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PubMed1997

Leukocyte-biomaterial interactions in the presence of Staphylococcus epidermidis: flow cytometric evaluation of leukocyte activation.

The adhesion of bacteria on a biomaterial surface is believed to be the first step in the development of biomaterial-related infection. The goal of this study was to investigate the mechanisms that permit adherent bacteria to persist on the surface of an implanted cardiovascular biomaterial. We hypothesized that circulating leukocytes are unable to adhere to the biomaterial surface under physiologic shear stress conditions, and this prevents them from interacting with adherent bacteria. To address this hypothesis, we investigated the adhesion profiles of Staphylococcus epidermidis and polymorphonuclear leukocytes (PMN), incubated under controlled shear stress conditions with the test biomaterial. We found that bacteria could adhere on the biomaterial surface, even when their concentration in the test medium was as low as 10(3) cfu/mL. At this concentration, the bacteria did not induce significant complement activation. PMN adhesion on the biomaterial surface was sensitive to shear stress and minimal at shear stress > 10 dynes/cm2. Low concentrations of bacteria could induce a significant increase in the expression of PMN adhesion molecules CD11b and CD11c. We conclude that the presence of bacteria induces PMN activation but does not increase PMN adhesion on biomaterial surfaces under physiologic shear stress conditions. This could be a major mechanism that protects adherent bacteria from PMN antibacterial activity.

Bacterial Adhesion↗

PubMed1998

Clonal expansion of Staphylococcus epidermidis strains causing Hickman catheter-related infections in a hemato-oncologic department.

The detailed analysis of 411 strains of coagulase-negative staphylococci (CoNS) obtained from 40 neutropenic hemato-oncologic patients (61 Hickman catheter episodes) on intensive chemotherapy is described. By random amplification of polymorphic DNA (RAPD) analysis, a total of 88 different genotypes were detected: 51 in air samples and 30 in skin cultures prior to insertion, 12 in blood cultures after insertion, and only 5 involved in catheter-related infections (CRI). Two RAPD genotypes of Staphylococcus epidermidis predominated, and their prevalence increased during patient hospitalization. At insertion, these clones constituted 11 of 86 (13%) CoNS isolated from air samples and 33 of 75 (44%) CoNS isolated from skin cultures. After insertion, their combined prevalence increased to 33 of 62 (53%) in catheters not associated with CRI and 139 of 188 (74%) in catheters associated with CRI (P = 0.0041). These two predominant S. epidermidis clones gave rise to a very high incidence of CRI (6.0 per 1,000 catheter days) and a very high catheter removal rate for CRI, 70%, despite prompt treatment with vancomycin. A likely source of S. epidermidis strains involved in CRI appeared to be the skin flora in 75% of cases. The validity of these observations was confirmed by pulsed-field gel electrophoresis (PFGE) of SmaI DNA macrorestriction fragments of blood culture CoNS isolates. Again, two predominant CoNS genotypes were found (combined prevalence, 60%). RAPD and PFGE yielded concordant results in 75% of cases. Retrospectively, the same two predominant CoNS clones were also found among blood culture CoNS isolates from the same hematology department in the period 1991 to 1993 (combined prevalence, 42%) but not in the period 1978 to 1982. These observations underscore the pathogenic potential of clonal CoNS types that have successfully and persistently colonized patients in this hemato-oncology department.

Air Microbiology↗

PubMed2005

Staphylococcus epidermidis polysaccharide intercellular adhesin production significantly increases during tricarboxylic acid cycle stress.

Staphylococcal polysaccharide intercellular adhesin (PIA) is important for the development of a mature biofilm. PIA production is increased during growth in a nutrient-replete or iron-limited medium and under conditions of low oxygen availability. Additionally, stress-inducing stimuli such as heat, ethanol, and high concentrations of salt increase the production of PIA. These same environmental conditions are known to repress tricarboxylic acid (TCA) cycle activity, leading us to hypothesize that altering TCA cycle activity would affect PIA production. Culturing Staphylococcus epidermidis with a low concentration of the TCA cycle inhibitor fluorocitrate dramatically increased PIA production without impairing glucose catabolism, the growth rate, or the growth yields. These data lead us to speculate that one mechanism by which staphylococci perceive external environmental change is through alterations in TCA cycle activity leading to changes in the intracellular levels of biosynthetic intermediates, ATP, or the redox status of the cell. These changes in the metabolic status of the bacteria result in the attenuation or augmentation of PIA production.

Acetic Acid↗

PubMed1988

[Pseudo- "toxic lens" syndrome over the course of 4 years caused by staphylococcus epidermidis endophthalmitis].

A 79-year-old female patient presented with recurrent severe iridocyclitis after extracapsular cataract extraction and implantation of a PMMA posterior chamber lens in 1983: In April, June, July, September, October and December 1984 as well as April and August 1985 and January and April 1987, the uveitis appeared without a hypopyon and was diagnosed as a "toxic lens syndrome". Consequently, IOL explantation followed by anti-inflammatory and - intermittently - immunosuppressive therapy was performed in October 1984 with a resulting improvement in vision. In June 1987, the patient presented with a full-blown bacterial endophthalmitis, which required immediate pars plana vitrectomy. Lens cortex material and capsule residues were removed. Staphylococcus epidermidis was cultured from the vitreous aspirate using enrichment techniques. Antibiotic topical and systemic therapy was performed according to the antibiotic sensitivity test and resulted in a complete cure of the intraocular inflammation.

Aged↗

PubMed2004

Polyurethanes loaded with antibiotics: influence of polymer-antibiotic interactions on in vitro activity against Staphylococcus epidermidis.

Acidic or basic polyurethanes were loaded with antibiotics to develop materials to prevent medical device-related infections. A correlation between polymer-antibiotic interactions and amount of drug absorbed by polymers and released over time was found. Since the employed antibiotics, i.e. amoxicillin, cefamandole nafate, rifampin and vancomycin, possessed at least an acidic group in their structural formula, the introduction of basic tertiary amines in the polyurethane side-chain resulted in an increased polymer ability to adsorb antibiotics. However, a stronger ionic interaction between this polymer and the antibiotics caused a release of lower amount of drug over time. Antibiotics released from polymers inhibited Staphylococcus epidermidis growth on agar. Antibiotic-loaded polyurethanes kept in water for increasing times were still able to show inhibition zones of bacterial growth. The antibacterial activity lasted up to 3 hours for amoxicillin, 24 hours for vancomycin, 8 days for cefamandole nafate and 8 months for rifampin.

Amoxicillin↗

PubMed1969

Effect of oxygen on heme and porphyrin accumulation from delta-aminolevulinic acid by suspensions of anaerobically grown Staphylococcus epidermidis.

The effect of various conditions on the accumulation of porphyrins and heme by resting suspensions of anaerobically grown cells of Staphylococcus epidermidis was examined. Anaerobically grown cells contain 10 to 15% of the amount of protoheme found in cells grown aerobically. Resting suspensions of anaerobically grown cells, when incubated aerobically in buffer with delta-aminolevulinic acid and glucose for 60 min, exhibited a fourfold increase in protoheme content. At high levels of delta-aminolevulinic acid, there was also a significant accumulation of porphyrins with the solubility and chromatographic properties of coproporphyrin and uroporphyrin. Protoporphyrin was not accumulated. When oxygen was excluded from the incubation mixture, accumulation of protoheme was prevented, but accumulation of coproporphyrin and total porphyrin was enhanced. Nitrate served as an electon acceptor as indicated by its reduction to nitrite; however, nitrate did not substitute for oxygen in causing the accumulation of protoheme. These results suggested that oxygen is required for one of the late steps of heme synthesis in S. epidermidis, possibly for the conversion of coproporphyrinogen to protoporphyrin. The inability of nitrate to substitute for oxygen suggests a role for molecular oxygen as a substrate rather than as an electron acceptor for heme synthesis.

Chromatography, Thin Layer↗

PubMed2006

Staphylococcus epidermidis adhesion on modified urea/urethane elastomers.

Block urea/urethane co-polymer films present elastomeric properties with the possible tuning of their surface properties within a wide range and are therefore considered relevant surfaces for possible medical applications. In particular, thin free standing films of urea/urethane elastomers with two soft segments, polypropylene oxide and more hydrophobic polybutadiene, develop multistable states with surface topography features with remarkable regularity. Moreover, complex surface structures may be obtained by UV radiation treatment followed by suitable mechanical action and also by extraction of the elastomer with a suitable solvent. In the present work, different modified elastomer samples were assayed for Staphylococcus epidermidis adhesion during 2 h and the extent of bacterial adhesion was evaluated by automatic cell enumeration. Bacterial adhesion assays demonstrate that the typical trend relating the increase in the number of adhered bacteria with the increase of the surface roughness does not hold for all materials. Results may be interpreted taking into account both the surface topography and the different types of micro-phase segregation of hydrophobic and hydrophilic parts of the elastomer.

Bacterial Adhesion↗

PubMed2003

Isolation of methicillin-resistant coagulase-negative staphylococci from patients undergoing continuous ambulatory peritoneal dialysis (CAPD) and comparison of different molecular techniques for discriminating isolates of Staphylococcus epidermidis.

Coagulase-negative staphylococci (CNS) have emerged as an important pathogen in nosocomial infections. About 80%-90% of CNS isolates associated with hospital infections are methicillin-resistant coagulase-negative staphylococci (MRCNS). The aims of this study were to screen for MRCNS isolates in the flora of a small population of patients undergoing continuous ambulatory peritoneal dialysis (CAPD) and to evaluate the discriminatory power of different molecular methods: pulsed-field gel electrophoresis (PFGE), mecA location, ClaI/mecA polymorphism and arbitrarily primed polymerase chain reaction (AP-PCR) for characterizing isolates of methicillin-resistant Staphylococcus epidermidis (MRSE). Seventy-nine CNS isolates were recovered from the 11 CAPD patients studied. Using a methicillin screening agar and a DNA specific mecA probe we verified that 30 of the 79 (38%) CNS isolates were resistant to methicillin (MRCNS). Twenty-two of the 30 MRCNS (73%) were MRSE, 7 (23%) methicillin-resistant S. haemolyticus (MRSH(ae)) and 1 (3%) methicillin-resistant S. hominis (MRSH(om)). All patients analyzed carried MRCNS in their flora, in one or more sites. Since CAPD patients have high risk for developing peritonitis, the colonization of these patients with MRCNS might represent an additional problem, due to the therapeutic restrictions imposed by these multiresistant isolates. A wide genetic diversity was verified when the PFGE of the MRSE isolates was analyzed. The 22 MRSE isolates displayed a total of 15 PFGE different patterns (11 PFGE types and 4 subtypes). The location of mecA in the SmaI-fragmented genome DNA did not bring any additional advantage for epidemiologic characterization of the isolates. The ClaI/mecA polymorphism was able to correctly discriminate 12 from the 15 PFGE patterns. In addition, the DNA of 20 MRSE isolates were used for AP-PCR typing. These isolates belonged to 14 PFGE patterns (11 types and 3 subtypes) and displayed 15 genotypes (for the association of PFGE, mecA location and ClaI/mecA polymorphism). A total of 17 different amplification patterns was verified using the primer 1. Only for 2 genotypes, strains having identical genetic backgrounds were further discriminated by AP-PCR (2 of 15 genotypes (87%) for AP-PCR and 1 of 15 genotypes for PFGE; (93%). Concluding, our results indicated that the AP-PCR can be an alternative and useful tool for monitoring and genotyping MRSE colonization and also to molecular characterizing MRSE outbreaks in hospitals.

Anti-Bacterial Agents↗

PubMed2001

A comparison of the adhesion of mammalian cells and Staphylococcus epidermidis on fibronectin-modified polymer surfaces.

The modification of polymer surfaces to mimic the extracellular matrix, and therefore, stimulate cell growth via receptor-mediated interactions, is one approach used to promote the integration of tissue-engineering scaffolds and biomaterials into the body. However, by optimizing surfaces for tissue integration it is likely that bacterial adhesion may also be affected, resulting in a greater risk of biomaterial-related infection. This could be detrimental to both the implant and the patient because biomaterial related infections are particularly resistant to host defenses and antibiotics. In this study, we analyzed the adhesion of a Staphylococcus epidermidis clinical isolate and 3T3 rat fibroblasts to tissue culture plastic coated with varying concentrations of fibronectin (Fn). Bacterial adhesion was always lower than tissue culture plastic and appeared to decrease with increasing Fn concentrations. Mammalian cell adhesion to Fn exceeded adhesion to tissue culture plastic but did not differ significantly over the range of protein concentrations or between 1 and 4 h of incubation. In most cases, the total surface coverage did not vary with time or Fn concentration, indicating that maximal cell adhesion and spreading occurred rapidly and at low protein concentrations. This study suggests that, by controlling the density of proteins or ligands on a surface, we can potentially optimize mammalian cell adhesion without stimulating bacterial adhesion, hence reducing the likelihood of infection.

3T3 Cells↗

PubMed2002

Effects on antibiotic resistance of Staphylococcus epidermidis following adhesion to polymethylmethacrylate and to silicone surfaces.

A number of studies appears to give emphasis to the role of prosthetic materials in determining microbial adherence and resistance to host defence and drug therapy. Aim of this study was to explore whether the direct contact with biomaterial substrata of different chemical nature could influence bacterial behaviour, determining possible changes in the bacteria population as far as antibiotic resistance is concerned. To this end, susceptibility to penicillin, erythromycin, clindamycin, cefamandole, imipenem, vancomycin, ciprofloxacin. ampicillin, cefazolin, trimethoprim-sulfamethoxazole, chloramphenicol, amikacin and netilmicin was evaluated in a methicillin-, gentamicin- and tobramycin-resistant Staphylococcus epidermidis strain, after in vitro adhesion to polymethylmethacrylate (PM MA) and to silicone elastomer. The susceptibility to antibiotics of both adherent bacteria and bacteria which, although exposed to the materials, had not undergone adhesion was measured as bacterial growth inhibition area onto a plate antibiogram. according to Kirby-Bauer and using an image analyser system. The results obtained suggest that the two test materials considered in this study were capable to condition bacterial behaviour. In particular. the adhesion onto PMMA surfaces induced a marked and significant decrease in susceptibility to the following beta-lactam antibiotics: cefamandole (32%), cefazolin (23%), imipenem (27%), ampicillin (31%). Moreover, PMMA caused a lower but significant reduction in resistance to vancomycin (15%), chloramphenicol (16%), amikacin (13%). netilmicin (13%), erythromycin (11%) trimethoprim-sulfamethoxazole (13%). In contrast, the adhesion onto silicone elastomer appeared to influence bacterial changes to a lesser extent and elicited a significant decrease in susceptibility only to cefazolin (10%) and amikacin (11). Further studies are required to thoroughly investigate the mechanisms of these variations, even though, also according to other authors, one of the best conceivable conclusions is that some material substrata can lead to selection of variant adhesive bacteria with increased antibiotic resistance.

Biocompatible Materials↗

PubMed2001

First results using automated epifluorescence microscopy to detect Escherichia coli and Staphylococcus epidermidis in WBC-reduced platelet concentrates.

BACKGROUND: Many methods have been tested for the detection of bacterial contamination in platelets. However, only those using molecular biology or cell culturing consistently detect contamination at levels below 10(5) bacteria per mL. This report describes the initial investigation into an alternative method that offers the possibilities of high sensitivity and rapid response while using available laboratory equipment and supplies. This method relies on a fluorescent nucleic acid stain, which preferentially stains bacteria but not platelets, and automated epifluorescence microscopy for rapid analysis. Measurements in WBC-reduced platelet concentrates (PCs) contaminated with bacteria are reported at concentrations between 10(3) and 10(6) bacteria per mL. STUDY DESIGN AND METHODS: Staphylococcus epidermidis or Escherichia coli was inoculated into aliquots of WBC-reduced PCs on Days 2 through 5 of storage. Bacterially inoculated and control PCs were stained, platelets and residual WBCs were lysed, and 200 microL of sample was filtered onto black polycarbonate filters. All preparations were done in triplicate. An automated epifluorescence microscope examined approximately 2 percent of the area of each filter and used image analysis to select the fluorescent particles that should be counted as bacteria. RESULTS: Samples containing 3 to 5 x 10(3) bacteria per mL produced about three times as many fluorescent particles classified as bacteria as the controls. Lower concentrations of S. epidermidis were detected because of higher fluorescence intensity. Simultaneous preparation of six samples requires about 35 minutes. Analysis of each prepared sample takes 10 minutes, for a total preparation and analysis time of about 95 minutes for 6 samples. CONCLUSION: Low concentrations (<5 x 10(3) bacteria/mL) of deliberately inoculated S. epidermidis or E. coli can be measured quickly in WBC-depleted PCs by using a fluorescent nucleic acid stain, differential lysis, and automated microscopy. Continued refinement of the method, studies employing other bacterial strains, and further validations of assay performance are warranted.

Automation↗

PubMedDate not supplied

Efficacy of antibiotic-lock technique with teicoplanin in staphylococcus epidermidis catheter-related sepsis during long-term parenteral nutrition.

BACKGROUND: The antibiotic-lock technique has been suggested to treat catheter-related sepsis in parenteral nutrition and to avoid catheter removal. METHODS: To determine the incidence of catheter-related sepsis, the bacteria involved, and the efficacy of the antibiotic-lock technique with teicoplanin, all patients (n = 263) undergoing parenteral nutrition from January 1997 to December 1999 in one center, with patients at the hospital (n = 209) and at home (54) were retrospectively studied. The antibiotic-lock technique with teicoplanin was systematically used in all suspected infections and maintained in staphylococcus epidermidis (SE) infections. RESULTS: A total of 21 of 263 patients had 34 infections (0.11/patient per year): 12 of 209 hospitalized and 9 of 54 home patients. A total of 10 of 34 infections were due to non-SE, and the catheter was immediately removed. The other 24 of 34 infections were due to SE; in 5 of 24, the catheter was removed after 48 hours of the antibiotic-lock technique because of persistent fever or thrombosis. A total of 5 of 12 patients had 2 or more infections on the same catheter. The antibiotic-lock technique prevented short-term catheter removal in these cases, but a second infection occurred within a median of 50 days. In 4 of 5 cases, a third infection occurred in a mean delay of 90 days so that the catheter was removed. In 3 of 5 patients, bacteria was analyzed with pulsed field gel electrophoresis, which showed that recurrent infections were due to the same strain in all cases. CONCLUSIONS: In this study, the incidence of catheter-related sepsis was low and mostly related to SE. Our results do not support the use of the teicoplanin antibiotic-lock technique in SE infections.

Anti-Bacterial Agents↗

PubMed2003

Characterization of poly(ethylene oxide) brushes on glass surfaces and adhesion of Staphylococcus epidermidis.

Poly(ethylene oxide) brushes have been covalently bound to glass surfaces and their presence was demonstrated by an increase in water contact angles from fully wettable on glass to advancing contact angles of 54 degrees, with a hysteresis of 32 degrees. In addition, electrophoretic mobilities of glass and brush-coated glass were determined using streaming potential measurements. The dependence of the electrophoretic mobilities on the ionic strength was analyzed in terms of a softlayer model, yielding an electrophoretic softness and fixed charge density of the layer. Brush-coated glass could be distinguished from glass by a 2-3-fold decrease in fixed charge density, while both surfaces were about equally soft. Adhesion of Staphylococcus epidermidis HBH276 to glass in a parallel plate flow chamber was extremely high and after 4 h, 19.0 x 10(6) bacteria were adhering per cm2. In contrast, the organisms did not adhere to brush-coated glass, with numbers below the detection limit, i.e. 0.1 x 10(6) per cm2. These results attest to the great potential of polymer brushes in preventing bacterial adhesion to surfaces.

Bacterial Adhesion↗

PubMed1981

Cross-reactions between Staphylococcus epidermidis and 23 other bacterial species.

By quantitative immunoelectrophoretic methods, 43 antigens were found in a mixture of sonicated preparations of four Staphylococcus epidermidis strains, using corresponding rabbit antiserum. Two of the antigens were identified as cell wall teichoic acid and a peptidoglycan antigen, respectively. Using this antigen/antibody reference system, cross-reactions between S. epidermidis antigens and antigens from other bacterial species were investigated. Fourteen of the S. epidermidis antigens cross-reacted with antigens from all S. aureus strains investigated. Only few cross-reactions were found between S. epidermidis and bacteria not belonging to the Micrococcaceae. The antigenic relatedness, expressed as a matching coefficient, seems promising for taxonomic work.

Antigens, Bacterial↗

PubMedDate not supplied

Staphylococcus epidermidis biotype 4: epidemiological conclusions from five different typing methods.

There is presently no accepted method for marking individual strains of Staphylococcus epidermidis. Consequently, five parameters for distinguishing such strains were examined and compared for their epidemiological efficacy: biotyping, serotyping, proteinase grouping, aminopeptidase profiles and antibiograms. Both biotyping and proteinase grouping were of limited use in identifying a particular strain, although they were helpful in initially categorizing such strains. Antibiograms were least useful because of similarities in susceptibility patterns among isolates. Serotyping and amino-peptidase profiles provided the best means of identifying an individual strain for epidemiological use. The applicability of these typing methods was demonstrated during a one year epidemiological study at a chronic disease hospital.

Aminopeptidases↗

PubMed2004

Electric current-induced detachment of Staphylococcus epidermidis biofilms from surgical stainless steel.

Biomaterial-centered infections of orthopedic percutaneous implants are serious complications which can ultimately lead to osteomyelitis, with devastating effects on bone and surrounding tissues, especially since the biofilm mode of growth offers protection against antibiotics and since removal frequently is the only ultimate solution. Recently, it was demonstrated that as a possible pathway to prevent infections of percutaneous stainless steel implants, electric currents of 60 to 100 microA were effective at stimulating the detachment of initially adhering staphylococci from surgical stainless steel. However, initially adhering bacteria are known to adhere more reversibly than bacteria growing in the later stages of biofilm formation. Hence, the aim of this study was to examine whether a growing Staphylococcus epidermidis biofilm can be stimulated to detach from surgical stainless steel by the use of electric currents. In separate experiments, four currents, i.e., 60 and 100 microA of direct current (DC) and 60 and 100 microA of block current (50% duty cycle, 1 Hz), were applied for 360 min to stimulate the detachment of an S. epidermidis biofilm that had grown for 200 min. A 100-microA DC yielded 78% detachment, whereas a 100-microA block current under the same experimental conditions yielded only 31% detachment. The same trend was found for 60 microA, with 37% detachment for a DC and 24% for a block current. Bacteria remaining on the surface after the current application were less viable than they were prior to the current application, as demonstrated by confocal laser scanning microscopy. In conclusion, these results suggest that DCs are preferred for curing infections.

Bacterial Adhesion↗

PubMed1993

An assay of Staphylococcus epidermidis biofilm responses to therapeutic agents.

Implant-associated infections tend to become persistent, resisting host defences and antibiotic therapy. Routine clinical laboratory testing of bacterial isolates in the fluid phase for antibiotic susceptibility may not be predictive of therapeutic outcomes and therefore a number of antibiotic regimes have been formulated empirically. The resistance of implant-associated infections has been ascribed to the production by bacteria, when adherent to the implant surface, of a shielding matrix of polymerized carbohydrates protecting the enclosed bacteria from immune defences and antibiotics. This complex of surface, bacteria and matrix is termed a biofilm. We describe a technique to evaluate the efficacy of antimicrobial agents directed against biofilm-forming strains of Staphylococcus epidermidis (a major cause of implant infections) utilizing standardized biofilm preparations formed on glass. The impact of the antimicrobial agents was assessed quantally using the end-point of permanent cessation of metabolic activity (cell death) of the entire biofilm determined by the loss of ability to reduce 2,3,5-triphenyltetrazolium chloride to a visible red intracellular precipitate of formazan. The relative rate of action of differing antimicrobial agents could be determined by the minimum period of exposure of the biofilm to the agents that is required to bring about sterilization, the clinically relevant marker. A wide range of antimicrobial substances could be evaluated, including chemical disinfectants, immunoreactive substances, antibiotics; singly and in combination; and the modifying effects of interaction with non-antibacterial therapeutic agents and other environmental factors. The technique is simple, inexpensive, reproducible and readily adaptable to the clinical situation where evaluation of therapeutic regimes for individual cases of prosthetic device-associated infection is required routinely with despatch and ease of interpretation.

Anti-Bacterial Agents↗

PubMed2004

Poly(ethylene glycol)-polyacrylate copolymers modified to control adherent monocyte-macrophage physiology: interactions with attaching Staphylococcus epidermidis or Pseudomonas aeruginosa bacteria.

The ability of various surface modifications of poly(ethylene glycol)-graft-polyacrylate (PEG-g-PA) copolymers (tethered adhesion peptides and fragments of monoclonal antibodies) to modulate monocyte-macrophage cell interactions with surface colonizing bacteria is reported. The PEG-g-PA copolymers were made to inhibit nonspecific protein and cellular adhesion. The copolymers were then covalently modified with either cell adhesion peptides (YRGDS, YEILDV, or YRGES) or fragments of antibodies to monocyte-macrophage integrin receptors (anti-VLA4, anti-beta(1), anti-beta(2), and anti-CD64), which are known to enhance macrophage adhesion and perhaps modulate their activation. Cytokine expression and phagocytosis response by surface adherent monocyte-macrophages to Staphylococcus epidermidis and Pseudomonas aeruginosa bacteria were quantified. The cytokine expression (interleukins 6 and 1 beta) of adherent macrophages in response to the modified polymers only and to bacterial challenges were quantified by dynamic ELISA assays. The adherent macrophage phagocytic response (oxidative burst) to various materials is compared to oxidative responses to both opsonized and nonopsonized S. epidermidis and P. aeruginosa bacteria. The efficiency of adherent macrophages to ingest and kill both species was determined using radiolabeled and fluorescent labeled bacterial cell ingestion studies as a function of the PEG-g-PA surface modification. Materials modified with adhesion peptides marginally enhanced (2x) macrophage attachment versus controls but, upon bacterial challenges, these materials predisposed adherent macrophages to overexpress proinflammatory cytokines and to exhibit a significant phagocytic response. Conversely, PEG-g-PA materials modified by fragments of monoclonal antibodies significantly enhanced (7x) macrophage adhesion but, upon bacterial challenge, "per cell" cytokine expression levels were reduced compared to peptide modified materials. Macrophages adhering to antibody fragment modified surfaces also exhibited sustained enhanced phagocytic response and higher bacterial killing efficiencies when compared with peptide modified materials.

Acrylic Resins↗