Biomaterial-centered Infection Research Articles

Antibacterial Analysis on Everolimus Grafted Biomaterial Surfaces using Polydopamine Intermediate Layer for Potential Use on Medical Devices

Biomaterial-Centered Infection (BCI) is a significant issue in the implantation of medical devices, primarily caused by the formation of bacterial biofilms on the device surface. One potential solution to address this problem is the use of antibacterial coatings. This study examines the effectiveness of everolimus as an antibacterial coating agent with polydopamine (PDA) as an intermediate layer. The commonly used biomaterial for medical scaffolds, poly(lactic acid) (PLA), was coated with everolimus after being submerged in PDA for 24 hours. The coated PLA was then subjected to antibacterial analysis, including culturing Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria in Luria-Bertani broth, bacterial count tests, and disc inhibition tests. The results showed that everolimus has antibacterial properties, but its efficacy varies with different types of bacteria adhering to the biomaterial surfaces. Specifically, the everolimus coating was found to be more effective in killing Gram-positive S. aureus. The absence of inhibition zones and the lack of further growth of both Gram-positive S. aureus and Gram-negative E. coli on the samples demonstrated the controlled release of everolimus, indicating the potential of the PDA layer in holding the everolimus release to the surrounding. Therefore, the study concludes that the formation of everolimus coating on biomaterial surfaces aided by the PDA layer, have significant potential in retarding bacterial colonies on medical devices.

Colonization and Infection of Indwelling Medical Devices by Staphylococcus aureus with an Emphasis on Orthopedic Implants.

The use of indwelling medical devices has constantly increased in recent years and has revolutionized the quality of life of patients affected by different diseases. However, despite the improvement of hygiene conditions in hospitals, implant-associated infections remain a common and serious complication in prosthetic surgery, mainly in the orthopedic field, where infection often leads to implant failure. Staphylococcus aureus is the most common cause of biomaterial-centered infection. Upon binding to the medical devices, these bacteria proliferate and develop dense communities encased in a protective matrix called biofilm. Biofilm formation has been proposed as occurring in several stages—(1) attachment; (2) proliferation; (3) dispersal—and involves a variety of host and staphylococcal proteinaceous and non-proteinaceous factors. Moreover, biofilm formation is strictly regulated by several control systems. Biofilms enable staphylococci to avoid antimicrobial activity and host immune response and are a source of persistent bacteremia as well as of localized tissue destruction. While considerable information is available on staphylococcal biofilm formation on medical implants and important results have been achieved on the treatment of biofilms, preclinical and clinical applications need to be further investigated. Thus, the purpose of this review is to gather current studies about the mechanism of infection of indwelling medical devices by S. aureus with a special focus on the biochemical factors involved in biofilm formation and regulation. We also provide a summary of the current therapeutic strategies to combat biomaterial-associated infections and highlight the need to further explore biofilm physiology and conduct research for innovative anti-biofilm approaches.

The effects of TGF-β1 on staphylococcus epidermidis biofilm formation in a tree shrew biomaterial-centered infection model

BackgroundTransforming growth factor-β1 (TGF-β1) has a wide range of biological functions. It antagonizes lymphocyte response, inhibits pro-inflammatory cytokines, and serves as a signal to turn off the immune response and inflammatory response. To study the correlation between TGF-β1 and T helper (Th)1/Th2 cytokine levels in tree shrews, and to explore the effects of different levels of TGF-β1 on central venous catheter (CVC)-centered Staphylococcus epidermidis biofilm formation in tree shrews.MethodsTree shrews were injected with different concentrations of TGF-β1, and venous blood was drawn after 48 h to measure the levels of Th1 and Th2 cytokines. A CVC was placed into the femoral vein, and TGF-β1 at different concentrations and PIA− (ATCC12228) and PIA+ (ATCC35984) standard strains of Staphylococcus epidermidis were injected into the tree shrews to establish a biomaterial-centered infection (BCI) model. After 72 h, the CVC was removed, and biofilm formation was detected using the API bacterial identification system, semi-quantitative biofilm formation assay, and scanning electron microscopy.ResultsIn the groups treated with TGF-β1 at different concentrations, the levels of Th1 cytokines interleukin-2 (IL-2), tumor necrosis factor (TNF), and interferon-γ (IFN-γ) were lower than those of normal group, while the levels of Th2 cytokines IL-6, IL-4 and IL-10 were higher than those of normal group. In the TGF-β1 groups at different concentrations, the positive rate of Staphylococcus epidermidis ATCC35984 biofilm formation was higher than that in non-TGF-β1 group, while there was no significant difference in the positive rate of Staphylococcus epidermidis ATCC12228 biofilm formation compared with that of the non-TGF-β1 group.ConclusionsTGF-β1 causes the imbalance of Th1/Th2 cytokines and Th1/Th2 shift in tree shrews, leading to Th1 cell-led decline in cellular immune function. TGF-β1 promotes PIA+ Staphylococcus epidermidis biofilm formation in the tree shrew BCI model, but it has no significant influence on PIA-Staphylococcus epidermidis biofilm formation on the surface of CVCs.

Dual-Function Antibacterial Micelle via Self-Assembling Block Copolymers with Various Antibacterial Nanoparticles.

Antibacterial biomaterials with kill-resist dual functions by combining multiple active components have been constructed, with a final aim at decreasing the incidence of biomaterial-centered infection. Self-assemblies of bactericidal ZnO or Ag–ZnO nanoparticles (NPs) with triblock copolymers, poly(ethylene glycol)-b-poly(3-hydroxybutyrate-co-3-hydroxyvalerate)–poly(ethylene glycol) (PEG–PHBV–PEG), showed a hydrophobic PHBV layer on NPs with PEG segments exposed outside via hydrogen bonding, resulting in long PEG (Mw = 2000) aggregation and short PEG (Mw = 1000) aggregation, respectively. These nanocomposite aggregations released ZnO or Ag–ZnO rapidly within initial few hours, and about 42–45% of NPs were left in the nanocomposites in deionized water for 16 d to improve the long-term antibacterial activity further. At the concentration below 50 μg/mL, the nanocomposite aggregation was cell-compatible with ATDC5 and showed sterilization rates over 91% against Escherichia coli and 98% against Staphylococcus aureus. Long PEG aggregation showed greater cell proliferation capacity than short PEG aggregation, as well as better bacterial resistance and bactericidal activity against both E. coli and S. aureus. The flexible self-assembling antibacterial NPs with antifouling block copolymers via adjusting the component ratio or the segment length have shown premise in the construction of the dual-function antibacterial materials.

Studies of the properties of CHG-Loaded alginate fibers for medical application

Biomaterial-centered infection (BCI) is an important cause affecting wound healing, and preventing infection can accelerate wound healing and improve patients' cooperation. Our objective was to establish a dressings model by sustainably releasing chlorhexidine gluconate (CHG) to develop an effective method to inhibit wound infection. In this study, CHG-loaded alginate fibers was fabricated by wet blending spinning technology. The drug entrapment was confirmed by scanning electron microscopy, fourier transform infrared spectra, differential scanning calorimetry, X-ray diffraction and optical microscope. The content of chlorhexidine gluconate in fibers was determined by high performance liquid chromatography. Analysis indicated that chlorhexidine gluconate was successfully encapsulated into the alginate fibers and fibrous surface folds and area increases due to the grafting. The thermal stability of co-spun fibers was improved with the increasing content of chlorhexidine gluconate. Co-spun fibers exhibited better antibacterial activity and stability in comparison with pure alginate fibers, indicating that alginate fibers grafted with chlorhexidine gluconate has wider application prospect in the field of biomedical dressings.

Bioinspired interface design modulates pathogen and immunocyte responses in biomaterial-centered infection combination therapy

Biologically inspired biomaterial interface design can regulate the behaviors of pathogens and immunocytes for “infection combination therapy”.

The Effect of Brominated Furanones on The Formation of Staphylococcus aureus Biofilm on PVC

To study the influence of brominated furanones on the formation of Staphylococcus aureus (SA) biofilm on PVC thus providing new avenues of research on the surface modification of materials and clinical treatment of biomaterial-centered infection. Three brominated furanones (furanone-1, furanone-2, and furanone-3) were coated on the surface of PVC material. Both the modified PVC materials and SA were co-cultivated together. To assess the thickness of bacterial biofilm and bacterium colony unit area on PVC materials, confocal laser scanning microscopy and scanning electron microscopy (SEM) were used to observe the surface structure of SA biofilm formation. All treatments were compared with the control group which was not coated with furanones. PVC materials coated with furanone-1 had an increase in bacterial biofilm as well as SA colony area when compared with control. However, there was no significant difference between treating with furanone-1 and furanone-3 (P > 0.05). The impact of different brominated furanones on SA biofilm formation on the surface of PVC materials is different, furanone-1 can promote the SA biofilm formation on the surface of PVC material.

Effect of Brominated Furanones on the Formation of Biofilm by Escherichia coli on Polyvinyl Chloride Materials

To study the influence of brominated furanones on the biofilm (BF) formation by Escherichia coli (E. coli) on polyvinyl chloride (PVC) material, and to provide new ways of surface modification of materials to clinically prevent biomaterial centered infection. Three brominated furanones, dissolved in ethanol, furanone-1(3,4-dibromo-5-hydroxyl-furanone), furanone-2(4-bromo-5-(4-methoxypheny)-3-(methylamino)-furanone), and furanone-3(3,4-dibromo-5,5-dimethoxypheny-2(5H)-furanone) with representative chemical structure, were coated on the surfaces of separate PVC materials (1 × 1 cm), respectively. The surface-modified PVC materials were incubated with E. coli and for controls, 75 % ethanol-treated PVC materials were used. This treatment played as control group. The cultivation incubations were for 6, 12, 18, and 24 h. The thickness of bacterial BF and bacterial community quantity unit area on the PVC materials was determined by confocal laser scanning microscopy (CLSM), and the surface structure of bacterial BF formation was examined by scanning electron microscopy (SEM). The results of CLSM indicated the thickness of bacterial BF and bacterial community quantity unit area on PVC materials treated with furanone-3 were significantly lower than that of control at all time points (P < 0.05), whereas, the differences between furanone-1 and furanone-2 groups and control group were not significantly different (P > 0.05). The results of SEM indicated that after 6 h incubation, the quantity of bacterial attachment to the surface of PVC material treated with furanone-3 was lower than the control group. By 18 h incubation there was completely formed BF structure on the surface of control PVC material. However, there was no significant BF formation on the surface of PVC material treated with furanone-3. The impact of different brominated furanones on SA biofilm formation on the surface of PVC materials are different, furanone-3 can inhibit E. coli biofilm formation on the surface of PVC material.

Enhancing antibacterial activity of surface-grafted chitosan with immobilized lysozyme on bioinspired stainless steel substrates

Bacterial infections have been widely recognized as a major cause of the failure of medical implants and devices. One promising strategy to reduce the incidence of infections is to impart the material surfaces with bactericidal function for inhibiting bacterial adhesion and biofilm formation. In this study, stainless steel (SS) surface was first activated by a biomimetic dopamine anchor to provide active amino groups, followed by covalently immobilizing chitosan (CS) with glutaraldehyde (GA) as a bifunctional linker. Hen egg white lysozyme, a natural defensive enzyme, was finally conjugated to the grafted chitosan to enhance biocidal functionality. The antibacterial assay results demonstrated substantial enhancement in bactericidal efficiency against Staphylococcus aureus (S. aureus) on the lysozyme-immobilized SS substrates under the neutral pH conditions as compared to the chitosan-grafted SS substrates. With the inherent advantages of robust anchoring ability of dopamine and specific functionality of lysozyme, the metallic substrates can be readily tailored with antibacterial property to combat biomaterial-centered infection for potential biomedical applications.

Bostrycin, a novel coupling agent for protein immobilization and prevention of biomaterial-centered infection produced by Nigrospora sp. No. 407

Bostrycin, a red antibacterial agent with tetrahydroanthraquinone structure, has been isolated from Nigrospora sp. No. 407. This study investigated the potential antibacterial and multifunctional properties of matrixes through immobilization of bostrycin on their surface for immobilization of protein and prevention of bacterial growth. Bostrycin was immobilized on nonwoven polypropylene (PP) fabric by a technique using glutaraldehyde and polyethyleneimine for the activation of the surface. Glucose oxidase immobilized on bostrycin-treated nonwoven PP fabric showed high activity. The immobilization process improved thermal stability of the enzymes. During repeated assay for 30 cycles, the enzyme activity dropped to only 70% of the initial activity. Both bostrycin-treated nonwoven PP fabric sample and subsequently immobilized glucose oxidase sample on the surface also still exhibited a bacteriostatic effect. This is the first study to show that bostrycin is a promising coupling agent for surface modification on matrix and its potential applications in protein immobilization and biomaterial-centered infection.

A novel total knee arthroplasty infection model in rabbits.

Objective An in vivo animal model was developed to test the efficacy of innovative therapies for the prevention of biomaterial centered infections caused by E. Staphylococcus. Methods Thir-ty-two New Zealand white rabbits were used in this study. After proper anesthesia, a knee prosthesis made by UHMWPE was cemented in the surface of tibia of knee. After 1 week of operation, the operated knee was inoculated with 0, 1×104, 1×106, and 1×108 colony forming units (CFU) of E. Staphylococcus. Animals were sacrificed after 1 week at which time tissues and biomaterial samples were examined for evidence of infec-tion. Joint tissue, implant and joint fluid were tested for the number of bacteria. The joint and implant sam-ple was examined by X-ray, MR, CLSM and pathological examination. Results A total of 32 knees were used for analysis. When saline was injected into the knee, 0 of the knees demonstrated evidence of biomate-rial centered infection. When 1×106 CFU of E. Staphylococcus was injected into the knee, all knees devel-oped a biomateriai centered infection. Evidence of septicemia (positive blood cultures) was found in a rabbit inoculated with 108 CFU of E. Staphylococcus. The infection rate was positively correlated with the amount of the E. Staphylococcus inoculated (R2=0.9939). Gross observation showed a great amount of abscess covered the surface of implant. The joint synovial membrane was erythrogenic. CLSM showed a lot of bacteria were adhesive to the surface of implant and there were films of ESS substance on the surface. Histological exami-nation showed an acute infection. The quantities analysis showed the number of bacteria in joint fluid is sig-nificantly high than that of on the surface of implant and in the joint tissue. Conclusion This rahbit knee infection model mimic clinical situation. It is a successful model and provides an effective method for the e-valuation of new treatment for biomaterial centered infection. Key words: Rabbits; Staphylococcal infections; Models; animal; Arthroplasty; replacement; knee

Reduction of Postoperative Spinal Implant Infection Using Gentamicin Microspheres

Three noncontiguous spinal implant sites in 1 rabbit were challenged with Staphylococcus aureus and local antibiotic prophylaxis was given with gentamicin in controlled-release microspheres (poly(lactic-coglycolic-acid) [PLGA]). Postoperative biomaterial-centered infection on and around the titanium rods was assessed using standard bacterial quantification essays. To assess surgical site and biomaterial-centered infection reduction with controlled release gentamicin from microspheres against S. aureus. A postoperative biomaterial-centered infection can be devastating after successful thoracolumbar spinal surgery and puts a high burden on patients, families, surgeons, and hospitals, endangering both our healthcare budget and our ability to perform challenging cases in patients with increasing numbers of comorbidities. Systemic antibiotics often do not reach "dead-space" hematomas where bacteria harbor after surgery, whereas local, controlled release gentamicin prophylaxis through PLGA microspheres showed favorable pharmacokinetics data to achieve local bactericidal concentrations for up to 7 days after surgery. A well published rabbit spinal implant model with systemic cephalosporin prophylaxis was challenged to create a baseline infection of approximately 70% in control sites. We then challenged 3 noncontiguous titanium rods inside the laminectomy defect with 10e6 colony forming units S. aureus and randomly treated 2 sites with gentamicin PLGA microspheres and 1 site with PLGA carrier only (control). Standard quantification techniques were used to assess biomaterial centered and soft tissue bacterial growth after 7 days. After establishing reliable infection rates in control sites, the therapeutic arm of the study was started. Surgical site infections were found in 75% of control sites, whereas gentamicin microspheres reduced the incidence down to 38% in the same rabbits. Biomaterial-centered infection was reduced from 58% to 23% only in all sites challenged with 10e6 S. aureus. Postoperative, biomaterial-centered infection was reduced at least 50% with intraoperative gentamicin microspheres in the face of systemic cephalosporin prophylaxis and high dose S. aureus in a laminectomy defect in rabbits. The data are statistically and clinically significant, and further animal testing is planned to confirm these results.

Silk-functionalized titanium surfaces for enhancing osteoblast functions and reducing bacterial adhesion

It would be ideal to have implants which can simultaneously inhibit bacterial adhesion and promote osteoblast functions. In this work, titanium surfaces were modified with poly(methacrylic acid) (P(MAA)) followed by immobilization of silk sericin. Firstly a trichlorosilane coupling agent, which is an atom transfer radical polymerization (ATRP) initiator, was immobilized on the oxidized titanium surface to facilitate the surface-initiated ATRP of methacrylic acid sodium salt (MAAS). The pendant carboxyl end groups of the grafted and partially protonated MAA chains were subsequently coupled with silk sericin via carbodiimide chemistry. The functionalized Ti surfaces were characterized by X-ray photoelectron spectroscopy, and assayed for osteoblast cell functions and bacterial adhesion. The covalently immobilized MAA brushes significantly reduce the adhesion of the two bacterial strains ( Staphylococcus aureus and Staphylococcus epidermidis) tested. The silk sericin-immobilized surfaces, at the same time, promote osteoblast cells' adhesion, proliferation, and alkaline phosphatase activity. Thus, the P(MAA) and silk sericin functionalized Ti surfaces have potential applications combating biomaterial-centered infection and promoting osseointegration.

Polyacrylamide brush coatings preventing microbial adhesion to silicone rubber

Silicone rubber is a frequently used biomaterial in biomedical devices and implants, yet highly prone to microbial adhesion and the development of a biomaterial-centered infection. Effective coating of silicone rubber to discourage microbial adhesion has thus far been impossible due to the hydrophobic character of its surface, surface deterioration upon treatment and instability of coatings under physiological conditions. Here we present a method to successfully grow polyacrylamide (PAAm) brushes from silicone rubber surfaces after removal of low molecular weight organic molecules (LMWOM), such as silane oligomers. PAAm brush coating did not cause any surface deterioration and discouraged microbial adhesion, even after 1-month exposure to physiological fluids. The method presented opens many new avenues for the use of silicone rubber as a biomaterial, without the risk of developing a biomaterial-centered infection.

Surface modification and properties of Bombyx mori silk fibroin films by antimicrobial peptide

The Bombyx mori silk fibroin films (SFFs) were modified by a Cecropin B ( CB) antimicrobial peptide, (NH 2)-NGIVKAGPAIAVLGEAAL-CONH 2, using the carbodiimide chemistry method. In order to avoid the dissolution of films during the modification procedure, the SFFs were first treated with 60% (v/v) ethanol aqueous solution, resulting a structural transition from unstable silk I to silk II. The investigation of modification conditions showed that the surface-modified SFFs had the satisfied antimicrobial activity and durability when they were activated by EDC·HCl/NHS solution followed by a treatment in CB peptide/PBS buffer (pH 6.5 or 8) solution at ambient temperature for 2 h. Moreover, the surface-modified SFFs showed the smaller contact angle due to the hydrophilic antimicrobial peptides coupled on the film surface, which is essential for the cell adhesion and proliferation. AFM results indicated that the surface roughness of SFFs was considerably increased after the modification by the peptides. The elemental composition analysis results also suggested that the peptides were tightly coupled to the surface of SFFs. This approach may provide a new option to engineer the surface-modified implanted materials preventing the biomaterial-centered infection (BCI).

A novel total knee arthroplasty infection model in rabbits

Infection of biomaterial implants is an expensive and devastating complication of orthopaedic surgery historically ranging from less than 1% in primary total knee arthroplasty (TKA) to 10% in revision TKA. An in vivo animal model was developed to test the efficacy of innovative therapies for the prevention of biomaterial centered infections caused by methicillin-resistant Staphylococcus aureus bacteria (MRSA). Twenty-two New Zealand White rabbits were used in this study. After proper anesthesia, a stainless-steel screw with a high molecular weight polyethylene (UHMWPE) washer was cemented in a defect created in the intra-articular, non-articulating portion of the lateral femoral condyle of each knee. After closure of the joint capsule, each knee was inoculated with 0, 10 2, 10 3, or 10 4 colony forming units (CFU) of MRSA. Animals were sacrificed after 7 days at which time joint aspirate, tissues and biomaterial samples were examined for evidence of infection. A total of 42 knees were used for analysis. When saline was injected into the knee, 0/10 of the knees demonstrated evidence of biomaterial centered infection (with the contralateral knee receiving 10 4 CFU MRSA). Four of 10 knees developed a biomaterial centered infection when 10 2 CFU MRSA was introduced. Seven out of 10 knees developed a biomaterial centered infection when either 10 3 or 10 4 CFU MRSA was injected. No evidence of septicemia (positive blood cultures) was found in any animal. This rabbit knee model utilizes commonly employed inexpensive orthopaedic implant materials in an in vivo milieu and provides an effective method for the evaluation of treatments for biomaterial centered infections.

Stability and effectiveness against bacterial adhesion of poly(ethylene oxide) coatings in biological fluids

Poly(ethylene oxide) (PEO) coatings have been shown to reduce the adhesion of different microbial strains and species and thus are promising as coatings to prevent biomaterial-centered infection of medical implants. Clinically, however, PEO coatings are not yet applied, as little is known about their stability and effectiveness in biological fluids. In this study, PEO coatings coupled to a glass substratum through silyl ether bonds were exposed for different time intervals to saliva, urine, or phosphate-buffered saline (PBS) as a reference at 37 degrees C. After exposure, the effectiveness of the coatings against bacterial adhesion was assessed in a parallel plate flow chamber. The coatings appeared effective against Staphylococcus epidermidis adhesion for 24, 48, and 0.5 h in PBS, urine, and saliva, respectively. Using XPS and contact-angle measurements, the variations in effectiveness could be attributed to conditioning film formation. The overall short stability results from hydrolysis of the coupling of the PEO chains to the substratum.

The Classic: Biomaterial-Centered Infection

The Classic: Biomaterial-Centered Infection

Positively charged biomaterials exert antimicrobial effects on gram-negative bacilli in rats

Biomaterial-centered infection is a much-dreaded complication associated with the use of biomedical implants. Although positively charged biomaterial surfaces stimulate bacterial adhesion, it has been suggested that surface growth of adhering Gram-negative bacilli is inhibited on positively charged surfaces . In the present paper, we determined the infection rate of differently charged poly(methacrylates) in rats. To this end, 2×10 6/cm 2 Escherichia coli O2K2 or 2×10 4/cm 2 Pseudomonas aeruginosa AK1 were seeded on glass discs coated with three differently charged poly(methacrylates) coatings in a parallel plate flow chamber. Three rats received six subcutaneous discs (two discs of each charge variant) seeded with E. coli, while three other rats received discs seeded with P. aeruginosa. The numbers of viable bacteria on the surfaces were determined 48 h after implantation. On 50% of all positively charged discs viable E. coli were absent, while the negatively charged discs were all colonized by E. coli. P. aeruginosa, however, were isolated from both positively and negatively charged discs. Probably, P. aeruginosa can circumvent the antimicrobial effect of the positive charge through the formation of extracellular polysacharides.

Adhesion to bile drain materials and physicochemical surface properties of Enterococcus faecalis strains grown in the presence of bile.

The aim of this study is to determine whether growth in the presence of bile influences the surface properties and adhesion to hydrophobic bile drain materials of Enterococcus faecalis strains expressing aggregation substance (Agg) or enterococcal surface protein (Esp), two surface proteins that are associated with infections. After growth in the presence of bile, the strains were generally more hydrophobic by water contact angles and the zeta potentials were more negative than when the strains were grown in the absence of bile. Nitrogen was found in lower surface concentrations upon growth in the presence of bile, whereas higher surface concentrations of oxygen were measured by X-ray photoelectron spectroscopy. Moreover, an up to twofold-higher number of bacteria adhered after growth in bile for E. faecalis not expressing Agg or Esp and E. faecalis with Esp on its surface. E. faecalis expressing Agg did not adhere in higher numbers after growth in bile, possibly because they mainly adhere through positive cooperativity and less through direct interactions with a substratum surface. Since adhesion of bacteria is the first step in biomaterial-centered infection, it can be concluded that growth in bile increases the virulence of E. faecalis.