Short-chain fatty acids (SCFAs) are potent immune modulators present in the gingival crevicular fluid. It is therefore likely that SCFAs exert a role in periodontal health and disease. To better understand how SCFAs can module inflammation, we screened acetic acid, propionic acid, and butyric acid for their potential ability to lower the inflammatory response of macrophages, gingival fibroblasts, and oral epithelial cells in vitro. To this end, RAW 264.7 and primary macrophages were exposed to LPSs from Porphyromonas gingivalis (P. gingivalis) with and without the SCFAs. Moreover, gingival fibroblasts and HSC2 oral epithelial cells were exposed to IL1β and TNFα with and without the SCFAs. We report here that butyrate was effective in reducing the lipopolysaccharide (LPS)-induced expression of IL6 and chemokine (C-X-C motif) ligand 2 (CXCL2) in the RAW 264.7 and primary macrophages. Butyrate also reduced the IL1β and TNFα-induced expression of IL8, chemokine (C-X-C motif) ligand 1 (CXCL1), and CXCL2 in gingival fibroblasts. Likewise, butyrate lowered the induced expression of CXCL1 and CXCL2, but not IL8, in HSC2 cells. Butyrate further caused a reduction of p65 nuclear translocation in RAW 264.7 macrophages, gingival fibroblasts, and HSC2 cells. Propionate and acetate partially lowered the inflammatory response in vitro but did not reach the level of significance. These findings suggest that not only macrophages, but also gingival fibroblasts and oral epithelial cells are susceptive to the anti-inflammatory activity of butyrate.

Periodontitis is an inflammatory process that is associated with caspase activity. Caspases could thus become molecular targets for the modulation of the inflammatory response to harmful factors, such as lipopolysaccharides (LPS) and TNFα. Here, the impact of the pan-caspase inhibitor Z-VAD-FMK (carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoro-methyl ketone) on the modulation of the LPS-induced inflammatory response of murine RAW 264.7 cells and primary macrophages was examined. Moreover, the inflammatory responses of human gingival fibroblasts, HSC2 oral squamous carcinoma cells and murine ST2 mesenchymal fibroblasts when exposed to TNFα were studied. Data showed that Z-VAD-FMK significantly lowered the inflammatory response of RAW 264.7 cells and primary macrophages, as indicated by the expression of IL1 and IL6. In murine ST2 mesenchymal fibroblasts, the TNFα-induced expression of CCL2 and CCL5 was significantly reduced. In human gingival fibroblasts and HSC2 cells, Z-VAD-FMK considerably reduced the TNFα-induced expression of CXCL8 and CXCL10. These findings suggest that pharmacological blocking of caspases in an inflammatory environment lowers the expression of cytokines and chemokines in periodontal cells.

The laser treatment and hydroxyapatite coating of dental implants are supposed to enhance osseointegration, but prior to preclinical testing, any negative impact on cell viability should be ruled out. This study aimed to evaluate the response of murine osteogenic cell lineage MC3T3-E1 and the bone marrow-derived stromal cells ST2 to surface modifications of machined titanium discs, e.g., laser treatment without and with hydroxyapatite coating, as well as sandblasting followed by acid etching. Scanning electron microscopy and the contact angle measurements revealed that laser treatment caused a honeycomb surface and higher wettability compared to a machined or sandblasting acid-etched surface. Hydroxyapatite coating, however, not only reduced the viability of MC3T3-E1 and ST2 cells but also provoked the expression and release of interleukin-6. These findings suggest that the laser treatment of titanium supports its hydrophilicity, but adding hydroxyapatite can reduce cell viability and induce the concomitant release of inflammatory cytokines.

Objective: Titanium surface modifications improve osseointegration in dental and orthopedic implants. However, soft tissue cells can also reach the implant surface in immediate loading protocols. While previous research focused on osteogenic cells, the early response of soft tissue cells still needs to be better understood. Material and methods: We have established a bioassay to this aim where human gingival fibroblasts, HSC2 oral squamous carcinoma cells, and murine bone marrow cells were cultured onto titanium discs or exposed to the respective supernatants for overnight. Modifications were double acid-etching (SLA), and coating with simulated body fluid (SBF) with or without odanacatib (ODN), a selective cathepsin K inhibitor reducing bone resorption. Results: Our findings indicate that direct contact with titanium discs, with all surface modifications, slightly reduces cell viability. Growing gingival fibroblasts on discs consistently showed a trend toward increased IL8 expression. In HSC2 cells, this setting significantly increased IL1 and IL8 expression, confirmed by the immunoassay. Murine bone marrow macrophages also showed an increase in IL1 and IL6 expressions. Supernatants of the respective discs failed to cause these changes. Although ODN coating inhibited cathepsin K, osteoclastogenesis remained unchanged. Conclusions: These findings suggest that titanium discs do not provide a favorable in vitro surface for oral soft tissue cells as they lose viability and respond with a moderately increased expression of inflammatory cytokines. Clinical relevance: The soft tissue surrounding a dental implant can impact rehabilitation success. Understanding how soft tissue cells respond to titanium surface is potentially relevant to understand clinical outcomes.

Objective: Characterize the newly formed bone tissue adjacent to implant surfaces. Methods: The implants were previously analyzed by scanning electron microscopy (SEM) and energy dispersion X-ray spectroscopy (EDS) and installed in the right tibia of 75 rats divided into polished, machined, and treated groups. After euthanasia on 3, 7, 15, 21, and 40 postoperative days, histological analysis was performed to evaluate bone-implant contact. Results: At topographic analysis on the SEM, the polish group had the smoothest and regular surface and roughness surface in the treated group. On the EDS analysis, all groups had no contamination on the superficies and similar compouds (Titanium, Aluminum, and Vanadium). In terms of quality, the process of osseointegration was similar across all groups. However, a quantitative analysis showed that mature bone formed around the implant threads, even in areas near the bone marrow, with significant difference at seven days between the polished and treated groups. Conclusion: The peri-implant repair pattern was similar among all surface types in all other timelines, with cortical bone formation throughout the titanium surface of the three implant groups. The acid-etched surface treatment group showed faster bone differentiation.