Periodontal Biofilm Research Articles

Inflammation indices in association with periodontitis and cancer.

Inflammation is a complex physiological process that plays a pivotal role in many if not all pathological conditions, including infectious as well as inflammatory diseases, like periodontitis and autoimmune disorders. Inflammatory response to periodontal biofilms and tissue destruction in periodontitis is associated with the release of inflammatory mediators. Chronic inflammation can promote the development of cancer. Persistence of inflammatory mediators plays a crucial role in this process. Quantification and monitoring of the severity of inflammation in relation to cancer is essential. Periodontitis is mainly quantified based on the severity and extent of attachment loss and/or pocket probing depth, in addition with bleeding on probing. In recent years, studies started to investigate inflammation indices in association with periodontal diseases. To date, only few reviews have been published focusing on the relationship between blood cell count, inflammation indices, and periodontitis. This review presents a comprehensive overview of different systemic inflammation indices, their methods of measurement, and the clinical applications in relation to periodontitis and cancer. This review outlines the physiological basis of inflammation and the underlying cellular and molecular mechanisms of the parameters described. Key inflammation indices are commonly utilized in periodontology such as the neutrophil to lymphocyte ratio. Inflammation indices like the platelet to lymphocyte ratio, platelet distribution width, plateletcrit, red blood cell distribution width, lymphocyte to monocyte ratio, delta neutrophil index, and the systemic immune inflammation index are also used in hospital settings and will be discussed. The clinical roles and limitations, relationship to systemic diseases as well as their association to periodontitis and treatment response are described.

Phage-based biocontrol of Porphyromonas gingivalis through indirect targeting.

Lytic phages are valuable agents for targeted elimination of bacteria in diverse applications. Nevertheless, lytic phages are difficult to isolate for some target pathogens. We offer proof of concept that this limitation may be overcome via indirect phage targeting, which involves knocking out species that interact closely with and benefit the primary problematic target bacteria. Our target (P. gingivalis) only forms a periodontal pathogenic biofilm if the pioneer colonizer (S. gordonii) offers its surface for P. gingivalis to attach. Phage predation of the co-adhesive S. gordonii significantly reduced abundance of the target pathogen by >99%, decreased the total biofilm biomass by >44%, and suppressed its production of cytotoxic metabolic byproducts. Thus, this research extends the scope of phage-based biocontrol for public health protection.

NIR-II triggered Cu(I) phosphide for chemodynamic and photothermal periodontitis treatment: Efficient reduction of bacterial co-aggregation

The synergy between chemodynamic therapy (CDT) and photothermal therapy (PTT) offers a promising antimicrobial strategy for periodontitis, yet faces challenges like complex material structure and limited NIR-I light penetration. Additionally, low endogenous H2O2 levels in biofilm and a focus on bacterial eradication over colonization prevention limit current treatments. To address these issues, we newly introduce a single-material system (Cu3P@PAH@Lox) that integrates dual functionalities to synergistically enhance antimicrobial effects and significantly reduce pathogen co-aggregation. This system utilizes PTT to increase local temperature, boosting •OH production in CDT while downregulating heat shock proteins to enhance PTT efficacy, forming a self-reinforcing feedback loop. Lactate oxidase (Lox) is employed to convert lactate—a metabolite in periodontal biofilm—into H2O2, further amplifying CDT's potential. In vitro Cu3P@PAH@Lox demonstrates a remarkable synergistic effect against dual-species biofilms by more than 2-log reduction of colony-forming unit. Moreover, Cu3P@PAH@Lox exhibits outstanding synergistic antibacterial performances to alleviate inflammation and destruction of tissue in vivo periodontitis model. Furthermore, the mechanism of pathogen co-aggregation disruption by PTT is verified via the Cbe-Ltp1-Ptk1-fimA signaling pathway. This single-material multimodal system we have herein demonstrated for the first time marks a significant advancement in periodontitis treatment, eradicating microbes and preventing bacterial colonization, offering a path to comprehensive periodontal care. Statement of significanceThe synergy between chemodynamic therapy (CDT) and photothermal therapy (PTT) has been considered a promising therapy for periodontitis. Yet, facing challenges, the complex material structure, limited NIR-I light penetration, low endogenous H2O2 level in biofilm, and a focus on bacterial eradication over colonization prevention are still insufficient. This study pioneers a unique, single-material system (Cu3P@PAH@Lox) that synergistically enhances antimicrobial effects and substantially curtails pathogen co-aggregation, advancing periodontitis therapy. By exploiting PTT to elevate local temperatures, thereby increasing hydroxyl radical production in CDT and concurrently suppressing heat shock proteins, the system establishes a potent, self-enhancing loop. Furthermore, lactate oxidase is innovatively utilized to convert lactate from periodontal biofilm into hydrogen peroxide, augmenting the efficacy of CDT. The introduction of Cu3P@PAH@Lox is poised to revolutionize periodontitis treatment, eliminating microbes and impeding bacterial colonization, thereby charting a course for comprehensive periodontal management.

Gold core@CeO2 halfshell Janus nanocomposites catalyze targeted sulfate radical for periodontitis therapy

The sulfate radical (SO4•-), known for its high reactivity and long lifespan, has emerged as a potent antimicrobial agent. Its exceptional energy allows for the disruption of vital structures and metabolic pathways in bacteria that are usually inaccessible to common radicals. Despite its promising potential, the efficient generation of this radical, particularly through methods involving enzymes and photocatalysis, remains a substantial challenge. Here, we capitalized on the peroxidase (POD)-mimicking activity and photocatalytic properties of cerium oxide (CeO2) nanozymes, integrating these properties with the enhanced concept of plasma gold nanorod (GNR) to develop a half-encapsulated core@shell GNRs@CeO2 Janus heterostructure impregnated with persulfate. Under near-infrared irradiation, the GNRs generate hot electrons, thereby boosting the CeO2's enzyme-like activity and initiating a potent reactive oxygen species (ROS) storm. This distinct nanoarchitecture facilitates functional specialization, wherein the heterostructure and efficient light absorption ensured continuous hot electron flow, not only enhancing the POD-like activity of CeO2 for the production of SO4•- effectively, but also contributing a significant photothermal effect, disrupting periodontal plaque biofilm and effectively eradicating pathogens. Furthermore, the local temperature elevation synergistically enhances the POD-like activity of CeO2. Transcriptomics analysis, as well as animal experiments of the periodontitis model, have revealed that pathogens undergo genetic information destruction, metabolic disorders, and pathogenicity changes in the powerful ROS system, and profound therapeutic outcomes in vivo, including anti-inflammation and bone preservation. This study demonstrated that energy transfer to augment nanozyme activity, specifically targeting ROS generation, constitutes a significant advancement in antibacterial treatment.

In-vitro effects of novel periodontal scalers with a planar ultrasonic piezoelectric transducer on periodontal biofilm removal, dentine surface roughness, and periodontal ligament fibroblasts adhesion

ObjectivesTo compare ultrasonic scaler prototypes based on a planar piezoelectric transducer with different working frequencies featuring a titanium (Ti-20, Ti-28, and Ti-40) or stainless steel (SS-28) instrument, with a commercially available scaler (com-29) in terms of biofilm removal and reformation, dentine surface roughness and adhesion of periodontal fibroblasts.Materials and methodsA periodontal multi-species biofilm was formed on specimens with dentine slices. Thereafter specimens were instrumented with scalers in a periodontal pocket model or left untreated (control). The remaining biofilms were quantified and allowed to reform on instrumented dentine slices. In addition, fibroblasts were seeded for attachment evaluation after 72 h of incubation. Dentine surface roughness was analyzed before and after instrumentation.ResultsAll tested instruments reduced the colony-forming unit (cfu) counts by about 3 to 4 log10 and the biofilm quantity (each p < 0.01 vs. control), but with no statistically significant difference between the instrumented groups. After 24-hour biofilm reformation, no differences in cfu counts were observed between any groups, but the biofilm quantity was about 50% in all instrumented groups compared to the control. The attachment of fibroblasts on instrumented dentine was significantly higher than on untreated dentine (p < 0.05), with the exception of Ti-20. The dentine surface roughness was not affected by any instrumentation.ConclusionsThe planar piezoelectric scaler prototypes are able to efficiently remove biofilm without dentine surface alterations, regardless of the operating frequency or instrument material.Clinical relevanceUltrasonic scalers based on a planar piezoelectric transducer might be an alternative to currently available ultrasonic scalers.

Selenium Nanoparticle Activity against S. mutans Biofilms as a Potential Treatment Alternative for Periodontitis.

The disruption of periodontal biofilms and prevailing antimicrobial resistance issues continue to pose a great challenge to the treatment of periodontitis. Here, we report on selenium nanoparticles (SeNPs) as a treatment alternative for periodontitis by determining their antibiofilm activity against S. mutans biofilms and the potential role of particle size in disrupting biofilms. SeNPs were synthesised via a reduction reaction. Various physicochemical characterisations were conducted on the NPs, including size and shape. The microbroth dilution method was used to conduct the biofilm and antibiofilm assay against S. mutans, which was analysed by absorbance. SeNPs displayed hydrodynamic sizes as low as 46 ± 4 nm at a volume ratio of 1:5 (sodium selenite/ascorbic acid) with good monodispersity and stability. Hydrodynamic sizes of SeNPs after resuspension in tryptic soy broth supplemented with 2.5% sucrose (TSB + 2.5% suc.) and incubated at 37 °C for 24 h, ranged from 112 to 263 nm, while the zeta potential values increased to greater than -11 mV. The biofilm assay indicated that S. mutans are weakly adherent, bordering on moderately adherent biofilm producers. The minimum biofilm inhibitory concentration (MBIC) was identified at 500 µg/mL. At a 1000 µg/mL concentration, SeNPs were able to inhibit S. mutan biofilms up to 99.87 ± 2.41% at a volume ratio of 1:1. No correlation was found between antibiofilm activity and particle size; however, antibiofilm activity was proven to be concentration-dependant. SeNPs demonstrate antibiofilm activity and may be useful for further development in treating periodontitis.

Oxygen enrichment mediated by calcium peroxide loaded gelatin methacrylate hydrogel eradicates periodontal biofilms

The low oxygen environment of the periodontal pocket favors pathogenic anaerobes' growth, biofilm formation, and quick recurrence after periodontal treatment. In contrast, oxygen is detrimental to anaerobes, such as Porphyromonas gingivalis (P. gingivalis), since they lack a complete anti-oxidation mechanism to detoxify the oxygen challenge. Therefore, consistently feeding pathogenic anaerobes with abundant oxygen would be an effective strategy to combat them. Here, we reported injectable oxygen-generating hydrogels as oxygen mediators to alleviate the local anaerobic environment and eliminate periodontal pathogens. Gelatin methacrylate (GelMA) hydrogels loaded with calcium peroxide (CPO) possessed excellent injectability and exhibited burst releases of oxygen within 24 h with a 40 % oxygen tension peak. CPO-GelMA hydrogels with CPO concentrations of 5, 10, and 15 % reduced 60, 99, and 89.9 % viable P. gingivalis, respectively. Five percentage CPO-GelMA hydrogel downregulated gingipain and fimA gene expression in P. gingivalis without resistance development. Moreover, the CPO-GelMA hydrogels remarkably prevented biofilm formation and eradicated both monospecies and multispecies bacterial biofilms. In conclusion, CPO-GelMA hydrogels exert remarkable antimicrobial and antibiofilm effects on subgingival biofilms, providing a promising strategy for periodontal treatment.

Use of over-the-counter mouthwashes as an additional measure in individual oral prophylaxis on adults with plaque-induced gingivitis: a double-blind, parallel, randomized controlled trial

BackgroundPlaque-induced gingivitis is a chronic inflammatory condition characterized by complete reversibility of tissue damage once the periodontal biofilm has been disorganised. The aim of this study was to evaluate the efficacy of two commercially available mouthwashes (MWs) versus a chlorhexidine (CHX) 0.12% MW in reducing gingival bleeding (GB) in adults with plaque-induced gingivitis.MethodsThe present study was a double-blind, parallel, randomized controlled trial involving 6492 gingival sites (i.e. 39 subjects × 28 teeth × 6 sites/tooth) aged 18–75 years. During a 2-week period, subjects were randomized to receive MWs: a control CHX 0.12% MW (group C, 1818 sites); a MW test containing CHX 0.09% + Citrox®/P complex (group CX, 2628 sites); a MW test based on natural compounds (group P, 2016 sites). GB was assessed at the inclusion visit (T0) and after 2 weeks of MW use (T1). Analyses of GB were compared between groups and then restricted to subjects with bleeding sites between 10 and 30% (moderate gingivitis) or ≥ 30% (severe gingivitis) at T0. Pairwise comparisons were made between groups and logistic regression was used to identify correlates of GB (T1).ResultsFor total bleeding site analysis, GB reduction between T0 and T1 ranged from 23% (C), 26% (CX) and 36% (P), respectively (all p < 0.05). Multiple comparison between groups showed that group C was significantly less effective (p < 0.05) than groups CX and P. Splitting the analysis, in patients with severe gingivitis (≥ 30% bleeding sites at T0), all MWs had a positive effect on GB with a reduction at T1 of 36% (C), 33% (CX) and 42% (P), respectively. While GB reduction between T0 and T1, was significant for all groups, the comparison among groups showed no significant difference between group C and CX, whereas the improvement was significant for group P. On the other hand, in adults with moderate gingivitis (< 30% bleeding sites at T0), only CX and P had a positive effect on GB reduction at T1(9% in CX and 2% in P, respectively), although the differences between the three groups were not significant.ConclusionThe daily use of MWs with natural components (groups P and CX) for 2 weeks should be considered positively as an adjunct to individual oral prophylaxis to reduce GB compared to the control MW containing CHX 0.12% (group C) in healthy adults with plaque-induced gingivitis. For subjects with severe gingivitis, it is advisable to first use natural MW (P) and then MW based on CHX 0.09% with natural components (CX), compared to MW with CHX 0.12% (C). For adults with moderate gingivitis, P and CX can be advisable, even if no definitive recommendations can be drawn.Trial RegistrationACTRN12622000215729, 07/02/2022.

Antibiofilm Activity of Probiotic Strains against Aggregatibacter actinomycetemcomitans

Aggregatibacter actinomycetemcomitans is a significant agent for periodontal disease. It develops biofilms on gingival plaque and promotes chronic pro-inflammatory response to periodontal tissues. As bacterial cells within the biofilm, this pathogen successfully eluded the human immune system by preventing the uptake of biofilm cells by phagocytic cells. Antibiotic-resistant biofilms might harm the environment, increase mortality, and add to the cost of healthcare. Anti-biofilms are necessary to prevent the growth of biofilms. It has been demonstrated that probiotics change how immune system cells produce cytokines. Therefore, the efficacy of five probiotic lactobacillus strains to prevent A. actinomycetemcomitans from growing bacteria and forming biofilms was evaluated. The organisms were incubated in anaerobic conditions at 37°C for a certain period. The antimicrobial test was done using the lawn method and measuring the size of the inhibition zone produced by the probiotic spots for each probiotic strain. Tetracycline antibiotic disc was used as a positive control to compare the size of the inhibition zone. The antibiofilm activity was performed using probiotic strains at their exponential and stationary phases against the exponential phase of A. actinomycetemcomitans. Our finding showed the probiotic lactobacillus could inhibit the growth of A. actinomycetemcomitans. The highest inhibition was demonstrated by L. johnsonii NBRC 13952. Besides, all probiotic lactobacillus used for this study significantly (p&lt;0.05) inhibited the A. actinomycetemcomitans biofilm using both stationary and exponential phases. In conclusion, probiotic lactobacillus might be a potential agent that can be used to counteract the periodontal pathogen biofilm. Further study should be done to elucidate the antibiofilm mechanism, and several trials of the probiotic lactobacilli against multiple species of biofilms might be helpful in evaluating their effectiveness.

Spatiotemporal monitoring of a periodontal multispecies biofilm model: demonstration of prebiotic treatment responses.

Biofilms are complex polymicrobial communities which are often associated with human infections such as the oral disease periodontitis. Studying these complex communities under controlled conditions requires in vitro biofilm model systems that mimic the natural environment as close as possible. This study established a multispecies periodontal model in the drip flow biofilm reactor in order to mimic the continuous flow of nutrients at the air-liquid interface in the oral cavity. The design is engineered to enable real-time characterization. A community of five bacteria, Streptococcus gordonii-GFPmut3*, Streptococcus oralis-GFPmut3*, Streptococcus sanguinis-pVMCherry, Fusobacterium nucleatum, and Porphyromonas gingivalis-SNAP26 is visualized using two distinct fluorescent proteins and the SNAP-tag. The biofilm in the reactor develops into a heterogeneous, spatially uniform, dense, and metabolically active biofilm with relative cell abundances similar to those in a healthy individual. Metabolic activity, structural features, and bacterial composition of the biofilm remain stable from 3 to 6 days. As a proof of concept for our periodontal model, the 3 days developed biofilm is exposed to a prebiotic treatment with L-arginine. Multifaceted effects of L-arginine on the oral biofilm were validated by this model setup. L-arginine showed to inhibit growth and incorporation of the pathogenic species and to reduce biofilm thickness and volume. Additionally, L-arginine is metabolized by Streptococcus gordonii-GFPmut3* and Streptococcus sanguinis-pVMCherry, producing high levels of ornithine and ammonium in the biofilm. In conclusion, our drip flow reactor setup is promising in studying spatiotemporal behavior of a multispecies periodontal community. IMPORTANCE Periodontitis is a multifactorial chronic inflammatory disease in the oral cavity associated with the accumulation of microorganisms in a biofilm. Not the presence of the biofilm as such, but changes in the microbiota (i.e., dysbiosis) drive the development of periodontitis, resulting in the destruction of tooth-supporting tissues. In this respect, novel treatment approaches focus on maintaining the health-associated homeostasis of the resident oral microbiota. To get insight in dynamic biofilm responses, our research presents the establishment of a periodontal biofilm model including Streptococcus gordonii, Streptococcus oralis, Streptococcus sanguinis, Fusobacterium nucleatum, and Porphyromonas gingivalis. The added value of the model setup is the combination of simulating continuously changing natural mouth conditions with spatiotemporal biofilm profiling using non-destructive characterization tools. These applications are limited for periodontal biofilm research and would contribute in understanding treatment mechanisms, short- or long-term exposure effects, the adaptation potential of the biofilm and thus treatment strategies.

An in vitro dynamic bioreactor model for evaluating antimicrobial effectiveness on periodontal polymicrobial biofilms: a proof-of-concept study.

The aim of this study was to investigate an in vitro dynamic bioreactor model by evaluating the antimicrobial effect of clinically relevant amoxicillin doses on polymicrobial microcosm biofilms derived from subgingival plaque. Biofilms from pooled subgingival plaque were grown for 108 hours in control and experimental dynamic biofilm reactors. Amoxicillin was subsequently infused into the experimental reactor to simulate the pharmacokinetic profile of a standard 500mg thrice-daily dosing regimen over 5 days and biofilms were assessed by live/dead staining, scanning electron microscopy, and quantitative polymerase chain reaction. Following establishment of the oral microcosm biofilms, confocal imaging analysis showed a significant increase in dead bacteria at 8hours (p = 0.0095), 48hours (p = 0.0070), 96hours (p = 0.0140), and 120hours (p<0.0001) in the amoxicillin-treated biofilms compared to the control biofilms. Nevertheless, viable bacteria remained in the center of the biofilm at all timepoints. Significant reductions/elimination in Campylobacter rectus, Tannerella forsythia, Aggregatibacter actinomycetemcomitans, and Peptostreptococcus anaerobius was observed among the amoxicillin-treated biofilms at the 96 and 120hour timepoints. A novel in vitro dynamic model of oral microcosm biofilms was effective in modeling the antimicrobial effect of a pharmacokinetically simulated clinically relevant dose of amoxicillin.

Oxygen-generating Hydrogels Disrupt Periodontal Biofilms by Local Oxygen Enrichment

Oxygen-generating Hydrogels Disrupt Periodontal Biofilms by Local Oxygen Enrichment

Effect of chlorhexidine mouthwashes on periodontal parameters and extrinsic tooth staining in orthodontic patients

Periodontal health and biofilm control are primordial to success in orthodontic treatment. This study aimed to evaluate the effect of chlorhexidine (CHX) mouthwashes on periodontal status and extrinsic tooth staining in orthodontic patients. Thirty-three patients of both sexes, aged 11-33 years, under orthodontic treatment with fixed appliances at<16 months, were randomly distributed into 2 groups. In the control group, patients received mechanical hygiene instruction, and in the experimental group, patients also used CHX wash twice a week for 60 days. The effectivity of the protocols was evaluated using the plaque, gingival, gingival bleeding, and discoloration indexes before the hygiene protocol was applied, 15, 30, and 60 days after protocol implementation. In the experimental group, there was a decrease in the plaque, gingival, and gingival bleeding indexes at the different evaluation periods (P<0.05). In addition, there was a significant difference in the discoloration index at 60 days compared with initial time points after implementing hygiene protocols in the experimental group (P<0.05). In contrast, there were no significant differences in plaque, gingival, gingival bleeding, and discoloration indexes in the control group at any time (P>0.05). CHX mouthwash administered 30 days, twice a week, significantly improved the periodontal status with mild brown staining. After this period, expressive extrinsic tooth staining was observed.

Chlorin-e6 conjugated to the antimicrobial peptide LL-37 loaded nanoemulsion enhances photodynamic therapy against multi-species biofilms related to periodontitis

In our previous studies, Chlorin-e6 (Ce6) demonstrated a significant reduction of microorganisms’ viability against multi-species biofilm related to periodontitis while irradiated with blue light. However, the conjugation of Ce6 and antimicrobial peptides, and the incorporation of this photosensitizer in a nanocarrier, is still poorly explored. We hypothesized that chlorin-e6 conjugated to the antimicrobial peptide LL-37 loaded nanoemulsion could inhibit a multi-species biofilm related to periodontitis during photodynamic therapy (PDT), the pre-treatment with hydrogen peroxide was also tested. The nanoemulsion (NE) incorporated with Ce6 was characterized regarding the physiochemical parameters. Images were obtained by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Later, the Ce6 and LL-37 incorporated in NE was submitted to UV–Vis analysis and Reactive Oxygen Species (ROS) assay. Finally, the combined formulation (Ce6+LL-37 in nanoemulsion) was tested against multi-species biofilm related to periodontitis. The formed nanoformulation was kinetically stable, optically transparent with a relatively small droplet diameter (134.2 unloaded and 146.9 loaded), and weak light scattering. The NE system did not impact the standard UV–VIS spectra of Ce6, and the ROS production was improved while Ce6 was incorporated in the NE. The combination of Ce6 and LL-37 in NE was effective to reduce the viability of all bacteria tested. The treatment with hydrogen peroxide previous to PDT significantly impacted bacterial viability. The current aPDT regimen was the best already tested against periodontal biofilm by our research team. Our results suggest that this combined protocol must be exploited for clinical applications in localized infections such as periodontal disease.- Nanoemulsion demonstrated to be an excellent nanocarrier for photodynamic application.- Chlorin-e6 incorporated in nanoemulsion showed great physicochemical and biophotonic parameters.- The combination of chlorin-e6 and LL-37 peptide in nanoemulsion is effective to eliminate periodontal pathogenic bacteria.- The treatment with hydrogen peroxide previous to PDT significantly impacted bacterial viability.

In vitro activity of hyaluronic acid and human serum on periodontal biofilm and periodontal ligament fibroblasts

ObjectivesA beneficial effect of cross-linked hyaluronic acid (cHA) on periodontal wound healing and regeneration has recently been demonstrated. The present in vitro study was designed to obtain deeper knowledge on the effect of cHA when applied in the gingival sulcus (serum-rich environment) during non-surgical periodontal therapy.Materials and methodsThe influence of cHA, human serum (HS), and cHA/HS on (i) a 12-species biofilm formation, (ii) the adhesion of periodontal ligament fibroblasts (PDLF) to dentine surface, (iii) the expression and secretion of interleukin-8, and (iv) the expression of receptors of HA in PDLF and gingival fibroblasts (GF) were evaluated.ResultsAt 4 h of biofilm formation, cHA and HS in combination (cHA/HS) slightly decreased the colony-forming unit counts in biofilm whereas the metabolic activity of biofilm was reduced in all test groups (cHA, HS, cHA/HS) vs. control. At 24 h, the quantity of biofilm was reduced in all test groups vs. untreated control. The test substances did not affect adhesion of PDLF to dentin. HS increased the expression of IL-8 by PDLF and GF which was partially downregulated by cHA. HS and/or cHA promoted the expression of the HA receptor RHAMM in GF but not in PDLF.ConclusionsIn summary, the present data indicate that serum neither negatively affect the activity of cHA against periodontal biofilm nor had any unwanted influence on the activity of PDLF.Clinical relevanceThese findings lend additional support for the positive effects of cHA on cells involved in periodontal wound healing, thus pointing to its potential use in non-surgical periodontal therapy.

HbA1c, And Blood Glucose, Changes When Treating Periodontal Disease with the Perio Protect Method™

Periodontal disease is a chronic wound resulting in host elevated local and systemic inflammatory markers in response to the microorganisms of the periodontal biofilm. Separately from its importance as an oral ailment, chronic periodontitis has gained relevance since it can develop into a systemic condition characterized by hyper-inflammation, disruption of the innate and adaptive immune system, and other system-wide alterations. There is compelling evidence that inflammation and chronic infection play an essential role in the development of inflammatory markers and of type 2 diabetes. Studies in humans suggest that circulating inflammatory marker levels may predict type 2 diabetes years in advance of the onset of the disease. Local and systemic expression of inflammatory cytokines, such as TNF-alpha and IL-6, increase in individuals with periodontitis. TNF-alpha and IL-6 impair intracellular insulin signaling, which may lead to insulin resistance and are significantly associated with cardiovascular events and stroke. This case study demonstrates treating periodontal disease with a direct medication delivery method (Perio Protect Method™) manages the patient’s periodontal disease and lowers daily blood glucose and HbA1c levels.

Evaluation of the antibacterial effect of hydroalcoholic extract of the galls of Quercus infectoria on Aggregatibacter actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans (Aa) plays a vital role in some destructive forms of periodontitis. While mechanical and chemical plaque control is the first step in periodontitis treatment, side effects of adjunctive chemical agents such as chlorhexidine (CHX) mouthwash have led to the application of natural alternatives with minimal side effects. Therefore, this study evaluated the antibacterial effect of the hydroalcoholic extract of Quercus infectoria (Qi) galls on Aa in vitro. The hydroalcoholic extract of Qi was obtained by the maceration method, and Aa bacterial strain was cultured. The inhibition zone diameter was measured through the agar well diffusion method. Also, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were determined by the broth microdilution method. All the experiments were repeated three times. 0.2% CHX was used as a control. The inhibition zone diameter of Aa increased with increasing concentration of Qi extract. While MIC and MBC values for CHX were 0.0039 and 0.0078 mg/mL, respectively, both MIC and MBC values of the Qi extract for this bacterium were similar, i.e., 2.5 mg/mL, which was significantly higherd. Since other in vivo studies have confirmed the other properties of this extract and its safety in terms of cytotoxicity and mutagenicity, hydroalcoholic extract of Qi may be used in mouthwashes or local delivery systems to affect periodontal biofilm.

Microfabrication approaches for oral research and clinical dentistry

Currently, there is a variety of laboratory tools and strategies that have been developed to investigate in-vivo processes using in-vitro models. Amongst these, microfabrication represents a disruptive technology that is currently enabling next-generation biomedical research through the development of complex laboratory approaches (e.g., microfluidics), engineering of micrometer scale sensors and actuators (micropillars for traction force microscopy), and the creation of environments mimicking cell, tissue, and organ-specific contexts. Although microfabrication has been around for some time, its application in dental and oral research is still incipient. Nevertheless, in recent years multiple lines of research have emerged that use microfabrication-based approaches for the study of oral diseases and conditions with micro- and nano-scale sensitivities. Furthermore, many investigations are aiming to develop clinically relevant microfabrication-based applications for diagnostics, screening, and oral biomaterial manufacturing. Therefore, the objective of this review is to summarize the current application of microfabrication techniques in oral sciences, both in research and clinics, and to discuss possible future applications of these technologies for in-vitro studies and practical patient care. Initially, this review provides an overview of the most employed microfabrication methods utilized in biomedicine and dentistry. Subsequently, the use of micro- and nano-fabrication approaches in relevant fields of dental research such as endodontic and periodontal regeneration, biomaterials research, dental implantology, oral pathology, and biofilms was discussed. Finally, the current and future uses of microfabrication technology for clinical dentistry and how these approaches may soon be widely available in clinics for the diagnosis, prevention, and treatment of relevant pathologies are presented.

Bovine Milk and Yogurt Affect Oral Microorganisms and Biofilms In-Vitro.

To evaluate the effect of bovine milk and yogurt on selected oral microorganisms and different oral biofilms. Milk was prepared from 0.5% fat (low-fat) and 16% fat (high fat) milk powder. For yogurt preparation, the strains Lactobacillus delbrueckii ssp. bulgarcius and Streptococcus thermophilus were added to the milk. Minimal inhibitory concentrations (MIC) and minimal microbiocidal concentrations (MMC) of the test compounds were measured against various microorganisms by the microbroth dilution technique. Cariogenic periodontal biofilms and one containing Candida were created on plastic surfaces coated with test substances. Further, preformed biofilms were exposed to the test substances at a concentration of 100% for 10 min and thereafter 10% for 50 min. Both colony forming units (cfu) and metabolic activity were quantified in the biofilms. Neither high-fat milk, low-fat milk nor casein inhibited the growth of any species. Yogurt and L. delbrueckii ssp. bulgaricus at low MIC and MMC suppressed the growth of Porphyromonas gingivalis and other bacteria associated with periodontal disease. High-fat yogurt decreased cfu in the forming periodontal biofilm by 90%. Both low- and high-fat yogurts reduced metabolic activity in newly forming and preformed periodontal and Candida biofilms, but not in the cariogenic biofilm. Yogurt and L. delbru eckii ssp. bulgaricus, but not milk, were bactericidal against periodontopathogenic bacteria. Yoghurt reduced the metabolic activity of a Candida biofilm and a periodontal biofilm. Yogurt and L. delbrueckii ssp. bulgaricus may have potential in prevention and therapy of periodontal diseases and Candida infections.

Evaluation of antimicrobial efficacy of immunoglobulin Y (IgY) against periodontal biofilm.

To determine the action of immunoglobulin Y (IgY) on supragingival microbiota and on subgingival microbiota in patients with gingivitis and periodontitis through microbial reduction assay. 40 systemically healthy patients were divided into two groups (gingivitis and periodontitis) with 20 patients per group. Supragingival and subgingival plaque samples were collected from each patient in Group I and Group II, respectively. Sample 1 and Sample 2 from each patient were immediately transferred into sterile Eppendorf tube 1 and tube 2 with microbial transport media, respectively. Both the tubes were then immediately transferred into an anaerobic jar and sent to the microbial facility. IgY was then added to these samples. All the samples were collected in duplicate vials to check the in vitro antimicrobial activity of microbes with IgY and without IgY. Microbial reduction percentage was calculated based on the colony count comparing the colonies with and without IgY. The mean CFUs in the gingivitis group with IgY samples was significantly lesser as compared to the periodontitis group. The mean CFUs in gingivitis and periodontitis group with IgY samples was significantly lesser as compared to those without IgY samples. IgY has a significant role in the reduction of bacterial count in supragingival and subgingival plaque samples. So, IgY when used as a local drug delivery agent or mouthwash, as an adjunct to scaling and root planing may reduce gingival and periodontal diseases but further studies showing its effect must be carried out to validate the same.