Biofilm Elimination Research Articles

Supragingival Biofilm: Toothpaste and Toothbrushes.

The formation of a physiological biofilm cannot be avoided under normal circumstances. However, the consequences of a supragingivally located biofilm, such as caries, gingivitis and, as a further effect, periodontitis, are relatively easy to avoid. The simplest and most common method used worldwide for the elimination of biofilm is periodic mechanical removal using a toothbrush or similar tools, such as chewing sticks or woods. This method was already used in ancient Egypt, and is still being used today, albeit advanced and improved with the help of toothpastes. Here we give a summary of the most common toothbrushes, highlighting their advantages and disadvantages. Furthermore, we provide an overview of the most common toothpastes, their ingredients, and functions. In addition, the ingredients will be critically evaluated and recommendations given for the use or non-use of certain ingredients for different target groups, such as children, healthy adults, or patients with special needs.

Combinatory Effect of ALA-PDT and Itraconazole Treatment for Trichosporon asahii.

Trichosporiosis is an opportunistic infection that includes superficial infections, white piedra, hypersensitivity pneumonitis, and invasive trichosporonosis. The effect of antifungal agents against these infections is largely weakened by drug resistance and biofilms-related virulence. Photodynamic therapy (PDT) is a new therapeutic approach developed not only to combat cancerous lesions but also to treat infectious diseases such as fungal infections. However, there are few studies on the antimicrobial mechanism of 5-aminolevulinic acid PDT (ALA-PDT) in treating Trichosporon. In this work, we explored the possibility of combining ALA-PDT with an antifungal agent to enhance the therapeutic efficacy of Trichosporon asahii (T. asahii) in a clinical setting and in vitro. The biofilms of T. asahii were constructed by a 96-well plate-based method in vitro. The planktonic and adherent T. asahii were exposed to different concentrations of photosensitizers and different light doses. After PDT treatment, counting colony-forming units and tetrazolium (XTT) reduction assay were used to estimate the antifungal efficacy. The minimal inhibitory concentration of itraconazole before and after PDT treatment was determined by the broth dilution method, and XTT viability assay was used to detect and evaluate the synergistic potential of ALA-PDT and itraconazole combinations in inhibiting biofilms. Scanning electron microscopy (SEM) was performed to assess the disruption of biofilms. Using combination therapy, we have successfully treated a patient who had a T. asahii skin infection. Further in vitro studies showed that the antifungal effect of ALA-PDT on planktonic and adherent T. asahii was dependent on the concentration of ALA and light dosages used. We also found that the sensitivity of both planktonic and biofilm cells to itraconazole were increased after ALA-PDT. Synergistic effect were observed for biofilms in ALA-PDT and itraconazole-combined treatment. The disruption of biofilms was confirmed by SEM, suggesting that ALA-PDT effectively damaged the biofilms and the destruction was further enhanced by ALA-PDT combination of antifungal agents. In conclusion, these data suggest that ALA-PDT could be an alternative strategy for controlling infections caused by Trichosporon. The combination therapy of ALA-PDT with itraconazole could result in increased elimination of planktonic cells and biofilms compared with single therapy. All these findings indicate that it could be a promising treatment against trichosporonosis. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

In Vitro Antimicrobial Activity of Fosfomycin, Vancomycin and Daptomycin Alone, and in Combination, Against Linezolid-Resistant Enterococcus faecalis.

IntroductionEnterococcus faecalis is a significant cause of nosocomial infections and is difficult to treat because of intrinsic and acquired resistance to many antibiotics. In addition, the emergence of linezolid-resistant E. faecalis (LZR-Efa) is reducing the choices available for anti-infective therapy. The aim of this study was to examine the in vitro antibacterial effects of fosfomycin (FM), vancomycin (VAN) and daptomycin (DAP), alone and in combination, against LZR-Efa.MethodsFive LZR-Efa strains and E. faecalis ATCC 29212 were studied. The antibacterial effects of FM, and of FM, VAN and DAP, were assessed using the time–kill assay. Biofilm formation and elimination were evaluated by crystal violet staining.ResultsWhen used at concentrations greater than 0.5 × MIC, FM did not produce dose-dependent effects against LZR-Efa isolates. The use of DAP (47.1 mg/L) alone, and FM (83 mg/L) combined with DAP (20.6 mg/L), produced a persistent inhibitory effect against both planktonic LZR-Efa isolates and those forming biofilms. In addition, FM and VAN combined with glucose-6-phosphate produced visible eradication effects against biofilms grown for 24 h, while DAP alone or combined with FM resulted in the best eradication activity against biofilms grown for 72 h prior to exposure.ConclusionThe use of FM combined with DAP provided the best potential therapeutic option for treating LZR-Efa infections out of those tested. In addition, the optimum treatment for biofilm elimination depended on the stage of biofilm formation.Electronic supplementary materialThe online version of this article (10.1007/s40121-020-00342-1) contains supplementary material, which is available to authorized users.

Self-Propelled Active Photothermal Nanoswimmer for Deep-Layered Elimination of Biofilm In Vivo

Increasing penetration of antibacterial agents into biofilm is a promising strategy for improvement of therapeutic effect and slowdown of the progression of antibiotic resistance. Herein, we design a near-infrared (NIR) light-driven nanoswimmer (HSMV). Under NIR light irradiation, HSMV performs efficient self-propulsion and penetrates into the biofilm within 5 min due to photothermal conversion of asymmetrically distributed AuNPs. The localized thermal (∼45 °C) and thermal-triggered release of vancomycin (Van) leads to an efficient combination of photothermal therapy and chemotherapy in one system. The active motion of HSMV increases the effective distance of photothermal therapy (PTT) and also improves the therapeutic index of the antibiotic, resulting in superior biofilm removal rate (>90%) in vitro. Notably, HSMV can eliminate S. aureus biofilms grown in vivo under 10 min of laser irradiation without damage to healthy tissues. This work may shed light on therapeutic strategies for in vivo treatment of biofilm-associated infections.

Ultrasmall AgNP-Impregnated Biocompatible Hydrogel with Highly Effective Biofilm Elimination Properties.

Ultrasmall silver nanoparticles (AgNPs; size < 3 nm) have attracted a great deal of interest as an alternative to commercially available antibiotics due to their ability to eliminate a wide range of microbial pathogens. However, most of these ultrasmall AgNPs are highly reactive and unstable, as well as susceptible to fast oxidation. Therefore, both the stability and toxicity remain major shortcomings for their clinical application and uptake. To circumvent these problems, we present a novel strategy to impregnate ultrasmall AgNPs into a biocompatible thermosensitive hydrogel that enables controlled release of silver alongside long-term storage stability and highly potent antibacterial activity. The advantage of this strategy lies in the combination of a homogenous dispersion of AgNPs in a hydrogel network, which serves as a sustained-release reservoir, and the unique feature of ultrasmall AgNP size, which provides an improved biofilm eradication capacity. The superior biofilm dispersion properties of the AgNP hydrogel is demonstrated in both single-species and multispecies biofilms, eradicating ∼80% of established biofilms compared to untreated controls. Notably, the effective antibacterial concentration of the formulation shows minimal toxicity to human fibroblasts and keratinocytes. These findings present a promising novel strategy for the development of AgNP hydrogels as an efficient antibacterial platform to combat resistant bacterial biofilms associated with wound infections.

Antimicrobial Properties of Nonantibiotic Agents for Effective Treatment of Localized Wound Infections: A Minireview.

Wounds present serious health problems in humans and animals. Importantly, if left untreated, wounds invariably lead to long-term morbidity. The inappropriate use and costs of antibiotics place significant challenges globally and affect the health budgets of many countries. Though some antibiotics are administered systemically, treatment of localized infections, in particular, chronic wound infections, does not need such therapy-this would minimize development of antibiotic resistance. Of these measures, nanoparticles of silver, ZnO, and gold seem to give promising results against common wound pathogens while having few limitations. Chemical components of essential oils, which are extracted from different plants, have been shown to act against common wound pathogens. Plant extracts have shown different mechanisms in biofilm elimination. Chlorhexidine and chlorine derivatives act as wound antiseptics. Attempts with biological agents such as maggots have also been shown to provide anti-infective as well as mechanical removal of wound debris. Honey, including those obtained from bees, has a wide coverage against wound pathogens. Glycerin and hypertonic saline are anti-infective through the concentration-dependent killing of pathogens. Hyperbaric oxygen acts against many wound pathogens, in particular anaerobes. This review is focused on nonantibiotic attempts for the cure of localized infections, in particular, chronic wounds with common wound pathogens.

Evaluation of Hydrogen Peroxide Vapour Biodecontamination Against Clinically Relevant Presentations of Multi-drug Resistant Biofilms

<h3>Background</h3> It has become acknowledged and accepted that microbiological contamination of surfaces within the healthcare environment can influence the rate of hospital acquired infection. Exploration of the presentation of microbiological organisms on healthcare surfaces has identified biofilms. Due to mechanisms such as the production of extracellular polymeric substances, enhanced protein and gene expression and co-ordinated support through quorum sensing, biofilms are more resistant to antimicrobial agents compared to planktonic phenotypes. Clinically relevant biofilms continue to exist on healthcare surfaces post high level terminal cleaning and research efforts are currently underway to investigate the link between the presence of biofilms within the patient environment and infection. <h3>Methods</h3> One high level disinfection technology used within the healthcare environment is hydrogen peroxide vapour (VH2O2). The presentation will introduce data from a 2-year government sponsored pilot evaluation to generate biofilms of clinically relevant microorganisms to be used in disinfectant evaluation and specifically to test the efficacy of a hydrogen peroxide vapour biodecontamination system. <h3>Results</h3> Biofilms of multi-drug resistant K.pneumoniae, S.aureus, A. baumannii, E.faecalis and P.aeruginosa with a microorganism density representative of the high loadings documented on clinical surfaces (averaging 7.15 Log10 CFU/cm2), were located in a 27m3 room, representative of a typical hospital side room. Biofilms were exposed to a standardised VH2O2 cycle, with timed removal of samples to determine the disinfection kinetics. The study included a desiccation stage to ensure biofilm formation experienced environmental stresses related to hospital surfaces. <h3>Conclusions</h3> Full elimination of all multi-drug resistant biofilms was achieved over triplicate repeats. Epifluorescence microscopy demonstrated massive disruption to biofilm colony formation and matrix structure as a result of oxidative damage. The presentation will discuss the applicability and implications of current mono-specie, planktonic-based standard disinfectant test methods and the need for standardised tests that better represent the challenge posed by biofilms.

Modeling the elimination of mature biofilms formed by Staphylococcus aureus and Salmonella spp. Using combined ultrasound and disinfectants

Biofilm formation by foodborne pathogens on food processing surfaces has contributed to numerous disease outbreaks and food recalls. We evaluated the following strategies for elimination of mature biofilm formed by Staphylococcus aureus and Salmonella spp. on stainless steel surfaces: acidic electrolyzed water (AEW), ozone water (OW), or ultrasound (40 kHz) alone, and combinations of ultrasound and disinfectants. The dynamics of elimination by combinations were determined using the Weibull and biphasic models. Treatment with AEW alone reduced the number of biofilm cells by approximately 3.0 log cfu/cm2, whereas less than 0.8 log cfu/cm2 of cells reduction was observed in biofilm exposed to OW or ultrasound alone, even with treatment for 20 min. The combination of AEW and ultrasound produced an obvious synergistic effect on biofilm reduction, achieving approximately 4.8 log cfu/cm2 reduction in Salmonella spp. biofilm. Interestingly, the biphasic model was a better fit than the Weibull model for the elimination process of mature biofilm formed by both pathogens and subjected to a combination of ultrasound and AEW, as determined by smaller values of the statistical parameters RMSE and AIC, although both models could evaluate the dynamic processes. Our findings indicated that a combination of ultrasound and AEW could effectively reduce the biofilm formed by pathogens on food contact surfaces, and that the biphasic model could predict the number of residual cells after biofilm exposure to this intervention approach.

A compact pulse-modulation air plasma jet for the inactivation of chronic wound bacteria: Bactericidal effects & host safety

Previously, a novel compact low-temperature plasma jet device with adjustable pulse-parameters was developed to use ambient air as a plasma source for the inactivation of chronic wound bacteria. Its adjustable burst-mode pulse was proved to be highly beneficial for the controllable generation of the desired reactive oxygen and nitrogen species. In this study, further bactericidal effects and its safety on human dermal fibroblasts (HDF) were investigated. The generated plasma inactivated Pseudomonas aeruginosa (PA) and Staphylococcus aureus (SA) effectively in a time- and dose-dependent manner. Quantification of colony-forming units from bacterial biofilms confirmed the elimination of biofilms formed by PA or SA. Next, HDF cells were plasma-treated and the morphology was observed using an inverted microscope and a scanning electron microscope. No particular morphological changes were observed on cell membrane and cell attachment. MTT assay of the HDF cells after plasma exposure also confirmed the functional enzymatic activity of the cells up to 5 min of treatment. Flow cytometric assay for annexin V and 7-AAD showed that plasma exposure did not significantly induce apoptosis or necrosis of the cell. Intracellular nitric oxide, reactive oxygen species, and Ki-67 protein, a marker for cell proliferation, were also measured by flow cytometry. Intracellular levels of nitric oxide significantly increased, while reactive oxygen species and Ki-67 protein showed no significant alteration after the plasma exposure. Taken together, we demonstrated that the newly developed adjustable-pulse low-temperature air plasma jet device was an effective tool for the elimination of bacteria especially on chronic wound, where the treatment is needed to be least invasive and safe for the surrounding tissue.

Antibiofilm Activity of Lactobacillus plantarum 12 Exopolysaccharides against Shigella flexneri.

In developing countries, Shigella flexneri is the most common enteric pathogen causing bacillary dysentery. Biofilm formation by S. flexneri can cause the emergence of antibiotic-resistant strains, which poses serious threats to food safety and human health. In this study, the effects of Lactobacillus plantarum 12 exopolysaccharides (L-EPSs) and S. flexneri exopolysaccharides (S-EPSs) on S. flexneri CMCC51574 biofilm formation were investigated. The results showed that L-EPS could decrease polysaccharide production in the extracellular polymeric matrix of S. flexneri and inhibit biofilm formation by S. flexneri L-EPS could decrease the minimum biofilm elimination concentration (MBEC) of antibiotics against S. flexneri biofilm and inhibit S. flexneri adhesion to and invasion into HT-29 cell monolayers, which might be ascribed to S. flexneri biofilm disturbance by L-EPS. In contrast, S-EPS exhibited the opposite effects compared to L-EPS. The monosaccharide composition analysis showed that L-EPS was composed of mannose, glucuronic acid, galactosamine, glucose, galactose, and xylose, with the molar ratio of 32.26:0.99:1.79:5.63:0.05:4.07, while S-EPS was composed of mannose, glucuronic acid, galactosamine, glucose, and galactose, with the molar ratio of 25.43:2.28:7.13:5.35. L-EPS was separated into the neutral polysaccharide L-EPS 1-1 and the acidic polysaccharide L-EPS 2-1 by ion-exchange chromatography and gel chromatography. L-EPS 2-1 exerted higher antibiofilm activity than L-EPS 1-1. The antibiofilm activity of L-EPS might be associated with its structure.IMPORTANCES. flexneri is a widespread foodborne pathogen causing food contamination and responsible for food poisoning outbreaks related to various foods in developing countries. Not only has biofilm formation by S. flexneri been difficult to eliminate, but it has also increased the drug resistance of the strain. In the present study, it was demonstrated that L-EPSs secreted by Lactobacillus plantrum 12 could inhibit S. flexneri biofilm formation on, adhesion to, and invasion into HT-29 cells. Also, L-EPSs could decrease the minimum biofilm elimination concentration (MBEC) of the antibiotics used against S. flexneri biofilm. Therefore, L-EPSs were shown to be bioactive macromolecules with the potential ability to act against S. flexneri infections.

Clean-in-place disinfection of dual-species biofilm (Listeria and Pseudomonas) by a green antibacterial product made from citrus extract

Food industries, which must ensure safety and quality of manufactured products, require effective and regular cleaning and disinfection processes. One of the most difficult issues to overcome is to ensure the elimination of biofilms that can generate contamination during food processing. Despite chemical disinfection is the most used strategy, it also requires to be optimised, reducing energy costs, time, and the environmental impact of these operations. In this work, the effectiveness of an environmental-friendly commercial disinfectant (Mico E-PRO®) made from food-grade citrus extracts has been addressed by two methods (i) in-vitro tests for evaluation of antimicrobial efficacy and (ii) a lab-simulated CIP system, where the efficacy was compared with NaClO 1%. In both cases, Mico E-PRO® shows that it is an effective product for the killing of E. coli (MBC = 625 ppm), L. monocytogenes (MBC = 625 ppm), S. enterica (MBC = 625 ppm) and P. aeruginosa (MBC = 156.25 ppm), being P. aeruginosa the most sensible to the disinfectant. The product also shows good bactericidal effect against a mature biofilm formed by L. innocua and P. putida – similar efficacy to that obtained with NaClO 1%. Overall, results demonstrated that Mico E-PRO® is a good option as a natural sanitiser for CIP systems, running as an efficient and safe alternative to the traditional chemicals-based disinfectants.

Effects of povidone-iodine composite on the elimination of bacterial biofilm.

Povidone-iodine (PVP-I) is well known as an antiseptic and exhibits extensive activity against various pathogens. However, due to its uniquely unpleasant nature, it cannot be used locally to deactivate various sinonasal pathogens. Therefore, we developed a PVP-I composite that blocks the unpleasant odor of PVP-I for use as a local antiseptic in the sinonasal cavity and evaluated its effect on bacterial biofilm's formation and elimination in in vivo and in vitro models. MTT, lactate dehydrogenase, and live/dead staining assay were performed to examine the cellular toxicity of PVP-I composites on the primary human nasal epithelial and RPMI 2650 cells. Crystal violet assay was performed to quantify bacterial biofilm after treating with various agents, including PVP-I and antibiotics. Hematoxylin-and-eosin staining, live/dead staining assay, and scanning electron microscopy were conducted to evaluate the effect of PVP-I on biofilm formation in a mice biofilm model. It was observed that the PVP-I composite did not have any significant toxic effect on the nasal epithelial cells. Furthermore, the PVP-I composite effectively inhibited the formation of bacterial biomass within a dose-dependent manner after 48 hours of incubation with Pseudomonas aeruginosa and Staphylococcus aureus. In mice, it effectively eliminated biofilm from the mucosa of the nasal cavity and maxillary sinus at the tested concentrations. The results of this study indicate that the PVP-I composite is a promising compound that could be used locally to prevent the formation of biofilms and to eliminate them from the sinonasal cavity.

Phages for Biofilm Removal.

Biofilms are clusters of bacteria that live in association with surfaces. Their main characteristic is that the bacteria inside the biofilms are attached to other bacterial cells and to the surface by an extracellular polymeric matrix. Biofilms are capable of adhering to a wide variety of surfaces, both biotic and abiotic, including human tissues, medical devices, and other materials. On these surfaces, biofilms represent a major threat causing infectious diseases and economic losses. In addition, current antibiotics and common disinfectants have shown limited ability to remove biofilms adequately, and phage-based treatments are proposed as promising alternatives for biofilm eradication. This review analyzes the main advantages and challenges that phages can offer for the elimination of biofilms, as well as the most important factors to be taken into account in order to design effective phage-based treatments.

Inactivation of Pseudomonas aeruginosa Biofilms by 405-Nanometer-Light-Emitting Diode Illumination.

Biofilm formation by Pseudomonas aeruginosa contributes to its survival on surfaces and represents a major clinical threat because of the increased tolerance of biofilms to disinfecting agents. This study aimed to investigate the efficacy of 405-nm light-emitting diode (LED) illumination in eliminating P. aeruginosa biofilms formed on stainless steel coupons under different temperatures. Time-dependent killing assays using planktonic and biofilm cells were used to determine the antimicrobial and antibiofilm activities of LED illumination. We also evaluated the effects of LED illumination on the disinfectant susceptibility, biofilm structure, extracellular polymeric substance (EPS) structure and composition, and biofilm-related gene expression of P. aeruginosa biofilm cells. Results showed that the abundance of planktonic P. aeruginosa cells was reduced by 0.88, 0.53, and 0.85 log CFU/ml following LED treatment for 2 h compared with untreated controls at 4, 10, and 25°C, respectively. For cells in biofilms, significant reductions (1.73, 1.59, and 1.68 log CFU/cm2) were observed following LED illumination for 2 h at 4, 10, and 25°C, respectively. Moreover, illuminated P. aeruginosa biofilm cells were more sensitive to benzalkonium chloride or chlorhexidine than untreated cells. Scanning electron microscopy and confocal laser scanning microscopic observation indicated that both the biofilm structure and EPS structure were disrupted by LED illumination. Further, reverse transcription-quantitative PCR revealed that LED illumination downregulated the transcription of several genes associated with biofilm formation. These findings suggest that LED illumination has the potential to be developed as an alternative method for prevention and control of P. aeruginosa biofilm contamination.IMPORTANCEPseudomonas aeruginosa can form biofilms on medical implants, industrial equipment, and domestic surfaces, contributing to high morbidity and mortality rates. This study examined the antibiofilm activity of 405-nm light-emitting diode (LED) illumination against mature biofilms formed on stainless steel coupons. We found that the disinfectant susceptibility, biofilm structure, and extracellular polymeric substance structure and composition were disrupted by LED illumination. We then investigated the transcription of several critical P. aeruginosa biofilm-related genes and analyzed the effect of illumination temperature on the above characteristics. Our results confirmed that LED illumination could be developed into an effective and safe method to counter P. aeruginosa biofilm contamination. Further research will be focused on the efficacy and application of LED illumination for elimination of complicated biofilms in the environment.

Effect of Dalbavancin on Staphylococcal Biofilms When Administered Alone or in Combination With Biofilm-Detaching Compounds

Microorganisms grown in biofilms are more resistant to antimicrobial treatment and immune system attacks compared to their planktonic forms. In fact, infections caused by biofilm-forming Staphylococcus aureus and Staphylococcus epidermidis are a large threat for public health, including patients with medical devices. The aim of the current manuscript was to test the effect of dalbavancin, a recently developed lipoglycopeptide antibiotic, alone or in combination with compounds contributing to bacterial cell disaggregation, on staphylococcal biofilm formation and elimination. We used real-time impedance measurements in microtiter plates to study biofilm growth dynamics of S. aureus and S. epidermidis strains, in the absence or presence of dalbavancin, linezolid, vancomycin, cloxacillin, and rifampicin. Further experiments were undertaken to check whether biofilm-detaching compounds such as N-acetylcysteine (NAC) and ficin could enhance dalbavancin efficiency. Real-time dose–response experiments showed that dalbavancin is a highly effective antimicrobial, preventing staphylococcal biofilm formation at low concentrations. Minimum biofilm inhibitory concentrations were up to 22 higher compared to standard E-test values. Dalbavancin was the only antimicrobial that could halt new biofilm formation on established biofilms compared to the other four antibiotics. The addition of NAC decreased dalbavancin efficacy while the combination of dalbavancin with ficin was more efficient than antibiotic alone in preventing growth once the biofilm was established. Results were confirmed by classical biofilm quantification methods such as crystal violet (CV) staining and viable colony counting. Thus, our data support the use of dalbavancin as a promising antimicrobial to treat biofilm-related infections. Our data also highlight that synergistic and antagonistic effects between antibiotics and biofilm-detaching compounds should be carefully tested in order to achieve an efficient treatment that could prevent both biofilm formation and disruption.

Ascorbic Acid Changes Growth of Food-Borne Pathogens in the Early Stage of Biofilm Formation.

Since bacterial biofilm may contribute to the secondary contamination of food during the manufacturing/processing stage there is a need for new methods allowing its effective eradication. Application of food additives such as vitamin C already used in food industry as antioxidant food industry antioxidants may be a promising solution. The aim of this research was evaluation of the impact of vitamin C (ascorbic acid), in a range of concentrations 2.50 µg mL−1–25.0 mg mL−1, on biofilms of Staphylococcus aureus, Escherichia coli, and Listeria monocytogenes strains isolated from food. The efficacy of ascorbic acid was assessed based on the reduction of optical density (λ = 595 nm). The greatest elimination of the biofilm was achieved at the concentration of vitamin C of 25.0 mg mL−1. The effect of the vitamin C on biofilm, however, was strain dependent. The concentration of 25.0 mg mL−1 reduced 93.4%, 74.9%, and 40.5% of E. coli, L. monocytogenes, and S. aureus number, respectively. For E. coli and S. aureus lower concentrations were ineffective. In turn, for L. monocytogenes the biofilm inhibition was observed even at the concentration of 0.25 mg mL−1. The addition of vitamin C may be helpful in the elimination of bacterial biofilms. Nonetheless, some concentrations can induce growth of the pathogens, posing risk for the consumers’ health.

Q-Switch Nd:YAG Laser-Assisted Elimination of Multi-Species Biofilm on Titanium Surfaces

(1) Background: The relatively high prevalence of peri-implantitis (PI) and the lack of a standard method for decontamination of the dental implant surface have pushed us to conduct further research in the field. Bacterial biofilms were found to play a primordial role in the etiology of PI. Therefore, the aim is to evaluate the efficacy of a laser-assisted elimination of biofilm protocol in the removal of a multi-species biofilm on titanium surfaces. (2) Methods: In total, 52 titanium discs (grade 4) were used. The study group consisted of 13 titanium disks contaminated with multi-species biofilms and subsequently irradiated with the laser (T + BF + L). The control groups consisted of the following types of titanium disks: 13 contaminated with multi-species biofilms (T + BF), 13 sterile and irradiated (T + L), 13 sterile and untreated (T). Q-Switch Nd:YAG laser Irradiation parameters were the following: energy density equal to 0.597 J/cm2 per pulse, power equal to 270 milliwatt per pulse, 2.4 mm of spot diameter, and 10 Hz repetition rate for pulse duration of six nanoseconds (ns). The laser irradiation was made during 2 s of total time in non-contact and at 0.5 mm away from the titanium disc surface. After treatment, presence of biofilms on the disks was evaluated by staining with crystal violet (CV), which was measured as optical density at six hundred thirty nm, and statistical analyses were done. (3) Results: the optical density values were 0.004 ± 0.004 for the study group T + BF + L, 0.120 ± 0.039 for group T + BF, 0.006 ± 0.003 for group T + L, and 0.007 ± 0.007 for group T. For the study group, laser treatment resulted in a total elimination of the biofilm, with mean values statistically significantly lower than those of contaminated titanium surfaces and similar to those of sterile titanium surfaces. (4) Conclusions: Our irradiation protocol provided a significant elimination of the multi-species biofilm on titanium surfaces. Laser treated titanium surfaces were biofilm-free, similar to the sterile ones.

Different contact lens case treatment methods for the elimination of pathogen biofilms associated with contact lens related microbial keratitis

Objective To investigate the biofilm-forming abilities of pathogens associated with contact lens related microbial keratitis and to compare the efficacies of different treatments in eliminating biofilms in contact lens cases (CLCs). Methods Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans biolm formation in polypropylene CLCs was examined by using a static biofilm formation model, which was incubated at 37 ℃ for 24 hours.According to the CLC treatment methods, the experimental groups were divided into a control group, a room temperature air-drying group, a contact lens care solution soaking group, a heat-drying group and a soaking-heating group.A pathogen colony counting-based serial dilution micro-counting method was applied to evaluate the biofilm elimination efficacies and pathogen killing rates of treatments. Results All four tested strains formed biofilms on the inner walls of the CLC, and the biomasses of S. aureus, P. aeruginosa, E.coli and C. albicans biofilm were (10.78±2.12), (9.19±0.57), (8.03±0.30), and (7.50±0.07)lg CFU/ml, respectively.The S. aureus biofilm biomass was significantly higher than those of the other strains (P<0.05). The biofilm biomasses of all the tested strains in the heat-drying and the soaking-heating groups were significantly lower than those in the control group (all at P<0.05); and the biofilm biomasses of S. aureus and E. coli in the soaking group were significantly lower than that in the control group (all at P<0.05). The heat-drying treatment resulted in a killing rate of (51.76±16.75)% for S. aureus, (68.63±4.43)% for P. aeruginosa, (83.51±13.97)% for E. coli, and (97.13±5.19)% for C. albicans, respectively (F=31.806, P<0.001). Significant differences were observed between the killing rates for each bacterium (all at P<0.05). The E. coli and C. albicans killing rates of the soak-heating treatment were (100.00±0.00) % and (97.79±7.67)%, respectively, and were significantly higher than (81.13±14.86)% of S. aureus and (74.22±11.91)% of P. aeruginosa (all at P<0.05). Conclusions Heating alone or combined with the use of contact lens care solution treatment can improve the pathogen killing rates and effectively eliminate the bacterial and fungal contaminations in CLCs. Key words: Microbial keratitis; Contact lens cases; Biofilm; Heating

A simplified in vitro model for investigation of the antimicrobial efficacy of various antiseptic agents to prevent peri-implantitis.

The biofilm formation by oral bacteria on the implant surface is one of the most remarkable factors of peri-implant infections, which may eventually lead to bone resorption and loss of the dental implant. Therefore, the elimination of biofilm is an essential step for the successful therapy of implant-related infections. In this work we created a basic in vitro model to evaluate the antibacterial effect of three widely used antiseptics.Commercially pure (CP4) titanium sample discs with sand blasted, acid etched, and polished surface were used. The discs were incubated with mono-cultures of Streptococcus mitis and Streptococcus salivarius. The adhered bacterial biofilms were treated with different antiseptics: chlorhexidine-digluconate (CHX), povidone-iodine (PI), and chlorine dioxide (CD) for 5 min and the control discs with ultrapure water. The antibacterial effect of the antiseptics was tested by colorimetric assay.According to the results, the PI and the CD were statistically the most effective in the elimination of the two test bacteria on both titanium surfaces after 5 min treatment time. The CD showed significant effect only against S. salivarius.Based on our results we conclude that PI and CD may be promising antibacterial agents to disinfecting the peri-implant site in the dental practice.

A noninvasive precise treatment strategy for implant-related infections based on X-ray-induced luminescent/photodynamic therapeutic multilayered device surface materials

Conventional prophylaxis and treatment of implant-related infections via devices’ surface biofilm elimination include extensive non-targeting antibiotics prescription, invasive surgical intervention, and periodic implant replacement, with their inevitable relapses being lifelong guaranteed leading to financial burdens and drug resistance risks for patients. Herein, we propose a novel, sustainable modality of noninvasive precise treatment strategy based on potential theranostic, multilayered Gd3+ (UV), Eu3+ (red) and Tb3+ (green) doped Y(PO3)3 X-ray-induced luminescent matrix surface materials cum a nanolayer coating of titanium dioxide with pH indicators on implantable medical devices, which simultaneously integrate X-ray-initiated pH-sensitive NIR luminescence for location detection of biofilm infection, and X-ray-excited, UV-triggered deep-tissue-localized antimicrobial photodynamic therapy. The Y(PO3)3 crystal composite structures allow doping of rare earth ions at rarely high concentrations and produce high emission intensity even under less-detrimental low-dose X-ray radiation. The feasibility of such precise treatment strategy has been substantiated through various pH and inactivation experiments. With the growing aging population, this work aims to provide a potentially new, patient-friendly, (i.e. surgery/antibiotics-free and lower-cost) proof-of-principle theranostic resolution for the increasing incidence of orthopaedic implant-related biofilm infections, from preventive device maintenance to precise curative eradication.