present a higher adhesion and proliferation of preosteoblastic cells and bacterial biofilm; (2) rough implant surfaces benefited the most by the plasma of argon treatment. Keywords: cell adhesion, implant surfaces, microbiologic growth, plasma of argon, protein adsorption, zirconium nitride