A poly(hydroxyethyl methacrylate)-Ag nanoparticle porous hydrogel for simultaneous in vivo prevention of the foreign-body reaction and bacterial infection.

The use of implants or indwelling medical devices has greatly enhanced the quality and efficacy of health care. However, foreign-body reactions (FBRs) and infections can lead to potential failure or removal of the devices, or increased morbidity and mortality of patients. Herein, we develop a silver nanoparticle (AgNP) loaded poly(hydroxyethyl methacrylate) hydrogel with spherical, interconnected 40 µm pores. The resulting hydrogels displayed good antibacterial properties regarding both gram positive bacteria (Staphylococcus aureus) and gram negative bacteria (Escherichia coli (E. coli)) in vitro and were highly efficient at inhibiting bacterial cell growth. Moreover, they exhibited an in vivo resistance to FBRs by reducing the immune responses, and completely prevented the formation of collagen capsules. Finally, in vivo studies of the E. coli infected mouse model demonstrated that the AgNP loaded porous hydrogels were highly efficient at resisting the bacterial FBRs and infections, while they promoted cell mitigation and infiltration. Findings from this work suggest that AgNP loaded porous hydrogels hold promise in various biomedical applications including in the new generation of implantable biomedical devices and tissue engineering scaffolds.

Do Bacteria and Biofilm Play a Role in Double-Capsule Formation around Macrotextured Implants?

BACKGROUND: The double capsule is a complication mostly described in aggressive macrotextured implants. Mechanical shear stress applied onto an immature periprosthetic capsule has been linked to their formation. The authors aim to demonstrate the role of bacterial phenotype and biofilm in the development of the double capsule. METHODS: Seven double capsules formed at the interface of macrotextured breast expander implants were studied using scanning electron microscopy. Two samples for each surface of the inner capsule layer (the prosthesis interface and the intercapsular space) were analyzed for bacteria cell size, bacterial density, and biofilm deposition. RESULTS: Although all routine bacterial cultures were negative, the prosthesis interface had both higher bacteria load and biofilm deposition compared with the intercapsular space (Mann-Whitney U test, p = 0.004 and p = 0.008, respectively). Moreover, bacteria cell sizes were significantly smaller at the prosthesis interface in six of seven samples. Comparison of bacteria density and biofilm dispersion showed an increase of biofilm extracellular matrix deposition over 2000 cells/mm (linear regression, p = 0.0025). These results indicate a common trend among bacteria species. CONCLUSIONS: Bacterial expression between the different surfaces of the double capsule displays significant differences; bacteria at the prosthesis interface are mostly in a biofilm state, whereas they demonstrate a planktonic phenotype at the intercapsular space. When a sufficient amount of bacteria are present at a specific location, quorum sensing may trigger a biofilm phenotypic switch in planktonic bacteria cells. Biofilm formation may alter capsule formation through immune response, thereby weakening capsule strength and facilitating extracellular matrix delamination and double-capsule formation. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, V.

Efficient Killing of Planktonic and Biofilm-Embedded Coagulase-Negative Staphylococci by Bactericidal Protein P128.

Coagulase-negative staphylococci (CoNS) are the major causative agents of foreign-body-related infections, including catheter-related bloodstream infections. Because of the involvement of biofilms, foreign-body-related infections are difficult to treat. P128, a chimeric recombinant phage-derived ectolysin, has been shown to possess bactericidal activity on strains of Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA). We tested the killing potential of P128 on three clinically significant species of CoNS, S. epidermidis, S. haemolyticus, and S. lugdunensis, under a variety of physiological conditions representing growing and nongrowing states. The MIC90 and minimum bactericidal concentration at which 90% of strains tested are killed (MBC90) of P128 on 62 clinical strains of CoNS were found to be 16 and 32 µg/ml (0.58 and 1.16 µM), respectively, demonstrating the bactericidal nature of P128 on CoNS strains. Serum showed a potentiating effect on P128 inhibition, as indicated by 4- to 32-fold lower MIC values observed in serum. P128 caused a rapid loss of viability in all CoNS strains tested. Persisters of CoNS that were enriched in the presence of vancomycin or daptomycin were killed by P128 at 1× the MIC in a rapid manner. Low concentrations of P128 caused a 2- to 5-log reduction in CFU in stationary-phase or poorly metabolizing CoNS cultures. P128 at low concentrations eliminated CoNS biofilms in microtiter plates and on the surface of catheters. Combinations of P128 and standard-of-care (SoC) antibiotics were highly synergistic in inhibiting growth in preformed biofilms. Potent activity on planktonic cells, persisters, and biofilms of CoNS suggests that P128 is a promising candidate for the clinical development of treatments for foreign-body-related and other CoNS infections.

Intraventricular Bone Wax as Cause of Recurrent Cerebrospinal Fluid Infection: A Neuroradiologic Pitfall.

BACKGROUND: Persistent or recurrent shunt infections can be secondary to predisposing factors, such as isolated compartments, cerebrospinal fluid leaks, or foreign bodies. CASE DESCRIPTION: A 5-year-old girl experienced several episodes of shunt infections. After careful reevaluation of all neuroradiologic records of the patient, a foreign body in the left frontal horn of the lateral ventricle was suspected. An endoscopic approach was used to identify and remove a small fragment of bone wax. CONCLUSIONS: To our knowledge, this is the first report of intraventricular bone wax causing persistent CSF infection. The diagnosis was difficult because wax resembles air on computed tomography and on magnetic resonance imaging, and so it had been not noticed for months. Only its persistence on several images raised the suspicion of foreign body inside the ventricular system.

Evaluation of a Novel Tc-99m Labelled Vitamin B12 Derivative for Targeting Escherichia coli and Staphylococcus aureus In Vitro and in an Experimental Foreign-Body Infection Model.

PURPOSE: Vitamin B12 (cyanocobalamin, Cbl) is accumulated by rapidly replicating prokaryotic and eukaryotic cells. We investigated the potential of a Tc-99m labelled Cbl derivative ([(99m)Tc]PAMA(4)-Cbl) for targeting infections caused by Escherichia coli and Staphylococcus aureus. In vitro binding assays were followed by biodistribution studies in a mouse model of foreign body infection. PROCEDURES: E. coli (ATCC 25922) and S. aureus (ATCC 43335) were used as test strains. [(57)Co]Cbl, [(67)Ga]citrate and [(99m)Tc]DTPA served as reference compounds. The in vitro competitive binding of [(57)Co]Cbl or [(99m)Tc]PAMA(4)-Cbl, and unlabeled Cbl, to viable or killed bacteria, was evaluated at 37 and 4 °C. A cage mouse model of infection was used for biodistribution of intravenous [(57)Co]Cbl and [(99m)Tc]PAMA(4)-Cbl in cage and dissected tissues of infected and non-infected mice. RESULTS: Maximum binding (mean ± SD) of [(57)Co]Cbl to viable E. coli was 81.7 ± 2.6 % and to S. aureus 34.0 ± 6.7 %, at 37 °C; no binding occurred to heat-killed bacteria. Binding to both test strains was displaced by 100- to 1000-fold excess of unlabeled Cbl. The in vitro binding of [(99m)Tc]PAMA(4)-Cbl was 100-fold and 3-fold lower than the one of [(57)Co]Cbl for E. coli and S. aureus, respectively. In vivo, [(99m)Tc]PAMA(4)-Cbl showed peak percentage of injected dose (% ID) values between 1.33 and 2.3, at 30 min post-injection (p.i.). Significantly higher retention occurred in cage fluids infected with S. aureus at 4 h and with E. coli at 8 h p.i. than in non-infected animals. Accumulation into infected cages was also higher than the one of [(99m)Tc]DTPA, which showed similar biodistribution in infected and sterile mice. [(57)Co]Cbl gradually accumulated in cages with peaks % ID between 3.58 and 4.83 % achieved from 24 to 48 h. Discrimination for infection occurred only in E. coli-infected mice, at 72 h p.i. [(67)Ga]citrate, which showed a gradual accumulation into cage fluids during 12 h, was discriminative for infection from 48 to 72 h p.i. (P < 0.05). CONCLUSION: Cbl displayed rapid and specific in vitro binding to test strains. [(99m)Tc]PAMA(4)-Cbl was rapidly cleared from most tissues and discriminated between sterile and infected cages, being a promising candidate for imaging infections in humans.

Fosfomycin-daptomycin and other fosfomycin combinations as alternative therapies in experimental foreign-body infection by methicillin-resistant Staphylococcus aureus.

The efficacy of daptomycin, imipenem, or rifampin with fosfomycin was evaluated and compared with that of daptomycin-rifampin in a tissue cage model infection caused by methicillin-resistant Staphylococcus aureus (MRSA). Strain HUSA 304 was used. The study yielded the following results for MICs (in µg/ml): fosfomycin, 4; daptomycin, 1; imipenem, 0.25; and rifampin, 0.03. The study yielded the following results for minimum bactericidal concentration (MBC) (in µg/ml): fosfomycin, 8; daptomycin, 4; imipenem, 32; and rifampin, 0.5. Daptomycin-rifampin was confirmed as the most effective therapy against MRSA foreign-body infections. Fosfomycin combinations with high doses of daptomycin and rifampin were efficacious alternative therapies in this setting. Fosfomycin-imipenem was relatively ineffective and did not protect against resistance.

Foreign-body response to sterile catheters is variable over 20 weeks.

We hypothesized that placement of sterile catheter material into the peritoneal cavity results in a foreign-body response that varies with exposure duration. Sterile medical Silastic catheter material was aseptically implanted into the abdomens of 42 anesthetized Sprague-Dawley rats. Controls (n = 18) underwent sham operations without catheter implantation. After 4, 8, or 20 weeks, the animals were anesthetized, the abdomen was opened, and the catheter material was recovered and processed to separate the cells adhering to the catheters. The cells, abdomen, and catheter material were all cultured to demonstrate sterility, and transport experiments were carried out. After euthanasia of the animals, abdominal wall tissue was examined for submesothelial thickness and vascular density, and immunohistochemistry (IHC) for cytokines was performed. Cells from the catheter material were processed for immunocytochemistry (ICC). The catheter, adherent cells, and abdomen were free of bacteria. Inflammatory changes in peritoneal thickness and angiogenesis were highest at 4 weeks and declined thereafter to 20 weeks. Transport of mannitol was higher at 4 weeks in treated animals than in controls, and albumin transport was higher at 8 weeks in treated animals than in controls. The IHC for cytokines demonstrated changes paralleling the structural alterations (p < 10(-5)). The ICC of the catheter cell layer demonstrated mesothelial cells, macrophages, fibroblasts, and T cells. Over 20 weeks, the foreign-body response to polymer catheters placed intraperitoneally in rats without injection of solution depends on exposure time, with an initial immune response evident at 4 weeks and decreasing thereafter.

Efficacy of high doses of daptomycin versus alternative therapies against experimental foreign-body infection by methicillin-resistant Staphylococcus aureus.

Since the currently approved dose of daptomycin (6 mg/kg of body weight/day) has been associated with clinical failures and resistance development, higher doses for some difficult-to-treat infections are being proposed. We studied the efficacy of daptomycin at high doses (equivalent to 10 mg/kg/day in humans) and compared it to that of reference and alternative treatments in a model of foreign-body infection with methicillin (meticillin)-resistant Staphylococcus aureus. In vitro studies were conducted with bacteria in the log and stationary phases. For the in vivo model, therapy with daptomycin at 100 mg/kg/day, vancomycin at 50 mg/kg/12 h, rifampin (rifampicin) at 25 mg/kg/12 h, or linezolid at 35 mg/kg/12 h was administered for 7 days. Antibiotic efficacy was evaluated using either bacteria from tissue cage fluids or those attached to coverslips. We screened for the emergence of linezolid- and rifampin-resistant strains and analyzed the surviving population from the daptomycin-treated group. Only daptomycin was bactericidal in both the log- and stationary-phase studies. Daptomycin (decrease in the log number of CFU per milliliter of tissue cage fluid, 2.57) and rifampin (decrease, 2.6 log CFU/ml) were better (P < 0.05) than vancomycin (decrease, 1.1 log CFU/ml) and linezolid (decrease, 0.9 log CFU/ml) in the animal model. Rifampin-resistant strains appeared in 60% of cases, whereas no linezolid resistance emerged. No daptomycin-resistant subpopulations were detected at frequencies of 10(-7) or higher. In conclusion, daptomycin at high doses proved to be as effective as rifampin, and the two were the most active therapies for this experimental foreign-body infection. These high doses ensured a profile of safety from the development of resistance.

[Long-term complications after injection of permanent tissue-fillers to the lips]. / Langtidskomplikationer efter injektion af permanent vaevsfylder i laeber.

We present two cases with signs of infection and granulomas seen years after injection of permanent fillers to the lips. These side effects are difficult to treat. They resemble an immunological foreign body response and late-onset infection as well as granuloma and scaring depending on the substance injected. Steroid injections are indicated in case of granuloma, and antibiotics should be used when infection occurs, but surgical excision can be required. It is important to emphasize this condition in order to give patients proper advice prior to treatment and to recognize symptoms.

The role of sigmaB in persistence of Staphylococcus epidermidis foreign body infection.

Staphylococcal biofilm formation depends on the transcription factor sigma(B). We further investigated the role of sigma(B) in biofilm formation and persistence in vitro and in vivo in a subcutaneous rat model. As expected, expression of all sigma(B) operon genes was transiently higher in the first 6 h of biofilm formation compared to planktonic bacteria, concurrent with a temporary upregulation of icaA and aap expression. However, we also observed a second upregulation of sigB expression in biofilm more than 2 days old without upregulation of icaA or aap. Biofilm formation by Staphylococcus epidermidis strains 8400 and 1457 was compared to that of isogenic mutants with inactivation of rsbU, of rsbUVW and of the entire sigma(B) operon. Both wild-type strains and the constitutively sigB-expressing rsbUVW mutant showed a strong biofilm-positive phenotype. The rsbUVW mutant biofilm was, however, thinner and more evenly spread than the wild-type biofilm. Inactivation of SigB in the rsbUVWsigB mutant or mutation of the positive regulator RsbU reduced both the number of sessile bacteria and polysaccharide intercellular adhesin (PIA) synthesis. These differences between the wild-types and their respective mutants appeared after 6 h in in vitro biofilms but only after 4 days in in vivo biofilms. Our results provide additional evidence for a role for sigma(B) in biofilm formation. They also suggest a role for sigma(B) in biofilm maturation and stability that is independent of PIA or accumulation-associated protein (Aap) and point to significant differences in the temporal development between in vitro and in vivo biofilms.

Histological study on acute inflammatory reaction to polyurethane-coated silicone implants in rats.

PURPOSE: Evaluating histologically the silicone peri-implant coated by polyurethane inflammation associated to the use of anti-microbial and bacterial contamination. METHODS: It was used 35 Wistar rats. The animals were divided in seven groups: I - Control; II - implant cavity contamination with 10 bacteria/ml; III - implant cavity contamination with 10 bacteria/ml; IV - implant cavity contamination with 10 bacteria/ml; V - identical contamination to group II and implant immersions in anti-microbial solution; VI - identical contamination in group III and implant immersions in the anti-microbial solution; VII - identical contamination of group IV and implant immersions in anti-microbial solution. It was evaluated morphometrically the peri-implant capsules after 30 days of introduction. RESULTS: The factors with more discriminating power were the giants cells of a strange body and the mononuclear. There was no correlation between the bacterial concentrations and the histological alterations. CONCLUSION: 1) The histological standard of the inflammatory reaction around the silicone implant coated with polyurethan is chronic granulomatosis type of a strange body; 2) There isn't correlation between concentration of Staphylococcus epidermidis and histological changes; 3) The use of anti-microbial solution decreased the mononuclear cell reactions, with the increase of giant cells in a strange body.

Histological study on acute inflammatory reaction to polyurethane-coated silicone implants in rats

PURPOSE: Evaluating histologically the silicone peri-implant coated by polyurethane inflammation associated to the use of anti-microbial and bacterial contamination. METHODS: It was used 35 Wistar rats. The animals were divided in seven groups: I - Control; II - implant cavity contamination with10 bacteria/ml; III - implant cavity contamination with 10 bacteria/ml; IV - implant cavity contamination with 10 bacteria/ml; V - identical contamination to group II and implant immersions in anti-microbial solution; VI - identical contamination in group III and implant immersions in the anti-microbial solution; VII - identical contamination of group IV and implant immersions in anti-microbial solution. It was evaluated morphometrically the peri-implant capsules after 30 days of introduction. RESULTS: The factors with more discriminating power were the giants cells of a strange body and the mononuclear. There was no correlation between the bacterial concentrations and the histological alterations. CONCLUSION: 1) The histological standard of the inflammatory reaction around the silicone implant coated with polyurethan is chronic granulomatosis type of a strange body; 2) There isn´t correlation between concentration of Staphylococcus epidermidis and histological changes; 3) The use of anti-microbial solution decreased the mononuclear cell reactions, with the increase of giant cells in a strange body.

OBJETIVO: Avaliar, histologicamente, a reação inflamatória aos implantes de silicone revestidos por poliuretano, com contaminação bacteriana, associada ou não ao uso de antimicrobianos. MÉTODOS: Utilizou-se 35 ratos Wistar. Os animais foram divididos em 7 grupos: I- Controle, II- contaminação da cavidade do implante com 10¹ bactérias/ml, III- contaminação da cavidade do implante com 10³ bactérias/ml, IV- contaminação da cavidade do implante com 10(5) bactérias/ml, V- contaminação idêntica ao grupo II e imersão dos implantes em solução antimicrobiana, VI- contaminação idêntica do grupo III e imersão dos implantes em solução antimicrobiana, VII- contaminação idêntica do grupo IV e imersão dos implantes em solução antimicrobiana. Avaliou-se morfometricamente as cápsulas peri-implantes após 30 dias da introdução. RESULTADOS: Os fatores com maior poder discriminante foram as células gigantes de corpo estranho e os mononucleares. Não houve correlação entre as concentrações bacterianas e as alterações histológicas. CONCLUSÕES: 1) O padrão histológico da reação inflamatória ao redor dos implantes de silicone revestidos com poliuretano é do tipo crônica granulomatosa de corpo estranho; 2) Não há correlação entre a concentração de bactérias Staphylococcus epidermidis e as alterações morfométricas; 3) O uso de solução antimicrobiana diminui a reação de células mononucleares, com aumento de células gigantes de corpo estranho.

Bacteriological analysis of meshes removed for complications after surgical management of urinary incontinence or pelvic organ prolapse.

The aim of this study is to examine the role of bacterial infection in complications following surgical management of urinary incontinence and genital prolapse using meshes. There were sixteen prostheses removed. Eight were monofilament polypropylene-knitted meshes, one was a silicone-coated polypropylene mesh, another was a collagen-coated polypropylene mesh, four were silicone-coated polyester meshes and two were polyester meshes. The most frequent cause for removal was symptomatic vaginal erosion (62%). Cultures were performed under aerobic, anaerobic and enrichment conditions. Infection was multimicrobial for 31% of meshes. When only one bacteria was found, it was Proteus mirabilis in 25% of cases. Forty-three per cent of bacterial quantifications were under 10(3) colony-forming units per millilitre. Bacterial contamination was found in all meshes, quantification was often low, and therefore, its exact role is not yet clear.

Influence of rifampin on capsule formation around silicone implants in a rat model.

This study investigated the effect of rifampin on the thickness of capsules around silicone implants by bactericidal activity against Stapylococcus epidermidis. Silicone blocks (1 x 1 cm) were placed into pockets created for each of the 40 rats included in the study. In group 1, the operation was performed under aseptic conditions. In group 2, standard S. epidermidis was inoculated into the pocket, whereas rifampin and S. epidermidis were applied in group 3. In group 4, only rifampin was applied topically on implants. After 12 weeks, the peri-implant capsules were removed and examined under a photomicroscope and a scanning electron microscope. The mean thickness of the capsules was 63.307 microm in group 1, 111.538 microm in group 2, 43.076 microm in group 3, and 30.384 mum in group 4. The differences between groups 2 and 3 and groups 2 and 4 were found to be statistically significant (p < 0.001). Rifampin appears to be an agent for preventing peri-implant capsule formation.

[Foreign body infections--biofilms and quorum sensing]. / Fremmedlegemeinfektioner--nyt om biofilm og quorum sensing.

Biofilms are structured consortia of bacteria embedded in self-produced polymer matrix. Biofilms are resistant to antibiotics, disinfectives and phagocytosis. The persistence of foreign body infections is due to biofilms. Chronic P. aeruginosa lung infection in cystic fibrosis patients is a biofilm. Bacteria in biofilms communicate by means of quorum sensing which activates genes for virulence factors. Biofilms can be prevented by antibiotic prophylaxis or early therapy or by quorum sensing inhibitors which make them susceptible to antibiotics and phagocytosis.

Metabolic activity of Staphylococcus epidermidis is high during initial and low during late experimental foreign-body infection.

Foreign-body infection (FBI) is notoriously resistant to eradication by antibiotic treatment. It is hypothesized that reduced bacterial metabolic activity contributes to this resistance. We examined the metabolic activity of Staphylococcus epidermidis in 204 samples recovered during in vitro foreign-body colonization and in 424 samples recovered during in vivo FBI in a rat model. Metabolic activity was measured by determining the amount of 16S rRNA per genome by quantitative PCR. The initial foreign-body-associated growth proved to be a metabolically active process, both in vitro and in vivo. The initial 16S rRNA content was similar to that observed during in vitro exponential-growth phase. However, during late in vivo FBI, a 114-fold (P << 0.0001) decrease in the 16S rRNA content was observed, indicating that there was markedly decreased metabolic activity. This decreased metabolic activity during late FBI can explain at least in part why such infections are so difficult to eradicate with conventional antibiotic treatment.