Streptococcus salivarius inhibits immune activation by periodontal disease pathogens.

BACKGROUND: Periodontal disease represents a major health concern. The administration of beneficial microbes has been increasing in popularity over efforts to manipulate the microbes using antimicrobial agents. This study determined the ability of Streptococcus salivarius to inhibit IL-6 and IL-8 production by gingival fibroblasts when activated by periodontal pathogens and their effect on the salivary microbiome. METHODS: Primary human gingival fibroblasts were challenged with Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum and a combination of all three. IL-6 and IL-8 cytokine release were measured. Using this same model, S. salivarius K12, M18 and different supernatant and whole-cell lysate fractions of S. salivarius K12 were administered to pathogen-induced fibroblasts. A patient study of healthy participants was also conducted to determine the effect S. salivarius K12 had on the native microbiome using 16S next generation sequence analysis. RESULTS: All pathogens tested induced a significant IL-6 and IL-8 response. S. salivarius K12 or M18, did not exhibit an increase in inflammatory cytokines. When either of the probiotic strains were co-administered with a pathogen, there were significant reductions in both IL-6 and IL-8 release. This effect was also observed when gingival fibroblasts were pre-treated with either S. salivarius K12 or M18 and then stimulated with the oral pathogens. Chewing gum containing S. salivarius K12 did not alter the salivary microbiome and did not increase inflammatory markers in the oral cavity. CONCLUSION: S. salivarius K12 and M18 prevented immune activation induced by periodontal disease pathogens. S. salivarius K12 did not alter the salivary microbiome or induce immune activation when administered as a chewing gum. These results warrant further study to determine if it may be an effective treatment in a model of periodontal disease.

Application of Novel 3,4-Dihydroxyphenylalanine-Containing Antimicrobial Polymers for the Prevention of Uropathogen Attachment to Urinary Biomaterials.

Introduction and Objective: Urinary catheters and stents are frequently prone to catheter-associated urinary tract infections (CAUTI) through biofilm formation. Several strategies have been evaluated in search of a stent coating to reliably prevent adherence of bacteria and biofilm. Previous in vivo and in vitro research with methoxylated polyethylene glycol 3,4-dihydroxyphenylalanine (DOPA) copolymer as a candidate coating showed promising results to reduce the bacterial attachment. We aimed to further enhance this antimicrobial activity by adding antimicrobial agents to newly synthesized DOPA-based copolymers. Materials and Methods: Building on our previous experience, novel copolymers were engineered based on DOPA. Quaternary ammonium groups and silver particles were added by cross-linking to increase the antimicrobial activity through both kill-by-contact and planktonic killing. After coating polyurethane sheets and measuring contact angles, all candidate coatings were challenged in vitro with an Escherichia coli culture. The most promising coatings were then further evaluated against a panel of seven clinically relevant uropathogens and planktonic killing, and microbial attachment was determined. Results: Initially, seven coatings were developed, referred to as Surphys 093-099. The most significant increase in contact angle was identified in Surphys-095 and -098. Surphys coatings S-094, S-095, and S-098 were cross-linked with silver and exhibited profound antimicrobial properties when challenged with E. coli. Further testing demonstrated S-095 to have antimicrobial efficacy against gram-positive and gram-negative bacteria at different silver-loading concentrations. The final coating, consisting of a 2 mg/mL solution of S-095 cross-linked with 0.25 mg/mL AgNO3, appeared to be highly bactericidal showing a ≥99.9% bacterial killing effect while remaining below cytotoxicity levels. Conclusions: We were able to engineer DOPA-based copolymers and add quaternary ammonium and silver particles, thus increasing the bactericidal properties of the coating. These coatings have exhibited a biologically significant ability to prevent uropathogens from attaching to biomaterials and represent a realistic opportunity to combat CAUTI.

Efficacy of Dentaq® Oral and ENT Health Probiotic Complex on Clinical Parameters of Gingivitis in Patients Undergoing Fixed Orthodontic Treatment: A Pilot Study.

OBJECTIVES: Probiotics act as a unique approach to maintaining oral health by supplementing the endogenous oral bacteria with additional naturally occurring beneficial microbes to provide defense against pathogens harmful to teeth and gingiva. The aim of this pilot study was to clinically evaluate the effects of probiotics on plaque accumulation and gingival inflammation in subjects with fixed orthodontics. METHODS: The pilot study was comprised of 15 healthy patients, aged 11 to 18 years, undergoing fixed orthodontic treatment. Patients used an all-natural, dissolving lozenge containing six proprietary probiotic strains (Dentaq® Oral and ENT Health Probiotic Complex)for 28 days. Gingival Index (GI) according to Löe-Silness and Plaque Index (PI) according to Quigley-Hein for all teeth were measured at baseline (Day Zero) and at the end of the probiotic regimen (Day 28). RESULTS: The mean baseline GI and PI scores within each patient decreased by 28.4% and 35.8%, respectively, by Day 28. Patients reported decreased tooth and gingival pain, decreased oral bleeding, and increased motivation to maintain proper oral hygiene over the course of the study. CONCLUSIONS: This pilot study provided preliminary support for the use of Dentaq Oral and ENT Health Probiotic Complex as a safe and effective natural health product for the reduction of plaque accumulation and gingival inflammation. The results demonstrate its potential therapeutic value and open the door for larger scale placebo-controlled clinical studies to verify these findings.

The Effect of Renal Cysts on the Fragmentation of Renal Stones During Shockwave Lithotripsy: A Comparative In Vitro Study.

PURPOSE: To assess the potential effect of simple renal cysts (SRC) on stone fragmentation during shockwave lithotripsy (SWL) in an in vitro model. MATERIALS AND METHODS: The in vitro model was constructed using 10% ordnance gelatin (OG). Models were created to mimic four scenarios: Model A-with an air-filled cavity (suboptimal for stone fragmentation); model B-without a cavity (normal anatomy); model C-with a 3-cm serum filled cavity (to represent a small SRC); model D-with a 4-cm serum filled cavity (to represent a larger SRC). SWL was applied to 24 standardized phantom stones (weight of 2±0.1 g) in each model using a standardized protocol. Stone fragments were retrieved, then dried overnight at room air temperature. Fragmentation coefficient (FC) was calculated for each stone, for fragments<4 mm and <2 mm. RESULTS: The OG in vitro model was robust enough for the proposed research. There was no fragmentation evident in model A as expected. The mean FC was 29.7 (±20.5) and 39.7 (±23.7) for <4 mm fragments (P=0.069) and 7.6 (±4.1) and 10.6 (±6.7) for <2 mm fragments (P=0.047), for noncystic and cystic models, respectively. The mean FC was 29.7 (±20.5), 38.8 (±26.2) and 40.7 (±21.3) for <4 mm fragments (P=0.213) and 7.6 (±4.1), 11.1 (±8) and 10.2 (±5.3) for <2 mm fragments (P=0.138), for models B, C, and D, respectively. CONCLUSION: Our in vitro experiment confirms better stone fragmentation associated with SWL in the presence of adjacent SRC.

Ureteral stent technology: Drug-eluting stents and stent coatings.

Ureteral stents are commonly used following urological procedures to maintain ureteral patency. However, alongside the benefits of the device, indwelling stents frequently cause significant patient discomfort (pain, urgency, frequency) and can become encrusted and infected. The importance of these sequelae is that they are not only bothersome to the patient but can lead to significant morbidity, urinary retention, ureteral damage, recurrent infections, pyelonephritis and sepsis. When these problems occur, stent removal or replacement alongside antibiotic, analgesic and/or other symptom-modifying therapies are essential to successfully treat the patient. In an attempt to prevent such morbidity, numerous approaches have been investigated over the past several decades to modify the stent itself, thereby affecting changes locally within the urinary tract without significant systemic therapy. These strategies include changes to device design, polymeric composition, drug-elution and surface coatings. Of these, drug-elution and surface coatings are the most studied and display the most promise for advancing ureteral stent use and efficacy. This article reviews these two strategies in detail to determine their clinical potential and guide future research in the area.

The influence of urinary pH on antibiotic efficacy against bacterial uropathogens.

OBJECTIVE: To determine the effects of pH on the activity of clinically relevant antibiotics against bacterial uropathogens. Numerous factors affect antibiotic efficacy within the urinary tract including pH. Because human urine can substantially vary from acidic (pH 4.5) to alkaline (pH 8) conditions and can be easily clinically manipulated, it would be a great advantage to better understand the role of pH in antibiotic treatment of urinary tract infection. MATERIALS AND METHODS: This in vitro study investigated the activity of 24 widely used antimicrobial agents against bacterial strains comprising 6 major uropathogenic species (Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterococcus faecalis, Staphylococcus saprophyticus, and Staphylococcus epidermidis) over the range of pH 5-8. Standard disk-diffusion and broth-microdilution assays were used. One-way analysis of variance was applied to determine significance (P <.05). RESULTS: For 18 of the 24 agents, pH was shown to play a significant role in overall inhibitory activity. Although most agents behaved similarly across most or all of the uropathogens tested, several only showed pH-dependent effects against certain organisms. The fluoroquinolones, co-trimoxazole, aminoglycosides, and macrolides all functioned optimally at alkaline pH, whereas the tetracyclines, nitrofurantoin, and many of the ß-lactams tested exhibited their highest activity under more acidic conditions. Sulfamethoxazole, oxacillin, amoxicillin and clavulanic acid, vancomycin, imipenem, and clindamycin were largely unaffected by pH. CONCLUSION: Clinicians should consider the urinary pH of their patients when treating urinary tract infection, especially in complicated scenarios. Future clinical investigations examining urinary pH and antibiotic efficacy may result in the application of decreased antibiotic dosages and regimen durations, potentially reducing antibiotic resistance development.

Selective target inactivation rather than global metabolic dormancy causes antibiotic tolerance in uropathogens.

Persister cells represent a multidrug-tolerant (MDT), physiologically distinct subpopulation of bacteria. The ability of these organisms to survive lethal antibiotic doses raises concern over their potential role in chronic disease, such as recurrent urinary tract infection (RUTI). Persistence is believed to be conveyed through global metabolic dormancy, which yields organisms unresponsive to external stimuli. However, recent studies have contested this stance. Here, various antibiotics that target different cellular processes were used to dissect the activity of transcription, translation, and peptidoglycan turnover in persister cells. Differential susceptibility patterns were found in type I and type II persisters, and responses differed between Staphylococcus saprophyticus and Escherichia coli uropathogens. Further, SOS-deficient strains were sensitized to ciprofloxacin, suggesting DNA gyrase activity in persisters and indicating the importance of active DNA repair systems for ciprofloxacin tolerance. These results indicate that global dormancy per se cannot sufficiently account for antibiotic tolerance. Rather, the activity of individual cellular processes dictates multidrug tolerance in an antibiotic-specific fashion. Furthermore, the susceptibility patterns of persisters depended on their mechanisms of onset, with subinhibitory antibiotic pretreatments selectively shutting down cognate targets and increasing the persister fraction against the same agent. Interestingly, antibiotics targeting transcription and translation enhanced persistence against multiple agents indirectly related to these processes. Conducting these assays with uropathogenic E. coli isolated from RUTI patients revealed an enriched persister fraction compared to organisms cleared with standard antibiotic therapy. This finding suggests that persister traits are either selected for during prolonged antibiotic treatment or initially contribute to therapy failure.

Urinary expression of novel tissue markers of kidney injury after ureteroscopy, shockwave lithotripsy, and in normal healthy controls.

BACKGROUND AND PURPOSE: Shockwave lithotripsy (SWL) and ureteroscopy (URS) are minimally invasive treatment alternatives for kidney stones. Although less invasive, SWL subjects the renal parenchyma to a high level of energy and the potential to cause renal injury. The ability to detect renal injury post-SWL in a reliable and noninvasive way would be clinically beneficial. Kidney injury molecule 1 (KIM-1) and N-acetyl-ß-D-glucosaminidase (NAG) are two proteins secreted by the kidney into the urine and have been found to be sensitive markers of acute kidney injury in transplant patients. The aim of this work was to measure urinary levels of KIM-1 and NAG in patients with kidney stone who were treated by SWL or URS and in nonstone volunteers. PATIENTS AND METHODS: Patients with kidney stones who were treated by SWL (n = 50) or URS (n = 10) were recruited. Voided urine samples were collected before and 2 to 3 hours after URS and SWL. In addition, further urinary specimens were collected 2 days and 2 weeks post-SWL treatment. Voided urine samples from healthy volunteers were also collected. RESULTS: Mean KIM-1 values were increased in patients with kidney stones when compared with volunteers. KIM-1 and NAG levels significantly increased post-SWL and returned to baseline within 2 weeks post-SWL. Poor kidney function was significantly associated with increased biomarker activity both in baseline and post-SWL measurements. There was no significant change in urinary KIM-1 and NAG concentrations before and after URS. CONCLUSIONS: Kim-1 and NAG levels significantly increased post-SWL treatment suggesting a potential role for these urinary markers in identifying patients at higher risk of tissue injury.

Persistence of the oral probiotic Streptococcus salivarius M18 is dose dependent and megaplasmid transfer can augment their bacteriocin production and adhesion characteristics.

Bacteriocin-producing probiotic Streptococcus salivarius M18 offers beneficial modulatory capabilities within the oral microbiome, apparently through potent inhibitory activity against potentially deleterious bacteria, such as Streptococcus pyogenes. The oral cavity persistence of S. salivarius M18 was investigated in 75 subjects receiving four different doses for 28 days. Sixty per cent of the subjects already had some inhibitor-producing S. salivarius in their saliva prior to probiotic intervention. Strain M18's persistence was dependent upon the dose, but not the period of administration. Culture analysis indicated that in some individuals the introduced strain had almost entirely replaced the indigenous S. salivarius, though the total numbers of the species did not increase. Selected subjects showing either high or low probiotic persistence had their salivary populations profiled using Illumina sequencing of the V6 region of the 16S rRNA gene. Analysis indicated that while certain bacterial phenotypes were markedly modulated, the overall composition of the oral microbiome was not modified by the probiotic treatment. Megaplasmids encoding bacteriocins and adhesion factors were transferred in vitro to generate a transconjugant S. salivarius exhibiting enhanced antimicrobial production and binding capabilities to HEp-2 cells. Since no widespread perturbation of the existing indigenous microbiota was associated with oral instillation and given its antimicrobial activity against potentially pathogenic streptococci, it appears that application of probiotic strain M18 offers potential low impact alternative to classical antibiotic prophylaxis. For candidate probiotic strains having relatively poor antimicrobial or adhesive properties, unique derivatives displaying improved probiotic performance may be engineered in vitro by megaplasmid transfer.

The use of triclosan eluting stents effectively reduces ureteral stent symptoms: a prospective randomized trial.

UNLABELLED: What's known on the subject? and What does the study add? Infection, encrustation and ureteral-stent-related symptoms (USRS) including pain, urgency and frequency are all major problems associated with stent use. No current ureteral stent or exogenously applied therapy adequately deals with these problems and antibiotic use is ineffective once a bacterial biofilm forms on the device. Triclosan is a broad spectrum antibacterial agent widely used in numerous healthcare products and has been previously shown to reduce inflammation on the skin and in the oral cavity. This study tested a triclosan-impregnated ureteral stent for its ability to reduce infection, encrustation and USRS. This study shows that while a triclosan-impregnated ureteral stent cannot reduce infection rates alone compared with antibiotic use, the stent can reduce several USRS including pain during indwelling. This study suggests that the triclosan eluting stent may have a role in treating patients, perhaps in combination with standard antibiotic therapy. OBJECTIVE: To evaluate the capacity of triclosan-loaded ureteral stents to reduce stent-associated bacterial attachment, biofilm formation and encrustation, thereby potentially reducing infection development and other device-related sequelae. PATIENTS AND METHODS: Twenty subjects requiring short-term stenting (7-15 days) were randomized to receive either a Percuflex Plus(®) non-eluting stent (control) or a Triumph(®) triclosan eluting stent. Control-stented subjects received 3 days of levofloxacin prophylaxis (500 mg once daily) while Triumph(®)-stented subjects did not. All subjects were assessed for positive urine and stent cultures, stent biofilm development and encrustation. Following device removal, each subject completed an analogue-scale symptom assessment questionnaire. RESULTS: Ureteral stenting was performed after nine ureteroscopic and one extracorporeal shock wave lithotripsy procedure in the control group and eight ureteroscopic and two shock wave lithotripsy procedures in the triclosan group. No significant differences were observed for culture, biofilm and encrustation between the two groups. Subjects in the triclosan group reported significant reductions in lower flank pain scores during activity (58.1% reduction, P = 0.017) and urination (42.6%, P = 0.041), abdominal pain during activity (42.1%, P = 0.042) and urethral pain during urination (31.7%, P = 0.049). CONCLUSIONS: In this study, the use of the Triumph(®) triclosan eluting stent had no marked impact on biofilm formation, encrustation or infection development in short-term stented patients. The Triumph(®) device led to significant reductions in several common ureteral-stent-related symptoms, supporting its use in this patient population.

Effects of subinhibitory concentrations of ciprofloxacin on Staphylococcus saprophyticus adherence and virulence in urinary tract infections.

BACKGROUND AND PURPOSE: Staphylococcus saprophyticus is a frequent cause of both uncomplicated and complicated urinary tract infections (UTI) in young females and has recently been established as the most prominent gram-positive uropathogen. Although the effects of subinhibitory concentrations of antimicrobials on numerous other pathogenic bacteria have been studied, little is known regarding how S saprophyticus responds under such conditions. MATERIALS AND METHODS: In this study, we investigated the effects of subminimum inhibitory concentrations (MIC) of ciprofloxacin (CIP) on S saprophyticus attachment to glass microscope slides, ureteral stent material, and T24 bladder cells, as well as its effects on S saprophyticus-induced proinflammatory cytokine expression in bladder cells. RESULTS: Adherence to glass microscope slides, ureteral stent material, and bladder cell monolayers were all significantly increased in the presence of sub-MIC levels of CIP. While the S saprophyticus challenge of T24 bladder cell monolayers significantly upregulated both interleukin (IL)-6 and IL-8 expression, sub-MIC CIP abrogated these effects, returning their secretion to control levels. CONCLUSIONS: Our results demonstrate that exposure to sub-MIC CIP increases S saprophyticus adherence to both abiotic and biotic surfaces including urinary device material and cultured bladder cells. In addition, low levels of this antimicrobial downregulate S saprophyticus-stimulated proinflammatory cytokine secretion in the bladder. These changes may make S saprophyticus more effective at colonizing the urinary tract and highlights the need for clinicians to consider the impact of subinhibitory concentrations of antimicrobials on bacteria when designing treatment strategies to manage UTI.

Third prize: the impact of fluid environment manipulation on shockwave lithotripsy artificial calculi fragmentation rates.

BACKGROUND AND PURPOSE: Studies have suggested that shockwave lithotripsy (SWL) stone fragmentation rates can be affected by characteristics of the fluid media surrounding the stone, although evidence to implicate the impact of urine specific gravity (SG) is limited and inconclusive. Our aim is to further explore the impact fluid media and SGs have on stone fragmentation using a variable focus lithotripter. MATERIALS AND METHODS: Artificial stones were presoaked for 24 hours in urine and then shocked in various fluid media including artificial urine (SG 1.010 control, 1.020, and 1.07), human pooled urine (HPU), degassed HPU, Pentastarch, 100% and 30% contrast, degassed 30% contrast, 100% ethanol, deionized water (dH(2)O), degassed dH(2)O, 5% glucose, Ringer lactate, 0.9% saline, glycerol, whole blood, and lubricating gel. After soaking, SWL using the Modulith SLX-F2 electromagnetic lithotripter was performed. Fragments were dried and sieved using a 4-mm diameter opening grid. Fragments >4 mm were weighed and fragmentation coefficients (FCs) calculated (pre-SWL weight - post-SWL weight)/(pre-SWL weight) × 100. Fifteen stones were shocked for each fluid group. RESULTS: Fluid type, viscosity, and degassing all significantly impacted stone fragmentation. While the solutions' SG, per se, did not appear to affect stone fragmentation, the use of degassed 30% contrast significantly improved stone destruction over the SG 1.010 artificial urine control (95.3% vs 71.4, P < 0.01). Furthermore, degassing improved comminution rates by increasing the number of completely fragmented stones (FC = 100%). Using degassed 30% contrast, 12/15 stones were completely fragmented, compared with only 2/15 in the control group (P = 0.007). Among the whole blood, glycerol, and lubricating gel groups, only 1/15, 0/15, and 1/15 stones reached 100% FC respectively in the narrow focus, possibly because of the detrimental impact of increased viscosity. CONCLUSIONS: Different fluid media can significantly affect FC in vitro. Among the various fluids tested, degassed 30% contrast significantly increased the FC and total number of completely fragmented stones.

Novel in vitro model for studying ureteric stent-induced cell injury.

OBJECTIVE: To develop a novel in vitro model for the study of bladder and kidney epithelial cell injury akin to stent movement, as ureteric stents are associated with urinary tract complications that can significantly add to patient morbidity. These sequelae may be linked to inflammation triggered by stent-mediated mechanical injury to the urinary tract. MATERIALS AND METHODS: T24 bladder and A498 kidney cell line monolayers were damaged mechanically by segments of either Percuflex Plus (PP) or Triumph (triclosan-eluting) stents (both from Boston Scientific Corporation Inc. Natick, MA, USA) and the resulting expression profiles of several pro-inflammatory cytokines and growth factors were analysed. RESULTS: After control injury using the PP stent, supernatants of both cell lines had significantly increased levels of interleukin (IL)-6, IL-8, basic fibroblast growth factor and platelet-derived growth factor BB, and A498 cells also had increased tumour necrosis factor alpha. In almost all cases, the presence of triclosan within the media abrogated the pro-inflammatory cytokine increases, while its effects on growth factors varied. CONCLUSION: This study suggests that stent-related symptoms in the bladder and kidney may be partially due to a local inflammatory response to epithelial damage caused by the presence and movement of the stent. Future stent design should take these inflammatory responses, with respect to physical injury, into consideration, using either more biocompatible materials or anti-inflammatory compounds such as triclosan.

Anti-adhesive coating and clearance of device associated uropathogenic Escherichia coli cystitis.

PURPOSE: A previous study showed decreased uropathogen adherence using a novel anti-fouling coating consisting of mussel adhesive protein mimics conjugated to poly(ethylene glycol). We assessed the ability of methoxy polyethylene glycol-dihydroxyphenylalanine (Nerites Corp. Ltd., Madison, Wisconsin) coated ureteral stents to resist bacterial adherence, infection development and encrustation in a rabbit model of uropathogenic Escherichia coli cystitis. MATERIALS AND METHODS: Sof-Flex stent curls that were uncoated and coated with 3 coatings, including Surphys 002, 008 and 009, respectively, and uncoated Percuflex Plus stents were inserted transurethrally into the bladder of 50 male New Zealand White rabbits (Charles River Laboratories, Montreal, Quebec, Canada), followed by instillation of uropathogenic E. coli strain GR12 (10(7) cfu). Urine was examined for bacteria on days 0, 1, 3 and 7, and for cytokine levels on day 7. On day 7 the animals were sacrificed. Stent curls and bladders were harvested for analysis. In a parallel experiment stents were challenged in vitro for 7 days with GR12 in human urine. RESULTS: Surphys 009 coated devices showed decreased urine and stent bacterial counts compared to those in controls. Eight of 10 rabbits in the Surphys 009 group had sterile urine by day 3 vs 1 in each control group (p = 0.013), while stent adherent organisms were decreased by more than 75%. While no statistical differences were found in encrustation and bladder inflammation across the groups, immune scoring was lowest in the uncoated Sof-Flex control and Surphys 009 groups (p = 0.030). CONCLUSIONS: Surphys 009 strongly resisted bacterial attachment, resulting in improved infection clearance over that of uncoated devices. However, this did not translate to decreased encrustation, which appeared to be independent of infection in this model.

Use of triclosan-eluting ureteral stents in patients with long-term stents.

BACKGROUND AND PURPOSE: Long-term use of ureteral stents is prevented by biofilm-related infection and encrustation mandating stent changes every few months. Triclosan is a broad-spectrum antimicrobial in numerous consumer and medical products and has been incorporated into a ureteral stent. We sought to determine the clinical effects of the triclosan-eluting stent in patients who needed long-term ureteral stenting. PATIENTS AND METHODS: Eight patients with long-term stents were enrolled prospectively. All received a control stent for 3 months along with preoperative and postoperative antibiotics. After 3 months, the control stent was removed, and a triclosan-eluting stent was placed for 3 months with no antibiotics administered. For both indwelling periods, urine cultures were obtained weekly and biweekly for the first and last 6 weeks, respectively, and antibiotics were prescribed when patients had both a positive urine culture and symptoms of urinary tract infection. On removal, stents were assessed for microorganisms and encrustation. RESULTS: Overall, similar microorganisms were isolated during each indwell period, although Staphylococcus and Enterococcus strains were isolated more frequently during control and triclosan stenting, respectively. Significantly fewer antibiotics were used during triclosan stenting, coinciding with a slightly higher number of positive urine cultures and significantly fewer symptomatic infections. No bacterial isolates developed antibiotic resistance during triclosan stent placement. CONCLUSIONS: Antibiotic use with control stents resulted in bacterial antibiotic resistance, which was not the case with the triclosan-eluting stents. Although triclosan-eluting stents did not show a clinical benefit in terms of urine and stent cultures or overall subject symptoms compared with controls, their use did result in decreased antibiotic usage and significantly fewer symptomatic infections. The triclosan-eluting stent alone is not sufficient to reduce device-associated infections in this difficult patient population.

Eliminating biofilm from ureteral stents: the Holy Grail.

PURPOSE OF REVIEW: Biofilms continue to be a major limiting factor in the long-term use of ureteral stents, promoting the development of chronic infections and antibiotic resistance and encrustation. Apart from stent removal in conjunction with antibiotic therapy, there is currently no treatment proven successful for completely eradicating a biofilm-related infection, highlighting the need for continued research in this area. RECENT FINDINGS: Research continues to focus mainly on biofilm prevention, specifically the development of novel coatings comprising antibacterial, antifouling or bacterial signalling compounds. Notably, all three strategies have generated candidate coatings showing recent success both in vitro and in vivo. SUMMARY: Despite the current lack of a completely biofilm-resistant device, coating or treatment strategy, continued research into the causation of bacterial biofilm formation and the continued development of novel antibacterial, antifouling and antibiofilm compounds is promising. Future work should be aimed at more rigorous testing of candidate coatings from both physical and bacterial challenge standpoints as well as increased in-vivo investigation via clinical trials.

The effects of triclosan on uropathogen susceptibility to clinically relevant antibiotics.

INTRODUCTION: Triclosan is a broad-spectrum antimicrobial agent currently used in numerous products including surgical scrubs and ureteral stents. Unfortunately, studies have shown triclosan resistance among several bacterial species. Our objective was to characterize resistance patterns of common uropathogens to triclosan and determine whether triclosan exposure would alter their susceptibility to common antibiotics. We hypothesized that triclosan exposure induces a metabolic stress rendering some bacterial strains more susceptible to other antibiotics. METHODS: Using largely clinical isolates comprising seven uropathogenic species, we conducted 24 hour growth experiments to determine triclosan minimal inhibitory concentrations (MIC) for each strain. Based upon these MICs, triclosan was added to agar plates at escalating sublethal concentrations and antibiotic disk diffusion assays were conducted using a range of clinically-relevant antibiotics. RESULTS: Varying susceptibility patterns were observed across all antibiotics studied. Several antibiotics demonstrated increased efficacy in conjunction with triclosan. The combined effect of triclosan with amoxicillin and gentamicin was superior when considering significant increases in susceptibility, with 6 (86%) and 5 (71%) of the 7 bacterial strains displaying enhanced sensitivity, respectively. The antimicrobial effects of nitrofurantoin and the fluoroquinolones were significantly enhanced for 4 (57%), 4 and 3 (42%) of the 7 pathogens, respectively. The two fluoroquinolones were the only antibiotics where susceptibility was negatively impacted (in one strain each) in combination with triclosan. CONCLUSIONS: The synergistic effects of triclosan and several antibiotics are consistent with a triclosan-dependent metabolic strain and/or membrane disruptive effect, and offers important insight into the combined use of antimicrobial compounds in clinical practice.

First prize: Novel uropathogen-resistant coatings inspired by marine mussels.

BACKGROUND AND PURPOSE: Success in the prevention of urinary device infections has been elusive, largely due to multiple bacterial attachment strategies and the development of urinary conditioning films. We investigated a novel anti-fouling coating consisting of mussel adhesive protein mimics conjugated to polyethylene glycol (mPEG-DOPA(3)) for its potential to resist conditioning film formation and uropathogen attachment in human urine. METHODS: Model TiO(2) -coated silicon disks ( approximately 75 mm(2)) were either coated with mPEG-DOPA(3) or left uncoated and sterilized using ethylene oxide gas. For bacterial attachment experiments, coated and uncoated surfaces were separately challenged with bacterial strains comprising six major uropathogenic species for 24 hours at 37 degrees C in human pooled urine. Starting inoculum for each strain was 10(5) CFU/mL and 0.5 mL was used per disk. Following incubation, the disks were thoroughly rinsed in phosphate buffered saline to remove non-adherent and weakly-adherent organisms and cell scrapers were employed to dislodge those that were firmly attached. Adherent bacteria were quantitated using dilution plating. Representative disks were also examined using scanning electron microscopy, energy dispersive x-ray analysis, and live/dead viability staining. RESULTS: The mPEG-DOPA(3) coating significantly resisted the attachment of all uropathogens tested, with a maximum >231-fold reduction in adherence for Escherichia coli GR-12, Enterococcus faecalis 23241, and Proteus mirabilis 296 compared to uncoated TiO(2) disks. Scanning electron microscopy and viability staining analyses also reflected these results and demonstrated the ability of the coating to resist urinary constituent adherence as well. CONCLUSION: Model surfaces coated with mPEG-DOPA(3) strongly resisted both urinary film formation and bacterial attachment in vitro. Future in vitro and in vivo studies will be conducted to assess whether similar findings can be demonstrated when these polymer coatings are applied to urologic devices.

Triclosan inhibits uropathogenic Escherichia coli-stimulated tumor necrosis factor-alpha secretion in T24 bladder cells in vitro.

BACKGROUND AND PURPOSE: Triclosan is an antimicrobial agent commonly used in consumer and medical products that inhibits bacterial fatty acid synthesis. In addition to its bactericidal effects, sublethal concentrations of triclosan reduce local inflammation, inhibit the growth of bacterial uropathogens, induce membrane stress, and inhibit P-fimbrial expression in uropathogenic Escherichia coli (UPEC). We tested whether sublethal concentrations of triclosan could reduce the adherence of UPEC to bladder and kidney cells and reduce the amount of the pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) produced by these cells during bacterial challenge in vitro. MATERIALS AND METHODS: Assays of bacterial growth, adhesion, and intracellularization were performed using UPEC GR12 incubated for 4 hours on monolayers of human T24 bladder cells or A498 kidney cells with various sublethal concentrations of triclosan. The expression profile of TNF-alpha from bladder cells was evaluated using ELISA. RESULTS: No significant decreases were observed in the adherence or invasion percentages of UPEC GR12 with either cell line when treated with sublethal amounts of triclosan. However, treatment with triclosan 0.5 microg/mL led to a significant decrease in the total number of UPEC GR12 recovered from T24 monolayers (P < 0.05). Importantly, a reduction in the expression of TNF-alpha by T24 cells was shown when UPEC GR12 was treated with triclosan (P < 0.05). CONCLUSIONS: Sublethal concentrations of triclosan did not inhibit the adhesion or intracellularization of UPEC into kidney or bladder cell lines but did significantly reduce the amount of TNF-alpha secreted by bladder cells. Therefore, the use of triclosan on ureteral stents may prove clinically beneficial, not only by inhibiting bacterial survival and growth within the urinary tract, but by reducing local inflammation as well.

In-vitro activity of triclosan-eluting ureteral stents against common bacterial uropathogens.

BACKGROUND AND PURPOSE: Ureteral stents are commonly used in urology today, but the biofilms that form on them within hours of placement may harbor bacteria that can result in infection or encrustation. Triclosan is an antimicrobial commonly used in consumer and medical products that inhibits bacterial fatty-acid synthesis. The bactericidal and bacteriostatic effect of a triclosan-eluting ureteral stent was tested against clinical isolates of common bacterial uropathogens in an in-vitro setting. MATERIALS AND METHODS: Triclosan eluted from a drug-loaded ureteral stent was suspended in artificial urine with bacterial pathogens (Escherichia coli C1214, Proteus mirabilis 296, Enterococcus faecalis 1131, Klebsiella pneumoniae 280, Staphylococcus aureus Newman, Pseudomonas aeruginosa AK1) to assess growth, virulence-promoter activity, and bacterial adherence to the stent. Generic stents were utilized as controls. RESULTS: Triclosan inhibited the growth of E. faecalis, K. pneumoniae, S. aureus, and P. mirabilis in a dose-dependent manner. Pseudomonas aeruginosa demonstrated significant resistance. Lower concentrations of triclosan downregulated E. coli virulence-factor promoters of outer membrane protein X and p-fimbriae. Triclosan stents had significantly fewer adherent viable bacteria than control stents. CONCLUSIONS: Triclosan-eluting ureteral stents inhibit the growth of common bacterial uropathogens and thus may reduce the incidence of urinary-tract infections and, potentially, encrustation. This drug-eluting stent provides both mechanical drainage of the upper urinary tract and local antibiosis.