Microbial Corrosion in Orthodontics.

Even with the exponential popularity of the contemporary clear aligners, the main stream of orthodontic practice still remains to be metal braces especially in adolescent age-group.1 Along with the advantages of metal braces such as lower cost, reduced friction, etc., there goes the disadvantages such as corrosion possibility, reduced esthetics, etc. Corrosion of orthodontic appliances is a widely researched topic.2-5 It is surprising to learn that microbially induced corrosion (MIC) has not been addressed in orthodontic literature till date. Microbial corrosion is an interesting arena which requires knowledge of both corrosion science and microbiology. The microorganisms capable of corrosion include various bacteria, fungi, and algae. The most common among them which has been widely indicated in MIC are the bacteria belonging to the sulfur cycle especially the sulfate-reducing bacteria (SRB). The connecting knot with orthodontics is the reported prevalence of these SRB in the oral cavity. SRB is prevalent in healthy individuals,6,7 patients associated with periodontitis6-11 and patients with gastrointestinal issues.12-14 The prevalence of SRB in the oral cavity has a greater clinical implication since the SRB have been proven to cause corrosion of stainless steel.15-24 There is literature attributing SRB as a potential cause in periodontal diseases7-11 as well as gastrointestinal diseases such as ulcerative colitis, inflammatory bowel diseases, and Crohn's disease.12 With its presence in the healthy oral environment already reported in the previous studies,6,7,25,26 it further emphasizes the absolute need to be researching on its corrosion possibility in the intra oral environment. The genus generally found intraorally was Desulfovibrio and Desulfobacter10 which is commonly regarded as the most "opportunistic" and ubiquitous group of sulfate reducers.6,7 There is an interesting literature on the inhibition of Desulfovibrio spp. by human saliva, the reason being quoted as salivary nitrate and nitrite.14 The mechanism behind the antimicrobial action of nitrate and nitrite is that they increase the oxidative stress on the bacteria.27 However, concentrations of salivary nitrate vary depending on the food intake, endogenous production, and salivary flow rate.28,29 Despite there exist natural inhibitors, the prevalence in oral cavity is high, 22% in healthy and 86% in patients associated with periodontitis.7 There is a predilection for the bacteria to grow when favorable conditions exist. Biofilms is one such favorable medium for the growth of SRB. Paster et al.26 identified SRB in biofilms of patients associated with refractory periodontitis, periodontitis, acute necrotizing ulcerative gingivitis (ANUG), and also in healthy subjects. Biofilm is a surface film composed of organic and inorganic saliva components that are colonized with microorganisms in extracellular polymeric substances adsorbed on all surfaces in the oral cavity.30 The oral biofilm formation is a complex process involving interspecies aggregation, which is surrounded by a cohesive matrix, forms a complex structure which in turn facilitates anaerobic growth. It is the intrinsic nature of oral biofilms which make the survival of facultative anaerobes such as SRB in the oral cavity possible. Literatures31-35 report that there are increased biofilm formations in orthodontic patients due to increased retentive areas caused by the brackets, ligatures, wires, mini implants, force components, and archwires. Bacteria in dental plaque function as a metabolically, functionally, and physically integrated community.36 The study by Mystkowska et al.37 mentioned that biofilm per se play a critical role in corrosion process by forming corrosive microcells. With time-dependent association, the microbes in the biofilm, along with saliva acting as an electrolyte and components from food, causes a decreased pH in the areas immediately under the biofilms. The decreased pH along with a change of oxygenation releases metal oxides and hydroxides from the metal surface ultimately leading to the corrosion of metallic structures.37-41 The initial roughness also acts in a vicious form promoting more biofilm adherence and the process repeats causing more corrosion. With the biofilm itself serving to initiate and propagate corrosion, the increased prevalence of SRB in patients associated with orthodontics treatment all the more increases the possibility of MIC of orthodontic materials.

Synergistic effect of high-mobility group box-1 and lipopolysaccharide on cytokine induction in bovine peripheral blood mononuclear cells.

High-mobility group box 1 (HMGB1) is one of the potent endogenous adjuvants released by necrotic and activated innate immune cells. HMGB1 modulates innate and adaptive immune responses in humans and mice by mediating immune cells crosstalk. However, the immuno-modulatory effects of HMGB1 in the bovine immune system are not clearly known. In this study, the effect of bovine HMGB1 alone or in combination with LPS on the expression kinetics of cytokines upon in vitro stimulation of bovine peripheral blood mononuclear cells (PBMCs) was investigated by quantitative PCR assay. The biological activity of bovine HMGB1 expressed in this prokaryotic expression system was confirmed by its ability to induce nitric oxide secretion in RAW 264.7 cells. The present results indicate that HMGB1 induces a more delayed TNF-α response than does LPS in stimulated PBMCs. However, IFN-γ, IFN-ß and IL-12 mRNA transcription peaked at 6 hr post stimulation after both treatments. Further, HMGB1 and LPS heterocomplex up-regulated TNF-α, IFN-γ and IL-12 mRNA expression significantly than did individual TLR4 agonists. The heterocomplex also enhanced the expression of TLR4 on bovine PBMCs. In conclusion, the data indicate that HMGB1 and LPS act synergistically and enhance proinflammatory cytokines, thereby eliciting Th1 responses in bovine PBMCs. These results suggest that HMGB1 can act as an adjuvant in modulating the bovine immune system and thus lays a foundation for using HMGB1 as an adjuvant in various bovine vaccine preparations.

Bacterial Ghosts of Escherichia coli Drive Efficient Maturation of Bovine Monocyte-Derived Dendritic Cells.

Bacterial ghosts (BGs) are empty cell envelopes derived from Gram-negative bacteria. They not only represent a potential platform for development of novel vaccines but also provide a tool for efficient adjuvant and antigen delivery system. In the present study, we investigated the interaction between BGs of Escherichia coli (E. coli) and bovine monocyte-derived dendritic cells (MoDCs). MoDCs are highly potent antigen-presenting cells and have the potential to act as a powerful tool for manipulating the immune system. We generated bovine MoDCs in vitro from blood monocytes using E. coli expressed bovine GM-CSF and IL-4 cytokines. These MoDCs displayed typical morphology and functions similar to DCs. We further investigated the E. coli BGs to induce maturation of bovine MoDCs in comparison to E. coli lipopolysaccharide (LPS). We observed the maturation marker molecules such as MHC-II, CD80 and CD86 were induced early and at higher levels in BG stimulated MoDCs as compared to the LPS stimulated MoDCs. BG mediated stimulation induced significantly higher levels of cytokine expression in bovine MoDCs than LPS. Both pro-inflammatory (IL-12 and TNF-α) and anti-inflammatory (IL-10) cytokines were induced in MoDCs after BGs stimulation. We further analysed the effects of BGs on the bovine MoDCs in an allogenic mixed lymphocyte reaction (MLR). We found the BG-treated bovine MoDCs had significantly (p<0.05) higher capacity to stimulate allogenic T cell proliferation in MLR as compared to the LPS. Taken together, these findings demonstrate the E. coli BGs induce a strong activation and maturation of bovine MoDCs.