Effects of N-acetyl-L-cysteine polysulfides on periodontitis in a mouse model.

BACKGROUND: Polysulfides are reported to be involved in various important biological processes. N-acetyl-l-cysteine polysulfide with 2 sulfane sulfur atoms (NAC-S2) regulates diverse toll-like receptor (TLR) signaling pathways. Here, we aimed to determine the role of NAC-S2 in periodontitis and explore the potential mechanism. METHODS: A periodontitis mouse model was established by ligating the subgingival between the first and second molars in wild-type, TLR4-/- , and Myd88-/- mice. RESULTS: NAC-S2 did not affect the proportion of macrophages (CD11b+ F4/80+ ) or neutrophils (CD11b+ GR-1+ ) in the bone marrow. Mechanically, lipopolysaccharides (LPS), Zymosan A, or poly I: C induced tumor necrosis factor (TNF), interleukin (IL)-6, and IL-1ß expression in bone marrow-derived macrophages (BMDMs) could be inhibited by NAC-S2. On the other hand, NAC-S2 suppressed the phosphorylation levels of IκB-α, p65, and IκB kinase (IKK)-ß induced by LPS in BMDMs, while LPS induced phosphorylation of ERK1/2, p38, and transforming growth factor ß-activated kinase 1 (TAK1) could not be affected by NAC-S2. In wild-type periodontitis mice, NAC-S2 administration decreased the cemento-enamel-junction-alveolar bone crest (CEJ-ABC) distance and the relative mRNA expression of TNF, IL-6, and IL-1ß, while such phenomena could not be observed in TLR4 deficiency or Myd88 deficiency mice. CONCLUSIONS: All of these results indicate that NAC-S2 ameliorates TLR4/NF-κB pathway mediated inflammation in mouse periodontitis model.

Beneficial Effects of Glycyrrhizin in Chronic Periodontitis Through the Inhibition of Inflammatory Response.

Elevated inflammatory cytokines and high mobility group box 1 (HMGB1) production are associated with chronic periodontitis (CP). Glycyrrhizin is the major constituent of Glycyrrhiza glabra. L. (Fabaceae) root with anti-inflammation activities. This study evaluated the effects of glycyrrhizin on CP. TNF-α-treated human periodontal ligament stem cell (hPDLSC) model was established, and was administrated with 1, 2 or 5 mM glycyrrhizin for 24 h. After treatment, the expression of HMGB1and inflammatory cytokines was monitored. Significantly increased HMGB1 (median: 5646.4, range: 1918.2-8233.7 vs median: 204.5, range: 98.7-283.6, pg/mL), TNF-α (median: 345.5, range: 161.0-567.9 vs median: 93.5, range: 58.1-159.3, pg/mL), IL-1ß (median: 2014.6, range: 209.5-4308.1 vs median: 224.5, range: 48.8-335.8, pg/mL) and IL-6 (median: 1223.6, range: 398.2-2183.8 vs median: 240.4, range: 105.2-400.5, pg/mL) were detected in gingival crevicular fluid from CP patients. Glycyrrhizin significantly prevented TNF-α-induced expression of HMGB1 (691.5 ± 136.4 vs 142.8 ± 57.3 pg/mL), IL-6 (388.1 ± 85.2 vs 189.4 ± 61.2 pg/mL) and IL-1ß (176.3 ± 47.2 vs 53.9 ± 25.7 pg/mL) in hPDLSC. In CP rats, glycyrrhizin significantly decreased HMGB1 (5795.6 ± 1121.5 vs 586.4 ± 436.8 pg/mL), TNF-α (421.8 ± 93.7 vs 87.9 ± 21.6 pg/mL), IL-6 (1423.8 ± 235.2 vs 622.6 ± 176.1 pg/mL) and IL-1ß (1562.8 ± 334.3 vs 733.5 ± 265.1 pg/mL) in gingival crevicular fluid. Glycyrrhizin suppresses inflammatory activities in CP rats and represents a promising molecule for controlling CP.

Fabrication and characterization of silk fibroin coating on APTES pretreated Mg-Zn-Ca alloy.

To delay the degradation of magnesium alloys, silk fibroin as a natural organic polymer coating was fabricated on a 3-amino-propyltriethoxysilane (APTES) pretreated Mg-Zn-Ca alloy. APTES pretreatment coated the surface of magnesium alloys with amino groups, which can bond with functional groups in silk fibroin to form a compact coating/substrate interface. The influences of the APTES concentration and drying temperature on the coating adhesion and interface were investigated to explore the optimal parameters in the fabrication process. The nanoporous silk fibroin films completely covered the APTES pretreated Mg-Zn-Ca surface, which reached a thickness of ~7 µm. The chemical states for the coated Mg-Zn-Ca alloy were compared to those of the bare Mg-Zn-Ca alloy and the APTES pretreated Mg-Zn-Ca alloy to illustrate the coating mechanism. During in vitro degradation and electrochemical measurements in simulated body fluid (SBF), the samples with the silk fibroin coating showed remarkably improved corrosion resistance and a slower degradation rate compared to those of the bare samples, suggesting that the silk fibroin coating was an effective protection coating for the substrates and can delay the degradation of magnesium alloys. Moreover, a model for the in vitro degradation was proposed. In vitro cell experiments confirmed the excellent biocompatibility of silk fibroin coated Mg-Zn-Ca structure.

Silk fibroin film-coated MgZnCa alloy with enhanced in vitro and in vivo performance prepared using surface activation.

Magnesium and its alloys have generated considerable interest as one of the most promising biodegradable metals for biomedical bone implants. However, the enormous challenges are to improve their rapid corrosion excessively as well as to endow them with biocompatibility and biosafety. Herein, we introduce a natural silk fibroin protein coating to control the corrosion resistance and enhance the biocompatibility of MgZnCa alloy. To obtain a robust and reliable coated structure, different surface-activation processes are employed to increase the available functional groups on MgZnCa surfaces before coating. Compared to oxygen plasma activation, our unique vacuum ultraviolet-ozone (VUV/O3) activation method is effective in realizing uniform silk fibroin films as a protective barrier on MgZnCa alloy surfaces, and the nanoscratch test verified the superior adhesion strength of the silk fibroin-coated magnesium alloy structure. Long-term immersion results combined with electrochemical tests showed the preferable in vitro anticorrosion behavior and a low degradation rate of coated Mg alloy (1/8 times that of uncoated Mg alloy). Cell adhesion and cytotoxicity tests demonstrated that silk fibroin-coated MgZnCa presented improved biocompatibility with bone marrow mesenchymal stem cells. An animal study involving silk fibroin-coated MgZnCa implanted on one side of a rabbit spine for 180 days showed remarkably improved in vivo corrosion resistance, with 1/18 times the degradation rate of uncoated MgZnCa. These results not only comprehensively confirmed the validity of the VUV/O3-activation method as a coating strategy but also implied the tremendous potential of the modified Mg alloy for application as a degradable biomedical implant material. STATEMENT OF SIGNIFICANCE: MgZnCa alloy is a promising material in clinical implantation. Silk fibroin (SF) is a natural organic material with biocompatibility and biodegradability. To date, the combination of SF and MgZnCa alloy has exhibited considerable prospects for orthopedic applications. The realization of a direct coating is an enormous challenge because strong chemical bonds cannot be easily formed between organic and inorganic materials. To solve this bottleneck, we proposed a unique vacuum ultraviolet-ozone (VUV/O3) surface-activation method for the first time to modify the Mg alloy surface before SF coating, which significantly enhanced both in vitro and in vivo performance, such as superior biocompatibility and remarkably improved corrosion resistance of magnesium alloys (∼1/18 the in vivo degradation rate of uncoated MgZnCa).