Introduction: The osteoconductive properties of bone grafting materials represent one area of research for the management of bony defects found in the fields of periodontology and oral surgery. It has been shown that surface wettability represents the crucial factor for osteoconductivity since the amount of growth factors and proteins on the material particles proportionally increase migration and adhesion of bone cells. Argon plasma treatment (PAT) and ultraviolet irradiation (UV) may increase the surface wettability and, consequently, improve the regenerative potential of the bone grafts Objective. The aim of the present in vitro study was to evaluate the effect of PAT and UV treatment on the osteoconductivity of bone grafts by assessing osteoblast adhesion and viability, surface topography, macrophage polarization and osteodifferentiation. Materials & Methodology: The following four frequently used bone grafts were selected for this study: biphasic calcium phosphate (BCP), synthetic hydroxyapatite (MG-HA), cancellous and cortical xenogenic bone matrices (CaBM, CoBM). Topographic analysis of bone grafts was obtained through X-Ray microtomography (µCT) and Scanning Electron Microscope (SEM). To characterize the biological response in vitro, human osteoblast cell lines (MG63, Saos-2) Mesenchymal Stem Cells (MSC) and macrophages (RAW 264.7) were used. To overcome the limitation of an in vitro model to simulate in vivo conditions, a bioreactor (LB2, IVTech) was used. In order to evaluate macrophage activation IL-1, IL-6, TNFα and TGFβ were analyzed. Differences between groups were analyzed using the ordinary one-way ANOVA with the Tukey’s multiple comparison test considering a p value <0,05. Results Surface analysis following treatments demonstrated minimal topographical differences when compared to controls. Plasma treatment significantly increased the number of adherent cells on all tested graft surfaces, which, interestingly, was not true for UV treatment. No difference among PAT, UV and controls on osteoblast viability was observed. Result suggest no difference in M1 and M2 macrophage polarization among the considered conditions, indicating that potential inflammatory reaction caused by Argon plasma is very low. Moreover, Argon of plasma was able to increase both RUNX2 and Collagen type-I levels compared to control condition, no difference was measured in UV treated samples. Conclusion: This study suggests that treatment of various bone grafting materials with PAT appears to enhance the osteoconductivity of bone substitutes by improving osteoblast adhesion and differentiation without affect cell viability. Macrophages are widely accepted as regulators of wound healing and play an important role in bone deposition and differentiation of mesenchymal progenitors. In this study that PAT and UV treatment do no exert inflammatory response. Treatment of bone grafts with PAT may result in faster osseointegration of the material itself owing to the faster cellular attachment and may favorably increase bone regeneration. Future animal studies are nevertheless warranted.
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