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This paper aims to review the evidence of the cellular activity on titanium samples exposed to Plasma of Argon (PoA) treatment. A systematic review was carried out based on the PRISMA statement by searching the Cochrane Library, PubMed, Web of Science, EMBASE and Scopus, up to October 2020. Papers were selected according to PICOS format that is: Population (P): osteoblasts, fibroblasts, gingival cells; Intervention (I): PoA disinfection treatment; Comparison (C): untreated controls; Outcome (O): cell culture; Setting (S): in vitro assays. The quality assessment was performed according to the CRIS Guidelines (Checklist for Reporting In vitro Studies). A total of 661 articles were found, of which 16 were included. The quality assessment revealed an overall poor quality of the studies analyzed. In vitro studies on the potential of PoA showed a potential effect in promoting higher cell adhesion and protein adsorption in the earliest times (hours). This outcome was not so evident when later stages of cell growth on the surfaces were tested and compared to the control groups. Only one study was conducted in vivo on a human sample regarding abutment cleaning. No meta-analysis was conducted because of the variety of experimental settings, mixed methods and different cell lines studied. PoA seems to be effective in promoting cell adhesion and protein adsorption. The duration of this effect remains unclear. Further evidence is required to demonstrate the long-term efficacy of the treatment and to support the use of PoA treatment in clinical practice.

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.

Objective: Plasma of argon treatment was demonstrated to increase material surface energy leading to stronger and faster interaction with cells. The aim of the present in vitro study was to test the effect of plasma treatment on different graft materials., Materials and Methods: Synthetic hydroxyapatite (Mg-HA), biphasic calcium phosphate (BCP), cancellous and cortical xenogeneic bone matrices (CaBM, CoBM) were used representing commonly used classes of bone substitute materials. Fifty serially numbered disks with a 10mm-diameter from each graft material were randomly divided into two groups: test group (argon plasma treatment) and control group (absence of treatment). Cell morphology (using pre-osteoblastic murine cells) and protein adsorption were analyzed at all samples from both the test and control group. Differences between groups were analyzed using the Mann-Whitney test setting the level of significance at p<0.05., Results: Plasma treatment significantly increased the protein adsorption at all samples. Similarly, plasma treatment significantly increased cell adhesion in all groups., Conclusions: Data confirmed that non-atmospheric plasma of argon treatment led to an increase of protein adsorption and cell adhesion in all groups of graft material to a similar extent., Clinical Relevance: Plasma of argon is able to improve the surface conditions of graft materials., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The rapid development and application of nanotechnology to biological interfaces has impacted the bone implant field, allowing researchers to finely modulate the interface between biomaterials and recipient tissues. In the present study, oxidative anodization was exploited to generate two alumina surfaces with different pore diameters. The former displayed surface pores in the mean range of 16-30 nm, while in the latter pores varied from to 65 to 89 nm. The samples were characterized by Field Emission Scanning Electron Microscopy (FESEM) and Energy Dispersive X-ray spectroscopy (EDX) analysis prior to being tested with pre-osteoblastic MC3T3-E1 cells. In vitro cell response was studied in terms of early cell adhesion, viability, and morphology, including focal adhesion quantification. Both the alumina samples promoted higher cell adhesion and viability than the control condition represented by the standard culture dish plastic. Osteogenic differentiation was assessed through alkaline phosphatase activity and extracellular calcium deposition, and it was found that of the two nano-surfaces, one was more efficient than the other. By comparing for the first time two nano-porous alumina surfaces with different pore diameters, our data supported the role of nano-topography in inducing cell response. Modulating a simple aspect of surface texture may become an attractive route for guiding bone healing and regeneration around implantable metals., Competing Interests: The authors declare no conflict of interest.

Treatments for enhancing surface energy were studied in an effort to create a more favorable environment for cell adhesion. Cold argon plasma (CAP) is able to improve titanium-cell contact, producing hydrophilic surfaces with higher wettability. The aim of this in vitro study was to estimate the early cell morphology after CAP treatment of different commercially available titanium surfaces. Surface wettability was significantly augmented in all the treated samples. The authors investigated how CAP affected the behavior of osteoblasts by evaluating the cell morphology outcome. Cell surface areas differed in a statistically significant way when plasma-treated samples were compared to the untreated ones. The positive effect of CAP was shown on smooth, moderately rough, and rough implant surfaces.

The aim of this in vitro study was to evaluate the early cell response and protein adsorption elicited by the argon plasma treatment of different commercially available titanium surfaces via a chair-side device. Sterile disks made of grade 4 titanium (n= 450, 4-mm diameter) with 3 surface topographies (machined, plasma sprayed, and zirconia blasted and acid etched) were allocated to receive 4 testing treatments (2% and 10% protein adsorption and cell adhesion with MC3T3-E1 and MG-63). Furthermore, the specimens were divided to undergo 1) argon plasma treatment (10 W, 1 bar for 12 min) in a plasma reactor, 2) ultraviolet (UV) light treatment for 2 h (positive control group), or 3) no treatment (control group). Pretreatment surface analyses based on a scanning electron microscope and profilometer images were also performed. Profilometric analysis demonstrated that the evaluated specimens perfectly suit the standard parameters. The use of argon plasma was capable of affecting the quantity of proteins adsorbed on the different surfaces, notwithstanding their roughness or topographic features at a low fetal bovine serum concentration (2%). UV light treatment for 2 h attained similar results. Moreover, both the plasma of argon and the UV light demonstrated a significant increase in the number of osteoblasts adherent at 10 min in all tested surfaces. Within its limitations, this in vitro study highlights the potential biological benefits of treating implant surfaces with plasma of argon or UV, irrespective of the roughness of the titanium surface. However, in vivo experiments are needed to confirm these preliminary data and settle the rationale of a treatment that might be clinically relevant in case of bone-reparative deficiencies. [ABSTRACT FROM AUTHOR]

Bone plays several physiological functions and is the second most commonly transplanted tissue after blood. Since the treatment of large bone defects is still unsatisfactory, researchers have endeavoured to obtain scaffolds able to release growth and differentiation factors for mesenchymal stem cells, osteoblasts and endothelial cells in order to obtain faster mineralization and prompt a reliable vascularization. Nowadays, the application of osteoblastic cultures spans from cell physiology and pharmacology to cytocompatibility measurement and osteogenic potential evaluation of novel biomaterials. To overcome the simple traditional monocultures in vitro, co-cultures of osteogenic and vasculogenic precursors were introduced with very interesting results. Increasingly complex culture systems have been developed, where cells are seeded on proper scaffolds and stimulated so as to mimic the physiological conditions more accurately. These bioreactors aim at enabling bone regeneration by incorporating different cells types into bio-inspired materials within a surveilled habitat. This review is focused on the most recent developments in the organomimetic cultures of osteoblasts and vascular endothelial cells for bone tissue engineering. [ABSTRACT FROM AUTHOR]